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

• DOI: 10.1007/s10295-019-02210-5
• Publication Date: 2019-07-09T12:53:31Z
• Authors: Miaomiao Wang; Wenjing Qi; Hongping Xu; Huimin Yu; Shuliang Zhang; Zhongyao Shen
• Keywords: D-Amino-Acid Oxidase; 0301 basic medicine; 03 medical and health sciences; Amino Acids; Peptides; Silicon Dioxide; Cephalosporins

Abstract 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.