Biotechnology for Biofuels | |
One-pot bioethanol production from cellulose by co-culture of Acremonium cellulolyticus and Saccharomyces cerevisiae | |
Enoch Y Park1  Kazuya Naruse1  Tatsuya Kato1  | |
[1] Laboratory of Biotechnology, Faculty of Agriculture, Department of Applied Biological Chemistry, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8017, Japan | |
关键词: Biorefinery; Saccharomyces cerevisiae; Acremonium cellulolyticus C-1; Biomass; Cellulase; Bioethanol; | |
Others : 798233 DOI : 10.1186/1754-6834-5-64 |
|
received in 2012-06-04, accepted in 2012-08-30, 发布年份 2012 | |
【 摘 要 】
Background
While the ethanol production from biomass by consolidated bioprocess (CBP) is considered to be the most ideal process, simultaneous saccharification and fermentation (SSF) is the most appropriate strategy in practice. In this study, one-pot bioethanol production, including cellulase production, saccharification of cellulose, and ethanol production, was investigated for the conversion of biomass to biofuel by co-culture of two different microorganisms such as a hyper cellulase producer, Acremonium cellulolyticus C-1 and an ethanol producer Saccharomyces cerevisiae. Furthermore, the operational conditions of the one-pot process were evaluated for maximizing ethanol concentration from cellulose in a single reactor.
Results
Ethanol production from cellulose was carried out in one-pot bioethanol production process. A. cellulolyticus C-1 and S. cerevisiae were co-cultured in a single reactor. Cellulase producing-medium supplemented with 2.5 g/l of yeast extract was used for productions of both cellulase and ethanol. Cellulase production was achieved by A. cellulolyticus C-1 using Solka-Floc (SF) as a cellulase-inducing substrate. Subsequently, ethanol was produced with addition of both 10%(v/v) of S. cerevisiae inoculum and SF at the culture time of 60 h. Dissolved oxygen levels were adjusted at higher than 20% during cellulase producing phase and at lower than 10% during ethanol producing phase. Cellulase activity remained 8–12 FPU/ml throughout the one-pot process. When 50–300 g SF/l was used in 500 ml Erlenmeyer flask scale, the ethanol concentration and yield based on initial SF were as 8.7–46.3 g/l and 0.15–0.18 (g ethanol/g SF), respectively. In 3-l fermentor with 50–300 g SF/l, the ethanol concentration and yield were 9.5–35.1 g/l with their yields of 0.12–0.19 (g/g) respectively, demonstrating that the one-pot bioethanol production is a reproducible process in a scale-up bioconversion of cellulose to ethanol.
Conclusion
A. cellulolyticus cells produce cellulase using SF. Subsequently, the produced cellulase saccharifies the SF, and then liberated reducing sugars are converted to ethanol by S. cerevisiae. These reactions were carried out in the one-pot process with two different microorganisms in a single reactor, which does require neither an addition of extraneous cellulase nor any pretreatment of cellulose. Collectively, the one-pot bioethanol production process with two different microorganisms could be an alternative strategy for a practical bioethanol production using biomass.
【 授权许可】
2012 Park et al.; licensee BioMed Central Ltd.
【 预 览 】
Files | Size | Format | View |
---|---|---|---|
20150117081410865.pdf | 6366KB | download | |
Figure 6. | 41KB | Image | download |
Figure 5. | 57KB | Image | download |
Figure 4. | 29KB | Image | download |
Figure 3. | 35KB | Image | download |
Figure 2. | 63KB | Image | download |
Figure 1. | 56KB | Image | download |
【 图 表 】
Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
Figure 6.
【 参考文献 】
- [1]Thurnheer T, Cook AM, Leisinger T: Co-culture of defined bacteria to degrade seven sulfonated aromatic compounds: efficiency, rates and phenotypic variations. Appl Microbiol Biotechnol 1988, 29:605-609.
- [2]Shim H, Shin EB, Yang ST: A continuous fibrous-bed bioreactor for BTEX biodegradation by a co-culture of Pseudomonas putida and Pseudomonas fluorescens. Adv Environ Res 2002, 7:203-216.
- [3]Parshina SN, Kijlstra S, Henstra AM, Sipma J, Plugge CM, Stams AJM: Carbon monoxide conversion by thermophilic sulphate-reducing bacteria in pure culture and in co-culture with Carboxydothermus hydrogenoformans. Appl Microbiol Biotechnol 2005, 68:390-396.
- [4]Laplace JM, Delgenes JP, Moletta R, Navarro JM: Ethanol production from glucose and xylose by separated and co-culture processes using high cell density systems. Proc Biochem 1993, 28:519-525.
- [5]Taniguchi M, Tohma T, Itaya T, Fujii M: Ethanol production from a mixture of glucose and xylose by co-culture of Pichia stipites and a respiratory-deficient mutant of Saccharomyces cerevisiae. J Ferment Bioeng 1997, 83:364-370.
- [6]Qian M, Tian S, Li X, Zhang J, Pan Y, Yang X: Ethanol production from diluted-acid softwood hydrolysate by co-culture. Appl Biochem Biotechnol 2006, 134:273-283.
