PRINT ISSN NO. : 2249-1570 ONLINE ISSN NO.: 2277-9396
INTERNATIONAL JOURNAL OF FOOD AND FERMENTATION TECHNOLOGY VOL. 2, NO. 2, DECEMBER 2012
Editor-in-Chief Prof. VK Joshi Professor and Head, Fermentation Technology Lab, Department of Food Science and Technology Dr YS Parmar University of Horticulture and Forestry Nauni, Solan, Himachal Pradesh, India E Mail:
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INTERNATIONAL JOURNAL OF FOOD AND FERMENTATION TECHNOLOGY
ABOUT THE JOURNAL The Journal publishes research papers on all the subjects related to the Food and Fermentation Technology. All aspects of food and fermentation technology would be considered. Papers on bio-technology, bio-chemical engineering, toxicological aspects having direct bearing or are related with food would be welcomed. R & D work related to the fermentation covering microbiology, bio-chemical aspects, genetics, indigenous fermented foods, toxicology or nutritive value would also be included. Articles highlighting the food standards and safety issues will be given special emphasis. Preparation and evaluation of alcoholic beverages would be an important aspect of the articles published. The review on any aspect of food processing, composition, nutrition, and fermentation would be considered. Management of food processing industrial waste would be an integral component of the papers published. The other aspects of food processing like low temperature preservation, dehydration, thermal processing, irradiation, emerging technologies viz., ohmic preservative, pulse electric field, high pressure preservation, enzymology, microbiological quality, food safety and standards, and food engineering, will also be considered. International Journal of Food and Fermentation Technology, a half yearly journal, publishes original research papers, short communications and review papers on topics which include in brief : • Food microbiology • Bio-chemical aspects of food • Genetic and genetically modified foods • Enology • Indigenous fermented foods • Toxicology, safety and quality • Food processing • Fermentation technology • Food engineering • Quality assurance • Food preservation • Food additives
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EDITORIAL BOARD Editor-in-Chief Prof. V K Joshi Professor and Head, Fermentation Technology Lab, Department of Food Science and Technology, Dr YS Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh, India
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Honorary Editor Dr (Mrs) Sunita Garg, IJNPR, NPARR and Wealth of India Division National Institute of Science Communication and Information Resources, CSIR, New Delhi, India
[email protected] Dr J K Gupta Department of Entomology and Apiculture Dr YS Parmar University of Horticulture and Forestry Nauni-Solan, Himachal Pradesh, India
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Dr N S Thakur Department of Food Science and Technology Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh, India
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Executive Editorial Board Dr (Mrs) Devina Vaidya Department of Food Science and Technology Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh, India
[email protected] Dr B L Attri Central Institute of Temperate Horticulture, Regional Station, Mukteshwar, Kumaun, Nainital, Uttarakhand, India.
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Dr Shashi Bhushan Division of Biotechnology, Institute of Himalayan Bio-resource Technology (CSIR), Palampur, Distt Kangra, Himachal Pradesh, India
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Dr Wamik Azmi Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla, Himachal Pradesh, India
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Dr Om Prakash Chauhan Fruits and Vegetables Technology Division Defence Food Research Laboratory Siddarthanagar, Mysore, India
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Dr Neerja S. Rana Department of Basic Sciences Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni-Solan, Himachal Pradesh, India
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Dr P S Panesar Department of Food Engineering and Technology SL Institute of Engineering and Technology, Longowal, Punjab, India
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EDITORIAL ADVISORY BOARD Dr Sumit Arora ( Dairy Science) Dairy Chemistry Division, National Dairy Research Institute Karnal, Haryana, India
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Dr Eveline Bartowsky (Wine Microbiology) The Australian Wine Research Institute P.O. Box 197, Glen Osmond, Australia.
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Prof Tek Chand Bhalla (Food Fermentation & Enzyme Tech.) Department of Biotechnology, Himachal Pradesh University, Shimla, Himachal Pradesh, India
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Dr L. Rebordinos ( Food Microbiology) Laboratorio de Microbiología y Genética. Facultad de Ciencias del Mar y Ambientales. Universidad de Cádiz. Polígono del río San Spain.
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Dr M C Pandey, (Meat and Meat products) Department of Freeze Drying and Animal Product Technology, Defense Food Research Laboratory, Mysore, India
[email protected] Dr V M Pratape, (Grain Science and Technology), Department of Grain Science and Technology, Central Food Technological Research Institute, Mysore, India
[email protected] Dr Pura Naik J (Plantation Crops) Division of Plantation, Spices and Flavour Tech., Central Food Technological Research Institute, Mysore, India
[email protected] Professor Pradeep Khanna (Mushroom Production) College of Basic Science, PAU Ludhiana, Punjab, India
[email protected] Dr Y S Dhaliwal (Food and Nutrition) Department of Food Science and Nutrition, College of Home Science, CSK HPKV, Palampur, Himachal Pradesh, India
[email protected] Dr R S Singh ( Food Fermentation and Enzyme Technology) Department of Biotechnology, Punjabi University, Patiala, Punjab, India.
[email protected] Dr S S Kanwar Department of Microbiology, CSK HPKV, Palampur, Distt Kangra Himachal Pradesh, India
[email protected] Dr RC Ray (Food Fermentation) Principal Scientist (Microbiology) Regional Centre of Central Tuber Crops Research Institute Dumuduma Housing Board, Bhubaneswar, Orissa, India Dr. Rintu Banerjee (Microbial Technology) Microbial Biotechnology and Downstream Processing Laboratory, Agricultural & Food Engineering Department Indian Institute of Technology, Kharagpur,West Bengal, India rin_
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Dr Aline Lonvaud (Wine and Brandy) Faculty of Enology, University Victor Segalen Bordeaux 2, France.
[email protected] Dr Creina S. Stockley ( Wine and Health) The Australian Wine Research Institute Australia.
[email protected] Dr M. Remedios Marín (Wine Modelling) Universidad Publica de Navarra Nafarroako Unibertsitste Publikoa Area de Tecnologia de Alimentos Universidad Publica de Navarra Campus Arrosadia Pamplona, (Navarra), Spain.
[email protected] Dr Philippe Jeandet (Sparkling Wine) Laboratory of Enology and Applied Chemistry, Unité de Recherche sur la Vigne et le Vin de Champagne, Research Unit N°2069 University of Reims, Faculty of Science, France.
[email protected] Dr Vasudeo P. Zambare Sequence Biotech. Pvt. Ltd. Nashik, Maharashtra
[email protected] Dr Luca Cocolin (Wine Microbiology) Dipartimento di Scienze degli Alimenti, Università degli studi di Udine, Facoltà di Agraria, via Marangoni, Udine, Italy.
[email protected] Dr Ginés Navarro (Wine Fermentation) Departamento de Química Agrícola, Geología y Edafología, Facultad de Química. Universidad de Murcia. Campus Universitario de Espinardo, Murcia. Spain.
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Dr R K Gupta (Statistics) Department of Basic Sciences Dr. Y S Parmar University of Horticulture and Forestry, Nauni, Solan, India.
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Dr S K Patyal Department of Entomology and Apiculture, DR. Y S Parmar University of Horticulture and Forestry, Nauni, Solan
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Contents International Journal of Food and Fermentation Technology Vol. 2 No. 2, December, 2012
Review paper Mechanism and manifestation of bacterial quorum sensing in food environment Suja Senan and Jashbhai B. Prajapati
103
Taste producing components in fish and fisheries products: A review Mohammed Golam Sarower, Abul Farah Md. Hasanuzzaman, Bhabananda Biswas and Hiroki Abe
113
Antimicrobial, antioxidant and phyto-chemicals from fruit and vegetable wastes: A review V. K. Joshi, Ashwani Kumar and Vikas Kumar
123
Research Paper Effect of freeze-drying and storage on β-carotene and ascorbic acid stability of mango milk shake K. Jayathilakan, Khudsia Sultana, M.C. Pandey and K. Radhakrishna
137
Physico-chemical and sensory evaluation of wines from different citrus fruits of Himachal Pradesh V. K. Joshi, Vikas Kumar and Ashwani Kumar
145
Assessment of stability and biopreservative effect of recombinant pediocin CP2 B. Kumar, P. P. Balgir and B. Kaur
149
Production of a herbal wine from Aloe vera gel and evaluation of its effect against common food borne pathogens and probiotics Neetika Trivedi, Praveen Rishi and Sanjeev Kumar Soni
157
Preparation and evaluation of wine from tendu (Diospyros melanoxylon L) fruits with antioxidants Umesh C. Sahu, Sandeep K. Panda, Uma B. Mohapatra and Ramesh C. Ray
167
Comparative studies on Bhatooru fermented with traditional inoculum (malera) and standard starter cultures Savitri and Tek Chand Bhalla
175
Sequential optimization of xylanase production by an indigenously isolated Acinetobacter species using solid state fermentation Mohammed S. Kanchawala, Mugdha N. Harmalkar and Madhavi R. Vernekar
185
Research Note Ethanol production efficiency of various species of cassava effluents using different inoculants Ekwere, Mercy R., Agha, N.C., Mepba, H. D and Igile, G.O.
193
Quality evaluation of guava-carrot jelly Jaydeep Singh and Suresh Chandra
197
Book Review
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From the Desk of Editor- in-Chief It is with great pleasure and a sense of fulfilment to state that we have successfully brought out two issues of inaugural volume of our journal “International journal of Food and Fermentation Technology” during 2011 to cater to the needs of food scientists, and R and D workers, and the academicians in the fields. The inaugural issue of the journal was released by the honourable Vice-chancellor of Dr YS Parmar university of Horticulture and Forestry, Nauni, Solan (India) Dr K.R.Dhiman on 26th June 2012 in a function held in the university to mark the beginning of the ICAR sponsored training on “Advances in Fruit and Vegetable Processing and Preservation” with me as a Course Director. I am equalty happly to inform you that the journal has received an overwhelming response from the scientists and academician from the field of food and fermentation technology.The journal during the past one year has published papers on various aspects of food technology and fermentation technology. It could be achieved only with the help of all the members of editorial board, the contributors and the publisher. I hope you would have relished the style, general get-up, cover page, coverage of the journal; scientific and technical contents, with an attractive of look A4 Size format. At the same time, we would welcome suggestions from the readers, contributors and peers of the field. Now, I am placing before you the first issue of Vol. 2 Number 2of the journal. I would like to express my deep gratitude to all those who have extended their help, guidance and co-operation in bringing out this issue. I siege this opportunity to request all the members of editorial board to exert more so that the journal gets high recognition among the peers and we are able to get the NAAS ratings to the journal. I am sure through your efforts we would get more number of papers of good quality. They should themselves contribute papers of high quality to encourage others to do so to the conceptual editorial I invite the members of editorial board to contribute liberally. Certainly, there would not be any page charges from them for this contribution. The scientists abroad can certainly contribute to improve the quality of this journal being in infancy. On behalf of the editorial board and the publisher, I assure the readers and the contributors that we would strive hard to ensure high level of scientific integrity, transparency, impartiality and accuracy in selection of articles and finally, production of the journal. I earnestly call upon all the scientists of food and fermentation technology to contribute their valuable research articles to the journal as a vehicle of transmission of their scientific findings. The contributors of review or mini-review can consult the editor- in- chief, member of the editorial board for the topic of review to contribute. At the same time, I must express very clearly that just because of some page charges we will not accept and publish substandard articles, but only peer reviewed and accepted papers would be published. I understand very well that the final scientific quality of any journal would be determined by the type of papers submitted to the journal so, the cooperation of all would be a pre-requisite in this regard and would be welcomed from the core of my heart.
V.K. Joshi
Conceptual Editorial
Modern Post-harvest Technology A Panacea for Developing Countries The Green Revolution and subsequent efforts made through the application of science and technology for increasing food production in India and world have brought self-reliance in food. With advent of advanced agriculture practices, the production of various crops have increased linearly. This phenomenon has been recorded not only from the developed countries but also in the developing nations of the world. At present, the world production of wheat is 653654525 MT, maize 840308214 MT, rice and paddy 696324394 MT, Buffalo milk, whole, fresh milk 92473371 MT, Cow milk, whole, fresh 600838992 MT, Indigenous cattle meat 63782689 MT, Indigenous pig meat 109100198 MT, Indigenous chicken meat 85860953 MT, Indigenous sheep meat 8689557 MT, fruits 609213509 MT, vegetables 965650533 MT. Out of these, milk, meat, fruits and vegetables are highly perishable crops. Food being a living commodity respire and therefore, is liable to be spoiled. The major causes of spoilage of food include spoilage by microorganisms, enzymes of microorganisms or the food. The non-enzymatic or purely chemical reactions and environmental conditions. After the crop is harvested, these factors lead to a considerable postharvest spoilage. In the developed countries, the quantam of spoilages is bare minimum but in the developing countries, like India. The postharvest losses are staggering high. These have been estimated to be as high as 25-30% leading to a loss of Rs. 52,000 crores annually. It may be astonishing but is a fact that the United Kingdom produces the amount of the fruit crop equal to what India wastes. The major difference lies in lack of proper infrastructure in the developing countries in contrast to the developed nations. Postharvest handling is the stage of crop production immediately following harvest. It largely determines the final quality and it includes cooling, cleaning, sorting and packing. After harvest, foods (e.g. fruits, vegetables, milk, meat, fish) are liable to accelerated physiological, chemical, and microbial processes that invariably lead to deterioration and loss of wholesomeness. It is
Figure 1: World production of various agricultural commodities Source: FAO, Stat: 2010
then, necessary to institute some measure of processing such as reduction in moisture content, denaturation of endogenous enzymes and microorganisms, or packaging in order to curtail the loss of perishables. Utilizing improved postharvest practices thus often results in reduced food losses, improved overall quality and food safety, and higher profits for growers and marketers. The most important goals of post-harvesting handling are to keep the product cool, to avoid moisture loss and slow down undesirable chemical changes, and avoiding physical damage such as bruising, to delay spoilage. The way the grains are stored after harvest becomes the major cause of contamination with fungi some of which are producer of toxin like aflatoxin. The fruit like apple after contamination has a toxin called patulin, if not stored properly. Sanitation is also an important factor, to reduce the possibility of pathogens that could be carried by fresh produce, for example, as residue from contaminated washing water. Consumption of such food results in the diseases of gastro-intestinal track which sometimes prove to be fatal. Postharvest loss reduction technology encompasses the usage of optimum harvest factors, reduction of losses in handling, packaging, transportation and storage with modern infrastructure machinery, processing into a wide variety of products, home scale preservation with low cost technology. Use of thermal processing, low temperature storage drying chemical and biological reactions coupled with other preservation techniques are applied to enhance the storability of the perishables. There are different constraints for postharvest management like: large number of small and marginal farmers with primitive system of cultivation, poor infrastructure in terms of handling, transport, storage, processing and marketing, lack of adequate quality systems and procedure like grading and sorting, hot and humid climates, cost of installation of post-harvest treatment facilities is expensive, lack of awareness training for the rural farmers and inconsistency in supply due to seasonality of the produce. Adoption of latest techniques could make available a large quantity of food by avoiding losses and provide quality food and nutrition, more raw materials for processing. Postharvest technology has the capability to meet food requirement of growing population. By adopting improper and inefficient methods of storage we are losing a substantial portion of production. If better methods of processing and storage are adopted, the losses could be reduced to a large extent. The developing countries need proper infrastructure for this and a strong will to adopt modern postharvest technology to save the wastage. Thus, it could certainty prove to be a panacea for the problems faced by the developing countries.
