MmcBC in Pelotomaculum thermopropionicum represents a novel group of prokaryotic fumarases.
The overall amino-acid sequence of MmcBC in Pelotomaculum thermopropionicum was substantially homologous (33%) to fumarase A in Escherichia coli, although its possible subunit structure was different from known fumarases and it lacked the fumarate-lyase signature sequence. Here, MmcBC in E. coli is expressed and characterized. The purified enzyme catalyzed reversible conversion of fumarate to L-malate at an optimum temperature of 70 degrees C. Its molecular size was 64.2 kDa, indicating that it consisted of one MmcB and one MmcC. EPR spectra revealed that it had an oxygen-sensitive [4Fe-4S] cluster. We propose that MmcBC represents a novel group of prokaryotic fumarases.
Shimoyama
Takefumi
T
Marine Biotechnology Institute, Heita, Kamaishi, Iwate, Japan. takefumi.shimoyama@mbio.jp
Rajashekhara
Eranna
E
Ohmori
Daijiro
D
Kosaka
Tomoyuki
T
Watanabe
Kazuya
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005649:Fumarate Hydratase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005650:Fumarates / Q000378:metabolism; D007700:Kinetics; D008293:Malates / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010459:Peptococcaceae / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D001426:Bacterial Proteins; D005650:Fumarates; D008293:Malates; D011994:Recombinant Proteins; C030298:malic acid; D005649:Fumarate Hydratase