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Cholesterol oxidase ChoL is a critical enzyme that catalyzes the conversion of diosgenin to 4-ene-3-keto steroids in Streptomyces virginiae IBL-14

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Abstract

Diosgenin transformation was studied in Streptomyces virginiae IBL-14, a soil-dwelling bacterium with diosgenin-degrading capacity. All of the derivatives isolated were identified as 4-ene-3-keto steroids. We cloned ChoL, a fragment of a cholesterol oxidase from S. virginiae IBL-14, and used gene-disruption techniques to determine its function in the oxidation of diosgenin to 4-ene-3-keto steroids. Subsequently, the entire open reading frame of ChoL was cloned by chromosome walking, and the His6-tagged recombinant protein was overproduced, purified, and characterized. ChoL consisted of 1,629 nucleotides that encoded a protein of 542 amino acids, including a 34-residue putative signal peptide at the N-terminal. ChoL showed 85% amino acid similarity to ChoA from Streptomyces sp. SA-COO. This enzyme can also oxidize other steroids such as cholesterol, sitosterol, and dehydroepiandrosterone, which showed higher affinity (K m = 0.195 mM) to diosgenin. The catalytic properties of this enzyme indicate that it may be useful in diosgenin transformation, degradation, and assay.

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Acknowledgements

This study was funded by the National Natural Science Foundation of China (project no. 20777016), the National Basic Research Program of China (project no. 2009CB724703), and the National High Technology Research and Development Program (“863” Program) of China (project no. 2008AA02Z209).

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China

    Bo Li, Wei Wang, Feng-Qing Wang & Dong-Zhi Wei

  2. East China University of Science and Technology, P.O.B. 311, Shanghai, 200237, China

    Feng-Qing Wang & Dong-Zhi Wei

Authors
  1. Bo Li
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  2. Wei Wang
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  3. Feng-Qing Wang
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  4. Dong-Zhi Wei
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Correspondence to Feng-Qing Wang or Dong-Zhi Wei.

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Li, B., Wang, W., Wang, FQ. et al. Cholesterol oxidase ChoL is a critical enzyme that catalyzes the conversion of diosgenin to 4-ene-3-keto steroids in Streptomyces virginiae IBL-14. Appl Microbiol Biotechnol 85, 1831–1838 (2010). https://doi.org/10.1007/s00253-009-2188-0

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