DICP OpenIR
Subject Area物理化学
Novel helper factors influencing recombinant protein production in Pichia pastoris based on proteomic analysis under simulated microgravity
Jie Huangfu1; Qi, Feng1,3; Liu, Hongwei2; Zou, Hanfa2; Ahmed, Muhammad Saad1; Li, Chun1; ChunLi
KeywordRecombinant Protein Strengthen Production Proteomics Analysis Simulate Microgravity (Smg) Pichia Pastoris
Source PublicationAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY
2015
DOI10.1007/s00253-014-6175-8
Volume99Issue:2Pages:653-665
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiotechnology & Applied Microbiology
WOS Research AreaBiotechnology & Applied Microbiology
WOS KeywordSHEAR MODELED MICROGRAVITY ; GENE-EXPRESSION ; SECRETION ; BIOSYNTHESIS ; RESPONSES ; STRESS
AbstractMicrogravity 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.
Language英语
WOS IDWOS:000348770900010
Citation statistics
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/143792
Collection中国科学院大连化学物理研究所
Corresponding AuthorChunLi
Affiliation1.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
GB/T 7714
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.
APA Jie Huangfu.,Qi, Feng.,Liu, Hongwei.,Zou, Hanfa.,Ahmed, Muhammad Saad.,...&ChunLi.(2015).Novel helper factors influencing recombinant protein production in Pichia pastoris based on proteomic analysis under simulated microgravity.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,99(2),653-665.
MLA Jie Huangfu,et al."Novel helper factors influencing recombinant protein production in Pichia pastoris based on proteomic analysis under simulated microgravity".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 99.2(2015):653-665.
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