期刊
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
卷 43, 期 10, 页码 1429-1439出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s10295-016-1817-8
关键词
Recombinant protein secretion and glycosylation; Pichia pastoris; Simulate microgravity (SMG) effects; Helper genes; AHA1 and YAP1
资金
- National Natural Science Foundation of China [21425624, 21376028, 21566032]
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, beta-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.
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