Enantioselectivity of recombinant Rhizomucor miehei lipase in the ring opening of oxazolin-5(4H)-ones.
Enantioselectivity of enzyme catalysis is often rationalized via active site models. These models are constructed on the basis of comparing the enantiomeric excess of product observed in a series of reactions which are conducted with a range of homologous substrates, typically carrying various side chain substitutions. Surprisingly the practical application of these simple but informative 'pocket size' models has been rarely tested in genetic engineering experiments. In this paper we report the construction, purification and enantioselectivity of two recombinant Rhizomucor miehei lipases which were designed to check the validity of such a model in reactions of ring opening of oxazolin-5(4H)-ones.
Turner
N A
NA
Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA, and Schools of Chemistry and Biological Sciences, Exeter University, Exeter EX4 4QD, UK.
Gaskin
D J
DJ
Yagnik
A T
AT
Littlechild
J A
JA
Vulfson
E N
EN
D020134:Catalytic Domain; D003001:Cloning, Molecular; D003198:Computer Simulation; D005656:Fungal Proteins; D006860:Hydrogen Bonding; D007700:Kinetics; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010080:Oxazoles / Q000737:chemistry / Q000378:metabolism*; D010081:Oxazolone; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020103:Rhizomucor / Q000201:enzymology*; D013237:Stereoisomerism
D005656:Fungal Proteins; D010080:Oxazoles; D011994:Recombinant Proteins; D010081:Oxazolone; D008049:Lipase