Naslov
Increased yield of enzymatic synthesis by chromatographic selection of differentN‐ glycoforms of yeast invertase

Autori
Andjelković, Uroš ; Gudelj, Ivan ; Klarić, Thomas ; Hinneburg, Hannes ; Vinković, Marijana ; Wittine, Karlo ; Dovezenski, Nebojša ; Vikić‐ Topić, Dražen ; Lauc, Gordan ; Vujčić, Zoran ; Josić, Djuro

Izvornik
Electrophoresis (0173-0835) 13 (2021); 33026663, 28

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
enzyme stability ; glycoform separation ; monolithic supports ; N-glycosylation ; organic solvent

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Biotehnologija

POVEZANOST RADA