ENZYMATIC PROCESS FOR THE PREPARATION OF CEPHALOSPORANIC 7$b(g)-(4-CARBOXYBUTANAMIDE) ACID BY MEANS OF THE MODIFIED ENZYME D-AMINOACID OXIDASE OF $i(TRIGONOPSIS VARIABILIS) PRODUCED IN $i(ESCHERICHIA COLI)

摘要

Enzymatic process for the preparation of cephalosporanic 7 beta -(4-carboxybutanamide) acid by using the modified enzyme D-aminoacid oxidase of Trigonopsis variabilis produced in Escherichia coli. The process for the expression of the enzyme comprises: (I) isolating the DNA corresponding to the gene which codes for the enzyme D-aminoacide 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 Escherichia coli (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 Escherichia coli with the resulting recombinant plasmide (VI) cultivating the transformed cells of Escherichia coli and (VII) recovering the enzyme D-aminoacid oxidase of the former cultivation operation through affinity chromatography.