title: The purification and properties of cyclohexanone oxygenase from Nocardia globerula CL1 and Acinetobacter NCIB 9871.
abstract: 1. Cyclohexanone oxygenases from Norcardia globerula CL1 and Acinetobacter NCIB 9871 have been purified 12-fold and 35-fold respectively and each gives a single symmetrical sedimentation peak in the ultracentrifuge and a single protein band on 2.25 nm average pore radius polyacrylamide gels. 2. The enzyme from N. globerula has a molecular weight of 53000 while that from Acinetobacter has a molecular weight of about 59000. Each is a single polypeptide chain with one mole of bound FAD per mole of protein that does not dissociate during purification. Acidification of the Acinetobacter enzyme in the presence of (NH4)2SO4 releases the bound FAD and yields native apoenzyme from which the active holoenzyme can be reconstituted. The apparent dissociation constant for the FAD is 40 nM.
mesh_terms: D000150:Acinetobacter / Q000201:enzymology*; D000596:Amino Acids / Q000032:analysis; D003460:Crystallization; D005486:Flavin Mononucleotide / Q000494:pharmacology; D005182:Flavin-Adenine Dinucleotide / Q000494:pharmacology; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000302:isolation & purification / Q000378:metabolism; D008970:Molecular Weight; D009615:Nocardia / Q000201:enzymology*; D010101:Oxygen Consumption; D011485:Protein Binding; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013055:Spectrophotometry, Infrared; D013056:Spectrophotometry, Ultraviolet; D013438:Sulfhydryl Compounds / Q000032:analysis
chemical_list: D000596:Amino Acids; D013438:Sulfhydryl Compounds; D005182:Flavin-Adenine Dinucleotide; D005486:Flavin Mononucleotide; D006899:Mixed Function Oxygenases