Immobilization of glutaryl-7-aminocephalosporanic acid acylase on silica gel and enhancement of its stability

Seung Won Park, Jeewon Lee, Suk In Hong, Seung Wook Kim

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Glutaryl-7-aminocephalosporanic acid (GL-7-ACA) acylase is an enzyme that converts GL-7-ACA to 7-aminocephalosporanic acid, a starting material for semisynthetic cephalosporin antibiotics. In this study, optimal conditions for the immobilization of GL-7-ACA acylase were determined by experimental observations and statistical methods. The optimal conditions were as follows: 1.1 M phosphate buffer (pH 8.3) as buffer solution, immobilization temperature of 20°C, and immobilization time of 120 min. Unreacted aldehyde groups were quenched by reaction with a low-molecular-weight material such as L-lysine, glycine, and ethanolamine after immobilization in order to enhance the activity of immobilized GL-7-ACA acylase. The activities of immobilized GL-7-ACA acylase obtained by using the low-molecular-weight materials were higher than those obtained by immobilized GL-7-ACA acylase not treated with low-molecular-weight materials. In particular, the highest activity of immobilized GL-7-ACA acylase was obtained using 0.4% (v/v) ethanolamine. We also investigated the effect of sodium cyanoborohydride in order to increase the stability of the linkage between the enzyme and the support. The effect on operational stability was obvious: the activity of immobilized GL-7-ACA acylase treated with 4% (w/w) sodium cyanoborohydride remained almost 100% after 20 times of reuse.

Original languageEnglish
Pages (from-to)185-198
Number of pages14
JournalApplied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology
Volume104
Issue number3
DOIs
Publication statusPublished - 2003 Mar 1

Keywords

  • Ethanolamine
  • Glutaryl-7-aminocephalosporanic acid acylase
  • Immobilization
  • Sodium cyanoborohydride
  • Statistical method

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Bioengineering

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