Abstract
The microbial biotransformation is a robust procedure in developing steroids and fungi are practical tools in this process; therefore, the fungal modification of testosterone by Penicillium pinophilum was investigated. The three prominent metabolites, including 14α-hydroxyandrost-4-en-3,17-dione (II), 14α-hydroxytestosterone (III), and 11α-hydroxytestosterone (IV), were isolated and characterized by chromatographic and spectroscopic methods. The time course profile showed that the content of the metabolites II and III began to decrease after 96 and 24 h, respectively. In comparison, the content of the metabolite IV remained stable after 24 h. In silico studies showed that the probability of binding to the androgen receptor remains high for all three metabolites. However, the probability of binding to the estrogen receptors α and β increased for metabolite IV but decreased for metabolite III. Penicillium pinophilum as a potentially viable biocatalyst could hydroxylate C-11α and C-14α positions and oxidize the C-17β hydroxyl group to 17-ketone in testosterone molecule.
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The datasets used and/or analyzed during the present study are available from corresponding author on reasonable request.
References
Al-Aboudi A, Mohammad MY, Musharraf SG, Choudhary MI, Atta-ur-Rahman (2008) Microbial transformation of testosterone by Rhizopus stolonifer and Fusarium lini. Nat Prod Res 22:1498–1509. https://doi.org/10.1080/14786410802234528
Andryushina VA, Voishvillo NE, Druzhinina AV, Stytsenko TS, Yaderets VV, Petrosyan MA, Zeinalov OA (2013) 14α-Hydroxylation of steroids by mycelium of the mold fungus Curvularia lunata (VKPM F-981) to produce precursors for synthesizing new steroidal drugs. Pharm Chem J 47:103–108. https://doi.org/10.1007/s11094-013-0905-6
Bartmańska A, Dmochowska-Gładysz J, Huszcza E (2005) Steroids’ transformations in Penicillium notatum culture. Steroids 70:193–198. https://doi.org/10.1016/j.steroids.2004.11.011
Cabeza MS, Gutiérrez EB, García GA, Avalos AH, Hernández MA (1999) Microbial transformations of testosterone to 5alpha-dihydrotestosterone by two species of Penicillium: P. chrysogenum and P. crustosum. Steroids 64:379–384. https://doi.org/10.1016/s0039-128x(98)00115-9
Faramarzi MA, Aghelnejad M, Tabatabaei Yazdi M, Amini M, Hajarolasvadi N (2008a) Metabolism of androst-4-en-3,17-dione by the filamentous fungus Neurospora crassa. Steroids 73:13–18. https://doi.org/10.1016/j.steroids.2007.06.008
Faramarzi MA, Badiee M, Yazdi MT, Amini M, Torshabi M (2008b) Formation of hydroxysteroid derivatives from androst-4-en-3,17-dione by the filamentous fungus Mucor racemosus. J Mol Catal b: Enzym 50:7–12. https://doi.org/10.1016/j.molcatb.2007.09.017
Fernandes P, Cabral JMS (2006) Biotransformations. In: Kristiansen B, Ratledge C (eds) Basic biotechnology, 3rd edn. Cambridge University Press, Cambridge, pp 579–626. https://doi.org/10.1017/CBO9780511802409.026
Fernandes P, Cruz A, Angelova B, Pinheiro H, Cabral J (2003) Microbial conversion of steroid compounds: recent developments. Enzyme Microb Technol 32:688–705. https://doi.org/10.1016/S0141-0229(03)00029-2
Ghasemi S, Mohajeri M, Habibi Z (2014) Biotransformation of testosterone and testosterone heptanoate by four filamentous fungi. Steroids 92:7–12. https://doi.org/10.1016/j.steroids.2014.09.002
Hanson JR, Nasir H, Parvez A (1996) The hydroxylation of testosterone and some relatives by Cephalosporium aphidicola. Phytochemistry 42:411–415. https://doi.org/10.1016/0031-9422(95)00968-X
Hegazy ME, Mohamed TA, ElShamy AI, Mohamed AE, Mahalel UA, Reda EH, Shaheen AM, Tawfik WA, Shahat AA, Shams KA, Abdel-Azim NS, Hammouda FM (2015) Microbial biotransformation as a tool for drug development based on natural products from mevalonic acid pathway: a review. J Adv Res 6:17–33. https://doi.org/10.1016/j.jare.2014.11.009
Heidary M, Habibi Z (2016) Microbial transformation of androst-4-ene-3,17-dione by three fungal species Absidia griseolla var. igachii, Circinella muscae and Trichoderma virens. J Mol Catal B Enzym 126:32–36. https://doi.org/10.1016/j.molcatb.2016.01.007
Holland HL, Dore S, Xu W, Brown FM (1994) Formation of 5 alpha steroids by biotransformation involving the 5 alpha-reductase activity of Penicillium decumbens. Steroids 59:642–647. https://doi.org/10.1016/0039-128x(94)90020-5
