Expression Vector Adaptation of Valencene-producing Saccharomyces cerevisiae and Optimization of Fermentation Carbon and Nitrogen Sources

doi:10.13560/j.cnki.biotech.bull.1985.2019-0564

Abstract

Abstract: Valencene,a terpenoid existing in various citrus fruits,was widely used in perfume,soap,food,and beverage industry. However,the concentration of valencene in nature is extremely low,and current strategy to obtain valencene is complicated and expensive,thus construction of cell factory for valencene biosynthesis will be a more efficient and environmentally friendly. Saccharomyces cerevisiae was chosen as host to construct cell factory in this study. Firstly,the valencene synthase from Callitropsis nootkatensis（CnVS）was introduced into S. cerevisiae strain,resulting in the preliminary synthesis of valencene,and the initial yield of this strain was 4.16 mg/L. Then the gene erg9 and rox1 in Mevalonate（MVA）pathway of S. cerevisiae were knocked out using CRISPR/Cas9 system,improving carbon flux towards valencene synthesis. The results of different carbon/nitrogen source concentrations in fermentation indicated that the high accumulation of cell growth may not be conducive to valencene accumulation. Finally,the influences of different CnVS expression vectors on valencene yield were examined and the highest yield of 17.54 mg/L was obtained,which was 4.2 times higher than the original strain.

Key words: Saccharomyces cerevisiae, valencene, gene knockout, expression vector

CHEN He-feng, ZHU Chao-yi, LI Shuang. Expression Vector Adaptation of Valencene-producing Saccharomyces cerevisiae and Optimization of Fermentation Carbon and Nitrogen Sources[J]. Biotechnology Bulletin, 2020, 36(1): 209-219.

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