A Regioselective Tryptophan 5-Halogenase Is Involved in Pyrroindomycin Biosynthesis in Streptomyces rugosporus LL-42D005
| Autor: | Bojan Bister, Karl-Heinz van Pée, Susanne Zehner, José A. Salas, Carmen Méndez, Roderich D. Süssmuth, Alexander Kotzsch |
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| Rok vydání: | 2005 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Magnetic Resonance Spectroscopy Mutant Genes Fungal Genetic Vectors Molecular Sequence Data Clinical Biochemistry Pseudomonas fluorescens Biochemistry Gene Expression Regulation Enzymologic chemistry.chemical_compound Affinity chromatography Biosynthesis Drug Discovery Amino Acid Sequence Cloning Molecular DNA Fungal Molecular Biology Chromatography High Pressure Liquid Indole test chemistry.chemical_classification Pharmacology biology Reverse Transcriptase Polymerase Chain Reaction Tryptophan General Medicine biology.organism_classification Streptomyces Anti-Bacterial Agents Enzyme chemistry Mutation Cosmid Molecular Medicine Electrophoresis Polyacrylamide Gel Spectrophotometry Ultraviolet Macrolides Oxidoreductases Plasmids |
| Zdroj: | Chemistry & Biology. 12(4):445-452 |
| ISSN: | 1074-5521 |
| DOI: | 10.1016/j.chembiol.2005.02.005 |
| Popis: | Summary The antibiotic compound pyrroindomycin B contains an indole ring chlorinated in the 5 position. The indole ring is probably derived from tryptophan, and thus primers derived from conserved regions of tryptophan halogenases were used to amplify and clone a DNA fragment that was then used to isolate a tryptophan 5-halogenase gene ( pyrH ) from a cosmid library of the pyrroindomycin producer Streptomyces rugosporus LL-42D005. A gene disruption mutant in the tryptophan 5-halogenase gene no longer produced pyrroindomycin B, but still produced pyrroindomycin A, the nonhalogenated derivative. The halogenase gene could be overexpressed in Pseudomonas fluorescens BL915 ΔORF1 and was purified to homogeneity by immobilized metal chelate ion affinity chromatography. Chlorinating and brominating activities with tryptophan as a substrate were detected in cell-free extracts and for the purified enzyme. |
| Databáze: | OpenAIRE |
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