酵母表面展示体系的构建及在纤维素降解中的应用<sup>*</sup>

doi:10.13523/j.cb.2112044

中国生物工程杂志

2022, Vol. 42

Issue (5): 91-99 DOI: 10.13523/j.cb.2112044

综述

酵母表面展示体系的构建及在纤维素降解中的应用^*

刘明珠¹,张良¹,郭芳¹,李春^1,^2,³,冯旭东^1,^**(

)

1 北京理工大学化学与化工学院化学工程系生物化工研究所医药分子科学与制剂工程工业和信息化部重点实验室北京 100081
2 清华大学化学工程系工业生物催化教育部重点实验室北京 100084
3 清华大学合成与系统生物学研究中心北京 100084

Construction of Yeast Surface Display System and Application in Cellulose Degrading

LIU Ming-zhu¹,ZHANG Liang¹,GUO Fang¹,LI Chun^1,^2,³,FENG Xu-dong^1,^**(

)

1 Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering, Ministry of Industry and Information Technology, Institute of Biochemical Engineering, Department of Chemical Engineering, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 100081, China
2 Key Lab for Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China
3 Center for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China

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摘要：

纤维素是来源广泛且储量较大的低成本可再生资源,但其结构致密难以利用。目前降解纤维素需要多种纤维素酶协作,而游离纤维素酶成本高、难以重复利用等问题限制了其广泛应用。利用酵母表面展示技术,可以将多个纤维素酶分别与锚定蛋白融合后共展示在细胞表面,从而构建酵母表面展示纤维素酶体系。这一体系可高效降解纤维素,一方面可以充分发挥表面展示的优点,如易回收、稳定性好、操作简单、成本低;另一方面可以将纤维素有效地降解为葡萄糖,并具有代谢产生物乙醇的潜力。阐述了酵母表面展示体系的构建原则,总结了影响展示体系效率的因素,介绍了这一技术在降解纤维素中的应用,为构建高效酵母表面展示纤维素酶体系及其他多酶体系提供参考。

关键词： 酵母表面展示; 纤维素酶; 锚定蛋白

Abstract:

Cellulose is a low-cost and renewable resource with abundant reserves, but it is difficult to be used because of its compact structure. Currently, the degradation of cellulose requires the cooperation of a variety of cellulases, but the high cost and the difficulty of re-use of free cellulase have limited its wide application. Yeast surface display technology can display multiple cellulase enzymes on the cell surface after being fused with the anchoring protein, so that a yeast surface display cellulase system could be constructed. This system can degrade cellulose efficiently. On the one hand, it has the advantages of surface display, such as easy recycling, good stability, simple operation, and low cost; on the other hand, it can also degrade cellulose into glucose effectively, and has the potential to produce bioethanol by metabolism. The paper summarizes the construction principles of a yeast surface display system, and divides yeast surface display systems into direct display systems and indirect display systems according to the different anchoring methods of exogenous proteins. The direct display system is that the exogenous protein is directly connected to the cell through the anchor protein. Indirect display systems are linked to cells through exogenous proteins by means of scaffolds. Factors affecting the efficiency of the display system mainly include cell type, anchor protein, scaffold, environmental factors and promoter type. The surface display system consists of GPI system, Pir system and FlO1 system according to different anchor proteins. The yeast surface display cellulase system can ferment cellulose to produce ethanol, and a lot of progress has been made so far, which provides a reference for the construction of an efficient yeast surface display cellulase system and other multi-enzyme systems. The review describes the construction principles of a yeast surface display system, summarizes the influence of major factors on the display system, and introduces the application of this technology in the degradation of cellulose. It provides guidance for the construction of high-efficiency yeast surface display of cellulase and other multi-enzyme systems.

Key words: Yeast surface display Cellulase Anchor protein

收稿日期: 2021-12-19 出版日期: 2022-06-17

ZTFLH:

Q814

基金资助: *北京市科技新星计划(Z191100001119099)

通讯作者: 冯旭东 E-mail: xd.feng@bit.edu.cn

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引用本文:

刘明珠,张良,郭芳,李春,冯旭东. 酵母表面展示体系的构建及在纤维素降解中的应用^*[J]. 中国生物工程杂志, 2022, 42(5): 91-99.

