Biosynthesis of (−)-5-Hydroxy-equol and 5‑Hydroxy-dehydroequol from Soy Isoflavone, Genistein Using Microbial Whole Cell Bioconversion

• Published in: ACS chemical biology 2017-11, Vol.12 (11), p.2883-2890
• Main Authors: Lee, Pyung-Gang, Kim, Joonwon, Kim, Eun-Jung, Lee, Sang-Hyuk, Choi, Kwon-Young, Kazlauskas, Romas J, Kim, Byung-Gee
• Format: Article
• Language: English
• Subjects: Actinobacteria - enzymology; Actinobacteria - genetics; Actinobacteria - metabolism; Biosynthetic Pathways; Biotransformation; Equol - genetics; Equol - metabolism; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression; Genistein - metabolism; Isoflavones - genetics; Isoflavones - metabolism; Oxidoreductases - genetics; Oxidoreductases - metabolism; Phytoestrogens - metabolism; Racemases and Epimerases - genetics; Racemases and Epimerases - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - metabolism
• ISSN: 1554-8929; 1554-8937
• DOI: 10.1021/acschembio.7b00624

Equols are isoflavandiols formed by reduction of soy isoflavones such as daidzein and genistein by gut microorganisms. These phytoestrogens are of interest for their various biological effects. We report biosynthesis from genistein to (−)-5-hydroxy-equol in recombinant E. coli expressing three reductases (daidzein reductase DZNR, dihidrodaidzein reductase DHDR, tetrahydrodaidzein reductase THDR) and a racemase (dihydrodaidzein racemase, DDRC) originating from the gut bacterium, Slackia isoflavoniconvertens. The biosynthesized 5-hydroxy-equol proved as an optically negative enantiomer, nonetheless it displayed an inverse circular dichroism spectrum to (S)-equol. Compartmentalized expression of DZNR and DDRC in one E. coli strain and DHDR and THDR in another increased the yield to 230 mg/L and the productivity to 38 mg/L/h. If the last reductase was missing, the intermediate spontaneously dehydrated to 5-hydroxy-dehydroequol in up to 99 mg/L yield. This novel isoflavene, previously not known to be synthesized in nature, was also detected in this biotransformation system. Although (S)-equol favors binding to human estrogen receptor (hER) β over hERα, (−)-5-hydroxy-equol showed the opposite preference. This study provides elucidation of the biosynthetic route of (−)-5-hydroxy-equol and measurement of its potent antagonistic character as a phytoestrogen for the first time.