Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports

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Mladen Mihailović
Jovana Trbojević-Ivić
Katarina Banjanac
Nenad Milosavić
Dušan Veličković
Milica Carević
Dejan Bezbradica

Abstract

In this study, two commercial supports (Eupergit® C and Purolite® A109) were chemically modified in order to introduce thiosulfonate groups, which could subsequently exclusively react with the cysteine residues on the surface of enzymes. Thereafter, the maltase from Saccharomyces cerevisiae was immobilized onto the obtained thiosulfonate-activated supports, resulting in high expressed enzymatic activities (around 50 %), while on the other hand, immobilization on unmodified supports yielded expressed activities less than 5 %. Moreover, protein loadings up to 12.3 mg g-1 and immobilized activities up to 3580 IU g-1 were achieved by employment of these thiosulfonate supports. Desorption experiments, performed on samples taken during immobilization, proved that immobilization on the thiosulfonate supports was the first step of fast adsorption onto the supports and the formation of covalent bonds between the thiosulfonate groups and the thiol groups of cysteine represented a second slower step. More importantly, although enzyme coupling occurred via cov­alent bond formation, the performed immobilization proved to be reversible, since it was shown that 95 % of the immobilized activity could be detached from the support after treatment with a thiol reagent (β-mercaptoethanol). Thus, the support could be reused after enzyme inactivation. 

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How to Cite
[1]
M. Mihailović, “Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports”, J. Serb. Chem. Soc., vol. 81, no. 12, pp. 1371–1382, Dec. 2016.
Section
Biochemistry & Biotechnology
Author Biographies

Mladen Mihailović, Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade

Department of Biochemical Engineering and Biotechnology

Jovana Trbojević-Ivić, Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12, 11000 Belgrade

Department of Biochemistry

Katarina Banjanac, Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade

Department of Biochemical Engineering and Biotechnology

Nenad Milosavić, Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12, 11000 Belgrade

Department of Biochemistry

Dušan Veličković, Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12, 11000 Belgrade

Department of Biochemistry

Dejan Bezbradica, Department of Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, 11000 Belgrade

Department of Biochemical Engineering and Biotechnology

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