非离子表面活性剂对生物丁醇发酵的影响
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摘要
传统的丙酮-丁醇发酵的产物浓度过低(丁醇终浓度约为1.3 wt%),导致后期分离成本过高,从而影响了该过程的经济性,限制了其工业化进程。本文研究了高添加量的小分子非离子表面活性剂对生物丁醇发酵的影响。以吐温80为例,实验表明,当表面活性剂添加量超过其临界胶束浓度后,丁醇发酵的终浓度会随着表面活性剂添加量的增加而增加。当添加量达到5 wt%时,丁醇终浓度可以达到1.6 wt%,远高于该菌种的抑制浓度(0.8 wt%)。为阐明表面活性剂的作用机理,实验考察了吐温80对丁醇的增溶效应以及对发酵菌体表面亲疏水性的影响。结果表明,吐温80对丁醇的增溶效果很小,而对菌体表面的亲疏水性有较明显的影响。
The low butanol concentration of acetone-butanol-ethanol fermentation causes uneconomical product recovery. In this work, the effect of small molecule non-ionic surfactants on butanol fermentation was evaluated, using laboratory stocks of Clostridium acetobutylicum ATCC 824. Non-ionic surfactants substantially increased butanol production when additive amount was higher than 1%(W/W). Butanol concentration reached 16.9 g/L with 5%(W/W) Tween 80 and 100 g/L glucose in a 5 L fermenter. It was found that surfactants micelle solubilization capacity to butanol was very limited, indicating that butanol could hardly enter the surfactants micelle. Butanol production improvement was probably caused by cell surface hydrophobicity change due to surfactants adsorption.
The low butanol concentration of acetone-butanol-ethanol fermentation causes uneconomical product recovery. In this work, the effect of small molecule non-ionic surfactants on butanol fermentation was evaluated, using laboratory stocks of Clostridium acetobutylicum ATCC 824. Non-ionic surfactants substantially increased butanol production when additive amount was higher than 1%(W/W). Butanol concentration reached 16.9 g/L with 5%(W/W) Tween 80 and 100 g/L glucose in a 5 L fermenter. It was found that surfactants micelle solubilization capacity to butanol was very limited, indicating that butanol could hardly enter the surfactants micelle. Butanol production improvement was probably caused by cell surface hydrophobicity change due to surfactants adsorption.
引文
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[17]Qureshi N,Blaschek HP.Butanol recovery from model solution/fermentation broth by pervaporation:evaluation of membrane performance.Biomass Bioenergy,1999,17(2):175–184.
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[20]Pursell MR,Mendes MA,Stuckey DC.Interactions between surfactants and biomass during liquid–liquid extraction.Biotechnol Prog,2009,25(6):1686–1694.
[21]Hua Z,Chen J,Lun S,et al.Influence of biosurfactants produced by Candida antarctica on surface properties of microorganism and biodegradation of n-alkanes.Water Res,2003,37(17):4143–4150.
[22]Arino S,Marchal R,Vandecasteele JP.Identification and production of a rhamnolipidic biosurfactant by a Pseudomonas species.Appl Microbiol Biotechnol,1996,45(1/2):162–168.
[23]Ahimou F,Jacques P,Deleu M.Surfactin and iturin A effects on Bacillus subtilis surface hydrophobicity.Enzyme Microb Technol,2000,27(10):749–754.
[24]Yang Y,Zhang LY,Shi ZP,Improvement of extractive fermentation performance of butanol by adding surfactacts into fermentation medium.Chin J Bioprocess Eng,2008,6(4):25–30(in Chinese).杨影,张龙云,史仲平,添加表面活性剂改善丁醇萃取发酵性能.生物加工过程,2008,6(4):25–30.
[25]Zhang YM,Maier WJ,Miller RM.Effect of rhamnolipids on the dissolution,bioavailability and biodegradation of phenanthrene.Environ Sci Technol,1997,31(8):2211–2217.
[26]Niu XL.Study on butanol extractive fermentation with oleyl alcohol and recovery product in extraction raffinate[D].Wuxi:Jiangnan University,2008(in Chinese).牛晓乐.以油醇为高效萃取剂的丁醇萃取发酵和回收萃余液中溶剂产物的研究[D].无锡:江南大学,2008.
[2]Swana J,Yang Y,Behnam M,et al.An analysis of net energy production and feedstock availability for biobutanol and bioethanol.Bioresour Technol,2011,102(2):2112–2117.
[3]Ranjan A,Moholkar VS.Biobutanol:science,engineering and economics.Int J Energy Res,2012,36(3):277–323.
[4]García V,P?kkil?J,Ojamo H.Challenges in biobutanol production:how to improve the efficiency?Renew Sust Energy Rev,2011,15(2):964–980.
[5]Kumar M,Gayen K.Developments in biobutanol production:new insights.Appl Energ,2011,88(6):1999–2012.
[6]Kharkwal S,Karimi IA,Chang MW,et al.Strain improvement and process development for biobutanol production.Recent Pat Biotechnol,2009,3(3):202–210.
