Co-aggregation of penicillin G acylase and polyionic polymers: An easy methodology to prepare enzyme biocatalysts stable in organic media

LORENA EVELYN WILSON SOTO, Andrés Illanes, Olga Abián, Benevides C.C. Pessela, Roberto Fernández-Lafuente, José M. Guisán

Research output: Contribution to journalArticlepeer-review

116 Scopus citations

Abstract

A novel type of biocatalyst that combines the good properties of cross-linked enzyme aggregates (CLEAs) and hydrophilic microenvironments has been developed. Dextran sulfate- and polyethyleneimine-coated CLEAs of penicillin acylase (CLEA-GDP) were prepared by adding the polymers of different sizes before the precipitation stage of the enzyme. This study presents the development and optimization of a protocol to produce such a biocatalyst using penicillin acylase as a model. Experiments show that CLEA-GDPs have a highly increased stability in organic media. The average half-life of the preparations was much higher than standard CLEA without a microenvironment (CLEA-G), (e.g., more than 25-fold) in the presence of dioxane. However, their thermal stability was not increased, which leads to the conclusion that the stability of CLEA-GDPs in organic media is due to the hydrophilic microenvironment that surrounds the protein enzyme more than to a conformational stiffening effect. This is further supported by solvation experiments that show a preferential hydration of CLEA when polymers are used to coat the enzyme. CLEA-GDPs are clearly better than other biocatalysts in terms of solvent stability.

Original languageEnglish
Pages (from-to)852-857
Number of pages6
JournalBiomacromolecules
Volume5
Issue number3
DOIs
StatePublished - May 2004
Externally publishedYes

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