Recombinat Production and Characterization of the SUMO-Tagged N-Glycosidase F Enzyme

Sercan Karav

doi:https://doi.org/10.29329/ijiasr.2024.1054.3

Original article | Open Access
International Journal of Innovative Approaches in Science Research 2024, Vol. 8(2) 70-78

Recombinat Production and Characterization of the SUMO-Tagged N-Glycosidase F Enzyme

Sercan Karav

pp. 70 - 78 | DOI: https://doi.org/10.29329/ijiasr.2024.1054.3

Published online: June 30, 2024 | Number of Views: 20 | Number of Download: 46

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Abstract

Glycans, a key component of cells, are abundant and diverse biopolymers found in covalent bonds of sucrose attached to proteins and lipids. They significantly contribute to mass and structural variation in biological systems. In order to characterize glycans, study their effects in various experiments, and comprehend their roles, it is necessary to isolate them from the proteins with which they are associated. Chemical methods and various enzymes separate glycans. The ease of application of chemical methods, despite their low cost, chemical methods ease of application affects the chemical structure of both glycans and the remaining part of the polypeptide during deglycosylation. Additionally, the high salt content of the separated glycans makes mass spectrometry analysis of these glycants difficult. For these reasons, the use of enzymes in glycan studies has increased in recent years. One of the most commonly used enzymes in this field, N-glycosidase F has a wide spectrum and the ability to successfully release various N-glycan structures from glycoprotein.

In this study, the PNGase F enzyme, secreted by Flavobacterium meningosepticum, was efficiently produced in a recombinant manner. The enzyme, which contains 314 amino acids, is the most effective method for removing N-glycan from glycoproteins.

Keywords: N-linked glycans, Glycoproteins, Glycosidases, Deglycosylation, In vivo cloning

How to Cite this Article

APA 6th edition
Karav, S. (2024). Recombinat Production and Characterization of the SUMO-Tagged N-Glycosidase F Enzyme . International Journal of Innovative Approaches in Science Research, 8(2), 70-78. doi: 10.29329/ijiasr.2024.1054.3

Harvard
Karav, S. (2024). Recombinat Production and Characterization of the SUMO-Tagged N-Glycosidase F Enzyme . International Journal of Innovative Approaches in Science Research, 8(2), pp. 70-78.

Chicago 16th edition
Karav, Sercan (2024). "Recombinat Production and Characterization of the SUMO-Tagged N-Glycosidase F Enzyme ". International Journal of Innovative Approaches in Science Research 8 (2):70-78. doi:10.29329/ijiasr.2024.1054.3.