[7]Li WJ,Chen CY,Cui HD,et al.Optimization of fermentation condition for Clostridium acetobutylicum.China Brew,2013,32(2):13–16(in Chinese).李伟佳,陈春燕,崔海娣,等.丙酮丁醇梭菌发酵条件优化研究.中国酿造,2013,32(2):13–16.
[8]Wang FQ,Yuan H,Xie H,et al.Mutation breeding of butanol high-yielding strain and its fermentation condition optimization.China Brew,2011,230(5):84–86(in Chinese).王风芹,原欢,谢慧,等.丁醇高产菌株诱变育种及发酵条件优化研究.中国酿造,2011,230(5):84–86.
[9]Mermelstein LD,Papoutsakis ET,Petersen DJ,et al.Metabolic engineering of Clostridium acetobutylicum ATCC 824 for increased solvent production by enhancement of acetone formation enzyme activities using a synthetic acetone operon.Biotechnol Bioeng,1993,42(9):1053–1060.
[10]Ishizaki A,Michiiwaki S,Crabbe E,et al.Extractive acetone-butanol-ethanol fermentation using methylated crude palm oil as extractant in batch culture of Clostridium saccharoperbutylacetonicum N1-4(ATCC 13564).J Biosci Bioeng,1999,87(3):352–356.
[11]Yang LR,Cen PL,Zhu ZQ.Study on acetone/butanol fermentation with in-situ solvent extraction.J Zhejiang Univ:Nat Sci Ed,1992,26(4):388–398(in Chinese).杨立荣,岑沛霖,朱自强.丙酮/丁醇间歇萃取发酵.浙江大学学报:自然科学版,1992,26(4):388–398.
[12]Ezeji TC,Qureshi N,Blaschek HP.Production of acetone,butanol and ethanol by Clostridium beijerinckii BA101 and in situ recovery by gas stripping.World J Microb Biot,2003,19(6):595–603.
[13]Ezeji TC,Qureshi N,Blaschek HP.Acetone butanol ethanol(ABE)production from concentrated substrate:reduction in substrate inhibition by fed-batch technique and product inhibition by gas stripping.Appl Microbiol Biot,2004,63(6):653–658.
[14]Banat FA,Al-Shannag M.Recovery of dilute acetone–butanol–ethanol(ABE)solvents from aqueous solutions via membrane distillation.Bioprocess Eng,2000,23(6):643–649.
[15]Elkina IB,Gilman AB,Ugrozov VV,et al.Separation of mineral acid solutions by membrane distillation and thermopervaporation through porous and nonporous membranes.Ind Eng Chem Res,2013,52(26):8856–8863.
[16]Li SY,Srivastava R,Parnas RS.Study of in situ1-butanol pervaporation from A-B-E fermentation using a PDMS composite membrane:validity of solution-diffusion model for pervaporative A-B-E fermentation.Biotechnol Progr,2011,27(1):111–120.
[17]Qureshi N,Blaschek HP.Butanol recovery from model solution/fermentation broth by pervaporation:evaluation of membrane performance.Biomass Bioenergy,1999,17(2):175–184.
[18]Liu GP,Wang W,Wu H,et al.Pervaporation performance of PDMS/ceramic composite membrane in acetone butanol ethanol(ABE)fermentation–PV coupled process.J Membrane Sci,2011,373(1/2):121–129.
[19]Liu GP,Wang W,Wu H,et al.Pervaporation membranes for biobutanol production.ACS Sustainable Chem,Eng,2013,2(4):546–560.
[20]Pursell MR,Mendes MA,Stuckey DC.Interactions between surfactants and biomass during liquid–liquid extraction.Biotechnol Prog,2009,25(6):1686–1694.
[21]Hua Z,Chen J,Lun S,et al.Influence of biosurfactants produced by Candida antarctica on surface properties of microorganism and biodegradation of n-alkanes.Water Res,2003,37(17):4143–4150.
[22]Arino S,Marchal R,Vandecasteele JP.Identification and production of a rhamnolipidic biosurfactant by a Pseudomonas species.Appl Microbiol Biotechnol,1996,45(1/2):162–168.
[23]Ahimou F,Jacques P,Deleu M.Surfactin and iturin A effects on Bacillus subtilis surface hydrophobicity.Enzyme Microb Technol,2000,27(10):749–754.
[24]Yang Y,Zhang LY,Shi ZP,Improvement of extractive fermentation performance of butanol by adding surfactacts into fermentation medium.Chin J Bioprocess Eng,2008,6(4):25–30(in Chinese).杨影,张龙云,史仲平,添加表面活性剂改善丁醇萃取发酵性能.生物加工过程,2008,6(4):25–30.
[25]Zhang YM,Maier WJ,Miller RM.Effect of rhamnolipids on the dissolution,bioavailability and biodegradation of phenanthrene.Environ Sci Technol,1997,31(8):2211–2217.
[26]Niu XL.Study on butanol extractive fermentation with oleyl alcohol and recovery product in extraction raffinate[D].Wuxi:Jiangnan University,2008(in Chinese).牛晓乐.以油醇为高效萃取剂的丁醇萃取发酵和回收萃余液中溶剂产物的研究[D].无锡:江南大学,2008.