Purification and partial characterization of the Pyrococcus horikoshii methylmalonyl-CoA epimerase.
Methylmalonyl-CoA epimerase (MCE) from the hyperthermophilic archaeon, Pyrococcus horikoshii, was expressed at high levels in Escherichia coli, purified, and partially characterized. The P. horikoshii MCE enzyme was a homodimer with an apparent molecular mass of 31,700 Da. The K(m) of the enzyme for methylmalonyl-CoA was 79 microM and the k(cat) was 240 s(-1). The P. horikoshii enzyme was extremely heat-stable and withstood boiling for 60 min without detectable loss in activity.
Department of Microbiology and Cell Science, University of Florida, Building 981, Room 1130, Gainesville, FL 32611, USA.
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D000214:Acyl Coenzyme A / Q000378:metabolism; D002845:Chromatography; D019281:Dimerization; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D021122:Protein Subunits / Q000737:chemistry; D043902:Pyrococcus horikoshii / Q000201:enzymology*; D019998:Racemases and Epimerases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature
D000214:Acyl Coenzyme A; D021122:Protein Subunits; D011994:Recombinant Proteins; C015357:methylmalonyl-coenzyme A; D019998:Racemases and Epimerases; C021581:methylmalonyl-coenzyme A racemase