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

  • Miaomiao Wang
  • Wenjing Qi
  • Hongping Xu
  • Huimin YuEmail author
  • Shuliang Zhang
  • Zhongyao Shen
Biocatalysis - Short Communication
  • 19 Downloads

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.

Keywords

Affinity-binding Immobilization Specific affinity peptide Mesoporous silica d-Amino acid oxidase Phage display 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of P. R. China (No. 21176143 and 21706145).

Supplementary material

10295_2019_2210_MOESM1_ESM.doc (336 kb)
Supplementary material 1 (DOC 336 kb)

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2019

Authors and Affiliations

  • Miaomiao Wang
    • 1
    • 2
  • Wenjing Qi
    • 1
    • 2
  • Hongping Xu
    • 1
    • 2
  • Huimin Yu
    • 1
    • 2
    • 3
    Email author
  • Shuliang Zhang
    • 1
    • 2
  • Zhongyao Shen
    • 1
    • 2
  1. 1.Department of Chemical EngineeringTsinghua UniversityBeijingPeople’s Republic of China
  2. 2.Key Laboratory of Industrial BiocatalysisTsinghua University, The Ministry of EducationBeijingPeople’s Republic of China
  3. 3.Center for Synthetic and Systems BiologyTsinghua UniversityBeijingPeople’s Republic of China

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