ausblenden:
Schlagwörter:
Acyl Coenzyme A/*chemistry
Cloning, Molecular
Cobamides/*chemistry
Escherichia coli/genetics
Intramolecular Transferases/*chemistry/physiology
Isomerases/*chemistry
Magnetic Resonance Spectroscopy
Models, Biological
Models, Chemical
Phenotype
Phylogeny
Recombinant Proteins/chemistry
Rhodobacter sphaeroides/*metabolism
Software
Substrate Specificity
Time Factors
Zusammenfassung:
Coenzyme B(12)-dependent mutases are radical enzymes that catalyze reversible carbon skeleton rearrangement reactions. Here we describe Rhodobacter sphaeroides ethylmalonyl-CoA mutase (Ecm), a novel member of the family of coenzyme B(12)-dependent acyl-CoA mutases, that operates in the recently discovered ethylmalonyl-CoA pathway for acetate assimilation. Ecm is involved in the central reaction sequence of this novel pathway and catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with a second enzyme that was further identified as promiscuous ethylmalonyl-CoA/methylmalonyl-CoA epimerase. In contrast to the epimerase, Ecm is highly specific for its substrate, ethylmalonyl-CoA, and accepts methylmalonyl-CoA only at 0.2% relative activity. Sequence analysis revealed that Ecm is distinct from (2R)-methylmalonyl-CoA mutase as well as isobutyryl-CoA mutase and defines a new subfamily of coenzyme B(12)-dependent acyl-CoA mutases. In combination with molecular modeling, two signature sequences were identified that presumably contribute to the substrate specificity of these enzymes.
