Enzymatic formation of a resorcylic acid by creating a structure-guided single-point mutation in stilbene synthase.
A novel C17 resorcylic acid was synthesized by a structure-guided Vitis vinifera stilbene synthase (STS) mutant, in which threonine 197 was replaced with glycine (T197G). Altering the architecture of the coumaroyl binding and cyclization pocket of the enzyme led to the attachment of an extra acetyl unit, derived from malonyl-CoA, to p-coumaroyl-CoA. The resulting novel pentaketide can be produced strictly by STS-like enzymes and not by Chalcone synthase-like type III polyketide synthases; due to the unique thioesterase like activity of STS-like enzymes. We utilized a liquid chromatography mass spectrometry-based data analysis approach to directly compare the reaction products of the mutant and wild type STS. The findings suggest an easy to employ platform for precursor-directed biosynthesis and identification of unnatural polyketides by structure-guided mutation of STS-like enzymes.
Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Center for Biotechnology and Interdisciplinary Studies, Troy, New York.
Namita
N
Bhan
Lingyun
L
Li
Chao
C
Cai
Peng
P
Xu
Robert J
RJ
Linhardt
Mattheos A G
MA
Koffas
D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D062385:Hydroxybenzoates / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D012118:Resorcinols / Q000737:chemistry / Q000378:metabolism*; D027843:Vitis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
D062385:Hydroxybenzoates; D012118:Resorcinols; C076950:alpha-resorcylic acid; D000217:Acyltransferases; C022674:stilbene synthase
novel non-natural polyphenols; resorcylic acid; stilbene synthase; structure-guided mutants; type III polyketides synthases