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High Density Cell Culture and Improvement of Glutathione Biosynthesis by Candida Utilis WSH 02-08

Author: ZhangWenYan
Tutor: ChenJian
School: Jiangnan University
Course: Fermentation Engineering
Keywords: Glutathione Utilis yeast High-density Fed - batch fermentation L- cysteine Mixed amino acids Peroxide Duress
CLC: TQ464.7
Type: Master's thesis
Year: 2006
Downloads: 294
Quote: 0
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Abstract


Glutathione (GSH) is a tripeptide having the biological activity of gamma-glutamyl group and a mercapto group, as a way to protect cells against ambient conditions irritation and stress of the important active tripeptide in vivo kinds of important physiological functions, in particular, plays a vital role for the maintenance of appropriate redox environment in vivo. Is widely used in clinical medicine, food industry, sports and other biological research. The Microbial Fermentation is one of the most common methods of GSH industrial production, glutathione is an intracellular product, the paper plant height accumulation of GSH utilis yeast (C. utilis WSH 02-08) starting strain, cells and GSH in high yield, high yield rate of the target, using a range of feasible options to increase cell density and GSH intracellular content, the main content of the study are as follows: (1) Glutathione is a cell within the product, to improve the cell density will help to improve the production of GSH. Constant speed fed and exponential feeding different culture of C. utilis WSH 02-08 fermentation synthesis of GSH. After 48 h exponential fed-batch culture, the dry cell weight (DCW) reached 69 g L-1, GSH production and intracellular GSH content have reached the maximum of 835 mg · L-1 and 1.41%, is to improve the cell density It is an ideal choice; (2) were investigated in the shake flask culture under the conditions of three consisting of GSH add L-glutamic acid, glycine and L-cysteine ​​precursor amino acid for the synthesis of GSH. The results show that when L-cysteine ​​is the key amino acids of GSH synthesis, when the concentration of 8 mmol · L-1, GSH concentration and intracellular GSH content was increased by 51% and 82%; (3) shake flask conditions, the three amino acids mixed added to improve the production and intracellular GSH content largest concentration of both 8 mmol · L-1, GSH concentration and intracellular GSH content were 295 mg · L-1 and 3.03%, was increased by 91% and 121%; (4) 7 L tank, study glucose fed-batch culture with added L-cysteine ​​binding cells to synthesize GSH. When a one-time add L-cysteine ​​concentration of 40 mmol · L-1, the intracellular GSH content up to 2.26%, the GSH production reached 1458 mg · L-1, to achieve a high cell density culture with high GSH intracellular unified content; (5) 7 L tank, when the one-time add mixed amino acid concentrations were 40 mmol · L-1, the intracellular GSH content up to reach 2.43% GSH yield reached 1568 mg L-1, GSH The average production rate of 27.5 mg · L-1 · h-1, so that the total amount of GSH and production strength has been greatly enhanced, and ultimately significantly increase the GSH yield of the target product;

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CLC: > Industrial Technology > Chemical Industry > Pharmaceutical chemical industry > Drug production of biological products > Amino acids,peptides,proteins
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