Biotechnology and Bioprocess Engineering

, Volume 15, Issue 3, pp 376–382 | Cite as

Post-immobilization of modified macromolecular reagents using assembled penicillin acylase for microenvironmental regulation of nanopores and enhancement of enzyme stability

  • Cheng Zhou
  • Shemin Zhu
  • Xiuming Wu
  • Bo Jiang
  • Tao Cen
  • Shubao Shen
Research Paper

Abstract

Penicillin acylase (PA) is known to regulate the microenvironment of nanospores. In this study, nanopores containing chemically-modified macromolecules co-assembled with immobilized PA were constructed. We also investigated the various types of functionalized mesocellular siliceous foams (MCFs) commonly used for the immobilization of PA by measuring the catalytic performance and stability of each PA preparation. Amino-MCF activated by p-benzoquinone was chosen as the optimum support for PA immobilization. Successful modification of macromolecules was verified by FT-IR and ultraviolet (UV) spectroscopy. The specific activity of PA co-assembled with dextran 10 k was 99.1 U/mg, which was 1.5-fold that of pristine immobilized PA, while the optimum pH was shifted to neutral. Compared to pristine immobilized and free PA, the optimum temperatures for the modified PA were 5 and 10°C higher, respectively. The residual activity of the ficoll derivative of PA after treatment at 50°C for 6 h was 70%, and this was later increased to 214.5% compared to that of pristine immobilized PA. The dextran 10 k derivative of PA exhibited 90.2% residual activity after 25 times of continuous use. The results show that chemically-modified macromolecules co-assembled with PA in amino-MCF provided a suitable microenvironment for enzyme stability.

Keywords

immobilized penicillin acylase covalent modification macromolecular crowding microenvironment modulation 

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Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg 2010

Authors and Affiliations

  • Cheng Zhou
    • 1
    • 2
  • Shemin Zhu
    • 1
    • 3
  • Xiuming Wu
    • 1
    • 2
  • Bo Jiang
    • 1
    • 2
  • Tao Cen
    • 1
    • 2
  • Shubao Shen
    • 1
    • 2
  1. 1.National Engineering Research Center for BiotechnologyNanjingChina
  2. 2.College of Biotechnology and Pharmaceutical EngineeringNanjing University of TechnologyNanjingChina
  3. 3.College of Material Science and EngineeringNanjing University of TechnologyNanjingChina

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