Guisan, J.M. and Sabuquillo, P. and Fernandez-Lafuente, R. and Fernandez-Lorente, G. and Mateo, C. and Halling, P.J. and Kennedy, D. and Miyata, E. and Re, D. (2001) Preparation of new lipases derivatives with high activity- stability in anhydrous media: adsorption on hydrophobic supports plus hydrophilization with polyethylenimine. Journal of Molecular Catalysis B: Enzymatic, 11 (4-6). pp. 817-824. ISSN 1381-1177Full text not available in this repository. (Request a copy from the Strathclyde author)
A novel method to prepare immobilized lipases derivatives is hereby proposed. Lipases are firstly adsorbed on supports having large internal surfaces covered by hydrophobic groups (e.g. polyacrylic resins covered by C18 moieties). Then, immobilized lipases are incubated in the presence of polyethyleneimine (PEI) at a pH value over the isoelectric point of the enzyme in order to cover the lipase surface with this polymer. In this way, we try to minimize all possible direct interactions between immobilized lipase and organic solvents when using these derivatives in anhydrous media. Lipases from Rhizomucor miehie (RML) and Candida rugosa (CRL) were immobilized according to the proposed protocol. These derivatives were very active and very stable when catalyzing esterifications and transesterifications in anhydrous media. For example, RML derivatives exhibited a very high synthetic activity (more than 1000 Units/g immobilized biocatalyst) even when catalyzing the esterification of lauric acid with octanol at water activity values very close to zero. On the contrary, covalently immobilized derivatives exhibited a much lower synthetic activity under similar conditions (less than 10 Units/g of immobilized biocatalyst), Moreover, these new RML derivatives preserve 100% activity after incubation for 3 days in anhydrous butanone in the presence of molecular sieves. Under the same conditions, commercial immobilized RML lost more than 90% of activity in less than 10 min.
|Keywords:||lipases, interfacial adsorption, enzymes in organic medium, artificial enzyme microenvironment, protein surface hydrophilization, sepabeads Selective adsorption, organic-solvents, immobilization, enantioselectivity, Chemistry, Biochemistry, Bioengineering, Catalysis, Process Chemistry and Technology|
|Subjects:||Science > Chemistry|
|Department:||Faculty of Science > Pure and Applied Chemistry|
|Depositing user:||Users 16 not found.|
|Date Deposited:||14 Mar 2006|
|Last modified:||13 May 2016 02:08|