纤维素水解液中抑制剂对纤维二糖利用菌株发酵能力的影响

摘要

木质纤维素在预处理的过程中会产生呋喃类、酚类和弱酸类物质,抑制菌株的生长和发酵。研究了典型抑制剂糠醛、苯酚和乙酸对可利用纤维二糖的工程酿酒酵母菌株生长和发酵的影响,考查了分别以纤维二糖和葡萄糖作为单一碳源时菌株对抑制剂的耐受能力。结果发现,糠醛对菌株的纤维二糖和葡萄糖利用能力的抑制作用随糠醛浓度增加而增大。低浓度(≤0.5 g/L)的苯酚对纤维二糖利用有一定的促进作用,高浓度(1.5 g/L)的苯酚会抑制菌株的生长和发酵。乙酸对菌株纤维二糖利用的影响最为显著,可以明显抑制菌株的生长、纤维二糖利用和乙醇生产,而一定浓度(0.5~4.0 g/L)的乙酸会促进葡萄糖的利用。研究中还发现,复合抑制剂对纤维二糖利用菌株的抑制作用强于单一抑制剂。

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	胡梦龙
	贺霖伟
	李炳志
	元英进

关键词：纤维素乙醇纤维二糖抑制剂纤维素水解液合成生物学

Abstract：

During the pretreatment of lignocellulosic materials for microbial fermentation, a wide range of inhibitors were generated, including furans, phenols and weak acids etc. These inhibitors would inhibit the cell growth and sugar utilization of fermentation microorganisms. In this study, three typical inhibitors, furfural, phenol and acetic acid, were used to investigate their influence on the genetically engineered Saccharomyces cerevisiae SyBE001603. The tolerance of strain to the three inhibitors when using cellobiose or glucose as the sole carbon source was evaluated. It indicated that furfural could inhibit both the cellobiose and glucose utilization capability of the strain, and more severe inhibition occured with higher furfural concentration. At a phenol concentration less than or equal to 0.5 g/L, the cellobiose utilization was facilitated, while at the phenol concentration of 1.5 g/L, the cell growth and cellobiose utilization were strongly inhibited. Acetic acid exhibited significant inhibition on the cell growth and the cellobiose utilization, and the cellobiose consumption rate and ethanol productivity were strongly reduced as the acetic acid concentration increased. However, when using glucose as the sole carbon source, acetic acid (0.5~4.0 g/L) would accelerate the glucose utilization. In addition, it was observed that the synergistic inhibition effects of multiple inhibitors were more severe than that of single inhibitor.

Keywords： cellulosic ethanol； cellobiose； inhibitor； lignocellulosic hydrolysate； synthetic biology

Received 2014-05-23;

Fund:

国家重点基础研究发展计划(973计划)(2013CB733600)国家自然科学基金项目(21020102040)资助。

Corresponding Authors: 李炳志,E-mail:bzli@tju.edu.cn。 Email: bzli@tju.edu.cn

About author: 胡梦龙(1990-),男,硕士研究生,主要从事纤维素乙醇方向的研究。

引用本文:

胡梦龙, 贺霖伟, 李炳志, 元英进.纤维素水解液中抑制剂对纤维二糖利用菌株发酵能力的影响[J]. 化学工业与工程, 2016,33(2): 77-84

Hu Menglong, He Linwei, Li Bingzhi, Yuan Yingjin.Influence of Inhibitors in Lignocellulosic Hydrolysates on Ethanol Fermentation of Cellobiose-Utilizing Saccharomyces cerevisiae Strain[J]. Chemcial Industry and Engineering, 2016,33(2): 77-84