Holland HL, Nguyen DH, Pearson NM (1995) Biotransformation of corticosteroids by Penicillium decumbens ATCC 10436. Steroids 60:646–649. https://doi.org/10.1016/0039-128x(95)00071-w
Hosseinabadi T, Vahidi H, Nickavar B, Kobarfard F (2015) Biotransformation of progesterone by whole cells of filamentous fungi Aspergillus brasiliensis. Iran J Pharm Res 14:919–924
Hu S, Genain G, Azerad R (1995) Microbial transformation of steroids: contribution to 14α-hydroxylations. Steroids 60:337–352. https://doi.org/10.1016/0039-128X(95)00006-C
Hunter AC, Rymer SJ, Dedi C, Dodd HT, Nwozor QC, Moghimi SM (2011) Transformation of structurally diverse steroidal analogues by the fungus Corynespora cassiicola CBS 161.60 results in generation of 8β-monohydroxylated metabolites with evidence in favour of 8β-hydroxylation through inverted binding in the 9α-hydroxylase. Biochim Biophys Acta Mol Cell Biol Lipids 1811:1054–1061. https://doi.org/10.1016/j.bbalip.2011.09.016
Hüttel W, Hoffmeister D (2011) Fungal biotransformations in pharmaceutical sciences. In: Hofrichter M (ed) Industrial applications X The Mycota: a comprehensive treatise on fungi as experimental systems for basic and applied research, vol 10. Springer, Heidelberg, pp 293–317. https://doi.org/10.1007/978-3-642-11458-8_14
Karpova NV, Andryushina VA, Stytsenko TS, Druzhinina AV, Feofanova TD, Kurakov AV (2016) A search for microscopic fungi with directed hydroxylase activity for the synthesis of steroid drugs. Appl Biochem Microbiol 52:316–323. https://doi.org/10.1134/S000368381603008X
Kolet SP, Niloferjahan S, Haldar S, Gonnade R, Thulasiram HV (2013) Biocatalyst mediated production of 6β,11α-dihydroxy derivatives of 4-ene-3-one steroids. Steroids 78:1152–1158. https://doi.org/10.1016/j.steroids.2013.08.004
Kolet SP, Haldar S, Niloferjahan S, Thulasiram HV (2014) Mucor hiemalis mediated 14α-hydroxylation on steroids: in vivo and in vitro investigations of 14α-hydroxylase activity. Steroids 85:6–12. https://doi.org/10.1016/j.steroids.2014.04.002
Kolšek K, Mavri J, SollnerDolenc M, Gobec S, Turk S (2014) Endocrine disruptome—an open source prediction tool for assessing endocrine disruption potential through nuclear receptor binding. J Chem Inf Model 54:1254–1267. https://doi.org/10.1021/ci400649p
Leresche JE, Meyer HP (2006) Chemocatalysis and biocatalysis (biotransformation): some thoughts of a chemist and of a biotechnologist. Org Process Res Dev 10:572–580. https://doi.org/10.1021/op0600308
Meena M, Zehra A, Dubey MK, Aamir M, Upadhyay RS (2017) Penicillium enzymes for the food industries. In: Gupta VK, Rodriguez-Couto S (eds) New and future developments in microbial biotechnology and bioengineering. Elsevier, Amsterdam, pp 167–186. https://doi.org/10.1016/B978-0-444-63501-3.00009-0
Nickavar B, Vahidi H, Eslami M (2019) An efficient biotransformation of progesterone into 11α-hydroxyprogesterone by Rhizopus microsporus var. oligosporus. Z Naturforsch C J Biosci 74:9–15. https://doi.org/10.1515/znc-2018-0092
Nicoletti R, Ciavatta ML, Buommino E, Tufano MA (2008) Antitumor extrolites produced by Penicillium species. Int J Biomed Pharm Sci 2:1–23
Panek A, Łyczko P, Świzdor A (2020) Microbial modifications of androstane and androstene steroids by Penicillium vinaceum. Molecules. https://doi.org/10.3390/molecules25184226
Peart PC, McCook KP, Russell FA, Reynolds WF, Reese PB (2011) Hydroxylation of steroids by Fusarium oxysporum, Exophiala jeanselmei and Ceratocystis paradoxa. Steroids 76:1317–1330. https://doi.org/10.1016/j.steroids.2011.06.010
Peart PC, Reynolds WF, Reese PB (2013) The facile bioconversion of testosterone by alginate-immobilised filamentous fungi. J Mol Catal B Enzym 95:70–81. https://doi.org/10.1016/j.molcatb.2013.05.025
Peart PC, Reynolds WF, Reese PB (2016) The generation of a steroid library using filamentous fungi immobilized in calcium alginate. J Mol Catal B Enzym 125:16–24. https://doi.org/10.1016/j.molcatb.2015.11.026
Perrone G, Susca A (2017) Penicillium species and their associated mycotoxins. In: Moretti A, Susca A (eds) Mycotoxigenic fungi. Humana Press, New York, pp 107–119. https://doi.org/10.1007/978-1-4939-6707-0_5
Petit P, Lucas E, Abreu L, Pfenning L, Takahashi J (2009) Novel antimicrobial secondary metabolites from a Penicillium sp. isolated from Brazilian cerrado soil. Electron J Biotechnol. https://doi.org/10.2225/vol12-issue4-fulltext-9
