Preparation and Characteristics of Sugarcane Low Alcoholic Drink by Submerged Alcoholic Fermentation Gan-Lin Chen, Feng-Jin Zheng, Bo Lin, Tian-Shun Wang & Yang-Rui Li
Sugar Tech An International Journal of Sugar Crops and Related Industries ISSN 0972-1525 Volume 15 Number 4 Sugar Tech (2013) 15:412-416 DOI 10.1007/s12355-013-0248-3
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Author's personal copy Sugar Tech (Oct-Dec 2013) 15(4):412–416 DOI 10.1007/s12355-013-0248-3
RESEARCH ARTICLE
Preparation and Characteristics of Sugarcane Low Alcoholic Drink by Submerged Alcoholic Fermentation Gan-Lin Chen • Feng-Jin Zheng • Bo Lin Tian-Shun Wang • Yang-Rui Li
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Received: 8 May 2013 / Accepted: 16 July 2013 / Published online: 2 August 2013 Ó Society for Sugar Research & Promotion 2013
Abstract A new low alcoholic drinks, was brewed from fresh sugarcane juice using different kinds of yeast, viz., wine yeast, Fali yeast and thermo-tolerant yeast by submerged alcoholic fermentation. The main characteristic parameters of fermented cane juice such as alcohol content, total sugars, total acids and reducing sugar were investigated during the process of alcoholic fermentation in order to evaluate the effect of yeasts on the quality of alcoholic drinks. The results showed that alcoholic fermentation period and the alcohol content of fermented cane juice ranged from 9 to 20 days and 2.8–4.8 %, respectively. With respect to sensory evaluation, the alcoholic drink obtained by using wine yeast was found suitable with yellow-brown colour, clear and brighter appearance, full wine and cane aromas and flavor. Its soft, mild, mellow and slightly sweet taste made it possible to be used as a beverage in dines. The results of this study concerning the use of sugarcane juice as one of raw materials could be promising for industrial fermentations of low alcoholic drinks production, and also provides a new approach to process sugarcane by-products.
G.-L. Chen B. Lin T.-S. Wang Y.-R. Li (&) Sugarcane Research Center, Chinese Academy of Agricultural Sciences/Sugarcane Research Institute/Guangxi Academy of Agricultural Sciences/Key Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture/Guangxi Key Laboratory of Sugarcane Genetic Improvement, Nanning 530007, Guangxi, China e-mail:
[email protected] F.-J. Zheng Agricultural Product Processing Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, Guangxi, China
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Keywords Sugarcane juice Alcoholic fermentation Low alcoholic drinks Yeast
Introduction Sugarcane is a field grown, thick, tough and tall perennial grass having stalks appear like bamboo, and is the most important sugar crop all around the world because it stores 15–18 % sucrose in its juicy stalks. In addition to making white sugar, sugarcane juice is highly beneficial to human health as it contains many amino acids such as aspartic acid, alanine and citric acid; vitamins A, B1, B2, B3, and C, niacin, riboflavin and essential nutrients such as calcium, phosphorus, manganese, zinc, etc. and especially the iron (9 mg/kg) (Huang et al. 2006; Legaz et al. 1990). Because of having such a nutritious contents, its juice is very important medicinally and used as a drink to prevent and heal sore throat, cold and flu, lowering glyceric index, fighting against prostration and breast cancer cells due to its alkaline nature, prolonging heat and physical activity, refreshing and energizing the body instantly as it is rich in carbohydrates, keeping the urinary flow clear and helps the kidneys to perform their functions smoothly and hastening recovery from jaundice. Sugarcane juice is also an excellent substitute for aerated drinks and Cola, and it is one amongst the sweet drinks suitable for diabetic patients also (Chen et al. 2011). Due to health benefits, there are many kinds of products exploited in China recently. The fresh cane juice drinks or compound juice with other fruit is the most popular one amongst them. Among the different substrates (apple fruit and alcohol) available for low alcohol drinks production, sugarcane, as a prime crop in China, is a good substrate owing to its high sugar content and easy
