caa a22 4500 60550217X CHVBK 20210128100604.0 cr unu---uuuuu 210128e20150501xx s 000 0 eng 10.1007/s00253-014-6263-9 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6263-9 Dye-linked d -amino acid dehydrogenase from the thermophilic bacterium Rhodothermus marinus JCM9785: characteristics and role in trans -4-hydroxy- l -proline catabolism [Elektronische Daten] [Takenori Satomura, Masaru Ishikura, Takashi Koyanagi, Haruhiko Sakuraba, Toshihisa Ohshima, Shin-ichiro Suye] A gene from the thermophilic Gram-negative bacterium Rhodothermus marinus JCM9785, encoding a dye-linked d-amino acid dehydrogenase homologue, was overexpressed in Escherichia coli, and its product was purified and characterized. The expressed enzyme was a highly thermostable dye-linked d-amino acid dehydrogenase that retained more than 80% of its activity after incubation for 10min at up to 70°C. When enzyme-catalyzed dehydrogenation of several d-amino acids was carried out using 2,6-dichloroindophenol as the electron acceptor, d-phenylalanine was the most preferable substrate among the d-amino acids tested. Immediately upstream of the dye-linked d-amino acid dehydrogenase gene (dadh) was a gene encoding a 4-hydroxyproline 2-epimerase homologue (hypE). That gene was successfully expressed in E. coli, and the gene product exhibited strong 4-hydroxyproline 2-epimerase activity. Reverse transcription PCR and quantitative real-time PCR showed that the six genes containing the dadh and hypE genes were arranged in an operon and were required for catabolism of trans-4-hydroxy-l-proline in R. marinus. This is the first description of a dye-linked d-amino acid dehydrogenase (Dye-DADH) with broad substrate specificity involved in trans-4-hydroxy-l-proline catabolism. Springer-Verlag Berlin Heidelberg, 2014 trans -4-hydroxy- l -proline catabolism nationallicence d -amino acid dehydrogenase nationallicence Dye-linked dehydrogenase nationallicence Thermophilic bacteria nationallicence Rhodothermus marinus nationallicence Satomura Takenori Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, 910-8507, Fukui, Japan aut Ishikura Masaru Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, 910-8507, Fukui, Japan aut Koyanagi Takashi Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, 910-8507, Fukui, Japan aut Sakuraba Haruhiko Department of Applied Biological Science, Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki-cho, 761-0795, Kita-gun, Kagawa, Japan aut Ohshima Toshihisa Department of Biomedical Engineering, Faculty of Engineering, Osaka Institute of Technology, Ohmiya, 5-16-1 Asahi-ku, 535-8585, Ohsaka, Japan aut Suye Shin-ichiro Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, 910-8507, Fukui, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/10(2015-05-01), 4265-4275 0175-7598 99:10<4265 2015 99 253 https://doi.org/10.1007/s00253-014-6263-9 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/s00253-014-6263-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Satomura Takenori Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, 910-8507, Fukui, Japan aut NATIONALLICENCE 700 1- Ishikura Masaru Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, 910-8507, Fukui, Japan aut NATIONALLICENCE 700 1- Koyanagi Takashi Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, 910-8507, Fukui, Japan aut NATIONALLICENCE 700 1- Sakuraba Haruhiko Department of Applied Biological Science, Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki-cho, 761-0795, Kita-gun, Kagawa, Japan aut NATIONALLICENCE 700 1- Ohshima Toshihisa Department of Biomedical Engineering, Faculty of Engineering, Osaka Institute of Technology, Ohmiya, 5-16-1 Asahi-ku, 535-8585, Ohsaka, Japan aut NATIONALLICENCE 700 1- Suye Shin-ichiro Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, 3-9-1 Bunkyo, 910-8507, Fukui, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/10(2015-05-01), 4265-4275 0175-7598 99:10<4265 2015 99 253