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Utilization of multiple substrates by butyrate kinase from Listeria monocytogenes.

Authors :
Sirobhushanam S
Galva C
Saunders LP
Sen S
Jayaswal R
Wilkinson BJ
Gatto C
Source :
Biochimica et biophysica acta. Molecular and cell biology of lipids [Biochim Biophys Acta Mol Cell Biol Lipids] 2017 Mar; Vol. 1862 (3), pp. 283-290. Date of Electronic Publication: 2016 Dec 06.
Publication Year :
2017

Abstract

Listeria monocytogenes, the causative agent of listeriosis, can build up to dangerous levels in refrigerated foods potentially leading to expensive product recalls. An important aspect of the bacterium's growth at low temperatures is its ability to increase the branched-chain fatty acid anteiso C15:0 content of its membrane at lower growth temperatures, which imparts greater membrane fluidity. Mutants in the branched-chain α-keto dehydrogenase (bkd) complex are deficient in branched-chain fatty acids (BCFAs,) but these can be restored by feeding C4 and C5 branched-chain carboxylic acids (BCCAs). This suggests the presence of an alternate pathway for production of acyl CoA precursors for fatty acid biosynthesis. We hypothesize that the alternate pathway is composed of butyrate kinase (buk) and phosphotransbutyrylase (ptb) encoded in the bkd complex which produce acyl CoA products by their sequential action through the metabolism of carboxylic acids. We determined the steady state kinetics of recombinant His-tagged Buk using 11 different straight-chain and BCCA substrates in the acyl phosphate forming direction. Buk demonstrated highest catalytic efficiency with pentanoate as the substrate. Low product formation observed with acetate (C2) and hexanoate (C6) as the substrates indicates that Buk is not involved in either acetate metabolism or long chain carboxylic acid activation. We were also able to show that Buk catalysis occurs through a ternary complex intermediate. Additionally, Buk demonstrates a strong preference for BCCAs at low temperatures. These results indicate that Buk may be involved in the activation and assimilation of exogenous carboxylic acids for membrane fatty acid biosynthesis.<br /> (Copyright © 2016 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1388-1981
Volume :
1862
Issue :
3
Database :
MEDLINE
Journal :
Biochimica et biophysica acta. Molecular and cell biology of lipids
Publication Type :
Academic Journal
Accession number :
27940001
Full Text :
https://doi.org/10.1016/j.bbalip.2016.12.001