Purification and properties of an amidase from Rhodococcus erythropolis MP50 which enantioselectively hydrolyzes 2-arylpropionamides.
An enantioselective amidase from Rhodococcus erythropolis MP50 was purified to homogeneity. The enzyme has a molecular weight of about 480,000 and is composed of identical subunits with molecular weights of about 61,000. The NH2-terminal amino acid sequence was significantly different from previously published sequences of bacterial amidases. The purified amidase hydrolyzed a wide range of aliphatic and aromatic amides, The highest enzyme activities were found with amides carrying hydrophobic residues, such as pentyl or naphthoyl. The purified enzyme converted racemic 2-phenylpropionamide, naproxen amide [2-(6-methoxy-2-naphthyl) propionamide], and ketoprofen amide [2-(3'-benzoylphenyl)propionamide] to the corresponding S-acids with an enantiomeric excess of >99% and an almost 50% conversion of the racemic amides. The enzyme also hydrolyzed different alpha-amino amides but without significant enantioselectivity.
Hirrlinger
B
B
Institüt fur Mikrobiologie, Universität Stuttgart, Germany.
Stolz
A
A
Knackmuss
H J
HJ
D000581:Amidohydrolases / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007660:Ketoprofen / Q000378:metabolism*; D008670:Metals; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009288:Naproxen / Q000378:metabolism*; D012240:Rhodococcus / Q000201:enzymology*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013237:Stereoisomerism; D013379:Substrate Specificity
D008670:Metals; D009288:Naproxen; D007660:Ketoprofen; D000581:Amidohydrolases