Construction of Yeast Surface Display System and Application in Cellulose Degrading

doi:10.13523/j.cb.2112044

China Biotechnology

2022, Vol. 42

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

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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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

Received: 19 December 2021 Published: 17 June 2022

ZTFLH:

Q814

Corresponding Authors: Xu-dong FENG E-mail: xd.feng@bit.edu.cn

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	Ming-zhu LIU
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	Xu-dong FENG

Cite this article:

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.

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

Table 1 The main components of the cell wall of Saccharomyces cerevisiae

Fig.1 The diagram of the construction of cellulase display on the surface of yeast cells (a) Direct display system (b) Indirect display system Where cohesin-dockerin is selected as an example

Fig.2 Typical yeast surface display system (a) GPI system. This system can display the N-terminus of the foreign protein, which is covalently linked to β-1, 6-glucan (b) Pir system. This system can display the N-terminus of the foreign protein, or insert the foreign protein into the Pir protein, which is covalently linked with β-1, 3-glucan (c) Flo1 system. This system can display the C-terminus or N-terminus of the foreign protein, which is non-covalently linked to β-1, 3-glucan

Table 2 Yeast surface-displayed cellulase system for ethanol fermentation


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