V K Joshi
Intl. J. of Food. Ferment. Technol. 2(2): 103-112, December, 2012
Review paper
Mechanism and manifestation of bacterial quorum sensing in food environment Suja Senan1* and Jashbhai B. Prajapati2 1
Assistant Professor, Department of Dairy Microbiology, SMC College of Dairy Science, Anand Agricultural University, Anand, India 2
Head, Department of Dairy Microbiology, SMC College of Dairy Science, Anand Agricultural, University, Anand , India
*
Email:
[email protected]
Paper no: 45
Received: 11 Oct, 2012
Received in revised form: 20 Nov , 2012
Accepted: 24 Nov, 2012
Abstract Research in the field of food science and technology has shown that bacteria can communicate with each other. This communication is population-density dependent and involves signalling molecules that can diffuse between the bacterial cells. The process of communication is called ‘quorum sensing’. Quorum sensing is now known to be a widespread phenomenon responsible for modulating many different activities in diverse Gram-negative and Gram-positive bacteria. This phenomenon has been found to be responsible in food spoilage and biofilm formation by food-related bacteria in the food industry in addition to pathogenesis. Understanding bacterial quorum-sensing systems can help in controlling the growth of undesirable food-related bacteria. Blocking the quorum-sensing signalling molecules in food related bacteria may possibly prevent quorum-sensing-regulated phenotypes responsible for food spoilage. With proper understanding of quorum sensing mechanism we can identify quorum-sensing inhibitors that could be used as food preservatives to enhance food safety and increase shelf life. ©2012 New Delhi Publishers. All rights reserved
Keywords: Communicate, signal, phenotype
Introduction Bacteria carry with them a myth that they are an unambitious unicellular entity whose single motto in life is to grow and divide ad infinitum in isolation. The evolved traits of collective social behavior and communication were believed to be only for multicellulars. This myth has been shattered with path breaking research confirming that bacterial species do coordinate communal behavior. It all began with a brilliant observation of growth kinetics of marine bioluminescent bacteria that the onset of exponential growth occurs without a lag but bioluminescence does not increase until midlogarithmic phase, probably due to an inhibitor in the medium (Farghaly, 1950). Another study concluded that the inhibitor
attached to the luciferase and the process is regulated by extracellular secreted signal components (Kempner and Hanson, 1968). The signal molecule was dubbed as ‘autoinducer’ and the desired response as ‘autoinduction’. In 1970, Nealson and colleagues reported that luminescence in the marine symbiont bacterium Vibrio fischeri that inhabit light organs of animals such as Euprymna scolopes, was produced only at high cell density. They were the first to propose the phenomenon of cell density-dependent autoinduction. The term “quorum sensing” specifically refers to the cell-density linked, coordinated gene expression in populations that experience threshold signal concentrations to induce a synchronized population response (Fuqua et al., 1994). There are many situations where the ability of a bacterial population to behave co-operatively could be highly
Senan and Prajapati
advantageous particularly in the contexts of conjugation, symbiosis, niche adaptation, production of secondary metabolites, combating the defense mechanisms of higher organisms and for facilitating population migration where the prevailing conditions have become unfavorable.
the autoinducing polypeptide (AIP) system. The 4th system, using autoinducer-2 (AI-2), is found in Gram positive as well as Gram-negative bacteria. It can be said that AI-2 is used for interspecies communication. The systems have a common flow of events (Sifri, 2008)
Bacterial language is chemical in nature using certain signaling molecules. They possess specific receptors which can detect these autoinducers. Upon binding of inducer to receptor, it activates transcription of certain genes, including those for inducer synthesis. When only a few bacteria are present, diffusion reduces the concentration of the inducer in the surrounding medium to almost zero. However, as the population grows the concentration of the inducer passes a threshold (quorate) and the receptor becomes fully activated. Activation of the receptor induces the up regulation of specific genes, causing all of the cells to begin transcription at approximately the same time.In this review, all these aspects have been described in context of food.
•
Production of small biochemical signal molecules by the bacterial cell;
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Release of the signal molecules, either actively or passively, into the surrounding environment;
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Recognition of the signal molecules by specific receptors once they exceed a threshold concentration
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Changes in gene regulation
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The autoinducer molecules important in quorum sensingare are summerized here. Autoinducer 1: Acyl homoserine lactones are the major group of autoinducer signals in gram-negative bacteria. They have a conserved homoserine lactone (HSL) ring with a variable acyl side chain. The length and saturation level of the acyl chains coupled to the presence or absence of oxo or hydroxyl substitutions at the C-3 position of the acyl chain provide variation and specificity for quorum-sensing communication in a mixed bacterial population (Shaw et al., 1997)
Decoding the language Signal molecules implicated in cell-to-cell communication are now known as autoinducers or quorum-sensing molecules, and their function is to regulate gene expression in other cells of the community, which, in turn, control a number of bacterial responses. Molecules implicated in quorum sensing (QS) possess the ability to induce their own production ,hence are known as ‘autoinducers’ (AI) some of the moleculars used by bacteria for quorum sensing are listed in Table 1. Cell-to-cell signaling systems are broadly grouped into 4 main categories. Two of these systems, which use autoinducer-1 (AI-1) and autoinducer-3 (AI-3), are found in Gram-negative cells, while the Gram-positive bacteria use a 3rd type of signaling system,
Autoinducer 2 has been discovered in many gram-negative bacteria as a global signal molecule for interspecies communication, as it is made by gram-positive as well as gramnegative bacteria (Bassler, 1999). AI-2 is identified it as a furanosyl borate diester. The proposed structure contains two fused five member rings containing one boron atom bridging the diester (Chen et al., 2002).
Table1: List of signal molecules used by bacteria for quorum sensing Organism
Signal molecule
Function
Vibrio fischeri Aeromonas hyrophila Aeromonas salmonicida Agrobacterium tumefaciens Burkholderia cepacia Chromobacterium violaceum Erwinia caratovora Pseudomonas aerofaciens Pseudomoans aeroginosa Rhizobium leguminosarum Serratia liquefaciens Yersinia pseudotuberculosis Rhodobacter sphaeroides
3-Oxo-C6-HSL N-butanoyl-HSL N-butanoyl-HSL 3-Oxo-C6-HSL N-octanoyl-HSL N-hexonoyl-HSL 3-Oxo-C6-HSL N-hexonoyl-HSL 3-Oxo-C6-HSL N-hexonoyl-HSL N-butanoyl-HSL N-octanoyl-HSL 7,8-cis-N-(Tetradecanoyl)-HSL
Bioluminescence Serine protease and metalloprotease production exoprotease Ti plasmid conjugal transfer Protease and siderophore porduction Hydrogen cyanide, antibiotics, Pectinase synthesis Phenazine antibiotic biosynthesis Virulence, biofilm formation. Rhizosphere genes Swarmer cell differentiation Bacterial aggregation and motility Prevents bacterial aggregation
Sources: Raina et al., 2009, Hammer and Bassler, 2003
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Mechanism and manifestation of bacterial quorum sensing in food environment
Auotinducer 3: Epinephrine/norepinephrine/AI-3 signaling is used as an interkingdom chemical signaling system between microbes and their hosts. This system is also exploited by pathogens to regulate virulence traits. In enterohemorrhagic E. coli (EHEC) O157:H7, it is essential for pathogenesis and flagella motility. These three signals activate expression of a pathogenicity island named Locus of Enterocyte Effacement (LEE), Shiga toxin, and the flagella regulon (Moreira and Sperandio, 2010) Cyclic dipeptides. Identified in strains of Pseudomonas these new signal molecules were the diketopiperazines (DKPs) cyclo (L-Ala-L-Val) and cyclo(L-Pro-L-Tyr). Autoinducing Peptides: Gram-positive species predominantly utilize small post translationally modified peptides for cell-to-
cell signaling. These are exported via ATP-binding cassette (ABC)-type transporters The genes encoding the precursor peptide, membrane bound sensor kinase protein, and the transporter machinery are usually located in a single gene cluster (Bassler, 2002). The LuxR/I signaling system: The luciferase operon in V. fischeri is regulated by two proteins, LuxI, responsible for the production of the AHL (acylhomoserine lactone ) and LuxR a transcription factor responsible for controlling gene expression in the presence of the autoinducer. The schematic view of the mode of action of the system is shown in Figure 1. AHLs have a conserved homoserine lactone ring connected through an amide bond to
Figure 1: The process of quorum induced bioluminescence by the LuxR/I system
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a variable acyl chain. Acyl chains vary in number of carbons from four to 18 and the third position may or may not be modified (carbonyl group, hydroxyl or fully reduced). Different acyl chains ensure that different AHLs will be recognized by different LuxR-type proteins. The LuxR/I system was the first one to be described in V. fischeri (Nealson et al., 1970). P. aeruginosa produces two AHLs, N-(3-oxododecanoyl)L-homoserine lactone (3OC12-HSL) and N-butanoylL-homoserine lactone (C4- HSL) (Pearson et al., 1995). These AHLs bind to and activate LasR and RlhR transcription factors, respectively (Parsek and Greenberg, 2000).
Quorum sensing in Gram-positive organisms Quorum sensing in Gram-positive organisms relies on autoinduction by small peptides, which interact with two component systems ultimately regulating gene transcription. These small peptides are usually products of oligopeptides that are cleaved and/or further modified before being exported from the bacterium by transporters. At threshold concentrations, the peptides are recognized by sensor kinases that initiate phosphous transfer to a response regulator. The peptides involved in Gram-positive QS are often specific for their cognate receptors. The QS system that S.aureus utilizes is one of the most studied systems in Gram-positive organisms. Currently, two quorum-sensing mechanisms have been explained in S. aureus. 1.
The first quorum-sensing system consists of a sevenamino-acid peptide autoinducer RAP (RNAIII activating peptide) and TRAP (target of RNAIIIactivating protein).
2.
Second quorum-sensing mechanism regulated by agr, (accessory gene regulator) locus, composed of two divergently transcribed units, namely RNAII and RNAIII whose transcription is under control of the P2 and P3 promoters respectively. The RNAII unit encloses four genes, namely agrB, agrD, agrC and agrA. The genes agrB and agrD are involved in the autoinducer production
The accessory gene regulator (Agr) system regulates toxin and protease secretion in staphylococci. At low cell density, the bacteria express proteins required for attachment and colonization, and as the cell density becomes higher, this expression profile switches to express proteins involved in toxin and protease secretion (Novick, 2003). Streptococcus pneumoniae was one of the original Gram positive systems characterized. The discovery of an ABC transporter that secreted the activator for competence, comA, revealed a class of ABC transporters that contained N-terminal
proteolytic domains (Zhou et al., 1995). This led to the characterization of the activator as Competence Inducing Factor (CSP). (Havarstein et al., 1996). The LuxS/AI-2 signaling system Vibrio harveyi QS system constitutes a mix between components of Gram-positive and Gram negative systems (Figure 2). Vibrio harveyi QS system constitutes a mix between components of Gram-positive and Gram negative systems (Figure 3 ). It has two QS systems: 1.
Autoinducer (AI-1) , an AHL, and is primarily involved in intraspecies signaling
2.
Autoinducer is a furanosyl borate diester involved in interspecies signaling
The structure of the 2 AI-2 signals are determined as 1.
furanosyl borate diester (BAI-2) used by V. harveyi to control luminescence
2.
furanone([2R,4SL]-2-methyl-2,3,3,4tetrahydroxytetrahydrofuran [R-THMF]) used by S. typhimurium AI-2 released by the bacterium accumulates in the cell’s environment
The only genes shown to be regulated by AI- 2 in other species encode for an ABC transporter in Salmonella typhimurium has system named Lsr (LuxSregulated), responsible for the AI-2 uptake (Taga et al., 2001). This ABC transporter is also present in E. coli and shares homology with sugar transporters (Figure 3a) The AI-3/epinephrine/norepinephrine signaling system Besides being used in bacterial interspecies signaling, AI-3 has an intrinsic role in interkingdom communication. AI-3 cross signals with the eukaryotic hormones epinephrine/ norepinephrine in an agonistic fashion (Sperandio et al., 2003). It was first found in as a compound found in spent media that activated the expression of genes involved in attachment of Entero Haemorrhagic E. coli (EHEC). The presence of AI-3 is also linked to the formation of attaching and effacing lesions by EHEC, loci of enterocyte effacement (LEE) operons located within the EHEC chromosome (Sperandio et.al 2003). Enterohemorrhagic E. coli colonizes the large intestine where it causes attaching and effacing (AE) lesions. The AE lesion is characterized by the destruction of the microvilli and the rearrangement of the cytoskeleton to form a pedestal-like structure, which cups the bacteria individually. Enterohemorrhagic E. coli senses AI-3 (produced by the normal
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Mechanism and manifestation of bacterial quorum sensing in food environment
Figure 2: The LuxS/AI-2 quorum-sensing system. (a) Vibrio harveyi (b) in Salmonella, E. coli
AI-2 receptor here is the LsrB (LuxS-regulated), periplasmic protein.
V. harveyi has two hybrid sensor kinases, LuxN and LuxQ sensing AI-1 and AI-2, respectively. LuxQ recognizes AI-2 complexed with its periplamic receptor LuxP.
↓ Upon binding to LsrB, AI-2 is transported inside the cell ↓ AI-2 is then phosphorylated by LsrK, and presumed to interact with LsrR.
↓ Upon sensing these signals, these kinases become phosphatases and the phosphorelay system (through LuxU and LuxO) is dephosphorylated.
↓ LsrR is a transcriptional repressor of the lsr operon.
↓ Consequently, LuxR mRNA is no longer degraded, and LuxR activates transcription of the luciferase operon.
↓ Upon complexing of Lsr to AI-2, LsrR no longer represses lsr transcription
Figure 3: Steps in the LuxS/AI-2 quorum-sensing system in V.harveyi
GI flora) and epinephrine/norepinephrine produced by the host to activate expression of the LEE genes and the flagella regulon (Sperandio et al., 2003). These signals are sensed by sensor kinases in the membrane of EHEC that relay this information through a complex regulatory cascade that activates the flagella regulon and the LEE pathogenicity island. The sensor for the flagella regulon is QseC that autophosphorylates in response to both epinephrine and
Figure 3a: Steps in the LuxS/AI-2 quorum-sensing system in E.coli
AI-3 and transfers its phosphate to the QseB response regulator, which in turn activates transcription of the flagella genes and itself (Clarke and Sperandio, 2005 ).
