Cloning and identification of a novel steroid 11α-hydroxylase gene from Absidia coerulea

Wang, Ruijie; Sui, Pengcheng; Hou, Xiangjiang; Cao, Tong; Jia, Longgang; Lu, Fuping; Singh, Suren; Wang, Zhengxiang; Liu, Xiaoguang

doi:10.1016/j.jsbmb.2017.04.006

Cited by 26 publications

(19 citation statements)

References 22 publications

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“…Hydroxylation at C-11 α position is important for the anti-bacterial activity of fusidic acid (Table 3), which is catalyzed by a cytochrome P450 monooxygenase FusB1. Thus far, only three steroid 11 α -hydroxylases CYP509C12, CYP5311B1 and 11 α -SH Aoch from Rhizopus oryzae , Aspergillus ochraceus and Absidia caerulea , respectiviely, have been reported26., 27., 28.. Phylogenetic analysis of FusB1 with other fungi-derived P450 enzymes involved triterpenoid/steroid modifications revealed that FusB1 form a cluster with CYP5311B1 and 11 α -SH Aoch , but not with steroid 11 α -hydroxylase CYP509C12, and CYP5150L8 involved in biosynthesis of ganoderic acid 29 and VidA/D/K for demethoxyviridin biosynthesis 30 (Supporting Information Fig.…”

Section: Discussionmentioning

confidence: 99%

Biosynthesis of clinically used antibiotic fusidic acid and identification of two short-chain dehydrogenase/reductases with converse stereoselectivity

Cao

et al. 2019

Acta Pharmaceutica Sinica B

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Fusidic acid is the only fusidane-type antibiotic that has been clinically used. However, biosynthesis of this important molecule in fungi is poorly understood. We have recently elucidated the biosynthesis of fusidane-type antibiotic helvolic acid, which provides us with clues to identify a possible gene cluster for fusidic acid ( fus cluster). This gene cluster consists of eight genes, among which six are conserved in the helvolic acid gene cluster except fusC1 and fusB1 . Introduction of the two genes into the Aspergillus oryzae NSAR1 expressing the conserved six genes led to the production of fusidic acid. A stepwise introduction of fusC1 and fusB1 revealed that the two genes worked independently without a strict reaction order. Notably, we identified two short-chain dehydrogenase/reductase genes fusC1 and fusC2 in the fus cluster, which showed converse stereoselectivity in 3-ketoreduction. This is the first report on the biosynthesis and heterologous expression of fusidic acid.

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Section: Discussionmentioning

confidence: 99%

Biosynthesis of clinically used antibiotic fusidic acid and identification of two short-chain dehydrogenase/reductases with converse stereoselectivity

Cao

et al. 2019

Acta Pharmaceutica Sinica B

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“…CYP509C12 from Rhizopus oryzae showed 11␣and minor 6␤-hydroxylation activities toward 11-deoxycortisol and testosterone (44). CYP5311B1 from Absidia coerulea was capable of hydroxylating 16,17␣-epoxyprogesterone at the 11␣ position (45). P450pra from Penicillium raistrickii was identified as a steroid hydroxylase with 15␣-hydroxylation activity toward D-ethylgonendione (13-ethyl-gon-4-ene-3,17-dione) (46).…”

Section: Discussionmentioning

confidence: 99%

Distinct Regioselectivity of Fungal P450 Enzymes for Steroidal Hydroxylation

Lü

Feng

Chen

et al. 2019

Appl Environ Microbiol

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In this study, we identified two P450 enzymes (CYP5150AP3 and CYP5150AN1) from Thanatephorus cucumeris NBRC 6298 by combination of transcriptome sequencing and heterologous expression in Pichia pastoris. The biotransformation of 11-deoxycortisol and testosterone by Pichia pastoris whole cells coexpressing the cyp5150ap3 and por genes demonstrated that the CYP5150AP3 enzyme possessed steroidal 7β-hydroxylase activities toward these substrates, and the regioselectivity was dependent on the structures of steroidal compounds. CYP5150AN1 catalyzed the 2β-hydroxylation of 11-deoxycortisol. It is interesting that they display different regioselectivity of hydroxylation from that of their isoenzyme, CYP5150AP2, which possesses 19- and 11β-hydroxylase activities. IMPORTANCE The steroidal hydroxylases CYP5150AP3 and CYP5150AN1 together with the previously characterized CYP5150AP2 belong to the CYP5150A family of P450 enzymes with high amino acid sequence identity, but they showed completely different regioselectivities toward 11-deoxycortisol, suggesting the regioselectivity diversity of steroidal hydroxylases of CYP5150 family. They are also distinct from the known bacterial and fungal steroidal hydroxylases in substrate specificity and regioselectivity. Biocatalytic hydroxylation is one of the important transformations for the functionalization of steroid nucleus rings but remains a very challenging task in organic synthesis. These hydroxylases are useful additions to the toolbox of hydroxylase enzymes for the functionalization of steroids at various positions.

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“…Real-time quantitative PCR was performed on Step One System (ABI, USA) to determine mRNA level of ptsG . The PCR reaction system (20 μl) and conditions were according to the method as described [ 25 ]. The Ct values were used to quantify the relative expression levels of ptsG by the 2 −ΔCt method with the constitutively expressed gapA , encoding glyceraldehyde-3-phosphate dehydrogenase A, as the internal control [ 26 ].…”

Section: Methodsmentioning

confidence: 99%

Improved ethanol productivity from lignocellulosic hydrolysates by Escherichia coli with regulated glucose utilization

et al. 2018

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BackgroundLignocellulosic ethanol could offer a sustainable source to meet the increasing worldwide demand for fuel. However, efficient and simultaneous metabolism of all types of sugars in lignocellulosic hydrolysates by ethanol-producing strains is still a challenge.ResultsAn engineered strain Escherichia coli B0013-2021HPA with regulated glucose utilization, which could use all monosaccharides in lignocellulosic hydrolysates except glucose for cell growth and glucose for ethanol production, was constructed. In E. coli B0013-2021HPA, pta-ackA, ldhA and pflB were deleted to block the formation of acetate, lactate and formate and additional three mutations at glk, ptsG and manZ generated to block the glucose uptake and catabolism, followed by the replacement of the wild-type frdA locus with the ptsG expression cassette under the control of the temperature-inducible λ pR and pL promoters, and the final introduction of pEtac-PA carrying Zymomonas mobilis pdc and adhB for the ethanol pathway. B0013-2021HPA was able to utilize almost all xylose, galactose and arabinose but not glucose for cell propagation at 34 °C and converted all sugars to ethanol at 42 °C under oxygen-limited fermentation conditions.ConclusionsEngineered E. coli strain with regulated glucose utilization showed efficient metabolism of mixed sugars in lignocellulosic hydrolysates and thus higher productivity of ethanol production.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0915-x) contains supplementary material, which is available to authorized users.

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Cloning and identification of a novel steroid 11α-hydroxylase gene from Absidia coerulea

Cited by 26 publications

References 22 publications

Biosynthesis of clinically used antibiotic fusidic acid and identification of two short-chain dehydrogenase/reductases with converse stereoselectivity

Biosynthesis of clinically used antibiotic fusidic acid and identification of two short-chain dehydrogenase/reductases with converse stereoselectivity

Distinct Regioselectivity of Fungal P450 Enzymes for Steroidal Hydroxylation

Improved ethanol productivity from lignocellulosic hydrolysates by Escherichia coli with regulated glucose utilization

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