Overexpressing target helper genes enhances secretion and glycosylation of recombinant proteins in Pichia pastoris under simulated microgravity

详细信息    查看全文

• 作者：Jie Huangfu ; Yinghua Xu ; Chun Li ; Jun Li
• 关键词：Recombinant protein secretion and glycosylation ; Pichia pastoris ; Simulate microgravity (SMG) effects ; Helper genes ; AHA1 and YAP1
• 刊名：Journal of Industrial Microbiology and Biotechnology
• 出版年：2016
• 出版时间：October 2016
• 年：2016
• 卷：43
• 期：10
• 页码：1429-1439
• 全文大小：1,113 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Chemistry
  Biotechnology
  Genetic Engineering
  Biochemistry
  Bioinformatics
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1476-5535
• 卷排序：43

文摘

In this study, the potential helper genes were identified through the data analysis of transcriptomic and proteomic profiling in recombinant Pichia pastoris cultured under simulated microgravity (SMG). Co-expressing of four genes PRX1, YAP1, AHA1, and YPT6, involved in the oxidative stress response and protein folding, exhibited promising helper factor effects on the recombinant protein yields in engineered P. pastoris, respectively. When two of the above genes were co-expressed simultaneously, β-glucuronidase (PGUS) specific activity was further increased by 30.3–50.6 % comparing with that of single helper gene, particularly when the oxidative stress response and protein folding genes were both present in the combinations. In addition, co-expressing co-chaperone AHA1 and transcription factor YAP1 not only enhanced PGUS secretion, but also affected its glycosylation. Thus, through deep “omics” analysis of SMG effects, our results provided combined impact of new helper factors to improve the efficacy of recombinant protein secretion and glycosylation in engineered P. pastoris.