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Isolation, molecular identification and statistical optimization of culture condition for a new extracellular cholesterol oxidase-producing strain using response surface methodology

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Abstract

The current work details the screening of about 100 isolates from various soil samples, from which 1 isolate was finally selected based on the productivity of cholesterol oxidase. Further biochemical identification tests and 16S rRNA gene sequencing identified this isolate as Streptomyces badius. A preliminary culture media optimization was carried out using the initial screening method of Plackett-Burman. Then, a Box-Behnken design was employed to investigate the optimum concentrations of medium components and interactive effects of main variables on cholesterol oxidase production. The regression analysis showed a significant coefficient of determination (R 2) value (91 %), which was in close agreement ensuring a satisfactory adjustment of the proposed model. Maximal enzyme production (2.38 U/mL, i.e., approximately more than 100 % activity in the basal medium) was obtained at: temperature 35 °C; Tween 20 0.1 %; pH 6.5 and yeast extract 0.15 %. This two-stage statistical approach provided rapid identification and integration of key medium parameters for Streptomyces sp., resulting in high cholesterol oxidase production.

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Acknowledgment

This work was supported by Research Council of Shiraz University of Medical Sciences, Shiraz, Iran.

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Authors and Affiliations

  1. Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, PO Box 71345-1583, Shiraz, Iran

    Zahra Moradpour, Abdollah Ghasemian, Azam Safari, Milad Mohkam & Younes Ghasemi

  2. Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, PO Box 71345-1583, Shiraz, Iran

    Zahra Moradpour, Abdollah Ghasemian, Azam Safari, Milad Mohkam & Younes Ghasemi

Authors
  1. Zahra Moradpour
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  2. Abdollah Ghasemian
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  3. Azam Safari
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  4. Milad Mohkam
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  5. Younes Ghasemi
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Correspondence to Abdollah Ghasemian.

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Moradpour, Z., Ghasemian, A., Safari, A. et al. Isolation, molecular identification and statistical optimization of culture condition for a new extracellular cholesterol oxidase-producing strain using response surface methodology. Ann Microbiol 63, 941–950 (2013). https://doi.org/10.1007/s13213-012-0547-z

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  • DOI: https://doi.org/10.1007/s13213-012-0547-z

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