LIU Ming-zhu,ZHANG Liang,GUO Fang,LI Chun,FENG Xu-dong. Construction of Yeast Surface Display System and Application in Cellulose Degrading. China Biotechnology, 2022, 42(5): 91-99.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2112044 或 https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I5/91

表1 酿酒酵母细胞壁的主要成分

图1 酵母表面展示纤维素酶体系示意图

图2 典型的酵母表面展示系统

表2 酵母表面展示纤维素酶体系用于发酵乙醇

[1]	Yamada R, Nakatani Y, Ogino C, et al. Efficient direct ethanol production from cellulose by cellulase- and cellodextrin transporter-co-expressing Saccharomyces cerevisiae. AMB Express, 2013, 3: 34. doi: 10.1186/2191-0855-3-34
[2]	Wachtmeister J, Rother D. Recent advances in whole cell biocatalysis techniques bridging from investigative to industrial scale. Current Opinion in Biotechnology, 2016, 42: 169-177. doi: S0958-1669(16)30140-9 pmid: 27318259
[3]	Smith M R, Gao H, Prabhu P, et al. Elucidating structure-performance relationships in whole-cell cooperative enzyme catalysis. Nature Catalysis, 2019, 2 (9): 809-819. doi: 10.1038/s41929-019-0321-8
[4]	Hossain S A, Rahman S R, Ahmed T, et al. An overview of yeast cell wall proteins and their contribution in yeast display system. Asian Journal of Medical and Biological Research, 2020, 5(4): 246-257. doi: 10.3329/ajmbr.v5i4.45261
[5]	Seo M J, Schmidt Dannert C. Organizing multi-enzyme systems into programmable materials for biocatalysis. Catalysts, 2021, 11(4): 409. doi: 10.3390/catal11040409
[6]	冯旭东, 李春. 酶的改造及其催化工程应用. 化学进展, 2015, 27(11): 1649-1657. doi: 10.7536/PC150419
	Feng X D, Li C. The improvement of enzyme properties and its catalytic engineering strategy. Progress in Chemistry, 2015, 27(11): 1649-1657. doi: 10.7536/PC150419
[7]	Salgaonkar M, Nadar S S, Rathod V K. Combi-metal organic framework (Combi-MOF) of α-amylase and glucoamylase for one pot starch hydrolysis. International Journal of Biological Macromolecules, 2018, 113: 464-475. doi: S0141-8130(17)34734-7 pmid: 29458106
[8]	Asghar A, Iqbal N, Noor T, et al. Efficient one-pot synthesis of a hexamethylenetetramine-doped Cu-BDC metal-organic framework with enhanced CO2 adsorption. Nanomaterials (Basel, Switzerland), 2019, 9(8): 1063.
[9]	Trobo Maseda L, H Orrego A, Guisan J M, et al. Coimmobilization and colocalization of a glycosyltransferase and a sucrose synthase greatly improves the recycling of UDP-glucose: glycosylation of resveratrol 3-O-β-D-glucoside. International Journal of Biological Macromolecules, 2020, 157: 510-521. doi: S0141-8130(20)32982-2 pmid: 32344088
[10]	Thygesen A, Oddershede J, Lilholt H, et al. On the determination of crystallinity and cellulose content in plant fibres. Cellulose, 2005, 12(6): 563-576. doi: 10.1007/s10570-005-9001-8
[11]	Yamada R, Hasunuma T, Kondo A. Endowing non-cellulolytic microorganisms with cellulolytic activity aiming for consolidated bioprocessing. Biotechnology Advances, 2013, 31(6): 754-763. doi: 10.1016/j.biotechadv.2013.02.007
[12]	Ellis G A, Klein W P, Lasarte-Aragonés G, et al. Artificial multienzyme scaffolds: pursuing in vitro substrate channeling with an overview of current progress. ACS Catalysis, 2019, 9(12): 10812-10869. doi: 10.1021/acscatal.9b02413
[13]	Bae J G, Kuroda K, Ueda M. Proximity effect among cellulose-degrading enzymes displayed on the Saccharomyces cerevisiae cell surface. Applied and Environmental Microbiology, 2015, 81(1): 59-66. doi: 10.1128/AEM.02864-14
[14]	Bischof R H, Ramoni J, Seiboth B. Cellulases and beyond: the first 70 years of the enzyme producer Trichoderma reesei. Microbial Cell Factories, 2016, 15(1): 106. doi: 10.1186/s12934-016-0507-6 pmid: 27287427
[15]	Oh E J, Jin Y S. Engineering of Saccharomyces cerevisiae for efficient fermentation of cellulose. FEMS Yeast Research, 2020, 20(1): foz089. doi: 10.1093/femsyr/foz089
[16]	Andreu C, del Olmo M L. Yeast arming systems: pros and cons of different protein anchors and other elements required for display. Applied Microbiology and Biotechnology, 2018, 102(6): 2543-2561. doi: 10.1007/s00253-018-8827-6
