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Discovery of a steroid 11[alpha]-hydroxylase from Rhizopus oryzae and its biotechnological applicationPetrič, Špela ...To overcome the chemically laborious stereo- and regioselective hydroxylation steps in the pharmaceutical production of corticosteroids and progestogens, certain fungal species, e.g. Rhizopus spp. ... and Aspergillus spp., are employed to perform the 11alpha-hydroxylation of the steroid skeleton, thereby significantly simplifying steroid drug production. Here we report for the first time the identification and expression of a fungal 11alpha-steroid hydroxylase, CYP509C12. The newly identified cytochrome P450, which is one of the 48 putative CYP genes in Rhizopus oryzae, was induced in the fungus by progesterone. By functionally expressing CYP509C12 in recombinant fission yeast, we were able to determine that its substrate spectrum includes progesterone as well as testosterone, 11-deoxycorticosterone, and 11-deoxycortisol, with the hydroxylations taking place predominantly at 11alpha and 6beta positions of the steroid ring system. To increase the 11alpha-hydroxylation activity of CYP509C12 in recombinant fission yeast, its natural redox partner, the R. oryzae NAD(P)H-dependent reductase, was coexpressed. The coexpression improved electron transfer to CYP509C12 and thusan increase in productivity from 246 to 300muMhydroxyPgd(-1) was observed,as well as a 7-fold increase of rate of hydroxyprogesterone formationwithin the linear phase of transformation. This newly developed strain displayed total bioconversion of progesterone into 11alpha-hydroxyprogesterone and small amounts of 6beta-hydroxyprogesterone within the first 6h of incubation with progesterone as substrate, hence demonstrating its potential for biotechnological application.Vir: Journal of biotechnology. - ISSN 0168-1656 (Vol. 150, št. 3, 2010, str. 428-437)Vrsta gradiva - članek, sestavni delLeto - 2010Jezik - angleškiCOBISS.SI-ID - 27654873
Avtor
Petrič, Špela |
Hakki, Tarek |
Bernhardt, Rita |
Žigon, Dušan |
Črešnar, Bronislava, 1950-2024
Teme
Rhizopus |
Enzymology |
Steroid Hydroxylases |
Cytochrome P-450 |
Schizosaccharomyces |
Enzymology |
Hydroxylation |
Adrenal Cortex Hormones |
Progesterone |
Hydroxyprogesterones |
Sequence Analysis |
Phylogeny |
Polymerase Chain Reaction |
Oligonucleotide Probes |
Dna, Complementary |
Gene Library |
Biotransformation |
Citokrom P-450 |
Steroidne hidroksilaze |
Rhizopus |
Hidroksilacija |
Schizosaccharomyces |
Hidroksiprogesteroni |
Zaporedje, analiza |
Filogeneza |
Polimerazna, verižna reakcija |
Oligonukleotidne sonde |
Progesteron |
Kortikosteroidi |
Genska knjižnica |
DNA komplementarna |
Biotransformacija
Vnos na polico
Trajna povezava
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Faktor vpliva
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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Povezave do osebnih bibliografij avtorjev | Povezave do podatkov o raziskovalcih v sistemu SICRIS |
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Petrič, Špela | 28887 |
Hakki, Tarek | |
Bernhardt, Rita | |
Žigon, Dušan | 03950 |
Črešnar, Bronislava, 1950-2024 | 08777 |
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