Volume 42, Issue 24 p. 2626-2636
Research Article

Increased yield of enzymatic synthesis by chromatographic selection of different N-glycoforms of yeast invertase

Uroš Andjelković

Corresponding Author

Uroš Andjelković

University of Belgrade–Institute of Chemistry, Technology and Metallurgy–National Institute of the Republic of Serbia, Belgrade, Serbia

Department of Biotechnology, University of Rijeka, Rijeka, Croatia

Correspondence: Dr. Uroš Andjelković, Department of Chemistry, Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Studentski trg 12–16, 11000 Belgrade, Serbia.

E-mail: [email protected]

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Ivan Gudelj

Ivan Gudelj

Genos Glycoscience Research Laboratory, Zagreb, Croatia

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Thomas Klarić

Thomas Klarić

Genos Glycoscience Research Laboratory, Zagreb, Croatia

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Hannes Hinneburg

Hannes Hinneburg

Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany

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Marijana Vinković

Marijana Vinković

NMR Centre, Ruđer Bošković Institute, Zagreb, Croatia

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Karlo Wittine

Karlo Wittine

Department of Biotechnology, University of Rijeka, Rijeka, Croatia

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Nebojša Dovezenski

Nebojša Dovezenski

Institute for Medical Research, University of Belgrade, Belgrade, Serbia

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Dražen Vikić-Topić

Dražen Vikić-Topić

NMR Centre, Ruđer Bošković Institute, Zagreb, Croatia

Department of Natural and Health Sciences, Juraj Dobrila University of Pula, Pula, Croatia

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Gordan Lauc

Gordan Lauc

Genos Glycoscience Research Laboratory, Zagreb, Croatia

Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia

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Zoran Vujčić

Zoran Vujčić

Faculty of Chemistry, University of Belgrade, Belgrade, Serbia

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Djuro Josić

Djuro Josić

Department of Biotechnology, University of Rijeka, Rijeka, Croatia

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First published: 07 October 2020
Citations: 2
Color online: See article online to view Figs. 1–6 in color.

Abstract

Invertases are glycosidases applied for synthesis of alkyl glycosides that are important and effective surfactants. Stability of invertases in the environment with increased content of organic solvent is crucial for increase of productivity of glycosidases. Their stability is significantly influenced by N-glycosylation. However, yeast N-glycosylation pathways may synthesize plethora of N-glycan structures. A total natural crude mixture of invertase glycoforms (EINV) extracted from Saccharomyces cerevisiae was subfractionated by anion-exchange chromatography on industrial monolithic supports to obtain different glycoforms (EINV1–EINV3). Separated glycoforms exhibited different stabilities in water-alcohol solutions that are in direct correlation with the amount of phosphate bound to N-glycans. Observed differences in stability of different invertase glycoforms were used to improve productivity of methyl β-d-fructofuranoside (MF) synthesis. The efficiency and yield of MF synthesis were improved more than 50% when the most stabile glycoform bearing the lowest amount of phosphorylated N-glycans is selected and utilized. These data underline the importance of analysis of glycan structures attached to glycoproteins, demonstrate different impact of N-glycans on the surface charge and enzyme stability in regard to particular reaction environment, and provide a platform for improvement of yield of industrial enzymatic synthesis by chromatographic selection of glycoforms on monolithic supports.

The authors have declared no conflict of interest.