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Construction method of recombinant plasmids for glutathione biosynthesis

A technology for glutathione and biosynthesis, applied in the field of bioengineering, can solve the problems of increased production cost of cysteine, toxic growth of Saccharomyces cerevisiae, bottleneck of glutathione, etc. simple steps

Active Publication Date: 2021-04-16
NANHUA UNIV
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Problems solved by technology

In actual production, the supply of precursor substances in glutathione synthesis is generally ensured by optimizing the medium (properly increasing the cysteine ​​content in it). Utilization is limited, and exogenous cysteine ​​is toxic to the growth of Saccharomyces cerevisiae; on the other hand, adding cysteine ​​increases production costs
At present, under the existing conditions, the ability of Saccharomyces cerevisiae to ferment and produce glutathione has reached a certain bottleneck

Method used

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  • Construction method of recombinant plasmids for glutathione biosynthesis
  • Construction method of recombinant plasmids for glutathione biosynthesis
  • Construction method of recombinant plasmids for glutathione biosynthesis

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Embodiment Construction

[0031] In order to facilitate the understanding of those skilled in the art, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings, and the contents mentioned in the embodiments are not intended to limit the present invention. It should be noted that if a certain step is not specified in the embodiments, it can be understood that the step is operated according to conventional experimental conditions, such as Sambrook and other molecular cloning experiment manuals (Sambrook J & Russell DW, Molecular Cloning: a Laboratory Manual, 2001) , or as recommended by the manufacturer's instructions.

[0032] It should be noted in advance that the PCR product recovery kit, gel recovery kit, and plasmid extraction kit used in the following examples are all commercially available Omega Bio-Tek products, pGAPZα A plasmid, used as the primary antibody The mouse anti-His antibody and the HRP-coupled goat anti-mouse antibody use...

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Abstract

The invention discloses a construction method of recombinant plasmids for glutathione biosynthesis, and relates to the technical field of bioengineering, the recombinant plasmids constructed by the method are saccharomyces cerevisiae double-gene co-expression plasmids, an OAS anabolism pathway of cysteine is introduced into saccharomyces cerevisiae cells through overexpression of serine O-acetyltransferase (SATase), synthesis of endogenous cysteine of saccharomyces cerevisiae can be promoted, the glutathione synthesis capacity of the saccharomyces cerevisiae in combination with overexpression of gamma-glutamylcysteine synthetase (GSH I) can be improved, and the growth speed of the saccharomyces cerevisiae in the logarithmic phase can be accelerated while the yield of the saccharomyces cerevisiae glutathione can be effectively improved.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for constructing a recombinant plasmid used for glutathione biosynthesis. Background technique [0002] Glutathione is the most abundant thiol-containing non-protein substance in eukaryotic cells. Under physiological conditions, more than 90% of glutathione exists as reduced GSH. Glutathione is commonly used as a pharmaceutical ingredient and is also widely used in the cosmetic and food industries. [0003] The production cost of synthesizing glutathione by chemical synthesis or enzymatic reaction is relatively high. Compared with chemically synthesized glutathione, glutathione with biological effects in organisms is L-type, while chemical synthesis will not Differential synthesis of L-GSH and D-GSH also increases the difficulty of later separation. Currently, the most economical glutathione production method in the industry is the fermentation method. Eukaryoti...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/81C12N15/64C12N15/54C12N15/52C12R1/865
Inventor 赵剑锋伍虹蓉高融
Owner NANHUA UNIV
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