Abstract
An open reading frame, rub52, has been identified as a gene encoding thymidine diphospho-glucose 2,3-dehydratase by sequence analysis of the rubradirin biosynthetic gene cluster of Streptomyces achromogenes var. nibradiris NRRL3061. The gene codes for a protein consisting of 458 amino acids with calculated molecular mass of 50862 Da. The gene was amplified and heterologously expressed in Escherichia coli as a soluble His-tagged fusion protein. C-2 deoxygenation functionality of thymidine diphospho-4-keto-6-deoxyglucose was assigned to the rub52 gene product from in vitro enzyme assay.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Cloning, Molecular
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Gene Expression Profiling / methods
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Gene Expression Regulation, Bacterial
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Gene Expression Regulation, Enzymologic / genetics*
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Glycosides / genetics
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Glycosides / metabolism
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Hydro-Lyases / biosynthesis
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Hydro-Lyases / genetics*
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Hydro-Lyases / metabolism*
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Molecular Sequence Data
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Naphthoquinones / metabolism*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Sequence Alignment
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Sequence Analysis, Protein
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Streptomyces / genetics*
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Streptomyces / metabolism*
Substances
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Glycosides
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Naphthoquinones
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Recombinant Fusion Proteins
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rubranitrose
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rubradirin
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Hydro-Lyases
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dTDPglucose 4,6-dehydratase