caa a22 4500 445818808 CHVBK 20180317145237.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s00792-010-0345-0 doi (NATIONALLICENCE)springer-10.1007/s00792-010-0345-0 Comparative study on dihydrofolate reductases from Shewanella species living in deep-sea and ambient atmospheric-pressure environments [Elektronische Daten] [Chiho Murakami, Eiji Ohmae, Shin-ichi Tate, Kunihiko Gekko, Kaoru Nakasone, Chiaki Kato] To examine whether dihydrofolate reductase (DHFR) from deep-sea bacteria has undergone molecular evolution to adapt to high-pressure environments, we cloned eight DHFRs from Shewanella species living in deep-sea and ambient atmospheric-pressure environments, and subsequently purified six proteins to compare their structures, stabilities, and functions. The DHFRs showed 74-90% identity in primary structure to DHFR from S. violacea, but only 55% identity to DHFR from Escherichia coli (ecDHFR). Far-ultraviolet circular dichroism and fluorescence spectra suggested that the secondary and tertiary structures of these DHFRs were similar. In addition, no significant differences were found in structural stability as monitored by urea-induced unfolding and the kinetic parameters, K m and k cat; although the DHFRs from Shewanella species were less stable and more active (2- to 4-fold increases in k cat/K m) than ecDHFR. Interestingly, the pressure effects on enzyme activity revealed that DHFRs from ambient-atmospheric species are not necessarily incompatible with high pressure, and DHFRs from deep-sea species are not necessarily tolerant of high pressure. These results suggest that the DHFR molecule itself has not evolved to adapt to high-pressure environments, but rather, those Shewanella species with enzymes capable of retaining functional activity under high pressure migrated into the deep-sea. Springer, 2010 Deep-sea nationallicence DHFR nationallicence High pressure nationallicence Molecular adaptation nationallicence Shewanella species nationallicence DHF : Dihydrofolate nationallicence DHFR : Dihydrofolate reductase nationallicence ecDHFR : DHFR from Escherichia coli nationallicence sb21DHFR : DHFR from Shewanella benthica strain DB21MT-2 nationallicence sb43992DHFR : DHFR from S. benthica strain ATCC43992 nationallicence sb6705DHFR : DHFR from S. benthica strain DB6705 nationallicence sfDHFR : DHFR from S. frigidimarina strain ACAM591 nationallicence sgDHFR : DHFR from S. gelidimarina strain ACAM456 nationallicence soDHFR : DHFR from S. oneidensis strain MR-1 nationallicence spDHFR : DHFR from S. putrefaciens strain IAM12079 nationallicence svDHFR : DHFR from S. violacea strain DSS12 nationallicence THF : Tetrahydrofolate nationallicence Murakami Chiho Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 739-8526, Higashi-Hiroshima, Japan aut Ohmae Eiji Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 739-8526, Higashi-Hiroshima, Japan aut Tate Shin-ichi Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 739-8526, Higashi-Hiroshima, Japan aut Gekko Kunihiko Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 739-8526, Higashi-Hiroshima, Japan aut Nakasone Kaoru Department of Biotechnology and Chemistry, School of Engineering, Kinki University, 739-2116, Higashi-Hiroshima, Japan aut Kato Chiaki Institute of Biogeosciences, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 237-0061, Yokosuka, Japan aut Extremophiles Springer Japan 15/2(2011-03-01), 165-175 1431-0651 15:2<165 2011 15 792 https://doi.org/10.1007/s00792-010-0345-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00792-010-0345-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Murakami Chiho Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 739-8526, Higashi-Hiroshima, Japan aut NATIONALLICENCE 700 1- Ohmae Eiji Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 739-8526, Higashi-Hiroshima, Japan aut NATIONALLICENCE 700 1- Tate Shin-ichi Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 739-8526, Higashi-Hiroshima, Japan aut NATIONALLICENCE 700 1- Gekko Kunihiko Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 739-8526, Higashi-Hiroshima, Japan aut NATIONALLICENCE 700 1- Nakasone Kaoru Department of Biotechnology and Chemistry, School of Engineering, Kinki University, 739-2116, Higashi-Hiroshima, Japan aut NATIONALLICENCE 700 1- Kato Chiaki Institute of Biogeosciences, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 237-0061, Yokosuka, Japan aut NATIONALLICENCE 773 0- Extremophiles Springer Japan 15/2(2011-03-01), 165-175 1431-0651 15:2<165 2011 15 792 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer