Enzymatic process for the preparation of cephalosporanic 7$b(g)-(4-carboxybutanamide) acid by means of the modified enzyme d-aminoacid oxidase of trigonopsis variabilis produced in escherichia coli

Abstract

Enzymatic process for the preparation of cephalosporanic 7&bgr;-(4-carboxybutanamide) acid by using the modified enzyme D-aminoacid oxidase of produced in The process for the expression of the enzyme comprises: (I) isolating the DNA corresponding to gene which codes for the enzyme D-aminoacid oxidase; (II) removing the intron which is contained in said gene; (III) inserting the DNA fragment obtained into the plasmide which is capable of replication in (IV) fusing at the extremity 5′ of the structural region of the gene a synthetic assembler which contains a nucleotide sequence which codes for six histidines; (V) transforming a strain of with the resulting recombinant plasmide; (VI) cultivating the transformed cells of and (VII) recovering the enzyme D-aminoacid oxidase of the former cultivation operation through affinity chromatography.