Repercussions of bacterial rendezvous In food spoilage In recent years, the detection of quorum sensing signals in
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spoiled food products has added a new dimension to study the process of food spoilage. Several proteolytic, lipolytic, chitinolytic, and pectinolytic activities associated with the deterioration of foods are regulated by quorum sensing. Several types of signaling molecules have been detected in different spoiled food products. Hence, disrupting the quorumsensing circuit can play a major role in controlling microbial gene expression related to human infection and food spoilage (Ragaert et al., 2007).Various signaling compounds, such as AI-1 and AI-2, have been detected in different food systems such as milk, meat, and vegetables (Bruhn et.al .,2004; Liu et.al., 2006; Pinto et.al., 2007). Milk: In Serratia proteamaculans strainB5a, the production of extracellular lipolytic and proteolytic enzymes is under AHLbased quorum-sensing system, which implies the involvement of quorum sensing in the spoilage of milk by Serratia spp. (Christensen et al., 2003), Similarly, the production of AHLs in both raw and pasteurized milk by the psychrotrophic bacteria Pseudomonas spp., Serratia spp., Enterobacter spp., and H. alvei shows that quorum sensing may play a role in the spoilage of milk and dairy products (Whitfield et al., 2000). Moreover, the detection of furanosyl BAI-2 signals in significant amounts in regular milk containing a low bacterial population (102 CFU/ mL) suggests the possible involvement of interspecies communication in milk spoilage (Lu et al., 2004). Nisin production in L. lactis has been studied extensively, and it has been shown that it acts as an autoinducer. It regulates its own production at the transcriptional level with the involvement of a two-component regulatory system (for a review, see Kleerebezem, M. 2004). An example of interspecies communication via QS among LAB is represented by L. plantarum NC8, in which not only plantaricin itself but also plantaricin-like peptides produced by other Gram-positive bacteria were shown to induce the production of plantaricin by L. plantarum. Fermented milk prepared by culturing the probiotic culture Leuconostoc mesenteroides, is a nutritious drink but at the same time a large number of spoilage microorganisms can grow in it when stored for a long time. The spoilage bacterial cultures are known to communicate with each other by release of signaling molecule which is described as quorum sensing. In a study of fermented milk, Pseudomonas was found to be responsible for spoiling the fermented milk. The signal molecule was identified as hexanoyl homoserine lactone. Two furanones namely 2(5H)-furanone and bromofuranone reduced acyl homoserine lactone and the shelf life was increased up to 9 days by reducing the spoilage bacterial cultures (Shobharani and Aggarwal, 2010). Meat: Jay et.al., in 2003 have reported the involvement of quorum sensing in the spoilage process of fresh meat products stored under aerobic refrigerated conditions and in the slime
formed on the meat surfaces. AHL signals, such as C4-HSL, 3oxo-C6-HSL, C6-HSL, C8-HSL, and C12-HSL, have been detected in aerobically chill-stored ground beef and chicken by the members of Pseudomonadaceae and Enterobacteriaceae (Liu et al., 2006). Additionally, food additives such as sodium propionate, sodium benzoate, sodium acetate and sodium nitrate may influence AI-2 production (Lu et al., 2004). In a study, Nychas et al., (2009) found that cellfree meat extract derived from spoiled minced pork meat stored aerobically at 5 and 20 °C contained QS signals. It was also observed, that the addition of cell-free meat extract from spoiled meat (containing QS signal molecules) to cultures of Pseud. fluorescens and Ser. marcescens resulted in an extension of the lag phase of Pseud. fluorescens but not of Ser. marcescens when compared to the control samples and in an increase of the metabolic activity for both the strains. The observed increase in metabolic activity was suggested to be related to the presence of some compounds in cell-free meat extract, including QS signal molecules (Nychas et al., 2009). Fruits and vegetables: Erwinia and Pseudomonas produce various pectinolytic enzymes, namely, pectin lyases, pectate lyase, polygalacturonase, and pectin methyl esterases, which are responsible for the spoilage of ready-to-eat vegetables, also produce a broad range of AHLs (mainly 3-oxo-C6-HSL and C6-HSL). Moreover, inoculation of bean sprouts with AHLproducing pectinolytic Pectobacterium carotovorum increased the rate of its spoilage (Rasch et al., 2005). The pectinolytic activity of Pseudomonadaceae or Enterobacteriaceae (mostly Erwinia spp.) growing to highcell densities (108 to 109 CFU g/L) in fruits and vegetables causes enzymatic browning, off-tastes, off-odors, and/or texture breakdown resulting in their spoilage. The pectinase of Erwinia carotovora is regulated by acylated homoserine lactone (Pirhonen et al., 1993), suggesting that rot is controlled by a quorum-sensing mechanism. In S. marcescens and S. liquefaciens secretion of several unrelated and potentially food-quality-relevant proteins such as the lipase LipA, the metalloprotease PrtA and the surface-layer protein (S-layer) SlaA (Riedel et al., 2001) is guided by QS.
In biofilms Food technologists and hygienists globally face the problem of presence of biofilms on foods and food contact surfaces, food processing environments including drains and floors, on fruits, vegetables, meat surfaces, and in low-acid dairy products often causing concerns for food safety. A defining feature of many biofilm-forming bacteria is the secretion of extracellular polymeric substances (EPS). The formation of biofilms begin with bacteria attaching on the surface, cell-to-cell aggregation and proliferation, exopolysaccharide matrix production,
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growth, maturation, and, finally, biofilm detachment or degradation. Quorum-sensing systems appear to be involved in all phases of biofilm formation. Many species, including the pathogen Pseudomonas aeruginosa, activate EPS production at high cell density (Hammer and Bassler, 2007). The foodborne pathogens E. coli O157:H7, Listeria monocytogenes, Yersinia enterocolitica, and Campylobacter jejuni are known to form biofilms on food surfaces and food contact equipment, leading to serious health problems, and economic losses (Kumar and Anand ,1998). Cells embedded in a biofilm are more resistant to cleaning agents and other antimicrobial substances, making them difficult to eradicate from processing equipment (Stoodley et al., 2002). Listeria monocytogenes biofilms were more resistant to cleaning agents and disinfectants including trisodium phosphate, chlorine, ozone, hydrogen peroxide, peracetic acid (PAA) and quaternary ammonium compounds (reviewed extensively by van Houdt et al., 2007). Bacteria in bioûlms show increased resistance towards various organic acids, ethanol and sodium hypochlorite. Resistance is attributed to different mechanisms: a slow or incomplete penetration of the biocide into the biofilm, an altered physiology of the biofilm cells, expression of an adaptive stress response by some cells, or differentiation of a small subpopulation of cells into persisting cells. Biofilms on processing equipment surface are a source of contamination for food products due to the continuous detachment of cells and spores from the biofilm. Moreover, biofilm formation may cause economic losses due to equipment failure or necessary extensive cleanup (Kumar and Anand, 1998). Microorganisms in bioûlms catalyze chemical and biological reactions causing metal corrosion in pipelines and tanks, and they can reduce the heat transfer efûcacy if bioûlms become sufûciently thick at plate heat exchangers and pipelines (Mittelman, 1998; Vieira et al., 1993). Most of the disinfectants used in cleaning are effective on free moving or planktonic bacteria. However, their effectiveness may decrease or may not be effective against bioûlm cells. If a microbial population faces high concentrations of an antimicrobial product, susceptible cells will be inactivated. However, some cells natural resistance or may acquire it mutation or genetic exchange. Such resistant bacteria can get into food contact surfaces and sometimes get into food too. Thus, there is a need to develop new control strategies especially the green strategies against biofilms. New control strategies are constantly emerging with main incidence in the use of biosolutions (enzymes, phages, interspecies interactions and antimicrobial molecules from microbial origin). Two questions need yet to be answered. (I) What parameters of a biofilm community inûuence the onset of quorum sensing (2) What are the functional consequences of quorum sensing in a bioûlm community.
In probiotics The probiotic research conducted over the past 20 years has resulted in a valuable source of data related to health beneficial effects of probiotics which are viable bacteria which when administered in adequate amounts confer to a health benefit to the host. Latest explanation of the beneficial effects of probiotics is quorum sensing. Quorum sensing regulates the virulence expression in probiotics which may interfere with the signalling system avoiding the onset of virulence in pathogenic bacteria. Symbiotic gut microorganisms release various soluble low molecular weight molecules of different chemical nature (surface and exogenous proteins, nucleases, lectins, peptides, amines, bacteriocines, fatty and amino acids, lactones, furanons, etc.). These molecules are able to sense environment, interact with corresponding cell surface, membrane, cytoplasm and nucleic acid receptors, to reply quickly and coordinately by induction of special sets of genes, to support stability of host genome and microbiome. General scheme of the intestinal bacterium-eucaryotic cells “cross-talk” can be summerized here : •
A quorum-sensing molecule is released by a bacterial population into the intestinal lumen, and is directly active on the intestinal epithelium.
•
Interactions between bacteria and the host cell surface require a direct contact.
•
“Modulins” are soluble factors produced by the bacteria that act directly on host cell functions.
•
Host cell biochemical pathways are specifically modified by these “modulins”,
•
Glycosylation modifications have an effect on the intestinal bacteria and on the intestinal epithelium defense mechanisms that help to fight against pathogens or virus infections (Freitas et al., 2003).
Further studies have nicely shown that several gut properties were primarily dependent on the cross talk between epithelium and bacteria (Lopez Boado et al., 2000). Probiotic L. acidophilus La-5 produces lactacin B when it senses live bacteria and the bacteriocin expression is controlled by an auto-induction mechanism involving the secreted peptide IP_1800 (Tabasco et al., 2009). In probiosies, the luxS gene appears to have a clear role in acidic stress response in probiotic lactobacilli easing their survival in gut (Moslehi-Jenabian et al., 2009). A recent report describes a bacterium isolated from the intestine gut of fish based on quorum sensing which has a probiotic characteristics by effectively reducing the amount of AHLs and the extracellular proteases activity of pathogen Aeromonas hydrophila (Chu et al., 2011)
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Figure 4: Strategies for quorum quenching
Gagging bacteria…… Quorum Quenching Interference with bacterial cell-to-cell signaling via the quorumsensing pathway to inhibit bacterial virulence and/or the development of biofilms is a new field of study with great potential applications. QS inhibitors are not involved in bacterial growth, inhibition of QS should not yield a strong selective pressure for development of resistance. A few examples of QSI are mentioned here. QSI, C-30, a derivative of a natural furanone was shown to inhibit QS and cause Pseudomonas biofilms to be susceptible to clearance by detergent and antibiotics when the biofilms were grown in its presence (Hentzer et al, 2003). Peterson et al (2008) recently showed that Apolipoprotein B, the major structural protein of lipoproteins, sequesters the QS signal AIP1, consequently inhibiting Agr-dependent virulence in MRSA isolates. Rasko et al (2008) carried out a HTS to identify a lead structure (N-phenyl-4-[[(phenylamino) thioxomethyl]amino]- benzenesulfonamide) LED209 which selectively blocked binding of signals (AI-3/ epinephrine and NE) to QseC, preventing QseC’s autophosphorylation, and consequently inhibiting QseC-mediated activation of virulence gene expression in EHEC O157. A remarkable study revealed by real-time RT-qPCR that the inhibition of virulence factors
regulation mechanisms by soluble molecules secreted by probiotics could represent an interesting way pathogenicity and virulence attenuation in Ps. aeruginosa nosocomial strains (Cotar et al., 2010). The anti-QS strategies developed so far have not yet been applied in a broad scale clinical trial and hence, it is difficult to assess their true potential and drawbacks at this stage. It is however certain that we need to expand our anti-microbial/anti-virulence targets and strategies. Pharmacologic interference of intercellular signaling can be envisioned at several steps in the quorum-sensing circuitry. Potential strategies include inhibiting receptor synthesis or function, reducing production or release of functional autoinducer, stimulating autoinducer degradation, or inhibiting autoinducer-receptor binding(Figure 4). In a study by Vattem and others (2007), bioactive dietary phytochemical extracts from common dietary fruit, herb, and spice extracts significantly inhibited quorum sensing in Chromobacterium violaceum. The extracts also inhibited swarming motility in pathogens EC O157:H7 and P. aeruginosa (PA-01), known to be modulated by quorum sensing. Vanilla, a widely used spice and flavour, can inhibit bacterial quorum sensing. Girennavar et al., (2008) have reported that natural furocoumarins from grapefruit juice act as the potent inhibitors of AI-1 and AI-2 activity and biofilm formation in S. typhimurium, EC, and P. aeruginosa. These results suggest that grapefruit juice can serve as a safe source of alternatives 110
Mechanism and manifestation of bacterial quorum sensing in food environment
to the halogenated furanones to develop strategies targeted at microbial quorum sensing. Thus, quorum-sensing inhibitors can also be used as food preservatives in selected foods where AHL-regulated traits are responsible for food spoilage (Bai and Rai, 2011). Biocontrol strategies that exploit bacterial QS provide an opportunity to •
Down regulate microbial activity and increase shelf life
•
Alter microbial activity such that survival of the targeted microorganism is unlikely.
The main advantages with this approach is: •
Does not leave open an opportunity for other undesirable microorganisms to colonize the niche
•
Down regulates the expression of enzymes such as proteases, thus limiting damage to the food.
•
Utilizes the bacteria’s own QS system against itself, thus decreases the possibility that the bacteria will adapt and become resistant to the QS inhibitor used.
Conclusion Far from being singular entities, it is now apparent that bacteria exist in multifaceted communities and are constantly communicating with each other. Microbes cooperate for nourishment, movement, virulence, iron acquisition, protection, quorum sensing, and production of multicellular biofilms or fruiting bodies. The study of bacterial quorum sensing has suggested several ideal targets for drug design as well as agricultural and food industrial applications. Food spoilage is the outcome of the biochemical activity of a microbial community attributed to quorum sensing (QS). Consequently, if QS is extensively elucidated it would be a futuristic tool in designing approaches for manipulating these communication systems, thereby reducing or preventing, for instance, spoilage reactions or even controlling the expression of virulence factors. The QS can be exploited for the benefit of food preservation and food safety. Thus, the communication between bacteria can be used as a medium for enhancing bacterial performance in food systems like fermentation. Conversely in actions where bacterial crosstalks lead to spoilage and biofilms, targeting the signaling molecules would be one of the “green” or “clean label” approach that is the need of the hour. References Bai, J., and Rai, V.R. 2011. Bacterial Quorum Sensing and Food Industry. ComprehensiveReviews inFood Scienceand Food Safety 10:184- 194.
