Construction of prototrophic glutathione-high-producing yeast strain mediated by CRISPR/Cas9.

Wenlong ZHOU; Liang TANG; Kai CHENG; Minzhi LIU; Yan YANG; Wei WANG

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Construction of prototrophic glutathione-high-producing yeast strain mediated by CRISPR/Cas9.

Author: Wenlong ZHOU ¹ ; Liang TANG ¹ ; Kai CHENG ¹ ; Minzhi LIU ¹ ; Yan YANG ¹ ; Wei WANG ¹
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1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Key Laboratory of Biosynthesis of Natural Products of National Health and Family Planning Commission, Institute of Materia Medica, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100050, China.
Publication Type:Journal Article
Keywords: CRISPR/Cas9; Saccharomyces cerevisiae; auxotrophy; glutathione
From: Chinese Journal of Biotechnology 2017;33(12):1999-2008
CountryChina
Language:Chinese
Abstract: Glutathione (GSH), a non-protein thiol product with various biological activities, has been widely used in pharmaceutical and food industries. Recently, genetic engineering becomes an important strategy for obtaining GSH-high-producing strains. However, auxotrophic selection markers used may result in reduced cell growth or GSH production. In the present study, clustered regularly interspaced short palindromic repeats (CRISPR) and Cas9-associated-system (CRISPR-Cas), in which gRNA expression constructs and homologous DNA fragments of target genes were co-transformed into Saccharomyces cerevisiae cells, was used for the construction of the prototrophic strain derived from the engineered auxotrophic strain W303-1b/FGP. As a result, the prototrophic strain W303-1b/FGPPT showed a significantly shorter culture cycle compared with the auxotrophic strain. Furthermore, chemically defined medium could be used to culture strain W303-1b/FGPPT that might have great interests in industrial fermentation.