The effect of 3-ketosteroid-Δ1-dehydrogenase isoenzymes on the transformation of AD to 9α-OH-AD by Rhodococcus rhodochrous DSM43269
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- 作者：Yang Liu ; Yanbing Shen ; Yuqian Qiao…
- 刊名：Journal of Industrial Microbiology and Biotechnology
- 出版年：2016
- 出版时间：September 2016
- 年：2016
- 卷：43
- 期：9
- 页码：1303-1311
- 全文大小：1,387 KB
- 刊物类别：Biomedical and Life Sciences
- 刊物主题：Chemistry
Biotechnology
Genetic Engineering
Biochemistry
Bioinformatics
Microbiology
Microbial Genetics and Genomics - 出版者：Springer Berlin / Heidelberg
- ISSN：1476-5535
- 卷排序：43
文摘
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.KeywordsRhodococcus rhodochrousBiotransformation3-Ketosteroid-Δ1-dehydrogenaseAndrost-4-ene-3,17-dione9α-Hydroxyandrost-4-ene-3,17-dione