Bassler, B.L. 1999. How bacteria talk to each other: regulation of gene expression by quorum sensing. Curr. Opin. Microbiol, 2:582–587. Bassler, B.L. 2002. Small talk: cell-to-cell communication in bacteria. Cell 109:421-424. Bruhn, J.B., Christensen, A.B., Flodgaard, L.R., Nielsen, K.F., Larsen T.O., Givskov, M. and Gram, L. 2004. Presence of acylated homoserine lactones (AHLs) and AHL-producing bacteria in meat and potential role of AHL in spoilage of meat. Appl Environ Microbiol, 70:4293–4302. Chen, X., Schauder, S., Potier, N., Van Dorssealaer, A., Pelczer, I., Bassler, B.L., and Hughson, F.M. 2002. Structural identiûcation of a bacterial quorum-sensing signal containing boron. Nature, 415:545–549. Christensen, A.B., Riedel, K., Eberl, L., Flodgaard, L.R., Molin, S., Gram, L. and Givskov, M. 2003. Quorum sensing-directed protein expression in Serratia proteamaculans B5a. Microbiology, 149:471–483. Chu W, Lu F, Zhu W. and Kang, C. 2011. Isolation and characterization of new potential probiotic bacteria based on quorum-sensing system. J Appl Microbiol. 110:202-208. Cotar AI, M C Chifiriuc, S Dinu, D Pelinescu, O Banu, V Lazar. 2010. Quantitative real-time PCR study of the influence of probiotic culture soluble fraction on the expression of Pseudomonas aeruginosa quorum sensing genes. Roumanian Archives Microb and Immu, 69:213- 223. Farghaly, A.H. 1950. Factors influencing the growth and light production of luminous bacteria. J. Cell. Comp. Physiol, 36:165–184. Freitas, M., Tavan, E., Thoreux, K., Cayuela, C., Sapin, C. and Trugnan, G., 2003. Lactobacillus casei DN-114 001 and Bacteroides hetaiotaomicron VPI-5482 inhibit rotavirus infection by modulating apical glycosylation pattern of cultured human intestinal HT29-MTX cells. Gastroenterology 124: A-475–A-476. Fuqua, W.C., Winans, S.C. and Greenberg, E.P. 1994. Quorum sensing in bacteria: the LuxR/LuxI family of cell density-responsive transcriptional regulators. J. Bacteriol, 176:269–275. Girennavar B., Cepeda, M.L., Soni, K.A., Vikram, A., Jesudhasan, P., Jayaprakash, G.K., Pillai, S.D. and Patil, B.S. 2008. Grapefruit juice and its furocoumarin inhibits autoinducer signaling and biofilm formation in bacteria. Intl J Food Microbiol, 125:204–208. Hammer, B.K., Bassler, B.L. 2007. Regulatory small RNAs circumvent the conventional quorum sensing pathway in pandemic Vibrio cholera, 104:11145-11149. Hammer, B., Bassler, B. 2003. Quorum sensing controls biofilm formation in Vibrio cholerae. Mol Microbiol, 50:101-104. Havarstein LS, Gaustad P, Nes IF and Morrison DA 1996 Identiûcation of the streptococcal competence-pheromone receptor. Mol Microbiol , 21:863–869 Hentzer M, Wu H, Andersen J B, Riedel K, Rasmussen TB, Bagge N, Kumar N, Schembri MA, Song Z, and Kristoffersen P, 2003. Attenuation of Pseudomonas aeruginosa virulence by quorum sensing inhibitors. EMBO J; 22:3803-3815. Jay JM, Vilai JP, and Hughes ME. 2003. Profile and activity of the bacterial biota of ground beef held from freshness to spoilage
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and physiological spoilage mechanisms during storage of minimally processed vegetables. Postharvest Biol Technol 44:185–194. Raina, S., De Vizio, D., Odell, M., Clements, M., Vanhulle, S., and Keshavarz, T. 2009. Microbial quorum sensing: a tool or a target for antimicrobial therapy? Biotechnol Appl Biochem, 54: 65-84. Rasch M, Rasmussen TB, Larsen T.O., Givskov M, and Gram L. 2005. Effect of quorum sensing inhibitors on food spoilage, 42–3 [abstract]. In: Institute of Food Technologists Annual Meeting. Chicago: Institute of Food Technologists. Rasko DA, Moreira CG, Li de R, Reading NC, Ritchie JM, Waldor MK, Williams N, Taussig R, Wei S., and Roth M, et al. 2008. Targeting QseC signaling and virulence for antibiotic development. Sci, 321:1078–1080. Riedel, K., Ohnesorg, T., Krogfelt, K. A., Hansen, T. S., Omori, K., Givskov, M. and Eberl, L. 2001. N-Acyl-L-homoserine lactonemediated regulation of the Lip secretion system in Serratia liquefaciens MG1. J Bacteriol 183: 1805–1809. Shaw, P.D., Ping, G., Daly, S. L., Cha, C., Cronan, J. E. Rinehart, Jr., K. L, and Farrand, S. K. 1997. Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography. Proc. Natl. Acad. Sci. 94: 6036-6041. Shobharani, P., Agrawal, R., 2010. Interception of quorum sensing signal molecule by furanone to enhance shelf life of fermented milk. Food Control, 21:61-69. Sifri, C.D. 2008. Quorum Sensing: Bacteria Talk Sense. Healthcare Epidemiology 47: 1070-1076. Sperandio V, Torres AG, Jarvis B, Nataro JP and Kaper JB. 2003. Bacteria-host communication: The language of hormones. Proc. Natl. Acad. Sci. USA 100: 8951–8956. Stoodley, P., Sauer K., Davies D.G., andCosterton JW. 2002. Biofilms as complex differentiated communities. Annu Rev Microbiol 56:187–209. Tabasco R, García-Cayuela T, Peláez C and Requena T. 2009. Lactobacillus acidophilus La-5 increases lactacin B production when it senses live target bacteria. Int J Food Microbiol. 132:109-116. van Houdt R, Moons P, Aertsen A, Jansen A, Vanoirbeek K, Daykin M, Williams P. and Michiels CW. 2007. Characterization of a luxI/luxR-type quorum sensing system and N-acylhomoserine lactone–dependent regulation of exo-enzyme and antibacterial component production in Serratia plymouthica RVH1. Res Microbiol., 158:150–158. Vattem DA, Mihalik K, Crixell SH and McLean RJC. 2007. Dietary phytochemicals as quorum sensing inhibitors. Fitoterapia, 78:302–310. Vieira, M. J., Melo, L. and Pinheiro, M. M. 1993. Bioûlm formation: hydrodynamic effects on internal diffusion and structure. Biofouling, 7:67-80. Whitfield, F.B., Jensen, N., Shaw, K.J. 2000. Role of Yersinia intermedia and Pseudomonas putida in the development of a fruity off-flavour in pasteurized milk. J Dairy Res., 67:561-159. Zhou, L, Hui, F.M. and Morrison, D.A. 1995. Characterization of IS1167, a new insertion sequence in Streptococcus pneumoniae. Plasmid. 33:127–138.
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Intl. J. of Food. Ferment. Technol. 2(2): 113-121, December, 2012
Review paper
Taste producing components in fish and fisheries products: A review Mohammed Golam Sarower*1, Abul Farah Md. Hasanuzzaman1, Bhabananda Biswas1 and Hiroki Abe2 1 2
Present address: Fisheries and Marine Resource Technology Discipline, Khulna University, Khulna 9208, Bangladesh Present address: Research Institute of Seafood Biochemistry, 3-23-20 Kamiigusa #205, Suginami, Tokyo 167-0023, Japan
*
Email:
[email protected]
Paper no: 46
Received: 15 Sept, 2012
Received in revised form: 19 Nov, 2012 Accepted: 14 Dec, 2012
Abstract The extractive constituents, which are known as taste active components of fisheries products in many research works have been reviewed..This review found glutamate, glycine, alanine, arginine, proline, valine, methionine, phenylalanine, tyrosine, inosine 5’-monophosphate (IMP), adenosine 5’monophosphate (AMP), guanosine 5’-monophosphate (GMP), trimethylamine, trimethylamine oxide (TMAO), glycine betaine, lactate, succinate as important contributors to the taste of raw and processed fisheries products. Sweet, salty, bitter, sour, and umami are the basic tastes, defined by these taste active components. Sweet taste is imparted by glycine, alanine, TMAO while bitter taste by arginine and other hydrophobic amino acids. Glutamate has a role in sour taste, and contributes to umami taste through synergetic effects in co-existence of IMP, GMP and AMP. The large amount of alanine or glutamate suppresses the sweetness effect of glycine through antagonistic effect. However, these taste-producing components vary with species, environments, various processing methods and relative quantities among them. ©2012 New Delhi Publishers. All rights reserved
Keywords: Fish, fisheries products, taste, umami
Fish and fisheries products with unique and diverse tastes are very popular and delicious foodstuffs to human being around the world. People of different cultures, countries and continents take these items for their daily food, and use as luxurious food item in various festivals. Fish, shrimps, prawns, crabs, and various mollusk species are consumed in fresh and various processed forms such as smoked, canned, marinated, dried, fermented, and condiments (e.g. fish sauce). These fresh and processed fisheries products are unique with its taste, texture and palatability, which are solely related to the contributions of extractive components. These taste-active components include several free amino acids such as glutamate, glycine, alanine, arginine, and nucleotides such as inosine 5’monophosphate (IMP), adenosine 5’-monophosphate (AMP), and guanosine 5’-monophosphate (GMP). Other than amino acids and nucleotides, several nucleosides and nucleic acid bases such as inosine, adenosine and uracil are found to be
taste-active in salted salmon eggs (Hayashi et al. 1990). Fuke and Konosu (1990) evaluated creatinine and lactate in dried skipjack, and succinate in short-necked clam as taste-active components. The contributions of extractive components to the sensory attributes and taste specificity of fish and fisheries products have been reported (Komata 1964; Konosu 1973; Hayashi et al. 1981; Konosu and Yamaguchi 1982; Konosu et al. 1988; Hayashi et al. 1990; Fuke and Konosu 1990, 1991; Fuke 1994; Shiau et al. 1996; Shirai et al. 1997 & 1996; Saito and Kunisaki 1998; Spurvey et al. 1998; Ninomiya 2002). These taste-active components differ with various processing methods, species, environments, and relative contents of taste-active components. Chiou et al. (2002) demonstrated the changes in extractive components such as adenosine 5’-triphosphate (ATP), -diphosphate (ADP), monophosphate (AMP), free
Sarower et al.
amino acids in the foot muscle of live small abalone (Haliotis diversicolor) during different storage temperatures. The amount of AMP, which varies with different cooking conditions, has been reported to be a factor related to the taste preference for the prawn muscle (Hatae et al., 1991) and soup of hard clam (Yamamoto and Kitao 1993). Fuke (1994) demonstrated that not only combinations of taste-active components but also the relative quantities among them are important in producing the specific flavor of specific seafood.
Composition of taste active components The specific taste of each food relies on extractive components which are defined as water-soluble, low molecular weight components, and classified into nitrogenous compounds (free and combined amino acids, nucleotides and related compounds, organic bases etc.) and non-nitrogenous compounds (sugars and organic acids) with the exception of vitamins, pigments and minerals. Noguchi et al., (1975), Miyazawa et al. ,(1979), and Hessel (1999) reported the effects of dipeptides on taste characteristics. According to Park et al., (2002b), peptides in fish sauce characterizing sour taste include Asp-Glu, Tyr-Pro and Val-Pro-Glu; peptides having rather flat taste are Gly-Pro-Orn-Gly, Asp-Phe, Glu-Phe and Glu-Met-Pro. Peptide having sweet taste is only Val-Pro and that having umami taste also includes only Asp-Met-Pro. Bitter peptides include Tyr-Pro-Orn, Val-Pro-Orn, Ala-Pro, Gly-Phe, Gly-Tyr and Phe-Pro (Table 1). Amino acids usually contribute a sour, bitter or sweet taste. Glycine and alanine impart pleasant sweet taste (Fuke and
Konosu 1991; Spurvey et al., 1998, Chen and Zhang 2007). Arginine is a bitter sensation producing amino acid with weak sweetness (Michikawa and Konosu 1995; Chen and Zhang 2007). Aspartate and glutamate have a sour taste but give umami taste in the presence of sodium salts (Yamaguchi et al. 1971). The umami taste of the Chinese mitten crab’s meat is significantly contributed by glutamate, IMP and AMP (Chen and Zhang 2007). According to Yamaguchi (1991), umami taste in association with monosodium glutamate (MSG) is elicited by glutamate. IMP and GMP are intensely flavor-enhancers of the umami taste, and have much stronger umami flavor enhanching capacitythan MSG (Yamaguchi et al., 1971).) AMP imparts umami taste to the muscle of squid (Kani et al. 2007; Shirai et al. ,1997). According to Lioe et al. (2004), phenylalanine and tyrosine are aromatic amino acids with a bitter taste, which are known to be important components of the savory taste of soy sauce in addition to glutamate. Phenylalanine is also found to enhance significantly the umami taste (Chen and Zhang 2007). The organic acids in fish and fisheries products are detected as lactic, acetic, malic, succinic acids, and fumarate (Storey and Storey 1983; Yamanaka et al. 1995; Itou et al., 2006; Kani et al. 2007). Glucose contributes a pleasant sweet taste to food (Chen and Zhang, 2007). Shirai et al., (1997) and Kani et al. (2007) reported trimethylamine oxide (TMAO) and trimethylamine (TMA) as taste-active components in squids. These TMAO and TMA contribute to sweetness and an agreeable characteristic squid flavor, respectively (Shirai et al. 1997). Glycine betaine is also identified as a taste-active
Table 1: Taste characteristics of oligopeptides identified in fish sauce and chemically synthesized (modified from Park et al. 2002b) Peptidesidentified Reported Tyr-Pro-Orn‡ Val-Pro-Orn‡ Gly-Pro-Orn-Gly‡ Ala-Pro† Asp-Glu† Asp-Pro‡ Asp-Phe† Glu-Pro‡ Glu-Phe‡ Gly-Phe† Gly-Tyr† Val-Pro† Tyr-Pro† Phe-Pro† Val-Pro-Glu²” Glu-Met-Pro²” Asp-Met-Pro²”
Flat Umami Sour Sour Flat Sour, bitter Bitter Bitter Flat Bitter Biter
Taste In the absence of NaCl
In the presence of 0.3% NaCl
Bitter Bitter Flat Bitter/Flat Sour/umami
Sweet/umami Umami/sweet Sweet/umami Sweet/umami Sweet/umami
Flat
Sweet/umami
Flat Bitter Bitter Sweet Sour Bitter Sour Flat Umami
Sweet/umami Sweet/umami Sweet Sweet/umami Sweet/umami Sweet/umami Sweet/umami Umami/sweet Sweet/umami
Concentration (mM) Reference value Tasted value
7.6
5
15-17 3 19 1.5
† Purchased from Bachem, Co.; ‡ Synthesized with a peptide synthesizer; ²” Synthesized by a liquid-phase method. 114
5 5 5 5 5
5 5 5 5 5 5 5 5 5
Taste producing components in fish and fisheries products: A review
component of G. borealis muscle (Shirai et al. 1997) and of Loliginidae squids (Kani et al. 2007). Michikawa et al. (1995) reported glycine betaine to impart only a weak sweetness but intensify thickness, fullness, and aftertaste of the synthetic extract of the whelk, Neptunea plycostata. Hayashi et al. (1978, 1979, 1981a,b) and Konosu et al., (1978) reported the contribution of sodium and chloride ions to the taste of snow crab. In addition to these, some inorganic ions such as Potassium and sodium could contribute salty taste to Chinese mitten crab meat flavor (Chen and Zhang 2007)
Taste evaluation Generally basic taste characteristics are classified as sweet, salty, bitter, sour and umami. The evaluation of taste-active components in relation to taste characteristics includes both omission and addition test. These tests with synthetic extracts simulating natural extracts are employed to identify the tasteactive components and to disclose their roles in producing the specific taste. In this type of tests, the triangle difference test is usually employed, but the paired difference test is also used to make sure the role of each taste-active component. The panel members are asked to answer to the difference between the synthetic extract and the synthetic extract from which one component or a group of components is omitted or added. At the same time, they are asked to evaluate the extracts usually by five-point rating scale for the items of basic tastes (sweetness, sourness, saltiness, bitterness and umami) and flavor characteristics (e.g., fullness, complexity, thickness, overall preference and so on), which are selected before sensory tests by an open panel discussion (Fuke and Konosu 1991). Apart from this commonly used evalution method, some other methods for fish taste preference have been addressed in the review by Kasumyan and Doving (2003).
Taste-active components of fish and fisheries products The taste-active components of fresh sea foods have been identified from abalone, scallop, short-necked clam, sea urchin, snow crab, salmon roe (Hayashi et al. 1990), Chinese mitten crab (Chen and Zhang 2007), Japanese spiny lobster and shovel-nosed lobster (Shirai et al., 1996), yellowtail (Kubota et al., 2002), and boreo-pacific gonate squid (Gonatopsis borealis) (Shirai et al., 1997), and other squids (Hatae et al. 1995a; Kagawa et al., 1999; Kani et al., 2007, 2008). There are also some research works on taste-active components in processed fisheries products such as dried skipjack (Fuke and Konosu 1991), fish sauce (Park et al., 2002a,b). Glutamate, proline, glycine, leucine, isoleucine, alanine, arginine, valine, methionine, IMP, GMP, AMP, TMAO and glycine betaine have been believed to be the taste active components in these fish and fisheries products.
