title: Overexpressing target helper genes enhances secretion and glycosylation of recombinant proteins in Pichia pastoris under simulated microgravity.
abstract: 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.
authors.affiliation: School of Life Science, Beijing Institute of Technology, Beijing, 100081, China.|China National Research Institute of Food and Fermentation Industries, Beijing, 100027, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Huangfu
authors.affiliation: School of Life Science, Beijing Institute of Technology, Beijing, 100081, China.
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Xu
authors.affiliation: School of Life Science, Beijing Institute of Technology, Beijing, 100081, China.
authors.forename: Chun
authors.initials: C
authors.lastname: Li
authors.affiliation: School of Life Science, Beijing Institute of Technology, Beijing, 100081, China. junlipitt@163.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
mesh_terms: D060846:Cell Engineering; D015870:Gene Expression; D006031:Glycosylation; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D017510:Protein Folding; D040901:Proteomics; D011994:Recombinant Proteins / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D018474:Weightlessness Simulation
chemical_list: D018832:Molecular Chaperones; D011994:Recombinant Proteins; D014157:Transcription Factors
keywords: AHA1 and YAP1; Helper genes; Pichia pastoris; Recombinant protein secretion and glycosylation; Simulate microgravity (SMG) effects