Enzymatic formation of a resorcylic acid by creating a structure-guided single-point mutation in stilbene synthase

TY - JOUR

T1 - Enzymatic formation of a resorcylic acid by creating a structure-guided single-point mutation in stilbene synthase

AU - Bhan, Namita

AU - Li, Lingyun

AU - Cai, Chao

AU - Xu, Peng

AU - Linhardt, Robert J.

AU - Koffas, Mattheos A.G.

N1 - Publisher Copyright: © 2014 The Protein Society.

PY - 2015/2/1

Y1 - 2015/2/1

N2 - 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.

AB - 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.

KW - novel non-natural polyphenols

KW - resorcylic acid

KW - stilbene synthase

KW - structure-guided mutants

KW - type III polyketides synthases

UR - http://www.scopus.com/inward/record.url?scp=84928898037&partnerID=8YFLogxK

U2 - 10.1002/pro.2600

DO - 10.1002/pro.2600

M3 - 文章

C2 - 25402946

AN - SCOPUS:84928898037

SN - 0961-8368

VL - 24

SP - 167

EP - 173

JO - Protein Science

JF - Protein Science

IS - 2

ER -