- [7]Golian H, Dumsday GJ, Stanley GA, Pamment NB: Evaluation of a recombinant Klebsiella oxytoca strain for ethanol production from cellulose by simultaneous saccharification and fermentation: comparison with native cellobiose-utilising yeast strains and performance in co-culture with thermotolerant yeast and Zymomonas mobilis. J Biotechnol 2002, 96:155-168.
- [8]Saddler JN, Chan MKH, Lous-Seize G: A one-step process for the conversion of cellulose to ethanol using anaerobic microorganisms in mono-and co-culture. Biotechnol Lett 1981, 3:321-326.
- [9]Lynd LR, Weimer PJ, van Zyl WH, Pretorious IS: Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 2002, 66:506-577.
- [10]Lynd LR, van Zyl WH, McBride JE, Laser M: Consolidated bioprocessing of cellulosic biomass: an update. Curr Opin Biotechnol 2005, 16:577-583.
- [11]Aro N, Pakula T, Penttila M: Transcriptional regulation of plant cell wall degradation by filamentous fungi. FEMS Microbiol Rev 2005, 29:719-739.
- [12]Gold ND, Martin VJJ: Global view of the Clostridium thermocellum cellulosome revealed by quantitate proteomic analysis. J Bacteriol 2007, 189:6787-6795.
- [13]Xu Q, Singh A, Himmel ME: Perspectives and new directions for the production of bioethanol using consolidated bioprocessing of lignocellulose. Curr Opin Biotechnol 2009, 20:364-371.
- [14]Fujita Y, Ito J, Ueda M, Fukuda H, Kondo A: Synergistic saccharification, and direct fermentation to ethanol, of amorphous cellulose by use of an engineered yeast strain codisplaying three types of cellulolytic enzyme. Appl Environ Microbiol 2004, 70:1207-1212.
- [15]Katahira S, Mizuike A, Fukuda H, Kondo A: Ethanol fermentation from lignocellulosic by a recombinant xylose- and cellooligosaccharide-assimilating hydrolysate yeast strain. Appl Microbiol Biotechnol 2006, 72:1136-1143.
- [16]Khaw TS, Katakura Y, Koh J, Kondo A, Ueda M, Shioya S: Evaluation of performance of different surface-engineered yeast strains for direct ethanol production from raw starch. Appl Microbiol Biotechnol 2006, 70:573-579.
- [17]Yanase S, Yamada R, Kaneko S, Noda H, Hasunuma T, Tanaka T, Ogino C, Fukuda H, Kondo A: Ethanol production from cellulosic materials using cellulase expressing yeast. Biotechnol J 2010, 5:449-455.
- [18]Yamanobe T, Mitsuishi Y, Takasaki Y: Isolation of cellulolytic enzyme producing microorganism, culture conditions and some properties of the enzymes. Agric Biol Chem 1987, 51:65-74.
- [19]Ikeda Y, Hayashi H, Okuda N, Park EY: Efficient cellulase production by the filamentous fungus Acremonium cellulolyticus. Biotechnol Progr 2007, 23:333-338.
- [20]Park EY, Naruse K, Kato T: Improvement of cellulase production in cultures of Acremonium cellulolyticus using pretreated waste milk pack with cellulase targeting for biorefinery. Bioresour Technol 2011, 102:6120-6127.
- [21]Prasetyo J, Zhu J, Kato T, Park EY: Efficient production of cellulase in the culture of Acremonium cellulolyticus using untreated waste paper sludge. Biotechnol Progr 2011, 1:104-110.
- [22]Park EY, Anh PN, Okuda N: Bioconversion of waste office paper to L(+)-lactic acid by filamentous microorganism Rhizopus oryzae. Bioresour Technol 2004, 93:77-83.
- [23]Ikeda Y, Park EY, Okuda N: Bioconversion of waste office paper to gluconic acid in a turbine blade reactor by the filamentous fungus Aspergillus niger. Bioresour Technol 2006, 97:1030-1035.
- [24]Prasetyo J, Naruse K, Kato T, Boonchird C, Harashima S, Park EY: Bioconversion of paper sludge to biofuel by simultaneous saccharification and fermentation using a cellulase of paper sludge origin and thermotolerant Saccharomyces cerevisiae TJ14. Biotechnol Biofuel 2011, 4:35. BioMed Central Full Text
- [25]Prasetyo J, Sumita S, Okuta N, Park EY: Response of cellulase activity in pH-controlled cultures of filamentous fungus Acremonium cellulolyticus. Appl Biochem Biotechnol 2010, 162:52-61.
- [26]Kansarn S: A novel concept for the enzymatic degradation mechanism of native cellulose by Acremonium cellulolyticus. Sch Electr Sci Res Rep 2002, 23:89-91. Shizuoka University Repository (SURE), 91. http://hdl.handle.net/10297/1453 webcite
- [27]Schneider WC: Phosphorous compounds in animal tissues. 1. Extraction and estimation of deoxypentose nucleic acid. J Biol Chem 1945, 161:293-295.
- [28]Ghose TK: Measurement of cellulase activities. International union of pure and applied chemistry. Pure Appl Chem 1987, 59:257-268.
- [29]Summer JB, Howell SF: A method for determination of saccharase activity. J Biol Chem 1935, 108:51-54.
- [30]Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the folin-phenol reagent. J Biol Chem 1951, 193:265-275.