Optimization of immobilization conditions of Thermomyces lanuginosus lipase on styrene-divinylbenzene copolymer using response surface methodology

AYBASTIER Ö., Demir C.

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, vol.63, pp.170-178, 2010 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 63
  • Publication Date: 2010
  • Doi Number: 10.1016/j.molcatb.2010.01.013
  • Journal Name: JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.170-178
  • Keywords: Thermomyces lanuginosus, Immobilization, Enzyme activity, Styrene-divinylbenzene, Response surface methodology, BIODIESEL FUEL PRODUCTION, CANDIDA-RUGOSA LIPASE, COVALENT IMMOBILIZATION, MICROBIAL LIPASE, OIL, BIOCATALYST, SELECTIVITY, IMPROVEMENT, ACTIVATION, PARAMETERS
  • Bursa Uludag University Affiliated: Yes

Abstract

Microbial lipase from Thermomyces lanuginosus (formerly Humicola lanuginosa) was immobilized by covalent binding on a novel microporous styrene-divinylbenzene polyglutaraldehyde copolymer (STY-DVB-PGA). The response surface methodology (RSM) was used to optimize the conditions for the maximum activity and to understand the significance and interaction of the factors affecting the specific activity of immobilized lipase. The central composite design was employed to evaluate the effects of enzyme concentration (4-16%, v/v), pH (6.0-8.0), buffer concentration (20-100 mM) and immobilization time (8-40h) on the specific activity. The results indicated that enzyme concentration, pH and buffer concentration were the significant factors on the specific activity of immobilized lipase and quadratic polynomial equation was obtained for specific activity. The predicted specific activity was 8.78 mu mol p-NP/mg enzyme min under the optimal conditions and the subsequent verification experiment with the specific activity of 8.41 mu mol p-NP/mg enzyme min confirmed the validity of the predicted model. The lipase loading capacity was obtained as 5.71 mg/g support at the optimum conditions. Operational stability was determined with immobilized lipase and it indicated that a small enzyme deactivation (12%) occurred after being used repeatedly for 10 consecutive batches with each of 24 h. The effect of methanol and tert-butanol on the specific activity of immobilized lipase was investigated. The immobilized lipase was almost stable in tert-butanol (92%) whereas it lost most of its activity in methanol (80%) after 15 min incubation. (C) 2010 Elsevier B.V. All rights reserved.