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学科主题: 物理化学
题名: Novel helper factors influencing recombinant protein production in Pichia pastoris based on proteomic analysis under simulated microgravity
作者: Jie Huangfu1;  Qi, Feng1, 3;  Liu, Hongwei2;  Zou, Hanfa2;  Ahmed, Muhammad Saad1;  Li, Chun1
通讯作者: ChunLi
关键词: Recombinant protein ;  Strengthen production ;  Proteomics analysis ;  Simulate microgravity (SMG) ;  Pichia pastoris
刊名: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
发表日期: 2015
DOI: 10.1007/s00253-014-6175-8
卷: 99, 期:2, 页:653-665
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biotechnology & Applied Microbiology
研究领域[WOS]: Biotechnology & Applied Microbiology
英文摘要: Microgravity and simulated microgravity (SMG) have quite significant effects on numerous microbial cellular processes. The effects of SMG on the production of recombinant proteins and transcription profiling in prokaryotic and eukaryotic expression host have been investigated. The present study showed that SMG significantly enhanced the specific productivities and activities of the reporter enzymes PGUS and AtXYN that were expressed in recombinant Pichia pastoris. Proteomic profiling revealed that 21 proteins were significantly up-regulated and 35 proteins were drastically down-regulated at the stationary phase, when the recombinant P. pastoris responded to SMG. Six strongly up-regulated genes, TPX, FBA, PGAM, ENO, SBA1, and AKR-E, involved in the oxidative stress response, methanol metabolism, glycolytic pathway, and protein folding, were selected to analyze their impacts on recombinant protein production by co-overexpression in the shaker flask fermentation. The co-overexpressed strains, particularly TPX, FBA, and PGAM, demonstrated promising results with approximately 2.46-fold, 1.58-fold, and 1.33-fold increases in the specific yields of PGUS compared to the control after 48 h of methanol induction, respectively. In the meantime, the corresponding PGUS specific activities were increased by 2.33-fold, 2.09-fold, and 1.32-fold, respectively. Thiol peroxidase (TPX), which is involved in the oxidative stress response, significantly influenced the transcriptional levels of the reporter gene PGUS. The present study provides valuable information for further exploration of the molecular mechanism of P. pastoris response to SMG and facilitates simulated microgravity for finding novel helper factors to rationally engineer the strains in normal fermentation by using proteomic studies.
关键词[WOS]: SHEAR MODELED MICROGRAVITY ;  GENE-EXPRESSION ;  SECRETION ;  BIOSYNTHESIS ;  RESPONSES ;  STRESS
语种: 英语
WOS记录号: WOS:000348770900010
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/143792
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Beijing Inst Technol, Sch Life Sci, Beijing 100081, Peoples R China
2.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
3.Fujian Normal Univ, Engn Res Ctr Ind Microbiol, Coll Life Sci, Fuzhou 350108, Peoples R China

Recommended Citation:
Jie Huangfu,Qi, Feng,Liu, Hongwei,et al. Novel helper factors influencing recombinant protein production in Pichia pastoris based on proteomic analysis under simulated microgravity[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2015,99(2):653-665.
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