1. Acyl-CoA Carboxylases (accD2 and accD3), Together with a Unique Polyketide Synthase (Cg-pks), Are Key to Mycolic Acid Biosynthesis in Corynebacterianeae Such as Corynebacterium glutamicum and Mycobacterium tuberculosis
- Author
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Kevin J. C. Gibson, Lothar Eggeling, Gurdyal S. Besra, Susumu Shioyama, Lynn G. Dover, Tadao Oikawa, Hermann Sahm, Alistair K. Brown, Karin Krumbach, and Roland Gande
- Subjects
chemistry [Polyketide Synthases] ,Time Factors ,Mutant ,Biochemistry ,Polymerase Chain Reaction ,Corynebacterium glutamicum ,metabolism [Plasmids] ,chemistry.chemical_compound ,chemistry [Malonyl Coenzyme A] ,chemistry [Lipids] ,Phylogeny ,chemistry.chemical_classification ,metabolism [Mycobacterium tuberculosis] ,biology ,acyl-CoA carboxylase ,Fatty Acids ,Lipids ,Malonyl Coenzyme A ,Blotting, Southern ,Phenotype ,Mycolic Acids ,Carbon-Carbon Ligases ,chemistry [Fatty Acids] ,Plasmids ,Genotype ,chemistry [Peptides] ,chemistry [Carbon-Carbon Ligases] ,Models, Biological ,Acyl-CoA ,Biosynthesis ,Polyketide synthase ,Lipid biosynthesis ,ddc:570 ,Escherichia coli ,Molecular Biology ,Fatty acid synthesis ,Cell Proliferation ,Models, Genetic ,Fatty acid ,Mycobacterium tuberculosis ,Cell Biology ,Lipid Metabolism ,Protein Structure, Tertiary ,chemistry ,metabolism [Corynebacterium glutamicum] ,metabolism [Mycolic Acids] ,Mutation ,biology.protein ,metabolism [Escherichia coli] ,Peptides ,Polyketide Synthases ,Gene Deletion ,Genome, Bacterial - Abstract
The Corynebacterianeae such as Corynebacterium glutamicum and Mycobacterium tuberculosis possess several unique and structurally diverse lipids, including the genus-specific mycolic acids. Although the function of a number of genes involved in fatty acid and mycolic acid biosynthesis is known, information relevant to the initial steps within these biosynthetic pathways is relatively sparse. Interestingly, the genomes of Corynebacterianeae possess a high number of accD genes, whose gene products resemble the beta-subunit of the acetyl-CoA carboxylase of Escherichia coli, providing the activated intermediate for fatty acid synthesis. We present here our studies on four putative accD genes found in C. glutamicum. Although growth of the accD4 mutant remained unchanged, growth of the accD1 mutant was strongly impaired and partially recovered by the addition of exogenous oleic acid. Overexpression of accD1 and accBC, encoding the carboxylase alpha-subunit, resulted in an 8-fold increase in malonyl-CoA formation from acetyl-CoA in cell lysates, providing evidence that accD1 encodes a carboxyltransferase involved in the biosynthesis of malonyl-CoA. Interestingly, fatty acid profiles remained unchanged in both our accD2 and accD3 mutants, but a complete loss of mycolic acids, either as organic extractable trehalose and glucose mycolates or as cell wall-bound mycolates, was observed. These two carboxyltransferases are also retained in all Corynebacterianeae, including Mycobacterium leprae, constituting two distinct groups of orthologs. Furthermore, carboxyl fixation assays, as well as a study of a Cg-pks deletion mutant, led us to conclude that accD2 and accD3 are key to mycolic acid biosynthesis, thus providing a carboxylated intermediate during condensation of the mero-chain and alpha-branch directed by the pks-encoded polyketide synthase. This study illustrates that the high number of accD paralogs have evolved to represent specific variations on the well known basic theme of providing carboxylated intermediates in lipid biosynthesis.
- Published
- 2004
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