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availability (Chen et al. 2011). From the industrial point of view for ethanol production, the sucrose-based substrates such as cane and sugar beet juices or molasses present many advantages including their relative abundance and renewable nature (Thomas and Kwong 2001; Taherzadeh 1999), In Brazil, sugar cane juice and sugar cane molasses are the substrates of choice for fuel ethanol because of its high sugar content and availability (Wheals et al. 1999). Sugarcane is also a prime economic crop in southern China (Liang et al. 2008). China is the third largest sugarcane and sugar producing country, and produced 10.51 million tons of cane sugar in 2011–2012 milling season. As the largest sugarcane producer of China, Guangxi Province produced 6.94 million tons of sugar in 2011–2012 milling season, accounted for 66.5 % of the total sugar production and almost 70 % of the cane sugar production in China. In addition to making cane sugar and direct drink, the sugarcane juice has high prospects for making alcoholic drinks, and during the recent years, because of the health fitness awareness campaigns, the processing of sugarcane juice for making soft drinks and other alcoholic drinks gained a major attention by researchers (Zhang et al. 2010; Li and Xiong 2009; Chen et al. 2011; Li et al. 2010; Beltran et al. 2008; Yu et al. 2009; Gong et al. 2011). In the present study, different kinds of yeasts were used to brew a new sugarcane low-alcoholic drink by submerged alcoholic fermentation of cane juice. During the fermentation processing, the alcohol content, total sugars, total acids and reducing sugar were investigated to evaluate the effect of yeasts on the quality of alcoholic drink, and the sensory evaluation indicators of colour and lustre, scent and fragrance, taste were presented for further developing of sugarcane drinks.
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Yeasts Almost ten kinds of different yeasts were selected and put to use for screening test and comparison test. Based on the descriptions and recommendations of previous trials, three kinds of yeast, of which the Fali yeast was provided by Harbin Mauri Yeast Co., Ltd., China, and the Wine yeast and Thermo-tolerant yeast were provided by Angel Yeast Co., Ltd., Hubei, China, were used in this experiments. Alcoholic Fermentation Based on our previous experimental results and recommendations, yeasts were weighted (for weight-to-capacity ratio, or w/v, viz. 0.90, 1.20 g/L), dissolved in 150–250 mL of water, and re-activated at 38–42 °C for 30 min. The fully activated yeasts were poured in a tank filled with 20 L of sugarcane juice and then mixed properly (Chen et al. 2011). The tank was sealed and left for fermentation at normal temperature in range of 20–25 °C. Analytical Methods During the period of alcoholic fermentation, the alcohol content (%), total sugars (%), total acids (%), reducing sugar (%) and other parameters of fermenting cane juice were measured using the methods as described in Analytical methods of wine and fruit wine of GB/T 15038-2006, State Standard of the People’s Republic of China (Anonymous 2007). Sensory evaluation of fermented cane juice for colour and luster, were evaluated by methods of GB/T 21172-2007/ ISO 11037-1999 (Anonymous 2008), sent and fragrance, evaluated by methods of GB 12313-1990/ISO 6564-1985 (Anonymous 1990), and taste, evaluated by methods of GB/ T 12312-2012/ISO 3972-1991 (Anonymous 2012), State Standard of the People’s Republic of China.
Materials and Methods Results and Discussion Sugarcane Juice Milling canes of sugarcane varieties ROC22 (bred in Taiwan) and GT series varieties bred in GXAAS, China were selected as materials. All damaged and spoiled materials were eliminated, and fresh healthy stalks were used. All stalks were crushed in a mechanical crusher. The fresh cane juice with 19°–21° brix was boiled for 10 min for sterilisation, siphoned off using a sterilisable plastic filter of diameter 5 mm, and then cooled at normal temperature in range of 20–25 °C. The cooled juice was diluted with an equal amount (for capacity-to-capacity ratio, or v/v, viz 1:1) of sterilised pure water and used for alcoholic fermentation (Chen et al. 2011).
