Biocatalytic synthesis of 3,4,5,3',5'-pentahydroxy-trans-stilbene from piceatannol by two-component flavin-dependent monooxygenase HpaBC.
HpaBC monooxygenase was previously reported to hydroxylate resveratrol to piceatannol. In this article, we report a novel catalytic activity of HpaBC for the synthesis of a pentahydroxylated stilbene. When Escherichia coli cells expressing HpaBC were incubated with resveratrol, the resulting piceatannol was further converted to a new product. This product was identified by mass spectrometry and NMR spectroscopy as a 5-hydroxylated piceatannol, 3,4,5,3',5'-pentahydroxy-trans-stilbene (PHS), which is a reportedly valuable biologically active stilbene derivative. We attempted to produce PHS from piceatannol on a flask scale. After examining the effects of detergents and buffers on PHS production, E. coli cells expressing HpaBC efficiently hydroxylated piceatannol to PHS in a reaction mixture containing 1.5% (v/v) Tween 80 and 100 mM 3-morpholinopropanesulfonic acid-NaOH buffer at pH 7.5. Under the optimized conditions, the whole cells regioselectively hydroxylated piceatannol, and the production of PHS reached 6.9 mM (1.8 g L(-1)) in 48 h.
a Faculty of Science and Engineering, Department of Applied Chemistry , Waseda University , Tokyo , Japan.
Toshiki
T
Furuya
b Health Science Research Center , Morinaga and Company Limited , Yokohama , Japan.
Masahiko
M
Sai
a Faculty of Science and Engineering, Department of Applied Chemistry , Waseda University , Tokyo , Japan.
Kuniki
K
Kino
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D055162:Biocatalysis; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D009025:Morpholines / Q000737:chemistry / Q000494:pharmacology; D011136:Polysorbates / Q000737:chemistry / Q000494:pharmacology; D000077185:Resveratrol; D012972:Sodium Hydroxide / Q000737:chemistry; D013267:Stilbenes / Q000378:metabolism* / Q000494:pharmacology
C473468:3,5,3',4',5'-pentahydroxy-stilbene; D001426:Bacterial Proteins; D003470:Culture Media; D009025:Morpholines; D011136:Polysorbates; D013267:Stilbenes; C008550:morpholinopropane sulfonic acid; D012972:Sodium Hydroxide; C041525:3,3',4,5'-tetrahydroxystilbene; D006899:Mixed Function Oxygenases; C026252:4-hydroxyphenylacetate 3-monooxygenase; D000077185:Resveratrol
HpaBC; hydroxylation; monooxygenase; resveratrol; stilbene