De Novo Biosynthesis of Oleanane-Type Ginsenosides in Saccharomyces cerevisiae Using Two Types of Glycosyltransferases from Panax ginseng,Journal of Agricultural and Food Chemistry

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De Novo Biosynthesis of Oleanane-Type Ginsenosides in Saccharomyces cerevisiae Using Two Types of Glycosyltransferases from Panax ginseng
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2022-02-11 , DOI: 10.1021/acs.jafc.1c07526
He Zhang _{1,

2} , Xin Hua _{2,

3} , Dongran Zheng _{1,

2} , Hao Wu _{2,

3} , Chuanwang Li _{1,

2} , Pan Rao _{1,

2} , Mengliang Wen ₄ , Yong-Eui Choi ₅ , Zheyong Xue _{1,

2} , Yu Wang _{2,

3} , Yuhua Li _{1,

2}

Affiliation

Oleanane-type ginsenosides are highly biologically active substances in Panax ginseng, a popular Chinese dietary plant. Lack of key enzymes for glycosylation reactions has hindered de novo synthesis of these bioactive molecules. We mined candidate glycosyltransferases (GTs) of the ginseng database by combining key metabolites and transcriptome coexpression analyses and verified their function using in vitro enzymatic assays. The PgCSyGT1, a cellulose synthase-like GT rather than a UDP-dependent glucuronosyltransferase (UGT), was verified as the key enzyme for transferring a glucuronosyl moiety to the free C3–OH of oleanolic acid to synthesize calenduloside E. Two UGTs (PgUGT18 and PgUGT8) were first identified as, respectively, catalyzing the glycosylation reaction of the second sugar moiety of C3 and the C28 in the oleanane-type ginsenoside biosynthetic pathway. Then, we integrated these GTs in combinations into Saccharomyces cerevisiae genome and realized de novo biosynthesis of oleanane-type ginsenosides with a yield of 1.41 μg/L ginsenoside Ro in shake flasks. This report provides a basis for effective biosynthesis of diverse oleanane-type ginsenosides in microbial cell factories.

更新日期：2022-02-11

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