Correlation of Alcohol Content and Total Sugars in Fermented Cane Juice During the alcoholic fermentation of sugarcane juice, total sugars involve sucrose and its monomers, fructose and glucose, existed in sugarcane juice, are important carbon sources for production of ethanol or low alcohol drinks (Liang et al. 2008). The fermentation process of sugarcane juice involves decomposition of carbohydrate source to produce alcohol and carbon dioxide using activated yeasts (Chen et al. 2011), and the quantity of alcohol content showed increasing gradually with the consumption of total sugars (Figs. 1, 2).
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Alcohol content (%)-
6.0 5.0
Wine yeast 0.9 g/L
1.2 g/L
Fali yeast 0.9 g/L Thermo-tolerant yeast 0.9 g/L
1.2 g/L 1.2 g/L
4.0 3.0 2.0 1.0 0.0 0
3
6
9
12
15
18
21
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27
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Time (d)
Fig. 1 Influence of different yeasts and doses on alcohol content of fermented juice
Fig. 3 Influence of different yeasts and doses on total acids of fermented juice Wine yeast 0.9 g/L Fali yeast 0.9 g/mL Thermo-tolerant yeast 0.9 g/L
1.6
Reducing sugar (%)-
1.4
1.2 g/mL 1.2 g/L 1.2 g/L
1.2 1.0 0.8 0.6 0.4 0.2 0.0
0
3
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9
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Time (d)
Fig. 2 Influence of different yeasts and doses on total sugars of fermented juice
It is clear from Fig. 1 that the actual fermentation period ranged from 9 to 20 days in case of different kinds of yeast during the process of alcoholic fermentation. Alcohol started to produce after 2 days of fermentation beginning, and gradually increased to the highest level from 3 to 9 days, depending on the yeast used; and finally became almost constant after 20 days of fermentation. In contrast, the total sugars content decreased rapidly until first 9 days and consumed completely in 9–20 days of fermentation, depending on the yeast used. In the actual alcoholic fermentation with yeast, such as wine yeast, the initial 2 days are called as pre-fermentation stage, during which the yeast cells adapted to atmosphere and breed (budding). Re-activating yeast by dissolving it in water could enhance the fermenting vitality of yeast and decrease this duration (Yadira et al. 2005). Generally, alcohol was not produced during this period. The period from 3rd to 9th days is called vigorous growing stage of fermentation. During this period, total sugars decrease rapidly and alcohol content increases constantly to its maximum, and the bubbles of CO2 come out on the surface and gives alcoholic fragrance (Sheehan 2009; Liang et al. 2008). The period after 10th day is the last stage of
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Fig. 4 Influence of different yeasts and dose on reducing sugar of fermented juice
fermentation, when sugar content is exhausted and very little change occurs in alcohol concentration. At this stage, the fermented cane juice looks clear and bright and brown or yellow in color. Three kinds of yeasts showed almost similar effects on fermentation process. However, the fermentation carried out using wine yeast could give up to 4.8 % alcohol content and the fermentation was completed within 8–18 days with 0.7 % residual sugars. Different doses of yeast (0.90, 1.20 g/L) did not show any difference in fermentation period or alcohol content except in case of Fali yeast where use of 1.20 g/L yeast resulted in higher alcohol content and shorter period of fermentation compared to 0.90 g/L dose of yeast (Fig. 1). Total Acids in Fermented Cane Juice The acidity of fermented cane juice should be appropriate in the process of alcoholic fermentation of cane juice, because higher acidity of fermented juice could be advantageous to inhibit propagation of harmful microorganisms, but will be disadvantageous to the activity of yeast at a certain extent. Conversely, lower acidity, such as
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Table 1 Sensory evaluation of different fermented cane juice Kinds
Wine yeast
Fali yeast
Thermo-tolerant yeast
Colour & luster
Yellow-brown, clear and brighter, no sediment and suspended substance
Brown, clear and brighter, no sediment and suspended substance
