MUTATED CEPHALOSPORIN HYDROXYLASE AND ITS APPLICATION IN DEACETYLCEPHALOSPORANIC ACID SYNTHESIS

摘要

The present invention relates to a mutant hydroxylase with increased activity and greater substrate specificity towards phenylacetyl-7-ADCA derivatives for the production of phenylacetyl-7-HACA derivatives, which carries one or more amino acid modification at residue positions when compared with the wild type hydroxylase from the following group of residues, Proline at position 7, Alanine at position 40, Threonine 51, Methionine at position 53, Glutamic acid at position 82, Arginine at position 91, Threonine at position 96, Glycine at position 108, Isoleucine at position 149, Valine at position 171, Alanine at position 177, Arginine at position 182, Methioinine at position 184, Isoleucine at position 193, Phenylalanine at position 195, Glutamine at position 209, Alanine at position 210, Valine at position 226, Methionine at position 233, Leucine at position 236, Alanine at position 237, Alanine at position 241, Valine at position 249, Arginine at position 250, Serine at position 251, Glycine at position 255, Glutamic Acid at position 258, Serine at position 260, Phenylalanine at position 267, Alanine at position 280, Valine at position 307 and Asparagine at position 313.. The invention further provides a process for the preparation of deacetyl cephalosporanic acid from the corresponding deacetoxy cephalosporanic acid using an enzyme of the present invention. The invention also provides the method for producing and processing of such enzymes.