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Genetic engineering approach for the production of rhamnosyl and allosyl flavonoids from Escherichia coli
Author(s) -
Simkhada Dinesh,
Lee Hei Chan,
Sohng Jae Kyung
Publication year - 2010
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.22782
Subject(s) - kaempferol , glycosylation , quercetin , escherichia coli , biotransformation , glycosyltransferase , biochemistry , chemistry , flavonoid , biosynthesis , glycoside , rhamnose , hydroxylation , biology , gene , enzyme , stereochemistry , polysaccharide , antioxidant
Abstract The main functions of glycosylation are stabilization, detoxification and solubilization of substrates and products. To produce glycosylated products, Escherichia coli was engineered by overexpression of TDP‐ L ‐rhamnose and TDP‐6‐deoxy‐ D ‐allose biosynthetic gene clusters, and flavonoids were glycosylated by the overexpression of the glycosyltransferase gene from Arabidopsis thaliana . For the glycosylation, these flavonoids (quercetin and kaempferol) were exogenously fed to the host in a biotransformation system. The products were isolated, analyzed and confirmed by HPLC, LC/MS, and ESI‐MS/MS analyses. Several conditions (arabinose, IPTG concentration, OD 600 , substrate concentration, incubation time) were optimized to increase the production level. We successfully isolated approximately 24 mg/L 3‐ O ‐rhamnosyl quercetin and 12.9 mg/L 3‐ O ‐rhamnosyl kaempferol upon feeding of 0.2 mM of the respective flavonoids and were also able to isolate 3‐ O ‐allosyl quercetin. Thus, this study reveals a method that might be useful for the biosynthesis of rhamnosyl and allosyl flavonoids and for the glycosylation of related compounds. Biotechnol. Bioeng. 2010;107: 154–162. © 2010 Wiley Periodicals, Inc.