Yellow-orange, clear and bright, no sediment and suspended substance
Scent & Full wine aromas and cane flavor, fragrance no odor
More wine aromas and cane flavor, no odor
Some wine aromas and cane flavor, no odor
Taste
Mild and mellow, slightly sweet, slightly astringent, soft and dry
Mild and mellow, sweet, bit and astringent, soft and dry, rounded and smoothly
Soft, mild and mellow, slightly sweet, slightly astringent, fresh and dry
near neutral, will be apt to reproduce and grow harmful microorganisms, which also could inhibit the activity of yeast (Chen 2009; Liang et al. 2008). The rate of increase in total acid content during pre-fermentation stage was very high and it slowed down and became constant during later stages of alcoholic fermentation. During initial 4 days, total acids content increased from 0.9 to 2.6 %, then increased little during next 5–20 days and stabilized gradually after 20 days. The total acids produced during the process ranged from 3.0 to 4.3 %. The highest acid content (4.3 %) was produced by Fali yeast (0.90 g/L). The ability of wine yeast to produce acid was found relatively stable and showed little difference at different doses (3.3 and 3.0 % at 0.90 and 1.20 g/L doses) (Fig. 3). Reducing Sugar in Fermented Cane Juice Reducing sugar of sugarcane juice, as a single sugar of carbohydrate nutrient, mainly includes glucose and fructose, which is another important parameter in the alcoholic fermentation of cane juice as shown in Fig. 4 (Chen et al. 2011). The reducing sugar content decreased gradually until exhausted in prolonged process of alcoholic fermentation. During initial 4 days (pre-fermentation stage), the reducing sugar decreased sharply due to their consumption in alcoholic fermentation. The rate of decrease in reducing sugar became slow during 5–20 days, and then stabilized later. At the end of alcoholic fermentation, the residual reducing sugar ranged from 0.05 to 0.10 %. The highest residual reducing sugar content (0.33 %) was found in juice fermented by using Fali yeast (0.90 g/L). Sensory Evaluation of Fermented Cane Juice The alcoholic drinks produced by using different kinds of yeast showed different sensory properties, such as colour and luster, scent and fragrance. Taste was evaluated by methods of GB/T China Standards or ISO Standards, which were mentioned as description and characterization (Table 1). The fermentation duration required by Fali yeast was found relatively shorter, i.e., the fermentation speed was faster compared to other yeasts. But the residual total
sugars content in juice fermented by using Fali yeast was higher than thermo-tolerant yeast and wine yeast. However, the cane juice fermented by using wine yeast was found better than other yeasts in regard to sensory evaluation, such as colour, lustre, fragrance and taste. It appears suitable with yellow-brown colour, clear and brighter, full wine and cane aromas and flavor. Its soft, mild, mellow and slightly sweet taste made it possible to be used as low alcohol drinks in dines.
Conclusions Sugar cane juice is the substrates of choice for fuel ethanol or alcohol drinks because of its high sugar content and availability (Wheals et al. 1999). Sucrose and its monomers, fructose and glucose, existed in sugarcane juice and molasses, are important carbon sources for production of several products such as ethanol or low alcohol drinks (Liang et al. 2008). In this study, the low alcohol drink obtained by alcoholic fermentation of sugarcane juice using suitable three kinds of yeast, which found to be up to 4.8 % of alcohol content, ranged from 3.0 to 4.3 % of total acids and 0.7 % residual sugar content. Our study suggests that sugarcane juice may be used commercially as a low alcohol drink by using submerged alcoholic fermentation by using suitable yeast species. Acknowledgments This study was supported by the International S&T Cooperation Program of China (Grant No. 2009DFA30820), the Guangxi Natural Science Foundation (2010GXNSFB013015), Fundamental Research Funds of GXAAS (Gui Nong Ke 2011YM26 & 2013YM02). The author would like to thank Dr. Manoj Kumar Srivastava, post-doctoral fellow, GXAAS (Nanning, China) for his assistance in the revision of this paper.
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