Dry entrapment of enzymes by epoxy or polyester resins hardened on different solid supports

• Published in: Enzyme and microbial technology Vol. 60; pp. 47 - 55
• Main Authors: Barig, Susann, Funke, Andreas, Merseburg, Andrea, Schnitzlein, Klaus, Stahmann, K.-Peter
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 10.06.2014
• Subjects: Adsorption; Ascomycota - enzymology; Ascomycota - genetics; Ashbya gossypii; Ashbya gossypii threonine aldolase; Aspergillus oryzae; Biocatalysis; Cross linked enzyme aggregates; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - genetics; Enzymes, Immobilized - metabolism; Epoxy resin; Epoxy Resins; Escherichia coli; Freeze Drying; Fungal Proteins - chemistry; Fungal Proteins - genetics; Fungal Proteins - metabolism; Glycine Hydroxymethyltransferase - chemistry; Glycine Hydroxymethyltransferase - genetics; Glycine Hydroxymethyltransferase - metabolism; Immobilization; Lipase - chemistry; Lipase - genetics; Lipase - metabolism; Molecular Weight; Polyester resin; Polyesters; Protein Structure, Quaternary; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Resins, Synthetic; Saccharomycetales - enzymology; Saccharomycetales - genetics; Substrate Specificity; Thermomyces lanuginosus; Thermomyces lanuginosus lipase; Immobilization; Epoxy resin; Ashbya gossypii threonine aldolase; Cross linked enzyme aggregates; Polyester resin; Thermomyces lanuginosus lipase
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/j.enzmictec.2014.03.013

•A dry entrapment immobilization method for enzymes was evolved.•Therefore water-free enzyme preparations were embedded in two different polymeric glues.•The method was proofed to be applicable on two very different enzymes, a lipase and a threonine aldolase. Embedding of enzymes was performed with epoxy or polyester resin by mixing in a dried enzyme preparation before polymerization was started. This fast and low-cost immobilization method produced enzymatically active layers on different solid supports. As model enzymes the well-characterized Thermomyces lanuginosus lipase and a new threonine aldolase from Ashbya gossypii were used. It was shown that T. lanuginosus lipase recombinantly expressed in Aspergillus oryzae is a monomeric enzyme with a molecular mass of 34kDa, while A. gossypii threonine aldolase expressed in Escherichia coli is a pyridoxal-5′-phosphate binding homotetramer with a mass of 180kDa. The enzymes were used freeze dried, in four different preparations: freely diffusing, adsorbed on octyl sepharose, as well as cross-linked enzyme aggregates or as suspensions in organic solvent. They were mixed with standard two-component resins and prepared as layers on solid supports made of different materials e.g. metal, glass, polyester. Polymerization led to encapsulated enzyme preparations showing activities comparable to literature values.