Structure and expression of cDNA for D-amino acid oxidase active against cephalosporin C from Fusarium solani.

摘要: D-Amino acid oxidase (DAO) was extracted and purified from cultured mycelia of Fusarium solani M-0718 (FERM P-2688). The enzyme able to oxidatively deaminate cephalosporin C 7-beta-(5-carboxy-5-oxopentanamido)cephalosporanic acid. Ninety-eight amino residues the F. DAO were determined by sequence analysis 9 peptides derived Acromobacter protease I digests protein. Complementary DNAs encoding isolated cDNA library hybridization with synthetic oligonucleotide probes corresponding partial sequences. Analysis nucleotide sequences clones revealed a 1,186-nucleotide 5'-terminal untranslated region 41 nucleotides, an open reading frame 1,083 nucleotides that encoded 361 acids, 3'-terminal 62 nucleotides. had 25% homology porcine kidney [EC 1.4.3.3] 37% Trigonopsis variabilis DAO. constructed plasmid overproduced in Escherichia coli. recombinant almost same molecular activity as native against C.