Abstract
We have cloned the gene that encodes a novel glucosyl transferase (AraGT) involved in rhamnosylation of the polyketide antibiotic Aranciamycin in Streptomyces echinatus. AraGT comprises two domains characteristic of bacterial glycosyltranferases. AraGT was synthesized in E. coli as a decahistidinyl-tagged polypeptide. Purified AraGT is dimeric, displays a T(mapp) of 30 degrees C and can glycosylate the aglycone of an Aranciamycin derivative as shown by liquid chromatography and mass spectrometry. The availability of functional AraGT will allow the generation Aranciamycin-based combinatorial libraries.
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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Anthracyclines / metabolism*
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Chromatography, Liquid
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Cloning, Molecular
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Escherichia coli / enzymology
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Glucosyltransferases / chemistry*
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Glucosyltransferases / genetics*
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Glucosyltransferases / isolation & purification*
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Glycosyltransferases / chemistry*
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Glycosyltransferases / genetics*
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Glycosyltransferases / isolation & purification*
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Mass Spectrometry
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Models, Biological
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Molecular Sequence Data
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Streptomyces / enzymology
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Streptomyces / genetics
Substances
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Anthracyclines
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Recombinant Proteins
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aranciamycin
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Glycosyltransferases
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Glucosyltransferases
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aranciamycin glucosyltransferase, Streptomyces echinatus