Purification, characterization, and application of a high activity 3-ketosteroid-Δ 1 -dehydrogenase from Mycobacterium neoaurum DSM 1381

摘要: Δ1-Dehydrogenation is one of the most important reactions for steroid drug modification. Numerous 3-ketosteroid-Δ1-dehydrogenases (KstDs) catalyzing this reaction were observed in various organisms. However, only a few have been characterized and used substrate conversion. In study, promising enzyme (KstD2) from Mycobacterium neoaurum DSM 1381 was purified characterized. Interestingly, KstD2 displayed high activity on range substrates, including 17α-hydroxypregn-4-ene-3,20-dione (17α-OH-P); androsta-4,9(11)-diene-3,17-dione (NSC 44826); 4-androstene-3,17-dione (AD). These performed under optimal conditions at 40 °C pH 8.0. Noteworthy, both stability sensitive to metal ions. After optimizing expression biocatalyst conditions, up 1586 U mg-1 intracellular KstD AD could be produced. Furthermore, associated conversion rate 99% with 30 g L-1 after 8 h. On other hand, we obtained 99%, 90%, over 80% 20 NSC 44826; 10 16,17α-epoxyprogesterone; 17α-OH-P or canrenone, respectively, 24 Sequence homology structural analyses indicated that residue R178 located unique short loop among cluster 2 crucial recognition which confirmed by mutagenesis. summary, study reports first purification characterization 2. Its remarkable properties deserve more attention potentially lead further industrial applications.