Establishing a Eukaryotic Pichia pastoris Cell-Free Protein Synthesis System
2020-06-18
发表期刊FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY (IF:4.3[JCR-2023],5.1[5-Year])
ISSN22964185
卷号8
发表状态已发表
DOI10.3389/fbioe.2020.00536
摘要

In recent years, cell-free protein synthesis (CFPS) systems have been used to synthesize proteins, prototype genetic elements, manufacture chemicals, and diagnose diseases. These exciting, novel applications lead to a new wave of interest in the development of new CFPS systems that are derived from prokaryotic and eukaryotic organisms. The eukaryotic Pichia pastoris is emerging as a robust chassis host for recombinant protein production. To expand the current CFPS repertoire, we report here the development and optimization of a eukaryotic CFPS system, which is derived from a protease-deficient strain P. pastoris SMD1163. By developing a simple crude extract preparation protocol and optimizing CFPS reaction conditions, we were able to achieve superfolder green fluorescent protein (sfGFP) yields of 50.16 ± 7.49 μg/ml in 5 h batch reactions. Our newly developed P. pastoris CFPS system fits to the range of the productivity achieved by other eukaryotic CFPS platforms, normally ranging from several to tens of micrograms protein per milliliter in batch mode reactions. Looking forward, we believe that our P. pastoris CFPS system will not only expand the CFPS toolbox for synthetic biology applications, but also provide a novel platform for cost-effective, high-yielding production of complex proteins that need post-translational modification and functionalization.
© Copyright © 2020 Zhang, Liu and Li.

关键词Cost effectiveness Recombinant proteins Synthetic biology YeastCell free protein synthesis Eukaryotic organisms Functionalizations Green fluorescent protein Novel applications Post-translational modifications Reaction conditions Recombinant protein productions
收录类别SCI ; SCIE ; EI
语种英语
资助项目Shanghai Pujiang Program[18PJ1408000] ; National Natural Science Foundation of China[31971348][31800720] ; Natural Science Foundation of Shanghai[19ZR1477200]
WOS研究方向Biotechnology & Applied Microbiology ; Science & Technology - Other Topics
WOS类目Biotechnology & Applied Microbiology ; Multidisciplinary Sciences
WOS记录号WOS:000546976900001
出版者Frontiers Media S.A.
EI入藏号20202708900643
EI主题词Cost effectiveness ; Recombinant proteins ; Synthetic biology ; Yeast
EI分类号Biotechnology:461.8 ; Organic Compounds:804.1 ; Food Products:822.3 ; Industrial Economics:911.2
WOS关键词TRANSCRIPTION-TRANSLATION SYSTEM ; IN-VITRO BIOSYNTHESIS ; PICHIA-PASTORIS ; CELL ; EXTRACT ; EXPRESSION ; DEGRADATION ; SEQUENCE ; YEAST
原始文献类型Article
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文献类型期刊论文
条目标识符https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/120213
专题物质科学与技术学院
物质科学与技术学院_PI研究组_李健组
物质科学与技术学院_硕士生
通讯作者Li, Jian
作者单位
School of Physical Science and Technology, ShanghaiTech University, Shanghai, China
第一作者单位物质科学与技术学院
通讯作者单位物质科学与技术学院
第一作者的第一单位物质科学与技术学院
推荐引用方式
GB/T 7714
Zhang, Lingkai,Liu, Wan-Qiu,Li, Jian. Establishing a Eukaryotic Pichia pastoris Cell-Free Protein Synthesis System[J]. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY,2020,8.
APA Zhang, Lingkai,Liu, Wan-Qiu,&Li, Jian.(2020).Establishing a Eukaryotic Pichia pastoris Cell-Free Protein Synthesis System.FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY,8.
MLA Zhang, Lingkai,et al."Establishing a Eukaryotic Pichia pastoris Cell-Free Protein Synthesis System".FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY 8(2020).
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