aro9 aro10基因过量表达对saccharomyces cerevisiae产3甲硫基丙醇代谢调控的影响 word格式.docxVIP

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aro9 aro10基因过量表达对saccharomyces cerevisiae产3甲硫基丙醇代谢调控的影响 word格式.docx

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aro9 aro10基因过量表达对saccharomyces cerevisiae产3甲硫基丙醇代谢调控的影响 word格式

Abstract3-(methylthio)-1-propanol is the characteristic aroma material of flavor liquor of sesame types in our country. It exists in wine, beer, soy sauce, tomato and potatoes. Its aroma threshold is lower and the suggested dosage in food is 0.1-10 mg/kg, Methionol has typical malt taste, sesame oil taste and cooked potato taste. In the study, the metabolic network of Saccharomyces cerevisiae was constructed. A strategy was developed based on metabolic flux analysis and a genetic strain was constructed with molecular biology technology. Then the effect on 3-(methylthio)-1-propanol was also investigated. The purpose of the study was to obtain the genetic engineering strain.Firstly, the influence of the transaminase (EC8) gene ARO9 cloning and overexpression on the metabolic flux analysis of 3-(methylthio)-1-propanol biosynthesis in S. cerevisiae INVSc1 was investigated. Results showed that the introduction of the recombinant plasmid pYES2-ARO9 containing ARO9 gene intoS. cerevisiae INVSc1 by using the lithium acetate method resulted in the overexpression of transaminase. And the level of methionol biosynthesis in the recombinant strain SC09-1 was significantly enhanced. Compared with the control strain incubated for 120 h, the yield of the recombinant strain was increased by 16.1%. These results clearly indicated the transaminase from S. cerevisiaes288c was a key rate-limiting enzyme in the biosynthesis of methionol, and the overexpression of ARO9 gene is helpful to the methionol biosynthesis.Secondly, the influence of decarboxylase (EC 2) gene ARO10 cloning and overexpression on the metabolic flux analysis of 3-(methylthio)-1-propanol biosynthesis in S. cerevisiae INVSc1 was investigated. Results showed that the introduction of the recombinant plasmid pYES2-ARO10 containing ARO10 gene into S. cerevisiae INVSc1 by using the lithium acetate method resulted in the overexpression of decarboxylase. And the level of methionol biosynthesis in the recombinant strain SC10-1