According to Konosu (1973), extractive components such as glutamate, glycine, AMP, and glycine betaine are the tasteactive components of abalone. Betaine, glycine and alanine impart sweetness and intensify umami as well. Chiou and Lai (2002) reported the contribution of glycine, glutamate and AMP to the taste of small abalone. Glycine, glutamate, alanine, valine, methionine, IMP and GMP are taste-active components in sea urchin. Valine is necessary for producing bitterness intrinsic of the gonad of sea urchin. Methionine was essential to afford the characteristic flavor, which changed to crab- or prawn-like flavor without methionine (Komata 1964; Komata et al. 1962). Alanine, glutamate, glycine, arginine, glycine betaine, AMP in snow and mud crab (Chiou and Huang 2003; Hayashi et al. 1981a), and cytosine monophosphate, GMP and sodium, potassium, chloride, and phosphate ions in snow crab (Hayashi et al., 1981a) are the taste-active components. Arginine, glycine, alanine, glutamate, IMP and AMP had strong taste impacts on the crab’s meat flavor. Glycine and alanine contributed major sweet taste, while glutamate, IMP and AMP contributed a strong umami taste (Chen and Zhang 2007). Konosu et al. (1987, 1988) and Watanabe et al. (1990) have reported the role of glutamate, glycine, alanine, arginine, AMP, sodium, potassium and chloride ions in the umami taste of scallop. The taste-active components in short-necked clam are glutamate, glycine, arginine, taurine (Tau), AMP, succinic acid, sodium, potassium and chloride ions (Fuke and Konosu 1990). Succinic acid has been reported to be one of the main tasteactive components in some seafood, such as clam (Spurvey et al., 1998). Glycine, alanine, proline, arginine, glutamate, serine, threonine, AMP, glycine betaine, TMA and TMAO, NaCl, KCl, and KH2PO4- have been identified as taste-active components of boreo pacific gonate squid. Threonine and serine have been known to be a taste-active component only in boreo pacific gonate squid (Shirai et al., 1997). Glycine, alanine, proline, arginine, glutamate, AMP, TMAO, glycine betaine, potassium, sodium, and chloride ions are identified as taste-active components in squid species, Sepioteuthis lessoniana (Kani et al., 2008). Taurine, proline, glycine, alanine, and arginine are the main free amino acids commonly found in the muscle of these four squid species Sepioteuthis lessoniana, Loligo bleekeri, L. edulis, and Todarodes pacificus (Kani et al., 2007). Shirai et al., (1996) identified IMP and phosphate ions as tasteactive substances in spiny lobster P. japonicus. Valine and methionine have been revealed to be taste-active only in sea urchin and shovel-nosed lobster (Shirai et al., 1996). Glutamate, IMP, inosine, adenosine, guanosine, uracil, trimethyl-amine, glucose, sodium, potassium, chloride and phosphate ions have been found to be taste-active in salted salmon eggs (Hayashi et al., 1990), and in yellowtail muscle (Kubota et al., 2002). In fish sauce, glutamate, threonine, alanine, valine, histidine,
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proline, tyrosine, cystine, methionine and pyroglutamate have been proven to have a major contribution to its taste (Park et al., 2002a). The most effective compound for recreating the characteristic flavor of fish sauce is glutamate, followed by pyroglutamate and alanine. Proline and methionine contribute to sweetness. Threonine, alanine and histidine have been found to be responsible for the characteristic fish sauce-like taste by the omission test (Park et al., 2002a). Taira et al., (2007) identified free amino acids, which are mainly accumulated in the mashes of flyingfish (Cypselurus agoo agoo), the small dolphinfish (Coryphaena hippurus), and the deep-sea smelt (Glossanodon semifasciatus), during fermentation as leucine, lysine, valine, and alanine, methionine and isoleucine. The deep-sea smelt was found to contain a large amount of glutamate. Glutamate, histidine, lysine, carnosine, IMP, inosine+hypoxanthine, creatinine, lactic acid, sodium, potassium and chloride ions are evaluated as taste-active components of dried skipjack (Fuke et al. 1989). Kodama (1913) first identified IMP as a responsible substance of the savory taste of “Katsuobushi” stock (dried and molded skipjack). According to Fuke and Konosu (1991), lactic acid has been evaluated to be a taste-active component in dried skipjack tuna.
Factors affecting taste characteristics Tastes are specific and varied with respect to species, different concentrations of taste-producing components in different parts of the body, chemical structure and relative portion of taste-active components, season and species-habitat as well. Variation in taste: It is notable that differences in the concentrations of these taste-active components lead to the characteristic tastes of different species. Kani et al., (2008) have demonstrated that saltiness and umami are weaker in Loligo sp. than in Sepioteuthis lessoniana but sweetness, sourness, and bitterness are evaluated to be stronger in L. bleekeri than in S. lessoniana. In Todarodes pacificus, umami taste is significantly stronger, while sweetness, sourness, and saltiness are all weaker than in S. lessoniana. There is also difference in taste of body portion due to varied contents of taste-active components such as free amino acids, nucleotide and related compounds, organic acids, and inorganic ions. The muscle of Loliginidae squid species, S. lessoniana, L. bleekeri, and L. edulis is considered to have a much sweeter taste than that of T. pacificus and G. borealis on account of the high contents of sweet amino acids, glycine, alanine, and proline. Compared with muscle, squid liver with high contents of taurine, glutamate, bitter amino acids, succinate, propionate, TMA, and glycine betaine, and with low contents of sweet amino acids, arginine, nucleotides, malate, and TMAO are characterized by a complicated taste containing umami,
bitterness, sourness, fishy flavor, and less sweetness. Liver may have more complicated flavor than muscle. If squid liver is mixed with muscle during cooking, it may give the muscle a more complicated taste than the sweetness and umami taste of muscle (Kani et al., 2007). They also reported higher phosphate ion in muscle than in liver; highest in the muscle of S. lessoniana, and lowest in that of T. pacificus. Diets with different composition may cause variation in taste specificity. Many studies showed the effect of different diets on muscle yield (Mercer et al. 1993) and proximate composition (Mercer et al., 1993; Watanabe et al. 1993; Britz and Hetch 1997), lipid composition (Dunstan et al., 1996), extractive components (Watanabe et al. 1993; Mai et al. 1994; Bewick et al., 1997), and glycogen (Watanabe et al., 1993) in the tissues of abalone. Chiou and Lai (2002) reported the differences in taste preference and the levels of extractive components and glycogen in cooked meats of small abalone Haliotis diversicolor fed gracilaria sp. and artificial diet. The contents of glutamate, glycine, and AMP are determined to vary considerably among muscle of abalone fed different diets (Chiou et al., 2001). In addition to diet, seasonal variation is also a factor causing changes in the composition of extractive components, and thus taste preferences of seafood. According to Konosu and Yamaguchi (1982), season is one of the factors influencing the contents of extractive components and glycogen of seafood. It has been reported that the extractive components and glycogen in oyster (Takaki and Simidu 1963; Sakaguchi and Murata 1989), ascidian (Watanabe et al., 1983, 1985; Park et al., 1990), clams (Hirano 1975; Chiou et al., 1996a, 1996b), scallop (Kawashima and Yamanaka 1996) and abalone (Weber 1970; Hirano 1975; Watanabe et al., 1992a; Hatae et al. 1995b; Hwang et al., 1997; Chiou et al., 2001) show marked seasonal changes, and that in most cases the season for higher palatability coincides with a higher contents of taste components. The levels of non-protein nitrogenous compounds in the fish and shellfish, such as milkfish (Chiou et al., 1995), ayu (Hirano and Suyama 1980), and yellowtail (Endo et al., 1974) are well determined to show seasonal variations. Chiou and Huang (2003) observed the increased total free amino acid and individual free amino acid such as glycine, alanine, and arginine in female crab (Scylla serrata) during August and November, while those in male crab in January, March, and August. The total ATP-related compounds in both crabs have been lower in the crabs collected from winter than in other seasons whereas higher glycine betaine in winter and early spring. Glycogen in female crab is reported to be higher in October and August, while in male crab muscle increased level of glycogen is observed from August through January, but decreased in spring. The levels of total taste active components including glycine, alanine, arginine, GMP, IMP,
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and AMP have been found to differ greatly in the muscle of puffer (Takifugu rubripes) between the season, and probably, the geographical location (Hwang et al., 2000). The level of these compounds is lower in the spawning season (MarchMay), but much higher from July to January when the puffer is more palatable. The storage condition and the methods of processing could change the level of taste-active components of fish and fisheries products, and thus influence the taste characteristics. Watanabe et al., (1992b) demonstrated slightly increased amount of total free amino acid in the muscle of disk abalone after one day of storage at 5 oC and 10 oC and after 5 days of storage at 0 oC, and thereafter decreased markedly. Wongso and Yamanaka (1996) have shown that the total amount of free amino acid in the adductor muscle of noble scallop increases gradually during storage at 0 oC and 5 oC prior to the initial decomposition stage. Chiou et al., (2002) have reported that the total amount of free amino acids in small abalone increases markedly during storage at 5 oC, 15 oC, and 25 oC. The dominant free amino acids, such as taurine, glutamate, glycine, alanine, and arginine are found to be increased after storage. The pH decreases and the volatile basic nitrogen and K-value increase during the storage in different temperature, while changes were prominent with the course of rising of temperature (Figure 1). Several reports have demonstrated that increased levels of extractive components after cooking can be attributed to flavor in abalone and kuruma prawn meats (Hatae et al., 1996; Matsumoto et al.,1991), as well as soup of hard clam (Yamamoto and Kitao 1993). During fermentation, increased free amino acids and organic acids are determined in fish sauces produced from three fish species, the flyingfish Cypselurus agoo agoo, the small dolphinfish Coryphaena hippurus, and the deepsea
smelt G. semifasciatus. It is found these products to have lower smell, saltiness, and bitterness, and higher sweetness and umami taste than a Vietnamese fish sauce, Nuoc mam (Taira et al., 2007). It has been reported that glutamate, aspartate, glycine, alanine, leucine, and isoleucine, and organic acids in fermented mackerel and rice product, narezushi, get increased which implicate the contribution of these components to the umami taste and the sour taste of narezushi (Itou et al., 2006). Itou and Akahane (2000) observed markedly increased glutamate, leucine, lysine, aspartate, and alanine in fermented mackerel with rice bran, heshiko. A lightly fermented mackerel with rice, sabazushi, is also reported to contain alanine, leucine, and valine abundantly (Chang et al., 1992). Intensification and/or suppression of taste: It has been known that the taste strength and quality of an original extract can be easily changed or improved by the addition and/or omission of extractive components. There is also synergistic or antagonistic effect observed among several taste-active components. The results from Kani et al., (2008) showed that the addition of glycine increases sweetness and umami tastes of squid. The incorporation of alanine, serine, and proline with glycine would lead to a further increased difference in the concentration of sweet amino acids in cooked abalone meats (Chiou and Lai 2002). Kani et al., (2008) have also reported that arginine intensifies the sweetness and suppresses the bitterness and umami of the squid (L. bleekeri) extract. Addition of chloride ion improves the taste of an unpalatable squid, T. pacificus. There is a synergistic effect between creatine and creatinine when they were found to be coexisting in fish sauce (Park et al., 2002a). Yamaguchi et al., (1971) reported a synergistic effect between MSG and IMP, GMP or AMP, which together in certain ratios produce a strong umami taste.
Figure 1: Changes in levels of (¡%) pH, (Ë%) volatile basic nitrogen (VBN), and (Ï%) K-value in the foot muscle of abalone during storage at different temperature (Chiou et al. 2002).
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According to Lioe et al. (2004, 2005) free aromatic amino acids such as phenylalanine and tyrosine play an important role in enhancing savory or umami taste at their sub-threshold concentrations in the presence of salt and free acidic amino acids. Kani et al., (2007) speculated improved umami taste of squid muscles through synergistic effect between AMP and glutamate. This effect provides the muscle with not only umami taste but also such taste sensory attributes as continuity, complexity, mouthfulness and palatability (Komata 1990; Fuke 1994). According to Konosu (1973), umami and characteristic flavor almost disappear when either glutamate or AMP is omitted from extract, whereas the absence of glycine results in reduced sweetness and umami taste. The sweetness of glycine may be masked by large amounts of alanine and/or glutamate (Park et al., 2002a). In the case of the omission of threonine, slightly increased umami taste is found but the overall taste decreases. Umami and sweetness decrease slightly and bitterness and astringent taste increase a little with the omission of alanine. Another taste-active component is valine, of which omission decreased umami and sweetness, and slightly increased sourness are reported. The omission of tyrosine and cystine impart a little sweetness to food taste (Park et al., 2002a). The bitterness of arginine can be masked by NaCl, glutamate, and/or AMP (Michikawa and Konosu 1995). The organic acid omitted solution show a clear decrease of umami and sweetness in fish sauce and a slight decrease of mouthfulness. The pyroglutamate-omitted solution showed a decrease of sweetness and gave a flat taste (Park et al., 2002a). The omission of potassium, sodium, and chloride ion is found to lead to a large change in taste of squid, including a decrease in sweetness, saltiness, and umami (Kani et al., 2008). Sodium ion is well known to intensify saltiness and sweetness and suppresses sourness and bitterness. If chloride ion is omitted from an extract, sweetness and umami tastes disappear, bitterness is significantly increased, and the quality of taste gets significantly declined in boiled snow crab meat (Hayashi et al., 1981b). Although glycogen is tasteless, it has been reported previously that the addition of glycogen to synthetic extracts of abalone and scallop improved flavor characteristics and overall taste preference (Konosu et al., 1988; Konosu 1973).
Conclusion In summary, the taste of fish and fisheries products as well as by-products is the function of the taste effects of many tasteactive components such as glutamate, glycine, alanine, arginine, proline, valine, methionine, phenylalanine, tyrosine, IMP, AMP, GMP, TMAO, TMA, glycine betaine, inosine, adenosine, uracil, creatine, lactate, succinate. The taste is also found to be varied with the existence of sodium, chloride,
potassium, and phosphate ions in fisheries products. Usually, glycine, alanine, TMAO and glycine betaine impart sweet taste in invertebrates while arginine contribute bitter test. Glutamate gives umami taste through synergetic effects in co-existence of IMP, GMP, AMP. On the other hand, there is also evidence of antagonistic effect among several taste-active components. Large amounts of alanine or glutamate mask the sweetness effect of glycine, and NaCl, glutamate, and/or AMP suppress the bitterness of arginine. Thus, the tastes can be modified by simply addition or omission of taste extractive components. However, the taste strength of taste-active components coincides well with species, environments, various processed methods, and proportion of taste-active components in the products. We believe that, though we recognize the taste of fish and fisheries products with five sensory flavour, still there are lots of scope to find out the modifications and even novelty of taste. We have shown in this short review that a little change in biochemical compositions, sex, storage time, temperature could change the taste of fish and fisheries products. The future works, therefore, should draw the scientists’ attention to work on it with the climate changes, genetic inheritance using advanced tools such as molecular applications in the cellular level and biochemistry of the taste-active components of fish and fisheries products.
