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Wu K, Li W, Song J, et al. Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System. Biochem Insights. 2015;8:1-8doi: 10.4137/BCI.S21580.
Wu, K., Li, W., Song, J., & Li, T. (2015). Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System. Biochemistry insights, 81-8. https://doi.org/10.4137/BCI.S21580
Wu, Ke, et al. "Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System." Biochemistry insights vol. 8 (2015): 1-8. doi: https://doi.org/10.4137/BCI.S21580
Wu K, Li W, Song J, Li T. Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System. Biochem Insights. 2015 Feb 16;8:1-8. doi: 10.4137/BCI.S21580. eCollection 2015. PMID: 25733914; PMCID: PMC4334050.
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Biochem Insights. 2015 Feb 16;8:1-8. doi: 10.4137/BCI.S21580. eCollection 2015.

Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System.

Biochemistry insights

Ke Wu, Wei Li, Jianrui Song, Tao Li

Affiliations

  1. College of Life Science, Sichuan Normal University, Chengdu, Sichuan, China.
  2. Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, USA. ; Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.

PMID: 25733914 PMCID: PMC4334050 DOI: 10.4137/BCI.S21580

Abstract

Cholest-4-en-3-one has positive uses against obesity, liver disease, and keratinization. It can be applied in the synthesis of steroid drugs as well. Most related studies are focused on preparation of cholest-4-en-3-one by using whole cells as catalysts, but production of high-quality cholest-4-en-3-one directly from cholesterol oxidase (COD) using an aqueous/organic two-phase system has been rarely explored. This study set up an enzymatic reaction system to produce cholest-4-en-3-one. We developed and optimized the enzymatic reaction system using COD from COX5-6 (a strain of Rhodococcus) instead of whole-cell biocatalyst. This not only simplifies and accelerates the production but also benefits the subsequent separation and purification process. Through extraction, washing, evaporation, column chromatography, and recrystallization, we got cholest-4-en-3-one with purity of 99.78%, which was identified by nuclear magnetic resonance, mass spectroscopy, and infrared spectroscopy. In addition, this optimized process of cholest-4-en-3-one production and purification can be easily scaled up for industrial production, which can largely decrease the cost and guarantee the purity of the product.

Keywords: biphasic system; cholest-4-en-3-one; enzymatic conversion; identification; production; purification

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