p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis

Glycosylated p-hydroxybenzoic acid methyl esters and structurally related phenolphthiocerol glycolipids are important virulence factors of Mycobacterium tuberculosis. Although both types of molecules are thought to be derived from p-hydroxybenzoic acid, the origin of this putative biosynthetic precu...

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Main Authors: Stadthagen, Gustavo, Kordula´kova, Jana, Griffin, R., Constant, Patricia, Bottova, Iveta, Barilone, Nathalie, Gicquel, Brigitte, Daffé, Mamadou, Jackson, Mary
Format: Article
Published: American Society for Biochemistry and Molecular Biology 2005
Online Access:https://eprints.nottingham.ac.uk/48706/
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author Stadthagen, Gustavo
Kordula´kova, Jana
Griffin, R.
Constant, Patricia
Bottova, Iveta
Barilone, Nathalie
Gicquel, Brigitte
Daffé, Mamadou
Jackson, Mary
author_facet Stadthagen, Gustavo
Kordula´kova, Jana
Griffin, R.
Constant, Patricia
Bottova, Iveta
Barilone, Nathalie
Gicquel, Brigitte
Daffé, Mamadou
Jackson, Mary
author_sort Stadthagen, Gustavo
building Nottingham Research Data Repository
collection Online Access
description Glycosylated p-hydroxybenzoic acid methyl esters and structurally related phenolphthiocerol glycolipids are important virulence factors of Mycobacterium tuberculosis. Although both types of molecules are thought to be derived from p-hydroxybenzoic acid, the origin of this putative biosynthetic precursor in mycobacteria remained to be established. We describe the characterization of a transposon mutant of M. tuberculosis deficient in the production of all forms of p-hydroxybenzoic acid derivatives. The transposon was found to be inserted in Rv2949c, a gene located in the vicinity of the polyketide synthase gene pks15/1, involved in the elongation of p-hydroxybenzoate to phenolphthiocerol in phenolic glycolipidproducing strains. A recombinant form of the Rv2949c enzyme was produced in the fast-growing non-pathogenic Mycobacterium smegmatis and purified to near homogeneity. The recombinant enzyme catalyzed the removal of the pyruvyl moiety of chorismate to form p-hydroxybenzoate with an apparent Km value for chorismate of 19.7Mandakcatvalueof0.102s1. Strong inhibition of the reaction by p-hydroxybenzoate but not by pyruvate was observed. These results establish Rv2949c as a chorismate pyruvate-lyase responsible for the direct conversion of chorismate to p-hydroxybenzoate and identify Rv2949c as the sole enzymatic source of p-hydroxybenzoic acid in M. tuberculosis.
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spelling nottingham-487062020-05-04T16:26:16Z https://eprints.nottingham.ac.uk/48706/ p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis Stadthagen, Gustavo Kordula´kova, Jana Griffin, R. Constant, Patricia Bottova, Iveta Barilone, Nathalie Gicquel, Brigitte Daffé, Mamadou Jackson, Mary Glycosylated p-hydroxybenzoic acid methyl esters and structurally related phenolphthiocerol glycolipids are important virulence factors of Mycobacterium tuberculosis. Although both types of molecules are thought to be derived from p-hydroxybenzoic acid, the origin of this putative biosynthetic precursor in mycobacteria remained to be established. We describe the characterization of a transposon mutant of M. tuberculosis deficient in the production of all forms of p-hydroxybenzoic acid derivatives. The transposon was found to be inserted in Rv2949c, a gene located in the vicinity of the polyketide synthase gene pks15/1, involved in the elongation of p-hydroxybenzoate to phenolphthiocerol in phenolic glycolipidproducing strains. A recombinant form of the Rv2949c enzyme was produced in the fast-growing non-pathogenic Mycobacterium smegmatis and purified to near homogeneity. The recombinant enzyme catalyzed the removal of the pyruvyl moiety of chorismate to form p-hydroxybenzoate with an apparent Km value for chorismate of 19.7Mandakcatvalueof0.102s1. Strong inhibition of the reaction by p-hydroxybenzoate but not by pyruvate was observed. These results establish Rv2949c as a chorismate pyruvate-lyase responsible for the direct conversion of chorismate to p-hydroxybenzoate and identify Rv2949c as the sole enzymatic source of p-hydroxybenzoic acid in M. tuberculosis. American Society for Biochemistry and Molecular Biology 2005-12-09 Article PeerReviewed Stadthagen, Gustavo, Kordula´kova, Jana, Griffin, R., Constant, Patricia, Bottova, Iveta, Barilone, Nathalie, Gicquel, Brigitte, Daffé, Mamadou and Jackson, Mary (2005) p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis. Journal of Biological Chemistry, 280 (49). 40699 -40706. ISSN 1083-351X http://www.jbc.org/content/280/49/40699 doi:10.1074/jbc.M508332200 doi:10.1074/jbc.M508332200
spellingShingle Stadthagen, Gustavo
Kordula´kova, Jana
Griffin, R.
Constant, Patricia
Bottova, Iveta
Barilone, Nathalie
Gicquel, Brigitte
Daffé, Mamadou
Jackson, Mary
p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis
title p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis
title_full p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis
title_fullStr p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis
title_full_unstemmed p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis
title_short p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis
title_sort p-hydroxybenzoic acid synthesis in mycobacterium tuberculosis
url https://eprints.nottingham.ac.uk/48706/
https://eprints.nottingham.ac.uk/48706/
https://eprints.nottingham.ac.uk/48706/