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

  • Biotechnology Methods - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

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.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (21425624, 21376028, and 21566032).

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    Authors and Affiliations

    1. School of Life Science, Beijing Institute of Technology, Beijing, 100081, China

      Jie Huangfu, Yinghua Xu, Chun Li & Jun Li

    2. China National Research Institute of Food and Fermentation Industries, Beijing, 100027, China

      Jie Huangfu

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    1. Jie Huangfu
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    3. Chun Li
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    Correspondence to Jun Li.

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    J. Huangfu and Y. Xu contributed equally to this study.

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    Huangfu, J., Xu, Y., Li, C. et al. Overexpressing target helper genes enhances secretion and glycosylation of recombinant proteins in Pichia pastoris under simulated microgravity. J Ind Microbiol Biotechnol 43, 1429–1439 (2016). https://doi.org/10.1007/s10295-016-1817-8

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