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A plasmid containing the genes encoding a fungal 17β-hydroxysteroid dehydrogenase, which ca-talyzes the reduction of 17-oxo group, and a mycobacterial glucose-6-phosphate dehydrogenase, which promotes the recycling of the essential coenzyme NAD(P)H, was constructed. Its constitutive expression in well-studied Mycolicibacterium neoaurum strains made it possible to increase the yield of C-17 hydroxysteroids significantly. In particular, recombinant strains created on the basis of M. neoaurum VKM Ac-1815D and M. neoaurum NRRL B-3805 ΔkstD exhibited predominant accumulation of testosterone, while the strain based on M. neoaurum VKM Ac-1816D accumulated 1(2)-dehydrotestosterone and testosterone simultaneously. [ABSTRACT FROM AUTHOR]