Journal of Applied Biological Chemistry
- Volume 43 Issue 3
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- Pages.136-140
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- 2000
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- 1976-0442(pISSN)
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- 2234-7941(eISSN)
A New Deoxyhexose Biosynthetic Gene Cluster in Streptomyces griseus ATCC10137: Heterologous Expression of dTDP-D-Glucose 4,6-Dehydratase Gene
- Kim, Sang Suk (Department of Biological Science, Institute of Bioscience and Biotechnology) ;
- Bang, Jung-Hee (Department of Biological Science, Institute of Bioscience and Biotechnology) ;
- Hyun, Chang-Gu (Department of Biological Science, Institute of Bioscience and Biotechnology) ;
- Kim, Joo-Woo (Department of Biotechnology, Central Labortory of Research, Dong-Wha Pharmaceutical Co. Ltd.) ;
- Han, Jae-Jin (Department of Biotechnology, Central Labortory of Research, Dong-Wha Pharmaceutical Co. Ltd.) ;
- Suh, Joo-Won (Department of Biological Science, Institute of Bioscience and Biotechnology)
- Received : 2000.05.30
- Published : 2000.09.30
Abstract
A novel 6-deoxyhexose biosynthetic gene cluster different from the one for the biosynthesis of streptomycin was isolated from Streptomyces griseus using specifically designed PCR primers to compare the sequence of known dTDP-glucose synthase genes. We cloned a 5.8-kb DNA from Streptomyces griseus ATCC10137, which contained the 4-ketoreductase homologue (grsB), dTDP-glucose synthase (grsD), and dTDP-glucose 4, 6-dehydratase (grsE) genes. Escherichia coli cultures containing plasmid of the PCR product which encoded the grsE region under the controUed T7 promoter were able to catalyze the formation of dTDP-4-keto-6-deoxy-D-glucose from TDP-glucose. The enzyme showed high substrate specificity, being specific to only dTDP-glucose that is known to be incorporated into secondary metabolites such as antibiotics.