• Title/Summary/Keyword: Actinosynnema pretiosum

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Statistical Approach to Development of Culture Medium for Ansamitocin P-3 Production with Actinosynnema pretiosum ATCC 31565

  • BANDI SRINIVASULU;KIM YOON JUNG;SA SOON OK;CHANG YONG-KEUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.930-937
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    • 2005
  • The Plackett-Burman design and the response surface method (RSM) with a central composite design (CCD) were employed to develop a culture medium for ansamitocin P-3 production using Actinosynnema pretiosum ATCC 31565. Among the 11 nutrients tested using the Plackett-Burman design, two carbon sources, sucrose and dextrin, and two nitrogen sources, polypeptone and yeast extract, were selected. Optimization of the concentrations of the selected nutrients was then performed using RSM with CCD. After two rounds of RSM, the optimum concentrations ($\%w/v$) of sucrose, dextrin, polypeptone, and yeast extract were identified as 4.5, 4.5, 0.16, and 0.89, respectively. The maximum ansamitocin P-3 titer was 45.2 mg/l with the optimized medium, which was about 6 times higher than that (7.315 mg/l) obtained with an $R_{2}YE$ medium before optimization.

Construction of asm2 Deletion Mutant of Actinosynnema pretiosum and Medium Optimization for Ansamitocin P-3 Production Using Statistical Approach

  • Bandi Srinivasulu;Kim Yoon-Jung;Chang Yong-Keun;Shang Guang-Dong;Yu Tin-Wein;Floss Heinz G.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1338-1346
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    • 2006
  • Ansamitocin P-3 is a potent antitumor agent produced by A. pretiosum. A deletion mutant of A. pretiosum was constructed by deleting the asm2 gene, a putative transcriptional repressor. The deletion mutant showed a 9-fold enhanced ansamitocin P-3 productivity. The response surface method with central composite design was employed to further optimize the culture medium composition for ansamitocin P-3 production by the deletion mutant. The concentrations of four medium ingredients, dextrin, maltose, cotton seed flour, and yeast extract, which have been reported as major components for ansamitocin production, were optimized through a series of flask culture experiments. The optimum concentrations of the selected factors were found to be dextrin 6.0%; maltose 3.0%; cotton seed flour 0.53%; and yeast extract 0.45%. The maximum titer of ansamitocin P-3 was 78.3 mg/l with the optimized composition, about 15-folds higher than the unoptimized titer of 5.0 mg/l obtained with YMG medium.