caa a22 4500 606212795 CHVBK 20210128101044.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s10295-014-1577-2 doi (NATIONALLICENCE)springer-10.1007/s10295-014-1577-2 Genetic differences in ksdD influence on the ADD/AD ratio of Mycobacterium neoaurum [Elektronische Daten] [Rili Xie, Yanbing Shen, Ning Qin, Yibo Wang, Liqiu Su, Min Wang] Mycobacterium neoaurum TCCC 11028 (MNR) and M. neoaurum TCCC 11028 M3 (MNR M3) significantly differ in the ratio of androst-1,4-diene-3,17-dione (ADD) to androst-4-ene-3,17-dione (AD) produced. The large fluctuations are related to the dehydrogenation activity of 3-ketosteroid-Δ1-dehydrogenase (KsdD). Analysis of the primary structure of KsdD showed that the Ser-138 of KsdD-MNR changed to Leu-138 of KsdD-MNR M3 because of C413T in the ksdD gene. This phenomenon directly affected KsdD activity. The effect of the primary structure of KsdD on dehydrogenation activity was confirmed through exogenous expression. Whole-cell transformation initially revealed that KsdD-MNR showed a higher dehydrogenation activity than KsdD-MNR M3. Then, ksdD gene replacement strain was constructed by homologous recombination. The results of steroid transformation experiments showed that the ability of the MNR M3ΔksdD::ksdD-MNR strain to produce ADD was improved and it returned to the similar level of the MNR strain. This result indicated that the ADD/AD ratio of the two M. neoaurum strains was influenced by the difference in ksdD. The mechanism by which residue mutations alter enzyme activity may be connected with the crystal structure of KsdD from Rhodococcus erythropolis SQ1. As a key amino acid residue in the active center position, Ser-138 played an important role in maintaining the active center in the hydrophobic environment of KsdD. This study may serve as a basis for future studies on the structural analysis and catalytic mechanism of dehydrogenase. Society for Industrial Microbiology and Biotechnology, 2015 Mycobacterium neoaurum nationallicence 3-ketosteroid-Δ1-dehydrogenase nationallicence Biotransformation nationallicence Androst-4-ene-3,17-dione nationallicence Androst-1,4-diene-3,17-dione nationallicence Xie Rili Tianjin, China aut Shen Yanbing Tianjin, China aut Qin Ning Tianjin, China aut Wang Yibo Tianjin, China aut Su Liqiu Tianjin, China aut Wang Min Tianjin, China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/4(2015-04-01), 507-513 1367-5435 42:4<507 2015 42 10295 https://doi.org/10.1007/s10295-014-1577-2 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-014-1577-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Xie Rili Tianjin, China aut NATIONALLICENCE 700 1- Shen Yanbing Tianjin, China aut NATIONALLICENCE 700 1- Qin Ning Tianjin, China aut NATIONALLICENCE 700 1- Wang Yibo Tianjin, China aut NATIONALLICENCE 700 1- Su Liqiu Tianjin, China aut NATIONALLICENCE 700 1- Wang Min Tianjin, China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/4(2015-04-01), 507-513 1367-5435 42:4<507 2015 42 10295