• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.906-911
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• 2003

The function of genes related to spectinomycin biosynthesis (spcD, speA, speB, spcS2) from Streptomyces spectabilis ATCC 27741, a spectinomycin producer, was analyzed. Each gene was subcloned from a spectinomycin biosynthetic gene cluster and overexpressed in E. coli BL21 (DE3) using pET vector. After incubating each purified protein with its possible substrates, the final products were analyzed using high-performance liquid chromatography (HPLC). From these results, spcD, speA, and speB have been identified to be dTDP-glucose synthase, myo-inositol monophosphatase, and myo-inositol dehydrogenase, respectively. In addition, the results suggest that the spcS2 gene product functions downstream of the speB gene product in the biosynthetic pathway of spectinomycin. Taken together, the present study elucidates the early steps of the biosynthetic pathway for 6-deoxyhexose (6-DOH) part (actinospectose) and aminocyclitol part (actinamine) of spectinomycin.

• Proceedings of the Microbiological Society of Korea Conference
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• 2001.11a
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• pp.146-156
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• 2001

The biosynthetic gene clusters for bluensomycin and spectinomycin were isolated and characterized from the bluensomycin producer, Streptomyces bluensis ATCC27420 and the spectinomycin producer, Streptomyces spectabilis ATCC27741, respectively. PCR primers were designed specifically to amplify a segment of dTDP-glucose synthase gene based on its conserved sequences of several actinomycete strains. By screening cosmid libraries using amplified PCR fragments, 30-kb and 45-kb DNA fragments were isolated from Streptomyces bluensis and Streptomyces spectabilis, respectively. Sequencing analysis of them revealed that each contains 15 open reading frames (ORFs). Some of these ORFs were turned out to be antibiotic resistance genes (blmA and speN), dTDP-glucose synthase genes (blmD and spcD), and dTDP-D-glucose 4,6-dehydratase genes (blmE and spcE), suggesting that the blm and spec gene clusters are likely involved in the biosynthesis of bluensomycin and spectinomycin, respectively.