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Inter mass optimization during insulin precursor fermentation

A technology of insulin precursor and fermentation process, which is applied in the field of yeast fermentation to produce recombinant porcine insulin precursor, which can solve the problems of low concentration and high industrial production cost, and achieve the effect of increasing concentration

Inactive Publication Date: 2007-09-05
SUNSHINE LAKE PHARM CO LTD
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  • Summary
  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

(2) Direct chemical synthesis from a single amino acid, which can be industrially produced at a high cost
In the process of producing recombinant porcine insulin precursor by yeast fermentation, Methylotrophic yeast Pichia Pastoris has been recognized as one of the excellent expression systems for the production of exogenous secreted proteins and intracellular proteins, and its robust Growth and some uniqueness have been developed into recombinant expression systems for commercial production of exogenous proteins, but the prior art (Invitrogen Yeast Fermentation Handbook) yeast fermentation has a low concentration (only 0.8g.L -1 ) deficiency

Method used

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Embodiment

[0021] Object: Pichia pastoris GS115 recombinant porcine insulin precursor fermentation process, the volume of the fermenter is 50L, the base material is 4% (w / v) glycerol as the batch fermentation stage, and the concentration of 50% (w / w) glycerol is continuously fed, To be cell density (OD 600 ) is about 300, stop adding glycerin, add 0.3% (w / v) methanol twice in batches, determine the initial methanol flow acceleration rate according to dissolved oxygen, and set the final OD 600 is 520, and the fermentation period T is 100h.

[0022] Control target: final OD of fermentation 600 for 520.

[0023] Specific steps are as follows:

[0024] (1) Waiting for OD 600 At about 300, stop adding glycerin. Additional 0.3% (w / v) methanol was added in two batches.

[0025] (2) Methanol is exhausted, dissolved oxygen rebounds, start to add methanol, and increase the supplementary acceleration rate, so that dissolved oxygen falls back to the lowest point of dissolved oxygen before the ...

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Abstract

A method for supplementing and optimizing feed during fermentation of insulin precursor is carried out by supplementing methanol to determine its fluid acceleration ratio, computing while adjusting for specific growth speed ration by difference of final fermented strain density and real-time strain density and fermentation period and controlling growth speed ratio. It has more protein concentration.

Description

technical field [0001] The invention belongs to a method for producing insulin, in particular to a method for producing recombinant porcine insulin precursor by yeast fermentation. Background technique [0002] The precursor of insulin, namely proinsulin, has a molecular weight of about 9000, contains 86 amino acids, and is a single-chain polypeptide. Insulin is a small molecular protein containing 51 amino acids, with a molecular weight of about 6000. Pig and human insulin differ only in one amino acid residue at the C-terminus of the B chain, the former being alanine and the latter being threonine, but the physiological effects of the two insulins are exactly the same. Insulin is the main hormone that promotes anabolism and regulates blood sugar stability, and is a specific drug for treating insulin-dependent diabetes. [0003] At present, several methods of producing human insulin used in the industry include: (1) direct extraction from human pancreas, but the supply of...

Claims

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Application Information

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IPC IPC(8): C12P21/02C12R1/84
Inventor 朱志钢郑强张富权谢俊杰唐新发
Owner SUNSHINE LAKE PHARM CO LTD
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