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Studies on Recombinant Human Insulin Expressed in Yeast Pichia Pastoris and Its Separation and Purification

Author: DongPeng
Tutor: ChenJinChun
School: Beijing University of Chemical Technology
Course: Fermentation Engineering
Keywords: Recombinant human insulin Pichia pastoris Ferment Separation and purification
CLC: TQ467
Type: Master's thesis
Year: 2006
Downloads: 424
Quote: 1
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Abstract


Diabetes and its complications are a serious hazard to human health . For patients with diabetes , insulin intervention as an alternative or complementary treatment is the most direct and effective way . The insulin drug after 80 years of development , from early animal pancreas extract animal insulin to biosynthetic human insulin and human insulin analogue , insulin drug has broad prospects for development . The laboratory of human proinsulin gene integrated into methanol nutritional yeast Pichia pastoris , constitute a genetically engineered bacteria , induced expression of recombinant human proinsulin . The first genetically engineered bacteria resuspended culture , SDS - PAGE and Western blotting test the expression of the protein molecular weight , consistent with the theoretical molecular weight , and insulin immunogenicity . The fermentation process of genetic engineering research concluded : Fermentation growth stage of the growth of bacteria and restrictive matrix glycerol residue concentration relationship with Monod relation to cell growth and maintenance coefficient m = , 0.007 g of / g · h the strains of cells in high - density culture is an excellent bacteria , has the potential for high- density culture ; optimized engineered bacteria fermentation conditions and basic salts medium formula based on the engineered bacteria induced strategy more than doubled the amount of protein . 10L fermentation tank on tank test using optimized shake flask culture conditions , the protein yield is more than five times the flask fermentation protein production . Proinsulin separated after the purification step has been optimized for the expression of recombinant human

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