Journal
ENZYME AND MICROBIAL TECHNOLOGY
Volume 60, Issue -, Pages 47-55Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.enzmictec.2014.03.013
Keywords
Immobilization; Epoxy resin; Polyester resin; Thermomyces lanuginosus lipase; Ashbya gossypii threonine aldolase; Cross linked enzyme aggregates
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Funding
- Brandenburg ministry for science and culture
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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 34 kDa, while A. gossypii threonine aldolase expressed in Escherichia coli is a pyridoxa1-5'-phosphate binding homotetramer with a mass of 180 kDa. 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. (C) 2014 Elsevier Inc. All rights reserved.
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