Stimulation of mycolic acid biosynthesis by incorporation of cis-tetracos-5-enoic acid in a cell-wall preparation from Mycobacterium smegmatis.
Mycolic acids are high molecular weight hydroxy fatty acids which are a covalently linked part of the cell wall structure of all mycobacteria and their biosynthetic pathways offer potential drug targets. Three good candidates, cis-tetracos-5-enoic acid and R or S trans-6-methyl-tetracos-4-enoic acids, for the key initial intermediates where mycolic acid biosynthesis might diverge from other metabolic pathways, were tested as possible substrates. A cell-wall preparation from Mycobacterium smegmatis, capable of mycolic acid synthesis, was developed to investigate the possible incorporation of these, and other 16 to 24 carbon acids into mycolic acids. The wall preparations were extracted with hexane and suspended in hexane/water (7:1, v/v), and in this low-water assay, only one of these acids, cis-tetracos-5-enoic acid, stimulated the incorporation of radioactive label from [1-14C]acetate into alpha- and alpha'-mycolic acids. The extraction method used did, however, abolish some enzyme activity and mycolic acid biosynthesis was not completely restored by cis-tetracos-5-enoate. The two methyl-branched acids did not enhance the amount of label in epoxymycolic acids. An initial key intermediate in the synthesis of alpha- and alpha'-mycolic acids has therefore been positively identified for the first time; intermediates in the initial stages of the biosynthesis of oxygenated mycolic acids such as epoxymycolates remain to be defined.
Wheeler
P R
PR
Department of Applied Biology, University of Hull, UK.
Besra
G S
GS
Minnikin
D E
DE
Ratledge
C
C
D051080:Acyl-Carrier Protein S-Malonyltransferase; D000217:Acyltransferases; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism; D005227:Fatty Acids / Q000096:biosynthesis; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology*; D009161:Mycobacterium / Q000187:drug effects* / Q000378:metabolism; D009171:Mycolic Acids / Q000737:chemistry / Q000378:metabolism*; D012997:Solvents; D013237:Stereoisomerism
D005227:Fatty Acids; D005229:Fatty Acids, Monounsaturated; D009171:Mycolic Acids; D012997:Solvents; C064583:tetracosenoic acid; D000217:Acyltransferases; D051080:Acyl-Carrier Protein S-Malonyltransferase