Molecular Cloning, Expression and Characterization of Oenococcus oeni Priming Glycosyltransferases.

Abstract:

:Oenococcus oeni is the main bacterial species that drives malolactic fermentation in wine. Most O. oeni strains produce capsular exopolysaccharides (EPS) that may contribute to protect them in the wine hostile environment. In O. oeni genome sequences, several genes are predicted to encode priming glycosyltransferases (pGTs). These enzymes are essential for EPS formation as they catalyze the first biosynthetic step through the formation of a phosphoanhydride bond between a hexose-1-phosphate and a lipid carrier undecaprenyl phosphate. In many microorganisms, mutations abolishing the pGT activity also abolish the EPS formation. We first made an in silico analysis of all the genes encoding putative pGT over 50 distinct O. oeni genome sequences. Two polyisoprenyl-phosphate-hexose-1-phosphate transferases, WoaA and WobA, and a glycosyltransferase (It3) were particularly examined for their topology and amino acid sequence. Several isoforms of these enzymes were then expressed in E. coli, and their substrate specificity was examined in vitro. The substrate specificity varied depending on the protein isoform examined, and several mutations were shown to abolish WobA activity but not EPS synthesis. Further analysis of woaA and wobA gene expression levels suggests that WoaA could replace the deficient WobA and maintain EPS formation.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Dimopoulou M,Claisse O,Dutilh L,Miot-Sertier C,Ballestra P,Lucas PM,Dols-Lafargue M

doi

10.1007/s12033-017-0021-z

subject

Has Abstract

pub_date

2017-08-01 00:00:00

pages

323-333

issue

8

eissn

1073-6085

issn

1559-0305

pii

10.1007/s12033-017-0021-z

journal_volume

59

pub_type

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