Development of tailor-made glycidyl methacrylate-divinyl benzene copolymer for immobilization of D-amino acid oxidase from Aspergillus species strain 020 and its application in the bioconversion of cephalosporin C.

Abstract A tailor-made glycidyl methacrylate-divinyl benzene (GMA-DVB) copolymer PC-3 was evolved by studying the effect of synthesis variables on binding and expression of d -amino acid oxidase (DAAO) from Aspergillus species strain 020. Almost quantitative binding (100%) and a high yield of immobilization per unit of enzyme loaded was achieved. Optimum pH, optimum temperature and K m for soluble and IME-DAAO preparations were 8.0–8.5 and 8.0; 30 and 37°C; and 0.73 and 2.78 mM for ceph C, respectively. The developed immobilized DAAO was used for the production of glutaryl-7-aminocephalosporanic acid from cephalosporin C (ceph C). A high conversion of >95% was achieved by using 3% (w/v) solution of ceph C, 48 U of DAAO per g of ceph C, keeping dissolved oxygen level above 50%, maintaining the pH between 7.6 and 7.8 and temperature at 24°C. The immobilized DAAO was used for 60 cycles in a stirred tank reactor.