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Miyazawa, K., Le, C., Ito, K. and Matsumoto, F. (1979). Studies on fish sauce. J. Facul. Appl.. Bio. Sci. Hiroshima Uni., 18: 5563. Ninomiya K. 2002. Umami: a universal taste. Food Rev. Int., 18(1): 23-38. Noguchi, M., Arai, S., Yamashita, M. Kato, H. and Fujimaki, M. 1975. Isolation and identification of acidic oligopeptides occurring in a flavor potentiating fraction from a fish protein hydrolysate. J. Agricul. Food Chem., 23:49-53. Park, C.K., Matsui, T., Watanabe, K., Yamaguchi, K. and Konosu, S. 1990. Seasonal variation of extractive nitrogenous components in ascidian Halocynthia roretzi tissues. Nippon Suisan Gakkaishi, 56: 1319-1330 (in Japanese). Park, J.N., Watanabe, T., Endoh, K.I., Watanabe, K. and Abe, H. 2002a. Taste-active components in a Vietnamese fish sauce. Fish. Sci., 68:913-920. Park, J.N., Ishida, K., Watanabe, T., Endoh,K.I., Watanabe, K., Murakami, M. and Abe, H. 2002b. Taste effects of oligopeptides in a Vietnamese fish sauce. Fish. Sci., 68:921928. Saito, M. and Kunisaki, N. 1998. Proximate composition, fatty acid composition, free amino acid contents, mineral contents, and hardness of muscle from wild and cultured puffer fish Takifugu rubripes. Nippon Suison Gakkaishi, 64:116-120. Sakaguchi, M. and Murata, M. 1989. Seasonal variations of free amino acids in oyster whole body and adductor muscle. Nippon Suisan Gakkaishi, 55: 2037-2041 (in Japanese). Shiau, C.Y., Pong, Y.J., Chiou, T.K. and Chai, T. 1996. Free amino acids and nucleotide-related compounds in milkfish (Chanos chanos) muscles and viscera. J. Agricul. Food Chem., 44: 2650-2653. Shirai, T., Hirakawa, Y., Koshikawa, Y., Toraishi, H., Terayama, M. and Suzuki, T. 1996. Taste components of Japanese spiny and shovel-nosed lobsters. Fish. Sci, 62:283-287. Shirai, T., Kikuchi, N., Matsuo, S., Uchida, S., Inada, H., Suzuki, T. and Hirano, T. 1997. Taste components of boreo pacific gonate squid Gonatopsis borealis. Fish. Sci., 63: 772–778. Spurvey, S., Pan, B.S. and Shahidi, F. 1998. Flavour of shellfish. In: ‘Flavor of meat, meat products, and seafoods’ F. Shahidi, Ed. p. 159-196 Blackie Academic & Professional, London. Storey, K.B. and Storey, J.M. 1983. Carbohydrate metabolism in cephalopod molluscs. In: ‘The Mollusca’ P.W. Hochachka, Ed. vol 1. p. 91-136 Academic Press, New York. Taira, W., Funatsu, Y. Satomi, M. Takano, T. and Abe, H. 2007. Changes in extractive components and microbial proliferation during fermentation of fish sauce from underutilized fish species and quality of final products. Fish. Sci., 73: 913-923. Takaki, I. and Simidu, W. 1963. Seasonal variation of chemical constituents and extractive nitrogenous in some species of shellfish. Nippon Suisan Gakkaishi, 29: 66-70 (in Japanese). Watanabe, K., Maezawa, H. and Konosu, S. 1983. Seasonal variation of extractive nitrogen and free amino acids in the muscle of ascidian Halocynthia roretzi. Nippon Suisan Gakkaishi, 49: 1755-1758 (in Japanese). Watanabe, K., Maezawa, H., Sato, M. and Konosu, S. 1985. Seasonal variation of extractive nitrogenous components in the muscle of ascidian Halocynthia roretzi. Nippon Suisan Gakkaishi,
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Intl. J. of Food. Ferment. Technol. 2(2): 123-136, December, 2012
Review paper
Antimicrobial, antioxidant and phyto-chemicals from fruit and vegetable wastes: A review V. K. Joshi, Ashwani Kumar and Vikas Kumar Department of Food Science and Technology, Dr Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh – 173 230, India *
Email:
[email protected]
Paper no: 47
Received: 04 Sept, 2012 Received in revised form: 01 Nov, 2012
Accepted: 03 Nov, 2012
Abstract Food processing industry including fruit and vegetable processing is the second largest generator of wastes into the environment only after the household sewage. The generation of biodegradable waste increased linearly with the growth and development of food processing industry. A huge amount of waste in the form of liquid and solid is produced in the fruit and vegetable processing industries is valuable but biodegradable natural resources with large economic potential. It causes pollution problem if not utilized or disposed-off properly. The waste obtained from fruit processing industry is extremely diverse due to the use of wide variety of fruits and vegetables, the broad range of processes and the multiplicity of the product. Different fruits and vegetable possess various quantities of waste. Waste product which is thrown into the environment has a very good antimicrobial and antioxidant potentiality. These are novel, natural and economic sources of antimicrobics and antioxidants, which can be used in the prevention of diseases caused by pathogenic microbes. These all benefits will open up as a scope for future utilization of the waste for therapeutic purpose. However, lack of pilot testing of the developed technologies, negative attitude of the industrialists and perhaps, less helping hand from the government sector are the major constraints in utilization of the waste. © 2012 New Delhi Publishers. All rights reserved
Keywords: Vegetable, utilization, environment,antioxidant,antimicrobial,waste, food processing.
Introduction The growing, processing and preparation of food result in the production of varying degree of waste material. The waste material may be in the form of leaf/straw, waste during harvesting, processing industry waste and after processing waste (Joshi and Devraj, 2008). The waste obtained from fruit processing industry is extremely diverse due to the use of wide variety of fruits and vegetables, the broad range of processes and the multiplicity of the product (William, 2005). Vegetables and some fruits yield between 25% and 30% of non-edible products (Ajila et al., 2010). The full utilization of horticultural produce is a requirement and a demand that needs to be met by countries wishing to implement low-waste technology in their agribusiness (Kroyer, 1995). Depending
on plant species, variety and tissue, high levels of healthprotecting antioxidants, such as vitamin C and E, phenolic compounds including phenyl-propanoids and flavonoids, and or carotenoids such as lycopene can be found. The waste materials such as peels, seeds and stones produced by the fruit and vegetable processing can be successfully used as a source of phytochemicals and antioxidants. The entire tissue of fruits and vegetables is rich in bioactive compounds, such as phenolic compounds, carotenoids, vitamins and in most cases, the wasted by products can present similar or even higher contents of antioxidant and antimicrobial compounds than the final produce can (Ayala-Zavala et al., 2010). The new aspects concerning the use of these wastes as byproducts for further ex ploitation on the production of food
Joshi et al.
additives or supplements with high nutritional value have gained increasing interest because these are highvalue products and their recovery may be economically attractive. The by-products represent an important source of sugars, minerals, organic acid, dietary fibre and phenolics which have a wide range of action which includes antitumoral, antiviral, antibacterial, cardioprotective and antimutagenic acti-vities (Jasna et al., 2009). Because of increasing threat of infectious diseases, the need of the hour is to find natural agents with novel mechanism of action. Natural products provide unlimited opportunities for new drug leads because of the unmatched availability of chemical diversity. Fruit and vegetable peels are thrown into the environment as agro waste which can be utilized as a source of antimicrobics. Utilisation of by-products is, however, limited due to the poor understanding of their nutritional and economic value (Schroeder, 1999).
According to Indian Agricultural Research Data Book (2012), the estimated area (In 000’HA) of fruits and vegetables is 6383 and 8495 and the production (In 000’MT) is 74878 and 146554 (Table 1). The extent of total losses in these commodities is approximately estimated as 20-30% of the total production, amounting to a loss of Rs. 30,000 crores per annum. Sliced apples produced 10.91% of pulp and seed (core) by-products and 89.09% of the final products. Peeled mandarins produced 16.05% of peels and 83.95% of final products. Diced papayas produced 6.51% of seeds, 8.47% of peels, 32.06% of unusable pulp (due to the lack of shape uniformity in a cube), and 52.96% of final products. Pineapples produced 9.12% of core, 13.48% of peels, 14.49% of pulp, 14.87% of top, and 48.04% of finished products. Mangoes produced 13.5% of seeds, 11% of peels, 17.94%
Table 1: Area and Production estimates for Horticulture crops (Area in 000' HA, Production in 000' MT and Productivity = MT/HA) 2008-09 Area
Production
2009-10 Pdy.
Area
Production
2010-11 Pdy.
Area
Production
Pdy.
Fruits Banana Mango Citrus Papaya Guava Apple Pineapple Sapota Grapes Pomegranate Litichi Others
709 2309 924 98 204 274 84 156 80 109 72 1083
26217 12750 8623 3629 2270 1985 1341 1308 1878 807 423 7234
37.0 5.5 9.3 37.1 11.1 7.2 16.0 8.4 23.6 7.4 5.9 6.7
770 2312 987 96 220 283 92 159 106 125 74 1105
26470 15027 9638 3913 2572 1777 1387 1347 881 820 483 7201
34.4 6.5 9.8 40.9 11.7 6.3 15.1 8.5 8.3 6.6 6.5 6.5
830 2297 846 106 205 289 89 160 111 107 78 1265
29780 15188 7464 4196 2462 2891 1415 1424 1235 743 497 7583
35.9 6.6 8.8 39.6 120 10.0 15.9 8.9 11.1 6.9 6.4 6.0
Fruits-Total
6101
68466
112
6329
71516
11.3
6383
74878
11.7
Vegetables Potato Tomato Onion Brinjal Tapioca Cabbage Cauliflower Okra Peas Sweet Potato Others
1828 599 834 600 280 310 349 432 348 124 2275
34391 11149 13565 10378 9623 6870 6532 4528 2916 1120 28006
18.8 18.6 16.3 17.3 34.3 22.1 18.7 10.5 8.4 9.0 123
1835 634 756 590 232 331 338 452 365 119 2332
36577 12433 12159 10165 8060 7281 6410 4803 3029 1095 31724
19.9 19.6 16.1 17.2 34.8 22.0 19.0 10.6 8.3 92 13.6
1863 865 1064 680 221 369 369 498 370 113 2083
42339 16526 15118 11896 8076 7949 6745 5784 3517 1047 27557
22.7 19.1 142 17.5 36.5 21.5 18.3 11.6 9.5 9.3 132
Veg.-Total
7981
129077
162
7985
133738
16.7
8495
146554
17.3
Source: Anonymous,2011-2012 124
Antimicrobial, antioxidant and phyto-chemical properties of fruit and vegetable wastes: A review
unusable pulp, and 57.56% of final products. It is emphasized that considerable amounts of fruit material are the by-products of the minimal processing, and the possibility of creating alternative processes to give added value to this wasted material must be considered (Ayala-Zavala et al., 2010). While according to FAO (2003), the total waste generated from fruits was estimated as 3.36 million tonnes (MT) out of the total production of 16.8 MT and particularly for banana it was 6.4 MT. India is producing 3 million tonnes of citrus fruits like mandarins, lime, lemon, and sweet orange. Citrus wastes are rich source of oil, pectin and variety of by-products. The failure or inability to salvage and reuse such materials economically results in the unnecessary waste and depletion of natural resources (Bhalerao et al., 1989). The extent of the waste produced and available from processing industries of some of the important fruits and vegetables, is given in Table 2 and 3. Table 2: Quantities of various fruit and vegetable processing wastes Commodity
Percent weight basis
Apple Apricot Grape fruit Orange Peach Pear Asparagus Bean, green Beet Broccoli Cabbage Carrot Cauliflower Peas Potatoes Spinach Sweet potato Tomato
12-47 8-25 3-58 3 11-40 12-46 3.2-30 5-20 7-4 20 5-25 18-52 8 6-79 5 10-40 15 5-25
Chemical composition The amount of pollution load and characteristics of the waste depend on the food being processed. Chemical composition of the wastes from fruits and vegetables show that it is a rich source of various nutrients. Some of these fruit and vegetable wastes are a rich source of vital constituents like carbohydrates, proteins, fats, minerals, fibres etc. Nutrient composition of some of the solid wastes from fruits and vegetables is given in the Table 4. The association between the diet rich in fruits and vegetables and a decreased risk of cardiovascular diseases and certain forms of cancer is supported by considerable epidemiological evidence (Ness and Powles, 1997; Riboli and Norat, 2003). Different studies have shown that free radicals present in the human organs cause oxidative damage to various molecules, such as lipids, proteins and nucleic acids, and are thus involved in the initiation phase of the degenerative diseases. Phenolic and other phytochemical antioxidants found in fruits and vegetables are capable of neutralising free radicals and may play a major role in the prevention of certain diseases (Kaur and Kapoor, 2001). Numerous studies have provided evidence for decreased risk of some chronic dis-eases e.g., some types of cancer, cardiovascular and neurodegenerative disorders with increased dietary intake of vegetables, fruits, teas, spices and other plant-based foods and supplements. The most abundant by-products of minimal processing of fresh-cut fruit and vegetable are peel and seed and those are reported to contain high amounts of phenolic compounds with antioxidant and antimicrobial properties (Shrikhande 2000; Muthuswamy et al., 2008; Tuchila et al., 2008).
Phytochemicals Plants synthesize a diverse array of secondary metabolites (phytochemicals) known to be involved in plant defence against microbial and fungal pathogens and insect pests, and in the last few decades several classes of phytochemicals have
Source: Gera IB and Kramer, 1969 Table 3: Fruits and vegetable processing wastes available in India Vegetable
Nature of waste
Mango Banana Citrus Pineapple Grapes Guavas Peas Tomato Potato Onion Apple
Peel, stones Peel Peel, rag and seed Skin, core Stem, skin and seeds Peel and core and seeds Shell Skin, core and seeds Peel Outer leaves Peel, pomace and seeds
Production(content)(tones)
Approx.waste(%)
Potentialquantities ofwaste (tones)
6987.7 2378.0 1211.9 75.7 565 565 107.7 464.5 2769.0 1102.0 1376.0
45 35 50 33 20 10 40 20 15 -
3144.4 832.3 6-06.0 24.7 68.3 90.3 415.3 412.0
Source: Gupta and Joshi, 2000. 125
Joshi et al.
Table 4: Composition of different fruit wastes (per 100g) Waste
Moisture(g)
Apple pomace Mango seed kernel Jack fruit (inner andouter portion Jack fruit seeds Jack seed flour Passion fruit peel Banana peel Sweet orange seeds Watermelon seeds Muskmelon seeds Pumpkin seeds Banana stem Central core Outer hardFibrous sheath Press juice from stem
Protein(g)
Fat (g)
Minerals(g)
Fibre(g)
Carbohy-drate (g)
_ 8.2 8.5 64.5 77.0 81.9 79.2 4.00 4.3 6.8 6.0
2.99 8.50 7.50 6.60 2.64 2.56 0.83 15.80 34.10 21.00 29.50
1.71 8.85 11.82 0.40 0.28 0.12 0.78 36.90 52.60 33.00 35.40
1.65 3.66 6.50 1.20 0.71 1.47 2.11 4.00 3.70 4.00 4.55
16.16 _ 30.77 1.50 1.02 5.01 1.72 14.00 0.80 30.00 12.00
17.35 74.49 14.16 25.80 18.12 _ 5.00 _ 4.50 _ 12.53
93.1 91.9 98.6
0.30 0.12 0.05
0.03 0.06 _
1.04 0.98 0.63
0.68 1.81 _
1.20 2.44 0.41
Source: Maini and Sethi, 2000.
been shown to help reduce the risk of various diseases e.g. cancer and coronary heart disease. Nowadays, there is a growing interest in finding phytochemicals as an alternative to synthetic substances, which are commonly used in the food, pharmaceutical and cosmetic industry. Epidemiological studies have pointed out that the consumption of fruits and vegetables imparts health benefits, e.g. reduced risk of coronary heart disease and stroke, as well as certain types of cancer. Apart from dietary fibre, these health benefits are mainly attributed to organic micronutrients such as carotenoids, poly-phenols, tocopherols, vitamin C and others. Flavonoids from fruits and vegetables probably reduce risks of diseases associated with oxidative stress, including cancer. Apples contain significant amounts of flavonoids with antioxidative potential. The products and byproducts obtained during the minimal processing of the fruits were analyzed for the phytochemical content and antioxidant status. It was found that the total phenolics and flavonoid contents were higher in the byproducts as compared to the final products, being more pronounced in mango seeds and peels. These compounds could be responsible for free radical inhibition activity. Several studies have shown that the content of phytochemical compounds is higher in peel and seeds with respect to the edible tissue. Gorinstein et al., (2001) found that the total phenolic compounds in the peels of lemons, oranges, and grapefruits were 15% higher than that of the pulp of these fruits. Peels from apples, peaches, pears as well as yellow and white flesh nectarines were found to contain twice the amount of total phenolic compounds as that contained in fruit pulp (Gorinstein et al., 2001). While the edible pulp of bananas (Musa paradisiaca) contains 232 mg/100 g of dry weight
phenolic compounds, this amount is about 25% of that present in the peel (Someya et al., 2002). Similarly, other studies have reported that pomegranate peels contain 249.4 mg/g of phenolic compounds as compared to only 24.4 mg/g phenolic compounds found in the pulp of pomegranates. Apple peels were found to contain up to 3300 mg/100 g of dry weight of phenolic compounds (Wolfe and Liu 2003). Grape seeds and skins, the byproducts of grape juice and white wine production, are also sources of several phenolic compounds, particularly mono, oligo, and polymeric proanthocyanidins (Shrikhande, 2000). It has been reported that the total phenolic compounds of seeds of several fruits, such as mangos, longans, avocados, and jackfruits, were higher than that of the edible product, and that the byproducts could be a valuable source of phytochemicals (Soong and Barlow, 2004). The peels and seeds of tomatoes are richer sources of phenolic compounds than the pulp of the tomatoes are. The phenolic compounds of 12 genotypes of tomatoes have been studied, and, in general, lower levels were found in the flesh, ranging from 9.2 to 27.0 mg/100 g, as compared to 10.4 to 40.0 mg/100 g in the peels (George et al., 2004). A similar observation was reported, and the total phenolic compounds (expressed as milligram of gallic acid equivalents per 100 g) of the skin, seeds, and pulp of tomatoes were found to be 29.1, 22.0, and 12.7 mg/100 g, respectively (Toor and Savage, 2005). It was also found that the peel byproduct of tomato cultivars (Excell, Tradiro, and Flavorine) had significantly higher levels of total phenolic compounds, total flavonoids, lycopene, ascorbic acid, and antioxidant activity as compared with the pulp and seeds (Toor and Savage, 2005). In general, there are up to 10-fold higher occur between the phenolic contents of byproducts than the
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Antimicrobial, antioxidant and phyto-chemical properties of fruit and vegetable wastes: A review
Table 5: Examples of functional food components Clasy’components
Source
Potential benefit
Beta-carotene
Various fruits
Lutein, Zeaxanthin
Citrus
Neutralizes free radicals which may damagecells; bolsters cellular antioxidant defences May contribute to the maintenance of healthy vision
Flavonoids Anthocyanidins
Berries, cherries, red grapes
Flavanols catechins, Apples, grapes epicatechins,procyanidins Flavanones Citrus foods Flavonols
Apples
Proanthocyanidins
Cranberries, apples, strawberries, grapes, wine, peanuts, cinnamon
Bolster cellular antioxidant defences; maycontribute to the maintenance of brain function May contribute to the maintenance of heart Health Neutralize free radicals which may damagecells; bolster cellular antioxidant defences Neutralize free radicals which may damagecells; bolster cellular antioxidant defences May contribute to the maintenance of urinary tract health and heart health
Source: Jasna et al., 2009.
pulp. Some of the examples of functional food componenets are shown in Table 5.
