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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]) |
ISSN | 22964185 |
卷号 | 8 |
发表状态 | 已发表 |
DOI | 10.3389/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. |
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 |
引用统计 | 正在获取...
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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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