Increased yield of enzymatic synthesis by chromatographic selection of different N-glycoforms of yeast invertase
Autori
Anđelković, UrošGudelj, Ivan
Klarić, Thomas
Hinneburg, Hannes
Vinković, Marijana
Wittine, Karlo
Dovezenski, Nebojša
Vikić-Topić, Dražen
Lauc, Gordan
Vujčić, Zoran
Josić, Đuro
Članak u časopisu (Recenzirana verzija)
,
Wiley-VCH GmbH
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
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 stabilityis 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 subfractionatedby anion-exchange chromatography on industrial monolithic supports to obtain different glycoforms (EINV1–EINV3). Separated glycoforms exhibited different stabilities in wateralcohol 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 impr...oved 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 inregard to particular reaction environment, and provide a platform for improvement of yield of industrial enzymatic synthesis by chromatographic selection of glycoforms on monolithic supports.
Ključne reči:
Enzyme stability / Glycoform separation / Monolithic supports / N-glycosylation / Organic solventIzvor:
Electrophoresis, 2020Izdavač:
- Wiley
Finansiranje / projekti:
- Proizvodnja, izolovanje i karakterizacija enzima i malih molekula i njihova primena u rastvornom i imobilizovanom obliku u biotehnologiji hrane, biogorivima i zaštititi životne sredine (RS-172048)
- Ministarstvo nauke, tehnološkog razvoja i inovacija Republike Srbije, institucionalno finansiranje - 200026 (Univerzitet u Beogradu, Institut za hemiju, tehnologiju i metalurgiju - IHTM) (RS-200026)
- HTP-GLYCOMET - Methods for high-throughput glycoproteomic analysis (EU-324400)
- Reinforcement of the Faculty of Chemistry, University of Belgrade, towards becoming a Center of Excellence in the region of WB for Molecular Biotechnology and Food research (EU-256716)
Napomena:
- This is the peer-reviewed version of the article: Anđelković, U., Gudelj, I., et. al., Electrophoresis, 2020, https://doi.org/10.1002/elps.202000092
- The published version: https://cer.ihtm.bg.ac.rs/handle/123456789/3732
Povezane informacije:
- Druga verzija
https://doi.org/10.1002/elps.202000092 - Druga verzija
https://cer.ihtm.bg.ac.rs/handle/123456789/3732
DOI: 10.1002/elps.202000092
ISSN: 0173-0835; 1522-2683
PubMed: 33026663
WoS: 000580477100001
Scopus: 2-s2.0-85092915655
Institucija/grupa
IHTMTY - JOUR AU - Anđelković, Uroš AU - Gudelj, Ivan AU - Klarić, Thomas AU - Hinneburg, Hannes AU - Vinković, Marijana AU - Wittine, Karlo AU - Dovezenski, Nebojša AU - Vikić-Topić, Dražen AU - Lauc, Gordan AU - Vujčić, Zoran AU - Josić, Đuro PY - 2020 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/3989 AB - 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 stabilityis 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 subfractionatedby anion-exchange chromatography on industrial monolithic supports to obtain different glycoforms (EINV1–EINV3). Separated glycoforms exhibited different stabilities in wateralcohol 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 inregard to particular reaction environment, and provide a platform for improvement of yield of industrial enzymatic synthesis by chromatographic selection of glycoforms on monolithic supports. PB - Wiley T2 - Electrophoresis T1 - Increased yield of enzymatic synthesis by chromatographic selection of different N-glycoforms of yeast invertase DO - 10.1002/elps.202000092 ER -
@article{ author = "Anđelković, Uroš and Gudelj, Ivan and Klarić, Thomas and Hinneburg, Hannes and Vinković, Marijana and Wittine, Karlo and Dovezenski, Nebojša and Vikić-Topić, Dražen and Lauc, Gordan and Vujčić, Zoran and Josić, Đuro", year = "2020", 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 stabilityis 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 subfractionatedby anion-exchange chromatography on industrial monolithic supports to obtain different glycoforms (EINV1–EINV3). Separated glycoforms exhibited different stabilities in wateralcohol 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 inregard to particular reaction environment, and provide a platform for improvement of yield of industrial enzymatic synthesis by chromatographic selection of glycoforms on monolithic supports.", publisher = "Wiley", journal = "Electrophoresis", title = "Increased yield of enzymatic synthesis by chromatographic selection of different N-glycoforms of yeast invertase", doi = "10.1002/elps.202000092" }
Anđelković, U., Gudelj, I., Klarić, T., Hinneburg, H., Vinković, M., Wittine, K., Dovezenski, N., Vikić-Topić, D., Lauc, G., Vujčić, Z.,& Josić, Đ.. (2020). Increased yield of enzymatic synthesis by chromatographic selection of different N-glycoforms of yeast invertase. in Electrophoresis Wiley.. https://doi.org/10.1002/elps.202000092
Anđelković U, Gudelj I, Klarić T, Hinneburg H, Vinković M, Wittine K, Dovezenski N, Vikić-Topić D, Lauc G, Vujčić Z, Josić Đ. Increased yield of enzymatic synthesis by chromatographic selection of different N-glycoforms of yeast invertase. in Electrophoresis. 2020;. doi:10.1002/elps.202000092 .
Anđelković, Uroš, Gudelj, Ivan, Klarić, Thomas, Hinneburg, Hannes, Vinković, Marijana, Wittine, Karlo, Dovezenski, Nebojša, Vikić-Topić, Dražen, Lauc, Gordan, Vujčić, Zoran, Josić, Đuro, "Increased yield of enzymatic synthesis by chromatographic selection of different N-glycoforms of yeast invertase" in Electrophoresis (2020), https://doi.org/10.1002/elps.202000092 . .