[17]	Liu X, Cheng J, Zhang G, et al. Engineering yeast for the production of breviscapine by genomic analysis and synthetic biology approaches. Nature Communications, 2018, 9: 448. doi: 10.1038/s41467-018-02883-z
[18]	Liang B, Li L, Tang X J, et al. Microbial surface display of glucose dehydrogenase for amperometric glucose biosensor. Biosensors and Bioelectronics, 2013, 45: 19-24. doi: 10.1016/j.bios.2013.01.050 pmid: 23454338
[19]	Tabañag I D F, Chu I M, Wei Y H, et al. The role of yeast-surface-display techniques in creating biocatalysts for consolidated BioProcessing. Catalysts, 2018, 8(3): 94. doi: 10.3390/catal8030094
[20]	Fan L H, Zhang Z J, Yu X Y, et al. Self-surface assembly of cellulosomes with two miniscaffoldins on Saccharomyces cerevisiae for cellulosic ethanol production. PNAS, 2012, 109(33): 13260-13265. doi: 10.1073/pnas.1209856109
[21]	Tanaka T, Matsumoto S, Yamada M, et al. Display of active beta-glucosidase on the surface of Schizosaccharomyces pombe cells using novel anchor proteins. Applied Microbiology and Biotechnology, 2013, 97(10): 4343-4352. doi: 10.1007/s00253-013-4733-0
[22]	Anandharaj M, Lin Y J, Rani R P, et al. Constructing a yeast to express the largest cellulosome complex on the cell surface. Proceedings of the National Academy of Sciences of the United States of America, 2020, 117(5): 2385-2394.
[23]	Yang X Y, Tang H T, Song M H, et al. Development of novel surface display platforms for anchoring heterologous proteins in Saccharomyces cerevisiae. Microbial Cell Factories, 2019, 18(1): 85. doi: 10.1186/s12934-019-1133-x
[24]	Yamada R, Taniguchi N, Tanaka T, et al. Cocktail delta-integration: a novel method to construct cellulolytic enzyme expression ratio-optimized yeast strains. Microbial Cell Factories, 2010, 9: 32. doi: 10.1186/1475-2859-9-32
[25]	Apiwatanapiwat W, Murata Y, Kosugi A, et al. Direct ethanol production from cassava pulp using a surface-engineered yeast strain co-displaying two amylases, two cellulases, and β-glucosidase. Applied Microbiology and Biotechnology, 2011, 90(1): 377-384. doi: 10.1007/s00253-011-3115-8 pmid: 21327413
[26]	Artzi L, Bayer E A, Moraïs S. Cellulosomes: bacterial nanomachines for dismantling plant polysaccharides. Nature Reviews Microbiology, 2017, 15 (2): 83-95. doi: 10.1038/nrmicro.2016.164 pmid: 27941816
[27]	Liang Y Y, Si T, Ang E L, et al. Engineered pentafunctional minicellulosome for simultaneous saccharification and ethanol fermentation in Saccharomyces cerevisiae. Applied and Environmental Microbiology, 2014, 80(21): 6677-6684. doi: 10.1128/AEM.02070-14
[28]	Fan S Q, Liang B, Xiao X X, et al. Controllable display of sequential enzymes on yeast surface with enhanced biocatalytic activity toward efficient enzymatic biofuel cells. Journal of the American Chemical Society, 2020, 142(6): 3222-3230. doi: 10.1021/jacs.9b13289
[29]	Cunha J T, Romaní A, Inokuma K, et al. Consolidated bioprocessing of corn cob-derived hemicellulose: engineered industrial Saccharomyces cerevisiae as efficient whole cell biocatalysts. Biotechnology for Biofuels, 2020, 13: 138. doi: 10.1186/s13068-020-01780-2
[30]	Camattari A, Goh A, Yip L Y, et al. Characterization of a panARS-based episomal vector in the methylotrophic yeast Pichia pastoris for recombinant protein production and synthetic biology applications. Microbial Cell Factories, 2016, 15(1): 139. doi: 10.1186/s12934-016-0540-5 pmid: 27515025
[31]	Liu X Y, Chi Z, Liu G L, et al. Inulin hydrolysis and citric acid production from inulin using the surface-engineered Yarrowia lipolytica displaying inulinase. Metabolic Engineering, 2010, 12(5): 469-476. doi: 10.1016/j.ymben.2010.04.004
[32]	Ni X M, Yue L X, Chi Z M, et al. Alkaline protease gene cloning from the marine yeast Aureobasidium pullulans HN2-3 and the protease surface display on Yarrowia lipolytica for bioactive peptide production. Marine Biotechnology (New York), 2009, 11( 1): 81-89.
[33]	Liu Z, Inokuma K, Ho S H, et al. Improvement of ethanol production from crystalline cellulose via optimizing cellulase ratios in cellulolytic Saccharomyces cerevisiae. Biotechnology and Bioengineering, 2017, 114(6): 1201-1207. doi: 10.1002/bit.26252