Antimicrobials The antimicrobial constituents are present in all parts of the plant viz. bark, stalks, leaves, fruits, roots, flowers, pods ,seeds, stems, latex, hull and fruit rind . The antimicrobial activities of a variety of naturally occurring phenolic compounds from different plant sources have been studied in detail (Burt 2004). Recent research has revealed that the fruit peels and seeds, such as grape seeds and peels, pomegranate peel and mango seed kernel (Kabuki et al., 2007) may potentially possess antimicrobial property. Various fruits (peel, flesh or seed) have been used in traditional medicine for stomach ache, sore eyes, fever, etc. Papaya has been shown to contain sulphydroxyl protease which can inhibit viralsor microbial infection (Rajashekhara et al.,1990). These compounds play an important role in fruits’ protection against pathogenic agents, penetrating the cell membrane of microorganisms, causing lysis. Phenolic compounds from spices such as gingeron, zingerone, and capsaicin have been found to inhibit the germination of bacterial spores (Burt 2004). Polyphenols contained in green tea (Camellia sinensis) combat against Vibrio cholerae O1,Streptococcus mutans and Shigella (Si et al., 2006). The fruit and vegetable peel extracts showed better antifungal activity than antibacterial activity; Gram-negative bacteria were more susceptible than Grampositive bacteria The most susceptible organism was fungi and Gram-negative K. pneumoniae. M. indica showed maximum and best antimicrobial activity. The antimicrobial
activity of an ethanol extract from mango seed kernels against food-borne pathogenic bacteria has also been reported. The mango extract was more effective against Gram-positive than Gram-negative bacteria, with a few exceptions (Kabuki et al., 2007). In addition, flavonoids have been reported to enhance the antibacterial, antiviral, or anticancer activities of compounds such as naringenin, acycloguanosine, and tamoxifen (Bracke et al., 1999). The mixture of phytochemical constituents in plant extracts can be an advantage due to the synergistic effect that the constituents may have (Bakkali et al., 2008). Citric, succinic, malic, acetic, and tartaric acids are commonly found in fruits and fresh-cut byproducts. They have been traditionally used in the food industry as preservative agents, attributing their antimicrobial efficacy to the pH changes of the treated media. In general, bacteria grow at a pH close to 6.5 to 7.5, but tolerate a pH range from 4 to 9. Yeasts are more tolerant to low pH values than bacteria are, whereas molds can grow in the widest pH range. One effective way of limiting microbial growth is increasing the acidity of a particular food by adding an acidic substance (Massilia et al., 2009). Acids attack cell walls, cell membranes, metabolic enzymes, protein synthesis systems, and the genetic material of microorganisms (Tripathi and Dubey, 2004). The usage of bioactive extracts as applied to fruit preservation is an alternative to chemical preservatives and helps to achieve consumer demand for fresh, nutritious and safe fruits, and vegetables that are free of synthetic additives. Some bioactive extracts have been proven to be effective antimicrobials and antioxidants; however, their addition to fruit may cause changes in sensorial attributes. For example, green tea extract (GT) has 127
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been evaluated as being able to act in the preservative treatment of fresh-cut lettuce. Different quality markers, such as respiration, browning, ascorbic acid, and carotenoid content were evaluated. Several GT concentrations (0.25, 0.5, and 1 g/ 100 ml) at different temperatures (20 °C and 50 °C) were tested. Optimal GT treatments (0.25 g/100 ml at 20 °C) were compared with chlorine (120 ppm at 20 °C). High GT concentrations (0.5 g/100 ml and 1.0 g/100 ml) to a large extent prevented ascorbic acid and carotenoid losses of 0.25 g/100 ml GT as did chlorine. However, GT enhanced the browning of the samples, probably as a result of the high polyphenol content of the treatment, though heat-shock reduced this negative effect. No significant differences were observed between chlorine and the optimal GT (0.25 g/100 mL at 20 °C) in the browning appearance and sensory properties. GT kept the antioxidant activity of the samples better than chlorine did. Ethanol extract of cinnamon bark (1% w/v) reduced the aerobic growth of bacteria inoculated fresh-cut apples significantly during storage at 6 °C up to 12 d. Catechin, chlorogenic acid, and phloridzin, 3 phenolic compounds that are abundant in apple processing byproducts, exhibited varying degree of inhibitory action toward the growth of tested food pathogenic and spoilage bacteria, fungi, and yeasts (Muthuswamy and Rupasinghe, 2007). However, it is important to note that these phenolics (except 25 mm phloridzin) did not inhibit the probiotic bacterium Lac. rhamnosus suggesting no or minimal threat to the beneficial colon microflora, if the phenolics are used as food additives at the desirable concentrations. Also these authors suggest that the major phenolic compounds of apple byproducts could find use as food additives, however, the regulatory aspects of the use of plant extracts as fresh-cut fruit additives must be contemplated. Bacterial infections remain an important problem for human health. The control of bacterial infections has been traditionally treated by inhibiting microbial growth using different types of
antibiotics. Therefore, the search of non toxic compounds which inhibit QS and so, the virulence of pathogenic bacteria can bring new alternatives for the treatment of bacterial infections in humans also notable are the antibacterial properties of berries. The cloudberry (Rubus chamaemorus), raspberry (Rubus idaeus), and bilberry (Vaccinium myrtillus) and crowberry (Empetrum nigrum) were effective against all of the bacterial strains tested. Bog bilberry (V. uliginosum) inhibited all the Gram-positive bacteria, but not Gram-negative E . coli, S . aureus, B . subtilis and M.luteus (Rauha et al., 2000). Essential oils from citrus offer the potential for all natural antimicrobials for use in improving the safety of organic or all natural foods (Joshi et al., 2011). Subba et al., (1967) determined that orange and lemon oil had in vitro antibacterial effects on Salmonella and other food-borne microorganisms. However, Fisher and Phillips (2006), on the other hand, found that Grampositive bacteria were more sensitive than Gram-negative in vitro. Seven citrus essential oils were screened by disc diffusion assay for their antibacterial activity against 11 serotypes/strains of Salmonella (Bryan et al., (2008).. The 3 most active oils were selected to determine the minimal inhibitory concentration (MIC) against the some Salmonella. Orange terpenes (C4), singles-folded d-limonene (C5), and orange essence terpenes (C6) all exhibited inhibitory activity against the Salmonella spp. On the disc diffusion assay orange terpens and d-limonene both had MICs of 1%. The most active compound, terpenes from orange essence, produced MIC that ranged from 0.125% to 0.5% against the 11 salmonella tested. (Table 6) The antimicrobial activity of some plant peels against microorganisms causing infection is summarised in Table 7. Mohamed et al., 1994 evaluated antimicrobial activity of extracts of ripe, unripe and leaves of guava (Psidium guajava); ripe, unripe and leaves of starfruit (Averrhoa carambola); ripe
Table 6: Mic (in percent, v/v) of orange oils against 11 Salmonella spp.
S. enteritidis 1773-92 S. senftenberg 43845 S. senftenberg 1402-94 S.tennessee 825-94 S. kentucky 1271-94 S. eidelberg 8326 S. enteritidis 13076 S. montevideo G4639 S. Michigan S. typhimurium (Copenhagen) S. stanfey H1256
C4
C5
C6
1 1 1 1 1 1 1 1 1 1 1
1 1 1 1 1 1 1 1 1 1 1
0.25 0.5 05. 0.5 0.25 0.25 0.13 0.25 0.25 0.5 0.5
Source: Bryan et al., 2008. 128
Antimicrobial, antioxidant and phyto-chemical properties of fruit and vegetable wastes: A review
while extractsfrom guava showed strong activity against Saccharomyces cerevisiae. Other than guava, ripe starfruit, rambai peel and rambutan peel showed potential for use against bacteria.
and unripe banana (Musa sapientum variety Montel); ripe and unripe papaya (Carica papaya); passionfruit (Passiflora edulis F. Flavicarpa) peel; two varieties of Lansium domesticum peel (langsat and duku); rambutan (Nephelium lappaceum) peel and rambai (Baccaurea motleyana) against Gram positive bacteria, Gram negative bacteria, yeast and fungi (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Lactobacillus bulgaricus; E. coli, Proteus vulgaricus, Pseudomonas aeruginosa, Salmonelli typhi; Saccharomyces cerevisiae, Candida lypolytica; Rhizopus spp., Aspergillus niger, and Chlamydomucor spp) by using both the filter paper disc diffusion and tube dilution assays. Extracts from ripe starfruit, guava leaves and rambai peel showed strong activity against all the bacteria tested, in most cases with activity stronger than 50µg streptomycin. Passionfruit peel, ripe and unripe guava showed activity against all the bacteria tested except E. coli. Rambutan peel too showed activity against all the bacteria tested except towards Pseudomonas aeruginosa. Most of the fruit wastes showed some activity towards bacteria but poor activity against yeast or fungi. Extracts from bananas, papayas, passionfruit peel, Lansium domesticum peels and rambutan peels showed activity against Candida lypolytica
Antioxidants There is evidence that chronic diseases, such as cancer and cardiovascular disease, may occur as a result of oxidative stress. Free radicals are endogenous initiators of degenerative processes, as they damage lipids, proteins and DNA, thus favouring development of a number of degen-erative diseases. The consumption of food rich in natural antioxidants, as well as food enriched with them, ensure the desirable antioxidant status and helps in prevention of the development of diseases caused by oxidative stress. The most publicized phytochemicals with antioxidant properties have been vitamin C, vitamin E, and beta-carotene (which the body converts into vitamin A). Evidence exists that vitamin E can help prevent atherosclerosis by interfering with the oxidation of low-density lipoproteins (LDL), a factor associated with increased risk of heart disease.
Table 7: Antimicrobial activity of some plant peels against some microorganisms causing infectious diseases Plant name
Extract
Microorganisms
Citrus grandis (Rutaceae)
Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Escherichia coli, Salmonella enteritidis
Citrus reticulata Blanco(Rutaceae)
Hexane, ethyl acetate, butanol, methanol, benzene: acetone Oil
Vitis vinifera(Vitaceae)
80% ethanol
Citrus reticulateBlanco(Rutaceae)
Flavonoid extract
Citrus acida Roxb. (Rutaceae)
Oil
Ficus carica L. (Moraceae)
Aqueous
Citrus bergamia Risso(Rutaceae)
Ethanolic fraction
Nephelium lappaceum L. (Sapindaceae)
Musa sapientum (Musaceae)
Chloroform, ethyl acetate, aqueous
Alternaria alternata , Rhizoctonia solani, Curvularia lunata, Fusarium oxysporum,Helminthosporium oryzae Staphylococcus aureus, Bacillus cereus Escherichia coli, Salmonella infantis, Campylobactercoli Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis,Salmonella typhimurium, Enterobacter cloacae Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Escherichia coli, Enterobacter aerogenes, Salmonella typhimurium, Aspergillus ficuum, Aspergillus niger, Aspergillus fumigatus, Aspergillus flavus, Fusarium saloni, Fusarium oxysporum, Pencillium digitatum,Candida utilis Bacillus cereus, Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli,Pseudomonas fluorescens Escherichia coli, Pseudomonas putida, Salmonella enterica, Listeria innocua, Bacillus subtilis, Staphylococcus aureus, Lactobacillus lactis, Sacharomyces cerevisiae Ether, methanol, aqueous Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi, Vibrio cholerae, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Salmonella enteritidis, Escherichia coli
Source: Chanda et al., 2010.
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The role of antioxidant phytochemicals in the prevention of these diseases has been mainly attributed to the prevention of LDL oxidation through a scavenging activity against peroxyl and hydroxyl radicals. Apple contains many different dietary phytonutrients with strong antioxidant capacities, such as phenolics, carotenoids, and vitamins, which may protect against free radicals. Apple peels have high concentrations of phenolic compounds and may assist in the prevention of chronic diseases. Phenolics are a much diversified group of secondary plant metabolites, which includes simple phenolic, phenolic acids (benzoic and cinnamic acid derivatives), lignans, lignins, coumarins, flavonoids, stilbenes, flavonolignans and tannins. Many of phenolic compounds have shown strong antioxidant properties as oxygen scavengers, peroxide decomposers, metal chelating agents, and free radical inhibitors. Besides antioxidant activity, phenolic compounds have a wide range of action which includes antitumoral, antiviral, antibacterial, cardioprotective, and antimutagenic activities. The conventional apple juice production (straight pressing of apple pulp or pressing after pulp enzyming) resulted in a juice poor in phenolics and with only 3–10% of the antioxidant activity of the fruit they were produced from. Polyphenols are one of the phytochemical groups whose ‘‘protective” properties include antioxidant, antimicrobial, anticancer and cardiovascular-protective activities. Different model systems were employed to evaluate the antioxidant properties of apple pomace polyphenols. The DPPH and superoxide ion radical scavenging activities of apple pomace polyphenols, and also their antioxidant property in the âcarotene/linoleic acid system were determined. The polyphenols examined were epicatechin, its dimer (procyanidin B2), trimer, tetramer and oligomer, quercetin glycosides, chlorogenic acid, phloridzin and 3-hydroxy-phloridzin. All the compounds showed strong antioxidant activities, and their DPPH-scavenging activities were 2-3 times and superoxide anion radical-scavenging activities were 10-30 times better than those of the antioxidant vitamins C and E. The total phenolics, total flavonoids, total flavan-3-ols, and some individual phenolic compounds contributed significantly to the antiradical activities of apple pomace. Flavonoids are polyphenolic antioxidants naturally present in vegetables, fruits, and beverages such as tea and wine. In vitro, flavonoids inhibit oxidation of low-density lipoprotein and reduce thrombotic tendency, but their effects on atherosclerotic complications in human beings are unknown (Hertog et al., 1993). The peel of Citrus fruit is a rich source of flavanones and many polymethoxylated flavones, which are very rare in other plants. Flavonoids in regularly consumed foods may reduce the risk of death from coronary heart disease in elderly men. The contents of polyphenols and tannins in fruit seed and peel are shown in Table 8.
