TY - JOUR
T1 - Molybdenum-containing membrane-bound formate dehydrogenase isolated from Citrobacter sp. S-77 having high stability against oxygen, pH, and temperature
AU - Nguyen, Nga T.
AU - Yatabe, Takeshi
AU - Yoon, Ki Seok
AU - Ogo, Seiji
N1 - Funding Information:
This work was supported by the World Premier International Research Center Initiative (WPI), grants-in-aid: 23655053 , 25620047 , 25248017 , and 24109016 (Scientific Research on Innovative Areas “Stimuli-responsive Chemical Species”) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan , and the Basic Research Programs CREST Type, “Development of the Foundation for Nano-Interface Technology” from Japan Science and Technology Agency (JST), Japan .
Publisher Copyright:
© 2014 The Society for Biotechnology, Japan.
PY - 2014/10/1
Y1 - 2014/10/1
N2 - Membrane-bound formate dehydrogenase (FDH) was purified to homogeneity from a facultative anaerobic bacterium Citrobacter sp. S-77. The FDH from Citrobacter sp. S-77 (FDHS77) was a monomer with molecular mass of approximately 150kDa. On SDS-PAGE, the purified FDHS77 showed as three different protein bands with molecular mass of approximately 95, 87, and 32kDa, respectively. Based on the N-terminal amino acid sequence analysis, the sequence alignments observed for the 87kDa protein band were identical to that of the large subunit of 95kDa, indicating that the purified FDHS77 consisted of two subunits; a 95kDa large subunit and a 32kDa small subunit. The purified FDHS77 in this purification did not contain a heme b subunit, but the FDHS77 showed significant activity for formate oxidation, determined by the Vmax of 30.4U/mg using benzyl viologen as an electron acceptor. The EPR and ICP-MS spectra indicate that the FDHS77 is a molybdenum-containing enzyme, displaying a remarkable O2-stability along with thermostability and pH resistance. This is the first report of the purification and characterization of a FDH from Citrobacter species.
AB - Membrane-bound formate dehydrogenase (FDH) was purified to homogeneity from a facultative anaerobic bacterium Citrobacter sp. S-77. The FDH from Citrobacter sp. S-77 (FDHS77) was a monomer with molecular mass of approximately 150kDa. On SDS-PAGE, the purified FDHS77 showed as three different protein bands with molecular mass of approximately 95, 87, and 32kDa, respectively. Based on the N-terminal amino acid sequence analysis, the sequence alignments observed for the 87kDa protein band were identical to that of the large subunit of 95kDa, indicating that the purified FDHS77 consisted of two subunits; a 95kDa large subunit and a 32kDa small subunit. The purified FDHS77 in this purification did not contain a heme b subunit, but the FDHS77 showed significant activity for formate oxidation, determined by the Vmax of 30.4U/mg using benzyl viologen as an electron acceptor. The EPR and ICP-MS spectra indicate that the FDHS77 is a molybdenum-containing enzyme, displaying a remarkable O2-stability along with thermostability and pH resistance. This is the first report of the purification and characterization of a FDH from Citrobacter species.
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U2 - 10.1016/j.jbiosc.2014.03.011
DO - 10.1016/j.jbiosc.2014.03.011
M3 - Article
C2 - 24751436
AN - SCOPUS:84921646116
SN - 1389-1723
VL - 118
SP - 386
EP - 391
JO - Journal of Bioscience and Bioengineering
JF - Journal of Bioscience and Bioengineering
IS - 4
ER -