title: An imprinted cross-linked enzyme aggregate (iCLEA) of sucrose phosphorylase: combining improved stability with altered specificity.
abstract: The industrial use of sucrose phosphorylase (SP), an interesting biocatalyst for the selective transfer of α-glucosyl residues to various acceptor molecules, has been hampered by a lack of long-term stability and low activity towards alternative substrates. We have recently shown that the stability of the SP from Bifidobacterium adolescentis can be significantly improved by the formation of a cross-linked enzyme aggregate (CLEA). In this work, it is shown that the transglucosylation activity of such a CLEA can also be improved by molecular imprinting with a suitable substrate. To obtain proof of concept, SP was imprinted with α-glucosyl glycerol and subsequently cross-linked with glutaraldehyde. As a consequence, the enzyme's specific activity towards glycerol as acceptor substrate was increased two-fold while simultaneously providing an exceptional stability at 60 °C. This procedure can be performed in an aqueous environment and gives rise to a new enzyme formulation called iCLEA.
authors.affiliation: Centre of Expertise for Industrial Biotechnology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Biosciences Engineering, Ghent University, Coupure Links 653, Ghent B-9000, Belgium.
authors.forename: Karel
authors.initials: K
authors.lastname: De Winter
authors.affiliation: Centre of Expertise for Industrial Biotechnology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Biosciences Engineering, Ghent University, Coupure Links 653, Ghent B-9000, Belgium.
authors.forename: Wim
authors.initials: W
authors.lastname: Soetaert
authors.affiliation: Centre of Expertise for Industrial Biotechnology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Biosciences Engineering, Ghent University, Coupure Links 653, Ghent B-9000, Belgium.
authors.forename: Tom
authors.initials: T
authors.lastname: Desmet
mesh_terms: D001644:Bifidobacterium / Q000201:enzymology*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D005964:Glucosyltransferases / Q000378:metabolism*; D005976:Glutaral / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry; D054802:Molecular Imprinting / Q000379:methods*
chemical_list: D004800:Enzymes, Immobilized; D005964:Glucosyltransferases; C025901:sucrose phosphorylase; D005990:Glycerol; D005976:Glutaral
keywords: CLEA; glucosyl glycerol; immobilization; imprinting; sucrose phosphorylase