Grape seed extract is a by-product derived from the grape seeds (Vitis vinifera) (from grape juice and wine processing) that is extracted, dried and purified to produce a polyphenolic compound rich extract (Lau and King, 2003).The extraction of crushed grape pomace with a mixture of ethyl acetate and water yielded phenolic compounds displaying antioxidant activities comparable to BHT in the Rancimat test. Catechin, picatechin, epicatechin gallate and epigallocatechin were the major constitutive units of grape skin tannins. Recent literature has evidenced antioxidant properties of GSE both in vivo and in vitro (Yilmaz and Toledo, 2004). The antioxidant properties of GSE are primarily due to flavonoids that can perform scavenging action on free radicals (superoxide, hydroxyl, and 1,1-diphenyl2-picrylhydrazyl (DPPH)), metal chelating properties, reduction of hydroperoxide formation and their effects on cell signalling pathways and gene expression (Jacob et al., 2008; Sato et al., 1996; Soobrattee et al., 2005). The presence of the functional group “–OH” in the structure and its position on the ring of the flavanoid molecule determine the antioxidant capacity. Addition of “–OH” groups to the flavonoid nucleus will enhance the antioxidant activity, while substitution by –OCH3 groups diminishes the antioxidant activity (Majo et al., 2008). Degree of polymerization of the procyanidins may also determine the antioxidant activity as the higher the degree of polymerization, the higher the antioxidant activity. Among the different parts of grape plant, grape seeds exhibit highest antioxidant activity followed by the skin and the flesh (Pastrana-Bonilla et al., 2003). The antioxidant potential of GSE is twenty and fifty fold greater. A new class of compounds, aminoethylthio-flavan-3-ol conjugates, has been obtained from grape pomace by thiolysis of polymeric proanthocyanidins in the presence of cysteamine. The antioxidant activity of the extracts obtained from grape by-products was analyzed by different in vitro tests: scavenging of the stable DPPH radical reactive •OH, O2 •- and of authentic peroxynitrite (ONOO-). The content of five phenolic constituents of biological interest: catechin and epicatechin in seeds and quercetin, rutin and resveratrol in skin extracts was investigated. All the five phenols investigated possessed strong antiradical activity. Quercetin, catechin and epicatechin showed maximum activity (respectively, IC DPPH• 50 5.5, 6.7 and 6.8 M, and IC ONOO” 50 48.8, 55.7 and 56.7 M). Recent reports indicate a wide range of biological activities, e.g. radioprotective effects, the prevention of cataract antihyperglycemic effects the enhancement of postprandial lipemia, the modulation of the expression of antioxidant enzyme systems, the inhibition of the protein kinease activity of the epidermal growth factor receptor, protective effects against oxidative damage in mouse brain cells , and anti-inflammatory effects. The high efficiency of natural phenolic extracts obtained from grape seeds as potent antioxidants was confirmed by the fact which encourages the
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Antimicrobial, antioxidant and phyto-chemical properties of fruit and vegetable wastes: A review
prospect of their commercialization as natural powerful antioxidants in foods in order to increase the shelf-life of food by preventing lipid peroxidation and protecting from oxidative damage. Many of the grape seed products are commercially available. Flavonoids from citrus that have been extensively studied for anti-oxidative, anti-cancer, anti-viral, and antiinflammatory activities, effects on capillary fragility, and an observed inhibition of human platelet aggregation. The citrus fruits possess another health benefit phytochemicals called limonoids, highly oxygenated triterpenoid. Citrus limonoids appear in large amounts in citrus juice and citrus tissues as water soluble limonoid glucosides or in seeds as water insoluble limonoid aglycones. Currently, limonoids are under investigation for a wide variety of therapeutic effects such as antiviral, antifungal, antibacterial, antineoplastic and antimalarial. Anthocyanins exhibit antioxidant activities and therefore, may contribute to the prevention of heart disease (Hou, 2003; Bagchi et al., 2004). Berries have been known to contain anthocyanin pigment abundantly and thus, have been used globally as a medicine or a source of health food/dietary supplement. Consequently, the antioxidant activity may be different among various berry extracts, in particular, the berry anthocyanin extracts in the commercial market (Nakajuma et al., 2004).
Extraction of anthocyanin from waste of different fruits, vegetables and flowers Waste from industrial processes, such as wine or juice production is an excellent source of anthocyanin pigment which could possibly be utilized as a food colourant. An
overview of the techniques employed in extracting the anthocyanin is made in Table 9. Absolute ethanol was used to facilitate subsequent concentration steps. Citric acid chelates techniques employed in extracting the anthocyanin. Metals may have an added protective effect throughout the processing of the spray dried powder. It is less corrosive than HCl and would still act to stabilize the anthocyanin structure in the cationic form by maintaining a low acid pH. The citric acid was added to the single strength extract such as that a 10 to 1 concentrate would have a pH of 3-3.2. This pH was chosen because the dried powders were used in colouring low acid food products. Different varities of grapes, as well as different extraction techniques, are used for the production of grape skin extract. Grape skin extract is available as a liquid or power and both versions are water soluble. The hue of the extract is pH and concentration dependent. While using methanol as a solvent, the methanol in the pigment solution can be removed by distillation and the resulting aqueous solution absorbed on an Amberlite C G-50 resin. The resin absorbs the anthocyanin and many of the impurities can be rinsed-off the column with water. Solvent extraction time, size of ground hulls, pH of extracting solvent, hull/ solvent ratio and concentration of SO2 in water were significant factors affecting yield of extracted anthocyanins. Ethanol-acetic acid-water was more effective extractant than acetic acid. Lycopene is much more widely distributed in fruits and vegetables than other pigments. It is predominantly found in
Table 8: Total polyphenols and tannin content in fruits seed and peel (mean ± SD) Fruit variety
Gooseberry Watermelon Apple (Idared) Apple (Sampion) Plum (Rcnkloda Ulena) Plum (W^gicrka zwykla) Melon (Galia) Red grapes (Alphonsc Lavallec) White grapes (Uva da Tavola) Lemon (Primofiori) Red grapefruit (Slar Ruby Citra) White grapefruit (Apemar) Grejpfrut biaty Kiwi fruit (Hayward) Orange (Midnight)
Total polyphenols mg catechin × 100 g”1 d.w.
Tannins mg catechin × lOOg’dw
Seeds
Peels
Seeds
Peels
800.7±30 969.3±16.4 345.0±32.1 702.5±8.3 436.8±4.2 147.3±3.0 57.2±2.6 9207.5±46.0 8220.2±60.3 158.8±0.7 222.5±14.5 205.5±6.1 102.0±2.5 212.0±84
698.7±11.9 335.7±20.8 1790.5±27.5 1613.7±11.3 334.0±8.7 578.8±13.8 466.5±8.8 5159.2±19.6 3794.5±32.9 966.2±16.5 557.7±10.9 528.8±12.5 1161.0±13.1 849.3±21.8
260.5±4.1 11.0±10.0 647.3±14.8 20.5±0.7 24.0±0.5 13.25±0.4 0 5577.2±26.1 3860.0±367.0 0 70.25±26.5 77.3±26.5 0 0
282.5±32.5 0 7420.7±90.2 1053.5±20.4 0 41.5±17.0 0 1410.3±88.0 937.2±35.0 0 0 61.25±17.3 136.0±4.0 0
Source: Chodak et al., 2007.
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chromoplast of the plant tissues and occur as the major pigment in red, fresh tomatoes, as well as in canned, condenced or processed tomato preparation.
The waste from fruits and vegetable processing industries being rich in polysaccharides (cellulose, hemicellulose and lignin) can be subjected to solid state fermentation (SSF) for the production of ethanol which has several uses (Badger and
Ethanol Table 9: Comparison of anthocyanin extraction techniques from different waste Type of waste
Extraction Techniques
Source
Fruits Grape Absolute ethanol (100%) with 0.1% HCl Grape Methanol with citric acid (0.01%) Concord grape 10LAbsolute ethanol (100%) with 0.01% citric acid Vitis vinifera var Methanol 1-M with HCl (99:1 v/v). Grenache noir Blue grapes Acidified methanol (75ml 3M HCl + 425 ml methanol) Concord grape Methanol acidified with 0.01 % citric acid Blueberries MeOH/ Formic acid/ Water (70/2/28) Bilberry, rabbiteye,90% ethanol with 0.1% H2SO4Filtrates collected after blueberry and centrifugation were applied to non-ionic polymeric blackcurrant absorbent. Then, elution with acidified ethanol (0.05% citric acid) Blackberries Methanol acidified with 0.1 % HCl Blackberries Liquid nitrogen powder + acetone: water (70/30 v/v) + acetone: chloroform at 1:2 v/v Black raspberry Dichloromethane-methanol (1:1 v/v) Tart cherry Homogenized with water at 10,000g for 10 min at 40C (Prunus cerasus) Strawberry Polyvinyl-polypyrrolidone (PVPP) resins with water were used for isolating anthocyanin. Then, anthocyanin were extracted from the resin by methanol with 0.1% HCl Cranberry Methanol with 0.03% HCl at a 5:1 solvent:pomace ratio Lychee Acidified ethanol (1.5 N HCl: 95% ethanol; 15:85 v/v) Fig Acetone and 0.1N NH-4OH (9:1 v/v). Re-extraction (Ficus carica L.) with1:1 (v/v) acetone + diethyl ether Vegetables Sweet potato Red radish cv. Fuego and Red fleshed potato tuber Red radish
Heidari et. al., 2006 Clydesdale et. al., 1978 Main et. al., 1978 Sarni-Manchado et. al.,1996 Thakur and Arya, 1989 Calvi and Francis, 1978 Gao and Mazza, 1994 Jun – ichiro Nakajima et. al.,2004
Julin et. al., 1992 Chiang and Wrolstad, 2005; Rodriguez-Soana and Wrolstad, 2001 Tian et.al., 2005. Amitabh et. al.,1992 Skrede et. al., 1992; Wrolstadand Putnam, 1969
Jackman et. al., 1996 Lee and Wicker,1991 Antoine et. al., 1976
1% HCl in water 1.Chemical purification :Acetone/ Chloroform2. Juice processing
Bassa and Francis, 1987 Guisti and Wrolstad, 1996a; Rodriguez-Soana et. al., 1999
Samples added with liquid nitrogen powder then added to 2L acetone/ Kg of skins. Futher re-extraction with acetone 30:70 (v/v). Filtrates combined with chloroform (1:2 acetone: chloroform v/v)
Hong and Wrolstad, 1990.
Flower Tradescantia pallida
0.1% HCl with water. Acid extract was purified using a cation exchange resin. Then, pigment were eluted from column with 0.1% HCl in methanol Purple sunflower 3 solvents system used i.e. (50:1:49) ethanol-acetic seeds acid-water(EAW), 0.01M acetic acid (AAc) or water containing SO2 Zebrina 0.1% HCl with water Roselle Water extraction (Hisbiscus sabdariffa) 132
Zulin et. al., 1992
Gao and Mazza,1996
Teh and Francis, 1988 Esselen and Sammy, 1973
Antimicrobial, antioxidant and phyto-chemical properties of fruit and vegetable wastes: A review
Broder, 1989, Jarosz, 1988). It can be used as a liquid fuel or liquid fuel supplement and as a solvent in many industries. Traditionally, alcohol is produced from liquid or liquid mash via submerged microbial fermentation. In recent years, there has been a considerable interest in the production of alcohol from food processing wastes such as apple pomace because of 1.) the rising energy costs of molasses ii) the negative cost of values of wastes as substrates. Apple pomace is not readily amenable to submerged microbial fermentation due to its nature. But solid state fermentation of apple pomace offers several advantages for ethanol production such as higher yield but has one difficulty of ethanol extraction from the solid materials. Different microorganisms have been used for the production of ethanol, predominantly yeast belonging to Saccharomyces cervisiae has been a micro- organism of choice (Joshi and Sandhu, 1996). A detailed process for production of ethanol from apple pomace has been been illustrated by Joshi et al. (1999). Natural fermentation of apple pomace was inferiour to the yeast inoculated fermentation for ethanol, crude and soluble proteins. The production of ethanol in natural fermentation was almost half than that of Saccharomyces cerevisiae fermented apple pomace. Partial aseptic and anaerobic conditions were provided to the solid state fermentation of apple pomace by addition of SO2 and found that addition of SO2 upto 200 ppm increased the ethanol content by Saccharomyces cerevisiae while it was 150 ppm for Candida utilis and Torula utilis (Hang et al., 1981). The original pH and the initial moisture content of apple pomace was found to be suitable for ethanol production, decreasing the pH or increasing the moisture content reduced the ethanol content. Fermentation time increased the ethanol production upto 96 hrs at 30oC and among the different nitrogen sources tried ammonium sulphate gave the highest ethanol production and Saccharomyces cerevisiae giving better response to it than Candida utilis and Torula utilis (Joshi and Devrajan, 2008). Addition of 0.4% of ammonium sulphate increased the ethanol yield. The combined effect of AMS and ZnSO4, however was detrimental to ethanol production but AMS alone gave better ethanol yield. It was found that all the yeast fermented apple pomace distillates contained methyl and butyl alcohols, and aldehyde. Saccharomyces cerevisiae fermented distillate had more desirable characteristics than those obtained from fermentation with other yeasts and thus, had potential for conversion into potable alcohol. The distillate obtained from Saccharomyces fermented apple pomace had more desirable characteristics. After fermenting apple pomace with Saccharomyces cerevisiae for ethanol production, four methods i.e. hydraulic pressing, direct distillation, steam distillation and vacuum distillation of fermented apple pomace were applied for separation of ethanol. Out of these, the steam
distillation was found to be the best as it induced minimum alteration in the fermented apple pomace (Devrajan, 1997, Joshi and Devrajan, 2008). Apple, pear and cherry wastes have also been utilized for production of ethanol. Waste from processing of orange can be employed for production of ethanol. Orange peels after enzymatic hydrolysis was found suitable to SSF by using Saccharomyces cerevisiae for ethanol production (Converti et al., 1989).
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Intl. J. of Food. Ferment. Technol. 2(2): 137-143, December, 2012
Research paper
Effect of freeze-drying and storage on β-carotene and ascorbic acid stability of mango milk shake K. Jayathilakan *, Khudsia Sultana, M.C.Pandey and K. Radhakrishna Freeze Drying and Animal Product Technology Division, Defence Food Research Laboratory, Siddarthanagar, Mysore –570011, India *
Email:
[email protected]
Paper no: 48
Received: 04 Oct, 2012
Received in revised form: 11 Nov, 2012 Accepted: 14 Nov, 2012
Abstract Freeze-drying is a process which retains the nutritional, sensorial and functional qualities of foods, together with extreme reduction in weight, high solubility, long shelflife at moderate temperature and the possibility to perform rehydration at any desired level. The application of this technique in the development of a Freeze dried mango milk shake powder using Badami mango (Mangifera indica L) pulp, milk (3.0g/100g fat, 8.5g/100g Solids Non Fat), nuts, flavourants and ascorbic acid to deliver RDA level of ascorbic acid and β-carotene was hence attempted. Optimized formulations were subjected to freeze drying for better storage stability. The water activity was found to be 0.22 and BET monolayer value 2.294g/100g solids. β-Carotene, ascorbic acid and oxidative rancidity profile monitored at ambient (28±2°C) and 37°C storage revealed no significant differences (p>0.01) at ambient temperature in ascorbic acid and thiobarbituric acid reactive substances (TBARS) but significant difference (p