Ramos-Ponce LM, Contreras-Esquivel JC, Saenz JM, Lara-Cisneros G, Garza-Garcia Y (2012) Study on production of mycophenolic acid by Penicillium pinophilum using response surface metodology. In: 2nd IEEE Portuguese meeting in bioengineering, Combria. https://doi.org/10.1109/ENBENG.2012.6331348
Sambyal K, Singh RV (2020) Production aspects of testosterone by microbial biotransformation and future prospects. Steroids 159:108651. https://doi.org/10.1016/j.steroids.2020.108651
Świzdor A, Panek A, Milecka-Tronina N (2017) Hydroxylative activity of Aspergillus niger towards androst-4-ene and androst-5-ene steroids. Steroids 126:101–106. https://doi.org/10.1016/j.steroids.2017.08.009
Tong WY, Dong X (2009) Microbial biotransformation: Recent developments on steroid drugs. Recent Pat Biotechnol 3:141–153. https://doi.org/10.2174/187220809788700157
Tweit RC, Muir RD, Dodson RM (1962) Microbiological transformations. XII. The substrate specificity of hydroxylations by a Penicillium sp., A.T.C.C. 12,556. J Org Chem 27:3654–3658. https://doi.org/10.1021/jo01057a061
Wu Y, Li H, Zhang XM, Gong JS, Rao ZM, Shi JS, Zhang XJ, Xu ZH (2015) Efficient hydroxylation of functionalized steroids by Colletotrichum lini ST-1. J Mol Catal B Enzym 120:111–118. https://doi.org/10.1016/j.molcatb.2015.07.003
Xu S, Li Y (2020) Yeast as a promising heterologous host for steroid bioproduction. J Ind Microbiol Biotechnol 47:829–843. https://doi.org/10.1007/s10295-020-02291-7
Yang B, Wang Y, Chen X, Feng J, Wu Q, Zhu D, Ma Y (2014) Biotransformations of steroids to testololactone by a multifunctional strain Penicillium simplicissimum WY134-2. Tetrahedron 70:41–46. https://doi.org/10.1016/j.tet.2013.11.039
Yildirim K, Kuru A (2016) The biotransformation of some steroids by Aspergillus sydowii MRC 200653. J Chem Res 40:78–81. https://doi.org/10.3184/174751916X14526064507450
Yildirim K, Gulsan F, Kupcu I (2010a) Biotransformation of testosterone and progesterone by Penicillium digitatum MRC 500787. Collect Czechoslov Chem Commun 75:675–683. https://doi.org/10.1135/cccc2009550
Yildirim K, Kupcu I, Gulsan F (2010b) Biotransformation of some steroids by Aspergillus wentii. Z Naturforsch C J Biosci 65:688–692. https://doi.org/10.1515/znc-2010-11-1209
Yildirim K, Uzuner A, Gulcuoglu EY (2010c) Biotransformation of some steroids by Aspergillus terreus MRC 200365. Collect Czechoslov Chem Commun 75:665–673. https://doi.org/10.1135/cccc2009545
Yildirim K, Uzuner A, Gulcuoglu EY (2011) Baeyer-Villiger oxidation of some steroids by Aspergillus tamarii MRC 72400. Collect Czechoslov Chem Commun 76:743–754. https://doi.org/10.1135/cccc2011008
Yildirim K, Saran H, Dolu OF, Kuru A (2013) Biotransformation of some steroids by Mucor hiemalis MRC 70325. J Chem Res 37:566–569. https://doi.org/10.3184/174751913X13745069090242
Yildirim K, Kuru A, Yılmaz Ş (2018) Biotransformation of testosterone by Ulocladium Chartarum MRC 72584. J Chem Res 42:444–446. https://doi.org/10.3184/174751918x15341764332783
Yildirim K, Kuru A, Yılmaz Ş (2019) Biotransformation of testosterone by Cladosporium sphaerospermum. Biocatal Biotransform 37:409–413. https://doi.org/10.1080/10242422.2019.1583747
Zhang H, Ren J, Wang Y, Sheng C, Wu Q, Diao A, Zhu D (2013) Effective multi-step functional biotransformations of steroids by a newly isolated Fusarium oxysporum SC1301. Tetrahedron 69:184–189. https://doi.org/10.1016/j.tet.2012.10.047
Acknowledgements
The authors acknowledge to Shahid Beheshti University of Medical Sciences for supporting this study as a part of Maryam Mehmannavaz’s PhD thesis that has been carried out at School of Pharmacy, Shahid Beheshti University of Medical Sciences.
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This study was supported by the Shahid Beheshti University of Medical Sciences, Tehran, Iran.
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MM performed the experiments, data acquisition and analysis, and wrote the first draft of the manuscript. BN designed the study, analyzed the data and edited the manuscript. All authors approved the final version of the manuscript.
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Mehmannavaz, M., Nickavar, B. Biotransformation of testosterone by the filamentous fungus Penicillium pinophilum. Arch Microbiol 204, 570 (2022). https://doi.org/10.1007/s00203-022-03191-3
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DOI: https://doi.org/10.1007/s00203-022-03191-3