[34]	Liu G L, Yue L X, Chi Z, et al. The surface display of the alginate lyase on the cells of Yarrowia lipolytica for hydrolysis of alginate. Marine Biotechnology, 2009, 11(5): 619-626. doi: 10.1007/s10126-009-9178-1
[35]	Yanase S, Hasunuma T, Yamada R, et al. Direct ethanol production from cellulosic materials at high temperature using the thermotolerant yeast Kluyveromyces marxianus displaying cellulolytic enzymes. Applied Microbiology and Biotechnology, 2010, 88(1): 381-388. doi: 10.1007/s00253-010-2784-z
[36]	Wang X D, Feng X D, Lv B, et al. Enhanced yeast surface display of β-glucuronidase using dual anchor motifs for high-temperature glycyrrhizin hydrolysis. AIChE Journal, 2019, 65(9): e16629.
[37]	Grimm A R, Sauer D F, Polen T, et al. A whole cell E. coli display platform for artificial metalloenzymes: poly(phenylacetylene) production with a rhodium-nitrobindin metalloprotein. ACS Catalysis, 2018, 8(3): 2611-2614. doi: 10.1021/acscatal.7b04369
[38]	Sato H, Hayashi T, Ando T, et al. Hybridization of modified-heme reconstitution and distal histidine mutation to functionalize sperm whale myoglobin. Journal of the American Chemical Society, 2004, 126(2): 436-437. doi: 10.1021/ja038798k
[39]	Onoda A, Fukumoto K, Arlt M, et al. A rhodium complex-linked β-barrel protein as a hybrid biocatalyst for phenylacetylene polymerization. Chemical Communications (Cambridge, England), 2012, 48(78): 9756-9758. doi: 10.1039/c2cc35165j
[40]	Grimm A R, Sauer D F, Davari M D, et al. Cavity size engineering of a β-barrel protein generates efficient biohybrid catalysts for olefin metathesis. ACS Catalysis, 2018, 8(4): 3358-3364. doi: 10.1021/acscatal.7b03652
[41]	Dong M S, Li T P, Li S H, et al. Expression and enzymatic characterization of rice α-galactosidase II displayed on yeast cell surface. Process Biochemistry, 2019, 81: 57-62. doi: 10.1016/j.procbio.2019.03.016
[42]	Dong M S, Gong Y, Guo J, et al. Optimization of production conditions of rice α-galactosidase II displayed on yeast cell surface. Protein Expression and Purification, 2020, 171: 105611. doi: 10.1016/j.pep.2020.105611
[43]	Inokuma K, Hasunuma T, Kondo A. Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and its original promoter. Biotechnology for Biofuels, 2014, 7(1): 8. doi: 10.1186/1754-6834-7-8
[44]	Baek S H, Kim S, Lee K, et al. Cellulosic ethanol production by combination of cellulase-displaying yeast cells. Enzyme and Microbial Technology, 2012, 51(6-7): 366-372. doi: 10.1016/j.enzmictec.2012.08.005
[45]	Tsai S L, DaSilva N A, Chen W. Functional display of complex cellulosomes on the yeast surface via adaptive assembly. ACS Synthetic Biology, 2013, 2(1): 14-21. doi: 10.1021/sb300047u
[46]	Kim S, Baek S H, Lee K, et al. Cellulosic ethanol production using a yeast consortium displaying a minicellulosome and β-glucosidase. Microbial Cell Factories, 2013, 12: 14. doi: 10.1186/1475-2859-12-14
[47]	Liu Z, Inokuma K, Ho S H, et al. Combined cell-surface display- and secretion-based strategies for production of cellulosic ethanol with Saccharomyces cerevisiae. Biotechnology for Biofuels, 2015, 8: 162. doi: 10.1186/s13068-015-0344-6
[48]	Fan L H, Zhang Z J, Mei S, et al. Engineering yeast with bifunctional minicellulosome and cellodextrin pathway for co-utilization of cellulose-mixed sugars. Biotechnology for Biofuels, 2016, 9: 137. doi: 10.1186/s13068-016-0554-6
[49]	Liu Z, Ho S H, Sasaki K, et al. Engineering of a novel cellulose-adherent cellulolytic Saccharomyces cerevisiae for cellulosic biofuel production. Scientific Reports, 2016, 6: 24550. doi: 10.1038/srep24550
[50]	Tabañag I D F, Chu I M, Wei Y H, et al. Ethanol production from hemicellulose by a consortium of different genetically-modified Sacharomyces cerevisiae. Journal of the Taiwan Institute of Chemical Engineers, 2018, 89: 15-25. doi: 10.1016/j.jtice.2018.04.029

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