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Affinity-binding immobilization of D-amino acid oxidase on mesoporous silica by a silica-specific peptide

Wang, Miaomiao, Qi, Wenjing, Xu, Hongping, Yu, Huimin, Zhang, Shuliang, Shen, Zhongyao
Journal of industrial microbiology & biotechnology 2019 v.46 no.11 pp. 1461-1467
D-amino-acid oxidase, Rhodosporidium toruloides, absorption, amino acids, cephalosporins, immobilized enzymes, industrial applications, peptide libraries, porous media, silica, thermal stability
Enzyme immobilization is widely used for large-scale industrial applications. However, the weak absorption through physical methods limits the recovery ability. Here, affinity-binding immobilization of enzymes was explored using a silica-specific affinity peptide (SAP) as a fusion tag to intensify the binding force between the enzyme and mesoporous silica (MPS) carrier. D-amino acid oxidase (DAAO) of Rhodosporidium toruloides was used as a model enzyme. The optimal screened SAP (LPHWHPHSHLQP) was selected from a M13 phage display peptide library and fused to the C-terminal of DAAO to obtain fused DAAOs with one, two and three SAP tags, respectively. The activity of DAAO–SAP–MPS was superior comparing with DAAO–2SAP–MPS and DAAO–3SAP–MPS; meanwhile DAAO–SAP–MPS shows 36% higher activity than that of DAAO–MPS. Fusion with one SAP improved the thermal stability with a 10% activity increase for immobilized DAAO–SAP–MPS compared to that of DAAO–MPS at 50 °C for 3 h. Moreover, the activity recovery of immobilized DAAO–SAP–MPS was 25% higher in operation stability assessment after six-batch conversions of cephalosporin to glutaryl-7-amino cephalosporanic acid than that of DAAO–MPS.