[1] Zhong C, Cao Y, Li B, et al. Biofuels in China: Past, present and future[J]. Biofuels, Bioproducts and Biorefining, 2010, 4(3): 326-342
[2] Brethauer S, Wyman C E. Review: Continuous hydrolysis and fermentation for cellulosic ethanol production[J]. Bioresource Technology, 2010, 101(13): 4 862-4 874
[3] Talebnia F, Karakashev D, Angelidaki I. Production of bioethanol from wheat straw: An overview on pretreatment, hydrolysis and fermentation[J]. Bioresource Technology, 2010, 101(13): 4 744-4 753
[4] Hahn-Hägerdal B, Galbe M, Gorwa-Grauslund M F, et al. Bio-Ethanol:The fuel of tomorrow from the residues of today[J]. Trends in Biotechnology, 2006, 24(12): 549-556
[5] Hasunuma T, Kondo A. Consolidated bioprocessing and simultaneous saccharification and fermentation of lignocellulose to ethanol with thermotolerant yeast strains[J]. Process Biochemistry, 2012, 47(9): 1 287-1 294
[6] Chauve M, Mathis H, Huc D, et al. Comparative kinetic analysis of two fungal β-glucosidases[J]. Biotechnology for Biofuels, 2010, 3(6):1-8
[7] Voutilainen S P, Puranen T, Siikaaho M, et al. Cloning, expression, and characterization of novel thermostable family 7 cellobiohydrolases[J]. Biotechnology and Bioengineering, 2008, 101(3): 515-528
[8] Galazka J M, Tian C, Beeson W T, et al. Cellodextrin transport in yeast for improved biofuel production[J]. Science, 2010, 330(6 000): 84-86
[9] Ha S J, Galazka J M, Kim S R, et al. Engineered Saccharomyces cerevisiae capable of simultaneous cellobiose and xylose fermentation[J]. Proceedings of the National Academy of Sciences, 2011, 108(2): 504-509
[10] Palmqvist E, Hahn-Hägerdal B. Fermentation of lignocellulosic hydrolysates. II: Inhibitors and mechanisms of inhibition[J]. Bioresource Technology, 2000,74(1): 25-33
[11] Ding M, Wang X, Yang Y, et al. Comparative metabolic profiling of parental and inhibitors-tolerant yeasts during lignocellulosic ethanol fermentation[J]. Metabolomics, 2012, 8(2): 232-243
[12] Parawira W, Tekere M. Biotechnological strategies to overcome inhibitors in lignocellulose hydrolysates for ethanol production: Review[J]. Critical reviews in Biotechnology, 2011, 31(1): 20-31
[13] Palmqvist E, Grage H, Meinander N Q, et al. Main and interaction effects of acetic acid, furfural, and p-hydroxybenzoic acid on growth and ethanol productivity of yeasts[J]. Biotechnology and Bioengineering, 1999, 63(1): 46-55
[14] Palmqvist E, Almeida J S, Hahn-Hägerdal B. Influence of furfural on anaerobic glycolytic kinetics of Saccharomyces cerevisiae in batch culture[J]. Biotechnology and Bioengineering, 1999, 62(4): 447-454
[15] Almeida J R, Modig T, Petersson A, et al. Increased tolerance and conversion of inhibitors in lignocellulosic hydrolysates by Saccharomyces cerevisiae[J]. Journal of Chemical Technology and Biotechnology, 2007, 82(4): 340-349
[16] Liu Z. Genomic adaptation of ethanologenic yeast to biomass conversion inhibitors[J]. Applied Microbiology and Biotechnology, 2006, 73(1): 27-36
[17] Allen S A, Clark W, McCaffery J M, et al. Furfural induces reactive oxygen species accumulation and cellular damage in Saccharomyces cerevisiae[J]. Biotechnology for Biofuels, 2010, 3(2): 1-10
[18] Klinke H B, Thomsen A, Ahring B K. Inhibition of ethanol-producing yeast and bacteria by degradation products produced during pre-treatment of biomass[J]. Applied Microbiology and Biotechnology, 2004, 66(1): 10-26
[19] Keweloh H, Weyrauch G, Rehm H J. Phenol-Induced membrane changes in free and immobilized Escherichia coli[J]. Applied Microbiology and Biotechnology, 1990, 33(1): 66-71
[20] Larsson S, Palmqvist E, Hahn-Hägerdal B, et al. The generation of fermentation inhibitors during dilute acid hydrolysis of softwood[J]. Enzyme and Microbial Technology, 1999, 24(3): 151-159
[21] Russell J B, Diez-Gonzalez F. The effects of fermentation acids on bacterial growth[J]. Advances in Microbial Physiology, 1997, 39: 205-234
[22] Kim H, Lee W H, Galazka J M, et al. Analysis of cellodextrin transporters from Neurospora crassa in Saccharomyces cerevisiae for cellobiose fermentation[J]. Applied Microbiology and Biotechnology, 2014, 98(3): 1 087-1 094
[23] Spiro S, Guest J R. Adaptive responses to oxygen limitation in Escherichia coli[J]. Trends in Biochemical Sciences, 1991, 16(8): 310-314
[24] Lian J, Li Y, Hamedirad M, et al. Directed evolution of a cellodextrin transporter for improved biofuel production under anaerobic conditions in Saccharomyces cerevisiae[J]. Biotechnology & Bioengineering, 2014, 111(8):1 521-1 531
[25] Ha S J, Galazka J M, Joong O E, et al. Energetic benefits and rapid cellobiose fermentation by Saccharomyces cerevisiae expressing cellobiose phosphorylase and mutant cellodextrin transporters[J]. Metabolic Engineering, 2013, 15:134-143
[26] Martinez A, Rodriguez M E, Wells M L, et al. Detoxification of dilute acid hydrolysates of lignocellulose with lime[J]. Biotechnology Progress, 2001, 17(2): 287-293