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LI Hui, WEN Shuping, HONG Wenrong. Construction of carbamoyltobramcin-producing engineering S.tenebrarius by disruption of aprK gene[J]. Journal of China Pharmaceutical University, 2013, 44(4): 368-373. DOI: 10.11665/j.issn.1000-5048.20130416
Citation: LI Hui, WEN Shuping, HONG Wenrong. Construction of carbamoyltobramcin-producing engineering S.tenebrarius by disruption of aprK gene[J]. Journal of China Pharmaceutical University, 2013, 44(4): 368-373. DOI: 10.11665/j.issn.1000-5048.20130416
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Construction of carbamoyltobramcin-producing engineering S.tenebrarius by disruption of aprK gene

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  • To construct a genetic engineering S. tenebrarius for production of carbamoyltobramycin. Recombinant plasmid pBK5 derivated from pKC1139 was constructed for disrupting gene aprK. pBK5 was introduced into S. tenebrariusTt-49 by conjugation. The single crossover mutant ST315 was obtained and cultivated for several rounds of sporulation in the absence of erythromycin. A desired double crossover mutant ST316 was achieved by PCR. TLC analysis indicated that ST316 produced high-yield carbamoyltobramycin, in which apramycin biosynthesis was blocked and the yield of carbamoyltobramycin was about 1 500 ug/mL.
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