Advantages of using non-isothermal bioreactors for the enzymatic synthesis of antibiotics: The penicillin G acylase as enzyme model

P. Travascio, E. Zito, A. De Maio, C. G P H Schroën, D. Durante, P. De Luca, U. Bencivenga, D. G. Mita

Research output: Contribution to journalArticlepeer-review

Abstract

A new hydrophobic and catalytic membrane was prepared by immobilizing Penicillin G acylase (PGA, EC.3.5.1.11) from E. coli on a nylon membrane, chemically grafted with butylmethacrylate (BMA). Hexamethylenediamine (HMDA) and glutaraldehyde (Glu) were used as a spacer and coupling agent, respectively. PGA was used for the enzymatic synthesis of cephalexin, using D(-)-phenylglycine methyl ester (PGME) and 7-amino-3-deacetoxycephalosporanic acid (7-ADCA) as substrates. Several factors affecting this reaction, such as pH, temperature, and concentrations of substrates were investigated. The results indicated good enzyme-binding efficiency of the pre-treated membrane, and an increased stability of the immobilized PGA towards pH and temperature. Calculation of the activation energies showed that cephalexin production by the immobilized biocatalyst was limited by diffusion, resulting in a decrease of enzyme activity and substrate affinity. Temperature gradients were employed as a way to reduce the effects of diffusion limitation. Cephalexin was found to linearly increase with the applied temperature gradient. A temperature difference of about 3°C across the catalytic membrane resulted into a cephalexin synthesis increase of 100% with a 50% reduction of the production times. The advantage of using non-isothermal bioreactors in biotechnological processes, including pharmaceutical applications, is also discussed.

Original languageEnglish
Pages (from-to)334-346
Number of pages13
JournalBiotechnology and Bioengineering
Volume79
Issue number3
DOIs
Publication statusPublished - Aug 5 2002

Keywords

  • Antibiotics
  • Cephalexin
  • Enzymatic synthesis
  • Grafted nylon membranes
  • Immobilized enzymes
  • Non-isothermal bioreactor
  • Penicillin G acylase

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

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