A method for stably producing porcine alpha interferon using recombinant Pichia pastoris

A technology of interferon alpha and Pichia pastoris, applied in the field of bioengineering, can solve the problems of accumulation of by-products of anaerobic metabolism, affecting the stability of fermentative production of interferon alpha in pigs, etc.

Active Publication Date: 2017-12-08
SHANGHAI ACAD OF AGRI SCI +1
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Problems solved by technology

In this way, the culture of Pichia pastoris is constantly switching back and forth between aerobic and anaerobic states, which easily causes the generation and accumulation of by-products of anaerobic metabolism, and ultimately seriously affects the normal induced expression of pig interferon alpha and the stability of fermentation production
[0004] So far, there are few reports on the generation and accumulation of metabolic byproducts in Pichia pastoris during high-density culture and their effects on the expression of target proteins during induction

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  • A method for stably producing porcine alpha interferon using recombinant Pichia pastoris
  • A method for stably producing porcine alpha interferon using recombinant Pichia pastoris
  • A method for stably producing porcine alpha interferon using recombinant Pichia pastoris

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[0023] The present inventor has found that pig interferon alpha cannot be expressed normally or fermentation performance is not good by studying whether and how the by-products of anaerobic metabolism accumulate when glycerol is fed, and the effect of accumulation of by-products of anaerobic metabolism on the induced expression of pig interferon alpha. The root cause of the stability, based on this, an improved DO-Stat glycerin feeding strategy was proposed. The method of the invention greatly improves the induced expression efficiency of the porcine alpha interferon and ensures the stability of fermentation.

[0024] In the present invention, "fermentation" refers to culturing the recombinant Pichia pastoris carrying the porcine interferon-alpha gene) to produce porcine-alpha interferon. More specifically, "fermentation" refers to a large-scale production process, such as a fermentation scale of 0.5-500 liters, such as a fermentation scale of 1 liter, 2 liters, 5 liters, 10 l...

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Abstract

The present invention relates to a method for stably producing porcine alpha interferon by using recombinant Pichia pastoris, and proposes an improved DO-Stat glycerin feeding strategy, the method of the present invention greatly improves the induced expression efficiency of porcine alpha interferon, The stability of fermentation is ensured.

Description

technical field [0001] The invention belongs to the technical field of bioengineering; more specifically, the invention relates to a method for stably producing porcine alpha interferon by using recombinant Pichia pastoris. Background technique [0002] Porcine α-interferon (porcine interferon-α, pIFN-α) is a veterinary drug with broad application prospects [Chinsangaram J et al., J Virol, 2001, 75(12):5498-5503]. Fermentation by recombinant Pichia pastoris is the main method for producing porcine interferon-alpha, which can be completely secreted in the fermentation broth. Based on the completion of the construction of recombinant bacteria and the optimization of the shake flask process, the research focus has shifted to the optimization of the fermenter-scale fermentation process. [0003] In recent years, studies on the optimization of porcine interferon-α induction process have shown that the high-efficiency induced expression of porcine interferon-α is directly related...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C12R1/84
Inventor 于瑞嵩史仲平李震高敏杰丁健董世娟朱于敏
Owner SHANGHAI ACAD OF AGRI SCI
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