type	Journal Article
authors	Kulikova VV, Revtovich SV, Bazhulina NP, Anufrieva NV, Kotlov MI, Koval VS, Morozova EA, Hayashi H, Belyi YF, Demidkina TV
title	Identification of O-acetylhomoserine sulfhydrylase, a putative enzyme responsible for methionine biosynthesis in Clostridioides difficile: Gene cloning and biochemical characterizations
journal	IUBMB Life
Activity	2.5.1.49
Family	2.5.1.49
sel	selected
ui	31359602
year	(2019)
volume	71
number	11
pages	1815-1823
keywords	Clostridioides difficile; O-acetylhomoserine sulfhydrylase; methionine biosynthesis
abstract	O-acetylhomoserine sulfhydrylase (OAHS) is a pyridoxal 5'-phosphate-dependent enzyme involved in microbial methionine biosynthesis. In this study, we report gene cloning, protein purification, and some biochemical characteristics of OAHS from Clostridioides difficile. The enzyme is a tetramer with molecular weight of 185 kDa. It possesses a high activity in the reaction of L-homocysteine synthesis, comparable to reported activities of OAHSes from other sources. OAHS activity is inhibited by metabolic end product L-methionine. L-Propargylglycine was found to be a suicide inhibitor of the enzyme. Substrate analogue Nγ -acetyl-L-2,4-diaminobutyric acid is a competitive inhibitor of OAHS with Ki = 0.04 mM. Analysis of C. difficile genome allows to suggest that the bacterium uses the way of direct sulfhydrylation for the synthesis of L-methionine. The data obtained may provide the basis for further study of the role of OAHS in the pathogenic bacterium and the development of potential inhibitors.
last changed	2020/02/17 09:39