Abstract
Filamentous fungus Rhizopus nigricans is used commercially for steroid 11α-hydroxylation reactions to prepare key steroid intermediates in the synthesis of a variety of steroid drugs. Although it has been previously established that the enzyme involved in steroid 11α-hydroxylation is a cytochrome P450 protein, the corresponding encoding gene remains elusive. In the present study, steroid 11α-hydroxylase activities in strain R. nigricans TCCC 41047 were shown to be expressed only under substrate induction. Transcriptome sequencing was conducted to establish the list of the cytochrome P450 genes (CYP) expressed in TCCC41047 under substrate induction. The results revealed that 14 CYP genes were expressed under induction conditions. Furthermore, qRT-PCR showed that the transcription of 3 CYP genes was highly induced by steroid substrate EP and thus identified as candidate genes for encoding the target 11α-hydroxylase gene and pave the way for the engineering of highly candidate genes will aid in the cloning and characterization of the target steroid 11α-hydroxylase gene and pave the way for the engineering of highly efficient production strains for preparing valuable steroid 11α-hydroxylation derivatives.
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References
Ayer DE, Schlage CA, Flynn GL (1976) Anti-inflammatory steroid. US Pat 3980 778
Salvador, Jorge AR, et al (2012) Anticancer steroids: linking natural and semi-synthetic compounds. Nat Prod Rep 30(2):324–374
Mahat SB, Garai S (1997) Advances in microbial steroid biotransformation. Steroids 62:332–345
Zeelen FJ (1990) Pharmaceutical chemistry of steroids. Elsevier, Amsterdam
Žakelj-Mavrič M, Belič I (1987) Hydroxylation of steroids with 11α-hydroxylase of Rhizopus nigricans. J Steroid Biochem 28(2):197–201
Sonomoto K, Hoq MM, Tanaka A, Sukui S (1983) 11β-Hydroxylation of cortexolone (Reichstein compound S) to hydrocortisone by Curvularia lunata entrapped in photo-cross-linked resin gels. Appl Environ Microb 45(2):436–443
Donova MV, Egorova OV (2012) Microbial steroid transformations: current state and prospects. Appl Microbiol Biotechnol 94:1423–1447
Breskvar K, Hudnik-Plevnik T (1981) Inducibility of cytochrome P-450 and of NADPH-cytochrome c reductase in progesterone treated filamentous fungi Rhizopus nigricans and Rhizopus arrhizus. J Steroid Biochem 14:395–399
Murray HC, Peterson DH (1952) Oxygenation of steroids by Mucorales fungi. US Pat 2602 769
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆CT method. Methods 25:402–408
Irrgang S, Schlosser D, Schmauder HP (1992) The steroid 15α-hydroxylase of Penicillium raistrickii i477 is inducible. Biotechnol Lett 14:33–38
Lin YY, Smith LL (1970) Microbial hydroxylations: VIII. Induction of steroid hydroxylases of Curvularia lunata by 19-nortestosterone analogs. Biochim Biophys Acta 218:526–531
Acknowledgments
This work was financially supported by a grant from the National High Technology Research and Development Program of China (863 Program) (No. 2011AA02A211).
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Zhao, J. et al. (2018). Transcriptomic Analysis of CYP Genes in Rhizopus nigricans and Identification of the Steroid 11α-Hydroxylase Candidate Genes. In: Liu, H., Song, C., Ram, A. (eds) Advances in Applied Biotechnology. ICAB 2016. Lecture Notes in Electrical Engineering, vol 444. Springer, Singapore. https://doi.org/10.1007/978-981-10-4801-2_3
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DOI: https://doi.org/10.1007/978-981-10-4801-2_3
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