References

Barsomian, G. D., Johnson, T. L., Borowski, M., Denman, J., Ollington, J. F., Hirani, S., McNeilly, D. S., & Rasmussen, J. R. (1990). Cloning and expression of peptide-N4-(N-acetyl-beta-D-glucosaminyl)asparagine amidase F in Escherichia coli. Journal of Biological Chemistry, 265(12), 6967–6972. https://doi.org/10.1016/S0021-9258(19)39245-2 [Google Scholar] [Crossref]
Bunyatratchata, A., Parc, A. L., De Moura Bell, J. M. L. N., Cohen, J. L., Duman, H., Arslan, A., Kaplan, M., Barile, D., & Karav, S. (2023). Release of bifidogenic N-glycans from native bovine colostrum proteins by an endo-β-N-acetylglucosaminidase. Enzyme and Microbial Technology, 162, 110138. https://doi.org/10.1016/j.enzmictec.2022.110138 [Google Scholar] [Crossref]
Duman, H., Kaplan, M., Arslan, A., Sahutoglu, A. S., Kayili, H. M., Frese, S. A., & Karav, S. (2021). Potential Applications of Endo-β-N-Acetylglucosaminidases From Bifidobacterium longum Subspecies infantis in Designing Value-Added, Next-Generation Infant Formulas. Frontiers in Nutrition, 8, 646275. https://doi.org/10.3389/fnut.2021.646275 [Google Scholar] [Crossref]
Freeze, H. H., & Varki, A. (1986). Endo-glycosidase F and peptide N-glycosidase F release the great majority of total cellular N-linked oligosaccharides: Use in demonstrating that sulfated N-linked oligosaccharides are frequently found in cultured cells. Biochemical and Biophysical Research Communications, 140(3), 967–973. https://doi.org/10.1016/0006-291X(86)90730-8 [Google Scholar] [Crossref]
Karav, S., Le Parc, A., Leite Nobrega De Moura Bell, J. M., Frese, S. A., Kirmiz, N., Block, D. E., Barile, D., & Mills, D. A. (2016). Oligosaccharides Released from Milk Glycoproteins Are Selective Growth Substrates for Infant-Associated Bifidobacteria. Applied and Environmental Microbiology, 82(12), 3622–3630. https://doi.org/10.1128/AEM.00547-16 [Google Scholar] [Crossref]
Karav, S., Parc, A. L., Moura Bell, J. M. L. N. D., Rouquié, C., Mills, D. A., Barile, D., & Block, D. E. (2015). Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. Enzyme and Microbial Technology, 77, 46–53. https://doi.org/10.1016/j.enzmictec.2015.05.007 [Google Scholar] [Crossref]
Krištić, J., Vučković, F., Menni, C., Klarić, L., Keser, T., Beceheli, I., Pučić-Baković, M., Novokmet, M., Mangino, M., Thaqi, K., Rudan, P., Novokmet, N., Šarac, J., Missoni, S., Kolčić, I., Polašek, O., Rudan, I., Campbell, H., Hayward, C., … Lauc, G. (2014). Glycans Are a Novel Biomarker of Chronological and Biological Ages. The Journals of Gerontology: Series A, 69(7), 779–789. https://doi.org/10.1093/gerona/glt190 [Google Scholar] [Crossref]
Loo, T., Patchett, M. L., Norris, G. E., & Lott, J. S. (2002). Using Secretion to Solve a Solubility Problem: High-Yield Expression in Escherichia coli and Purification of the Bacterial Glycoamidase PNGase F. Protein Expression and Purification, 24(1), 90–98. https://doi.org/10.1006/prep.2001.1555 [Google Scholar] [Crossref]
Mussar, K. J., Murray, G. J., Martin, B. M., & Viswanatha, T. (1989). Peptide: F: studies on the glycoprotein aminoglycan amidase from Flavobacterium meningosepticum. Journal of Biochemical and Biophysical Methods, 20(1), 53–68. https://doi.org/10.1016/0165-022X(89)90081-X [Google Scholar] [Crossref]
Nuck, R., Zimmermann, M., Sauvageot, D., Josić, D., & Reutter, W. (1990). Optimized deglycosylation of glycoproteins by peptide-N4-(N-acetyl-β-glucosaminyl)-asparagine amidase fromFlavobacterium meningosepticum. Glycoconjugate Journal, 7(4), 279–286. https://doi.org/10.1007/BF01073372 [Google Scholar] [Crossref]
Parc, A. L., Karav, S., Rouquié, C., Maga, E. A., Bunyatratchata, A., & Barile, D. (2017). Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows. PLOS ONE, 12(2), e0171477. https://doi.org/10.1371/journal.pone.0171477 [Google Scholar] [Crossref]
Pekdemir, B., Duman, H., Arslan, A., Kaplan, M., Karyelioğlu, M., Özer, T., Kayılı, H. M., Salih, B., Henrick, B. M., Duar, R. M., & Karav, S. (2022). Immobilization of a Bifidobacterial Endo-ß-N-Acetylglucosaminidase to Generate Bioactive Compounds for Food Industry. Frontiers in Bioengineering and Biotechnology, 10, 922423. https://doi.org/10.3389/fbioe.2022.922423 [Google Scholar] [Crossref]
Şahutoğlu, A. S., Duman, H., Frese, S. A., & Karav, S. (2020). Structural insights of two novel N-acetyl-glucosaminidase enzymes through in silico methods. TURKISH JOURNAL OF CHEMISTRY, 44(6), 1703–1712. https://doi.org/10.3906/kim-2006-19 [Google Scholar] [Crossref]
Sucu, B., Bayraktar, A., Duman, H., Arslan, A., Kaplan, M., Karyelioğlu, M., Ntelitze, E., Taştekin, T., Yetkin, S., Ertürk, M., Frese, S. A., Henrick, B. M., Kayili, H. M., Salih, B., & Karav, S. (2021). Recombinant Production of Bifidobacterial Endoglycosidases for N-glycan Release. Journal of Visualized Experiments, 173, 62804. https://doi.org/10.3791/62804 [Google Scholar] [Crossref]
Tarentino, A. L., Gomez, C. M., & Plummer, T. H. (1985). Deglycosylation of asparagine-linked glycans by peptide:N-glycosidase F. Biochemistry, 24(17), 4665–4671. https://doi.org/10.1021/bi00338a028 [Google Scholar] [Crossref]
Varki, A. (1993). Biological roles of oligosaccharides: All of the theories are correct. Glycobiology, 3(2), 97–130. https://doi.org/10.1093/glycob/3.2.97 [Google Scholar] [Crossref]

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