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Title 

Sequence analysis and expression of rubN2 as dTDP-glucose 4,6-dehydratase gene cloned from Streptomyces achromogenes var. rubradiris NRRL3061, rubradirin producer

Authors 

Jin Chul YooJi Young ChoiSung Cheol YunChun Gyu KimJung Joon LeeSook Bok KimJae Kyung Sohng

Issue Date 

2000

Citation 

Journal of Biochemistry Molecular Biology & Biophysics, vol. 4, no. 0, pp. 313-321

Keywords 

biosynthesisdTDP-glucose 4,6-dehydrataseNAD+rubradirinrubranitrosestreptomycesglucoseDNA sequenceexpression vector

Abstract 

Southern blot analysis of the genomic library of Streptomyces achromogenes var. rubradiris NRRL3061, a rubradirin producer revealed the presence of three genes homologous to dTDP-glucose 4,6-dehydratase gene. Only rubN2 is located closely to the locus for polyketide synthase gene and aminohydroxybenzoic acid synthase gene being involved in the biosynthesis of rubransarol moiety of rubradirin. The rubN2 was expressed in Escherichia coli BL21(DE3) using the T7 expression vector pRSET-B. The purified protein was a homodimer with a subunit of molecular weight of 39,000. Only dTDP-glucose was utilized as a substrate by the expressed enzyme. The K(m) for dTDP-glucose and V(max) values were determined to be 35 μM and 199 nmol/min/mg protein, respectively. The K(m) for NAD+ was 20 μM. DNA sequencing of rubN2 and biochemical studies of RubN2 suggest that the gene encode dTDP-glucose 4,6-dehydratase highly specific for dTDP-glucose and obligate biosynthesis of rubranitrose.

ISSN 

1025-8140

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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