title: Adenosylcobalamin-dependent methylmalonyl-CoA mutase from Propionibacterium shermanii. Active holoenzyme produced from Escherichia coli.
abstract: The linked structural genes coding for both subunits of adenosylcobalamin-dependent methylmalonyl-CoA mutase from the Gram-positive bacterium Propionibacterium shermanii have been altered by site-directed mutagenesis and placed under the control of an inducible phage-T7-specific plasmid promoter in Escherichia coli. Conditions have been found under which both alpha- and beta-subunits are produced in soluble form, in near 1:1 ratio, and assemble to form apo-mutase totalling about 5% of the total cellular protein. Methylmalonyl-CoA mutase purified from these cells could be readily converted into the holoenzyme by addition of adenosylcobalamin. The active holoenzyme apparently crystallizes in the same space group as an inactive corrinoid-containing form of the enzyme obtained previously.
authors.affiliation: Department of Biochemistry, University of Cambridge, U.K.
authors.forename: N
authors.initials: N
authors.lastname: McKie
authors.forename: N H
authors.initials: NH
authors.lastname: Keep
authors.forename: M L
authors.initials: ML
authors.lastname: Patchett
authors.forename: P F
authors.initials: PF
authors.lastname: Leadlay
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D003038:Cobamides / Q000494:pharmacology*; D003460:Crystallization; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D007535:Isomerases / Q000378:metabolism*; D008765:Methylmalonyl-CoA Mutase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D011424:Propionibacterium / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D003038:Cobamides; D011994:Recombinant Proteins; D007535:Isomerases; D008765:Methylmalonyl-CoA Mutase; C000913:cobamamide