Identification, function, and application of 3-ketosteroid Δ1-dehydrogenase isozymes in Mycobacterium neoaurum DSM 1381 for the production of steroidic synthons

Microbial Cell Factories
Ruijie ZhangBaoguo Zhang

Abstract

3-Ketosteroid-Δ1-dehydrogenase (KstD) is a key enzyme in the metabolic pathway for chemical modifications of steroid hormones. Only a few KstDs have thus far been characterized biochemically and applied for the production of steroidal pharmaceutical intermediates. Three KstDs, KstD1, KstD2, and KstD3, were identified in Mycobacterium neoaurum DSM 1381, and they shared up to 99, 85 and 97% amino acid identity with previously reported KstDs, respectively. In this paper, KstDs from M. neoaurum DSM 1381 were investigated and exemplified their potential application for industrial steroid transformation. The recombinant KstD2 from Bacillus subtilis exhibited higher enzymatic activity when 4-androstene-3,17-dione (AD) and 22-hydroxy-23, 24-bisnorchol-4-ene-3-one (4HP) were used as the substrates, and resulted in specific activities of 22.40 and 19.19 U mg-1, respectively. However, the specific activities of recombinant KstD2 from Escherichia coli, recombinant KstD1 from B. subtilis and E. coli, and recombinant KstD3, also fed with AD and 4HP, had significantly lower specific activities. We achieved up to 99% bioconversion rate of 1,4-androstadiene-3,17-dione (ADD) from 8 g L-1 AD after 15 h of fermentation using E. coli transformant B...Continue Reading

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Citations

Sep 10, 2020·Plants·Rituraj BatthHenrik Toft Simonsen
Mar 5, 2021·Microbial Biotechnology·Carmen Felpeto-SanteroJosé Luis García
Apr 7, 2021·Biotechnology Advances·Ali Rohman, Bauke W Dijkstra

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Methods Mentioned

BETA
PCR
enzymatic
electrophoresis

Software Mentioned

BLAST
GS De Novo Assembler
ClustalW
Glimmer
MEGA6
FgenesB
UGENE

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