Biosynthesis of chloro-beta-hydroxytyrosine, a nonproteinogenic amino acid of the peptidic backbone of glycopeptide antibiotics
| Autor: | Stefan Pelzer, Efthimia Stegmann, Daniel Bischoff, Oliver Puk, Claudia Kittel, Roderich D. Süssmuth, Stefan Weist, Wolfgang Wohlleben |
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| Rok vydání: | 2004 |
| Předmět: |
DNA
Bacterial medicine.drug_class Operon Mutant Molecular Sequence Data Genetics and Molecular Biology Glycopeptide antibiotic Biology Microbiology Mass Spectrometry chemistry.chemical_compound Biosynthesis Bacterial Proteins Cytochrome P-450 Enzyme System Nonribosomal peptide Vancomycin Actinomycetales medicine Peptide Synthases Molecular Biology chemistry.chemical_classification Genetic Complementation Test Sequence Analysis DNA Glycopeptide Amino acid Anti-Bacterial Agents Dihydroxyphenylalanine Complementation Biochemistry chemistry Genes Bacterial Gene Deletion |
| Zdroj: | Journal of bacteriology. 186(18) |
| ISSN: | 0021-9193 |
| Popis: | The role of the putative P450 monooxygenase OxyD and the chlorination time point in the biosynthesis of the glycopeptide antibiotic balhimycin produced by Amycolatopsis balhimycina were analyzed. The oxyD gene is located directly downstream of the bhp (perhydrolase) and bpsD (nonribosomal peptide synthetase D) genes, which are involved in the synthesis of the balhimycin building block β-hydroxytyrosine (β-HT). Reverse transcriptase experiments revealed that bhp , bpsD , and oxyD form an operon. oxyD was inactivated by an in-frame deletion, and the resulting mutant was unable to produce an active compound. Balhimycin production could be restored (i) by complementation with an oxyD gene, (ii) in cross-feeding studies using A. balhimycina JR1 (a null mutant with a block in the biosynthesis pathway of the building blocks hydroxy- and dihydroxyphenylglycine) as an excretor of the missing precursor, and (iii) by supplementation of β-HT in the growth medium. These data demonstrated an essential role of OxyD in the formation pathway of this amino acid. Liquid chromatography-electrospray ionization-mass spectrometry analysis indicated the biosynthesis of completely chlorinated balhimycin by the oxyD mutant when culture filtrates were supplemented with nonchlorinated β-HT. In contrast, supplementation with 3-chloro-β-HT did not restore balhimycin production. These results indicated that the chlorination time point was later than the stage of free β-HT, most likely during heptapeptide synthesis. |
| Databáze: | OpenAIRE |
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