Title

Expression And Characterization Of Rv0447C Product, Potentially The Methyltransferase Involved In Tuberculostearic Acid Biosynthesis In Mycobacterium Tuberculosis

Keywords

fatty-acid; methyltransferase; Mycobacterium tuberculosis; oleic-acid; S-adenosyl- l -methionine; tuberculostearic acid

Abstract

In this study, a previously uncharacterized gene (Rv0447c) of Mycobacterium tuberculosis, designated as an unknown fatty-acid methyltransferase (ufaA1), was cloned, expressed in Escherichia coli, and purified. The biochemical characterization of the purified protein (UfaA1) showed it to be a methyltransferase that catalyzes biosynthesis of the tuberculostearic acid (10-methylstearic-acid, TSA), a significant constituent lipid of the mycobacterial cell wall and a clinical marker of the disease. Here, we show that UfaA1 transfers the methyl group from S-adenosyl-l-methionine (SAM) to the double bond of oleic acid in phosphatidylethanolamine or phosphatidylcholine to produce TSA. Optimal activity was obtained between pH 7.0 and pH 8.0. The methyltransferase activity of UfaA1 was severely inhibited by S-adenosyl-l-homocysteine. The Km values for dioleyl phosphatidylethanolamine, SAM, and nicotinamide adenine dinucleotide phosphate were 14, 13, and 83 μM, respectively, with Vmax of 1.3-1.6 nmol/Min. These results identify the Rv0447c gene product of M. tuberculosis as the methyltransferase that catalyzes the biosynthesis of TSA. This provides new information in mycobacterial cell wall synthesis. © 2013 International Union of Biochemistry and Molecular Biology, Inc.

Publication Date

7-1-2013

Publication Title

Biotechnology and Applied Biochemistry

Volume

60

Issue

4

Number of Pages

412-416

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1002/bab.1112

Socpus ID

84883169483 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/84883169483

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