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Enhanced human lysozyme production by Pichia pastoris via periodic dissolved oxygen concentration control
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  • Luqiang Jia,
  • Teng Li,
  • Yixuan Wu,
  • Chunsen Wu,
  • Huaxiang Li,
  • Agen Huang
Luqiang Jia
Yangzhou University
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Teng Li
Yangzhou University
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Yixuan Wu
Yangzhou University
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Chunsen Wu
Yangzhou University
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Huaxiang Li
Yangzhou University
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Agen Huang
Yangzhou University
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Abstract

In human lysozyme (hLYZ) production by P. pastoris, severe reactive oxygen species (ROS) accumulation occurred during the cell growth phase when using DO-stat control mode, leading to a limited expression of hLYZ. In this study, a novel periodic dissolved oxygen concentration (DO) control strategy was proposed to solve these problems. This strategy intermittently/periodically switched the cultivation environment from “high DO-low glycerol concentration” (DO-stat control mode) to “low DO-high glycerol concentration” for 4 cycles. With the strategy: (1) the highest cell concentration (~120 g-DCW L-1) with healthy and functional skeleton was achieved at the end of cell growth phase by controlling ROS levels below 48.35 Fluorescence intensity g-DCW-1; (2) cells metabolic activities in the successive induction phase were largely enhanced; (3) hLYZ activity reached the highest level of 2.18×105 IU mL-1, which was about 2-fold than that obtained with the DO-stat control mode, when the same induction strategy was adopted.