Enhanced Biogas Production from Palm Oil Mill Effluent Supplemented with Untreated Oil Palm Empty Fruit Bunch Biomass with a Change in the Microbial Community
誌名
日本食品工学会誌 = Japan journal of food engineering
ISSN
13457942 Ali, A.A.Md. Zainudin, Md.H.Md. Idris, A.
著者
Baharuddin, A.S. Sulaiman, A. Matsui, T. Osaka, N. Oshibe, H. Hassan, Md.A. Shirai, Y.
巻/号
13巻3号
掲載ページ
p. 37-41
発行年月
2012年9月
農林水産省 農林水産技術会議事務局筑波事務所 Tsukuba Office, Agriculture, Forestry and Fisheries Research Council Secretariat
Japan Journal of Food Engineering, Vol. 13, No.3, pp. 37 - 41, Jun. 2012
000 Note 000 Enhanced Biogas Production from Palm Oil Mill Effluent Supplemented with Untreated Oil Palm Empty Fruit Bunch Biomass with a Change in the Microbial Community Ahmad Amiruddin Mohd ALI\ Mohd Huzairi Mohd ZAINUDIN2 , Azni IDRIS3, Azhari Samsu BAHARUDDIN3, Alawi SULAIMAN4, Toru MATSUI5, Noriko OSAKA5, Hiroshi OSHIBE5, Mohd Ali HASSAN2, and Yoshihito SHIRAIl, t Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, 2-4 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0196,Japan 2 Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia 3 Faculty of Engineering, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia 4 Faculty of Plantation and Agrotechnology, Universiti Teknologi MARA, 40450 Shah Alam, Selangor, Malaysia 5 Fundamental Technology Department, Technical Research Institute, Tokyo Gas Co., Ltd. 1-7-7 Suehiro-cho Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan
1
The biogas and biomethane production in a 50 litre closed stirred tank anaerobic bioreactor treating palm oil mill effluent (PO ME) supplemented with oil palm biomass in the form of oil palm empty fruit bunch (OPEFB) under mesophilic condition was evaluated. With OPEFB supplementation, the biogas and biomethane generation increased by 63% and 52%, respectively. During this process, we found changes in the OPEFB morphology and microbial community through microbiota analysis using 16S rRNA gene clone library method, after OPEFB was added, suggesting that the increased biogas and biomethane production would be due to enhanced lignocellulosic biomass degradation. Key words: Biogas, biomethane, microbial community, oil palm empty fruit bunch (OPEFB), palm oil mill effluent (POME)
1. Introduction
Biogas and biomethane fermentation of lignocellulosic biomass have previously been attempted but with varied success due to the rate limiting hydrolysis of the materials which affects both fermentation rate and extent [1]. Without any recourse to pretreatments, the presence of lignin is one of the major drawbacks in lignocellulosic fermentation, as it renders the lignocellulose resistant to biological and chemical degradation, thus hampering biogas and biomethane productivity [2, 3]. The palm oil industry represents the largest agro-economic sector in Malaysia. The iu.dustry produces abundant biomass wastes with 19.8 million tonnes of oil palm empty fruit bunch (OPEFB) and more than 50 million tonnes of palm oil mill effluent (POME) being generated from over 400 mills in Malaysia annually [4,5]. Biogas production from
(Received 17 May. 2012: accepted ll]uly. 2012)
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anaerobic fermentation of POME has been extensively studied [6-10]. Recently, co-digestion of POME with OPEFB pretreated with alkali, acid and steam were reported to increase biogas productivity in thermophilic PO ME fermentation [11, 12]. However, presently no study has been carried out to identify the changes in additional OPEFB and microbial consortia related to increased biogas and biomethane productivity in such systems. This study deals with the biogas and biomethane enhancement by OPEFB supplementation as well as the changes in the OPEFB morphology and microbial community during the fermentation. 2. Materials and methods 2.1 POME Raw PO ME was collected from Seri Ulu Langat Palm Oil Mill, Dengkil, Selangor, Malaysia, and preserved in < 4°C prior to use.
