Purification and characterization of methylmalonyl-CoA mutase from a methanol-utilizing bacterium, Methylobacterium extorquens NR-1.
High activity (about 50 mU x mg protein(-1)) of methylmalonyl-CoA mutase (82-95% apo-enzyme) was constantly found during the cell growth of a methanol-utilizing bacterium, Methylobacterium extorquens NR-1. The apo-enzyme was purified to homogeneity and characterized. The purified enzyme was colorless. An apparent Mr of M. extorquens NR-1 enzyme was calculated to be 150,000 +/- 5,000 by Superdex 200 HR gel filtration. SDS-polyacrylamide gel electrophoresis of the purified enzyme gave two protein bands with an apparent Mr of 85.000 +/- 2,000 and 70,000 +/- 2,000, indicating that the M. extorquens NR-1 enzyme is composed of two nonidentical subunits. NH2-terminal amino acid sequences of the small and large subunits of M. extorquens NR-1 enzyme showed no significant homology to those of the enzyme from other species. Some enzymological properties of the M. extorquens NR-1 enzyme were studied.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis / Q000737:chemistry; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007535:Isomerases / Q000302:isolation & purification / Q000378:metabolism; D000432:Methanol; D008765:Methylmalonyl-CoA Mutase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D020601:Methylobacterium extorquens / Q000201:enzymology*; D008970:Molecular Weight; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D000596:Amino Acids; D007535:Isomerases; D008765:Methylmalonyl-CoA Mutase; D000432:Methanol