p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis

Stadthagen, Gustavo; Kordula�kova, Jana; Griffin, R.; Constant, Patricia; Bottova, Iveta; Barilone, Nathalie; Gicquel, Brigitte; Daff�, Mamadou; Jackson, Mary

Abstract

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.

Citation

Stadthagen, G., Kordula´kova, J., Griffin, R., Constant, P., Bottova, I., Barilone, N., Gicquel, B., Daffé, M., & Jackson, M. (2005). p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis. Journal of Biological Chemistry, 280(49), https://doi.org/10.1074/jbc.M508332200

• Journal Article Type: Article
• Acceptance Date: Oct 6, 2005
• Publication Date: Dec 9, 2005
• Deposit Date: Dec 13, 2017
• Publicly Available Date: Dec 13, 2017
• Journal: Journal of Biological Chemistry
• Print ISSN: 0021-9258
• Electronic ISSN: 1083-351X
• Publisher: American Society for Biochemistry and Molecular Biology
• Peer Reviewed: Peer Reviewed
• Volume: 280
• Issue: 49
• DOI: https://doi.org/10.1074/jbc.M508332200
• Public URL: https://nottingham-repository.worktribe.com/output/703505
• Publisher URL: http://www.jbc.org/content/280/49/40699
• Additional Information: This research was originally published in the Journal of Biological Chemistry. Gustavo Stadthagen, Jana Korduláková, Ruth Griffin, Patricia Constant, Iveta Bottová, Nathalie Barilone, Brigitte Gicquel, Mamadou Daffé, Mary Jackson. p-Hydroxybenzoic acid synthesis in mycobacterium tuberculosis. J. Biol. Chem. 2005; Vol 280, no 49 : 40699-4076. © 2005 the American Society for Biochemistry and Molecular Biology.
• Contract Date: Dec 13, 2017