caa a22 4500 605499683 CHVBK 20210128100552.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00253-015-6563-8 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6563-8 Comparison of CYP106A1 and CYP106A2 from Bacillus megaterium  - identification of a novel 11-oxidase activity [Elektronische Daten] [Flora Kiss, Daniela Schmitz, Josef Zapp, Tobias Dier, Dietrich Volmer, Rita Bernhardt] The CYP106A subfamily hydroxylates steroids, diterpenes, and triterpenes in a regioselective and stereoselective manner, which is a challenging task for synthetic chemistry. The well-studied CYP106A2 enzyme, from the Bacillus megaterium strain ATCC 13368, is a highly promising candidate for the pharmaceutical industry. It shares 63% amino acid sequence identity with CYP106A1 from B. megaterium DSM319, which was recently characterized. A focused steroid library was screened with both CYP106A1 and CYP106A2. Out of the 23 tested steroids, 19 were successfully converted by both enzymes during in vitro and in vivo reactions. Thirteen new substrates were identified for CYP106A1, while the substrate spectrum of CYP106A2 was extended by seven new members. Finally, six chosen steroids were further studied on a preparative scale employing a recombinant B. megaterium MS941 whole-cell system, yielding sufficient amounts of product for structure characterization by nuclear magnetic resonance. The hydroxylase activity was confirmed at positons 6β, 7β, 9α, and 15β. In addition, the CYP106A subfamily showed unprecedented 11-oxidase activity, converting 11β-hydroxysteroids to their 11-keto derivatives. This novel reaction and the diverse hydroxylation positions on pharmaceutically relevant compounds underline the role of the CYP106A subfamily in drug development and production. Springer-Verlag Berlin Heidelberg, 2015 CYP106A1 nationallicence CYP106A2 nationallicence Cortisol nationallicence Cortisone nationallicence 11β-Hydroxysteroid dehydrogenation nationallicence Kiss Flora Institute of Biochemistry, Saarland University, Campus B 2.2, 66123, Saarbruecken, Germany aut Schmitz Daniela Institute of Biochemistry, Saarland University, Campus B 2.2, 66123, Saarbruecken, Germany aut Zapp Josef Institute of Pharmaceutical Biology, Saarland University, 66123, Saarbruecken, Germany aut Dier Tobias Institute of Bioanalytical Chemistry, Saarland University, 66123, Saarbruecken, Germany aut Volmer Dietrich Institute of Bioanalytical Chemistry, Saarland University, 66123, Saarbruecken, Germany aut Bernhardt Rita Institute of Biochemistry, Saarland University, Campus B 2.2, 66123, Saarbruecken, Germany aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/20(2015-10-01), 8495-8514 0175-7598 99:20<8495 2015 99 253 https://doi.org/10.1007/s00253-015-6563-8 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/s00253-015-6563-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kiss Flora Institute of Biochemistry, Saarland University, Campus B 2.2, 66123, Saarbruecken, Germany aut NATIONALLICENCE 700 1- Schmitz Daniela Institute of Biochemistry, Saarland University, Campus B 2.2, 66123, Saarbruecken, Germany aut NATIONALLICENCE 700 1- Zapp Josef Institute of Pharmaceutical Biology, Saarland University, 66123, Saarbruecken, Germany aut NATIONALLICENCE 700 1- Dier Tobias Institute of Bioanalytical Chemistry, Saarland University, 66123, Saarbruecken, Germany aut NATIONALLICENCE 700 1- Volmer Dietrich Institute of Bioanalytical Chemistry, Saarland University, 66123, Saarbruecken, Germany aut NATIONALLICENCE 700 1- Bernhardt Rita Institute of Biochemistry, Saarland University, Campus B 2.2, 66123, Saarbruecken, Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/20(2015-10-01), 8495-8514 0175-7598 99:20<8495 2015 99 253