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The effect of 3-ketosteroid-Δ1-dehydrogenase isoenzymes on the transformation of AD to 9α-OH-AD by Rhodococcus rhodochrous DSM43269

  • Genetics and Molecular Biology of Industrial Organisms
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Rhodococcus rhodochrous DSM43269 is well known for its 3-ketosteroid-9α-hydroxylases. However, the function of its 3-ketosteroid-Δ1-dehydrogenases (KSDD) remains unknown. This study compared the involvement of ksdds in the strain’s androst-4-ene-3,17-dione (AD) transformation via gene deletion. The conversion was performed using AD as substrate or directly with 9α-hydroxyandrost-4-ene-3,17-dione (9α-OH-AD). The single deletion of ksdd1 or ksdd3 did not appear to result in the accumulation of 9α-OH-AD, whereas the single mutant △ksdd2 could preserve this compound to some extent. To further compare the role of ksdds in this strain, double mutants were constructed. All ksdd2 mutants combined with ksdd1 and/or ksdd3 resulted in the accumulation of 9α-OH-AD, among which the double mutant △ksdd2,3 behaved similarly to the single mutant △ksdd2 in this process. The mutant that lacked both ksdd1 and ksdd3 was still displayed, with no effect on the degradation of 9α-OH-AD. The triple mutant △ksdd1,2,3 was then constructed and exhibited the same capability as △ksdd1,2, accumulating more 9α-OH-AD than △ksdd2,3 and △ksdd2. The transcription of KSDD1 and KSDD2 increased, whereas that of KSDD3 seemed to exhibit no change, despite the use of the inducer AD or 9α-OH-AD. Thus, only ksdd1 and ksdd2 were involved in the transformation of AD to 9α-OH-AD. ksdd2 had the main role, ksdd1 had a minor effect on 9α-OH-AD degradation, and ksdd3 did not exhibit any action in this course.

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Acknowledgments

This work was supported by the National High Technology Research and Development of China (2011AA02A211), National Natural Science Foundation of China (21276196, 21406167), Key Project of Chinese Ministry of Education (213004A), and Tianjin Programs for Science and Technology Development (15ZCZDSY00510).

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Authors and Affiliations

  1. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People’s Republic of China

    Yang Liu, Yanbing Shen, Yuqian Qiao, Liqiu Su, Can Li & Min Wang

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  1. Yang Liu
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  2. Yanbing Shen
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  3. Yuqian Qiao
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  4. Liqiu Su
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Correspondence to Yanbing Shen or Min Wang.

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Liu, Y., Shen, Y., Qiao, Y. et al. The effect of 3-ketosteroid-Δ1-dehydrogenase isoenzymes on the transformation of AD to 9α-OH-AD by Rhodococcus rhodochrous DSM43269. J Ind Microbiol Biotechnol 43, 1303–1311 (2016). https://doi.org/10.1007/s10295-016-1804-0

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  • DOI: https://doi.org/10.1007/s10295-016-1804-0

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