Purification and characterization of homodimeric methylmalonyl-CoA mutase from Sinorhizobium meliloti.

Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 0410427 Publication Model: Print-Electronic Cited Medium: Print ISSN: 0302-8933 (Print) Linking ISSN: 03028933 NLM ISO Abbreviation: Arch Microbiol Subsets: MEDLINE

Original Publication: Berlin, New York, Springer-Verlag.

Methylmalonyl-CoA Mutase/*chemistry ; Methylmalonyl-CoA Mutase/*isolation & purification ; Sinorhizobium meliloti/*enzymology; Apoenzymes/isolation & purification ; Chromatography, Gel ; Cobamides ; Dimerization ; Electrophoresis, Polyacrylamide Gel ; Enzyme Activation ; Holoenzymes/metabolism ; Kinetics ; Methylmalonyl-CoA Mutase/biosynthesis ; Methylmalonyl-CoA Mutase/metabolism ; Molecular Weight ; Protein Subunits ; Sinorhizobium meliloti/growth & development

High activity (>60 munit/mg protein) of 5'-deoxyadenosylcobalamin-dependent methylmalonyl-CoA mutase (EC 5.4.99.2) was constantly found during growth of a strain of the root-nodule-forming bacterium Sinorhizobium meliloti harboring an extra plasmid-encoded copy of the methylmalonyl-CoA-mutase-encoding bhbA gene. The enzyme was purified to homogeneity and characterized. The purified enzyme was found to be a colorless apo-form. The apparent molecular weight of the enzyme was calculated to be 165,000+/-5,000 by Superdex 200 HR gel filtration. SDS-PAGE of the purified enzyme resolved one protein band with an apparent molecular mass of 80.0+/-2.0 kDa, indicating that the S. meliloti enzyme is composed of two identical subunits. The NH(2)-terminal sequence was identical to that predicted from the bhbA nucleotide sequence. Monovalent cations were required for enzyme activity.

0 (Apoenzymes)
0 (Cobamides)
0 (Holoenzymes)
0 (Protein Subunits)
EC 5.4.99.2 (Methylmalonyl-CoA Mutase)
F0R1QK73KB (cobamamide)

Date Created: 20030705 Date Completed: 20030926 Latest Revision: 20161124

20250114

10.1007/s00203-003-0570-3

12844209