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Ahmad AmifUddin Mohd AU, Mohd Huz~ri Mohd ZAlNUDIN, Ami !DRlS, Azhari Samsu BAHARUDDlN, A1awi SUIAIMAN, TofU MATSUI, Noriko OSAKA, Hiroshi OSHIBE, MohdAli HASSAN, and Yoshihito SHIRAI
2.2 POME sludge
Matured and substrate-acclimatized POME sludge inoculum was obtained from the settling tank of a 500 m3 closed anaerobic digester located at FELDA Serting Hilir Mill Biogas Plant, Negeri Sembilan, Malaysia, where several studies on POME treatment were previously done [8, 10]. 2.30PEFB
Press-shredded OPEFB was obtained from Seri Ulu Langat Palm Oil Mill, Dengkil, Selangor, Malaysia. The OPEFB was then dried and ground using a hammer mill (Hsiangtai CW-1) and passed through a 2.0 mm sieve prior to use. 2.4 Methane fermentation set up
The system comprised of two 50 litre closed stirred bioreactors. One was used as a control experiment while the other was used for the test experiment. A 40 litre working volume was used with mesophilic condition (37 ±2°C) maintained throughout the experimental period. Wet gas meters (Shinagawa Seiki Co., OSK 14608) were used to record biogas flowrate. Fourty litres of PO ME sludge was fed into the bioreactors for the start up stage to stabilise the fermentation system and deplete all biogas that may be contributed from residual organic matter in the sludge inoculum. Four litres of raw PO ME was then fed into the control bioreactor. Simultaneously, 4 litres of raw POME and OPEFB was fed into the test bioreactor. TS level within the control bioreactor after POME feeding was 39 gil, i.e. 3.9%. The TS level was increased to 5.4% by adding OPEFB into the test bioreactor. The fermentation was continued until the biogas production from the control bioreactor had ceased.
clone library method. Firstly, DNA was extracted using DNA extraction kit (UltraClean Soil DNA Isolation Kit, MO-BIO). The DNA was extracted based on the manufacturer's instruction except for modification as described below. Firstly, 0.3 g of sample was added into 2 ml bead solution tube. After that, 500 ,LII of lysozyme (l00 mg/ml) was added into the same tube. The bead solution tube containing sample and el1jYmes was incubated at 37°C for 30 min. The subsequent steps were continued according to the manufacturer's instruction. Polymerase Chain Reaction (PCR) analysis was then conducted whereby the 16S rRNA fragments was amplified with primer 27F and 1492R. The PCR amplification and construction of 16S rRNA gene clone library were carried out based on methodology by Zhu et al. [14]. The partial 16S rRNA sequence were sent to First Base Laboratories Sdn. Bhd., a sequencing company for sequence determination. The 750R sequences primer was used for sequence analysis. Sequence similarity to closest relative searches were conducted by matching the sequence database at National Center for Biotechnology Information (NCBI) using nucleotidenucleotide Basic Local Alignment Search (BLASTn) and Ribosomal Database Project II (RDPII) using sequence match program. 3. Results and discussion 3.1 Biogas and biomethane productivity
Figure 1 shows that with only PO ME fed into the system, 152litres of biogas was produced. After POME with OPEFB was fed, the biogas generated was approximately 248 litres within the same duration, giving an increase in biogas generation by 63%. With OPEFB addition, 134 litres of biomethane was produced compared to 88 litres
2.5 Analytical methods
Composition of CH 4 and CO 2 gases were analysed using a gas chromatograph (GC) (TCD Shimadzu GC2014). Chemical oxygen demand (COD) analyses were conducted in accordance to the APHA Standard Methods [13]. All biogas and biomethane values are presented as values at Standard Temperature and Pressure (STP, i.e., O°C, 760 mmHg). OPEFB morphology was analysed using Scanning Electron Microscope (SEM) (Hitachi S-3400N).
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