The microbial biotransformation is a robust procedure in developing steroids and fungi are practical tools in this process therefore, the fungal modi cation of testosterone by Penicillium pinophilum was investigated. The three prominent metabolites, including 14α-hydroxyandrost-4-en-3,17-dione (II), 14αhydroxytestosterone (III), and 11α-hydroxytestosterone (IV), were isolated and characterized by chromatographic and spectroscopic methods. The time course pro le showed that the content of the metabolites II and III began to decrease after 24 h and 96 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.

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 24 h and 96 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.