The specificity of alpha-glucosidase from Saccharomyces cerevisiae differs depending on the type of reaction: hydrolysis versus transglucosylation
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2014
Authors
Dusan, VelickovicNenad, Milosavic
Dejan, Bezbradica
Bihelović, Filip
Segal, Ann Marie
Šegan, Dejan M.
Trbojević-Ivić, Jovana
Aleksandra, Dimitrijevic
Article (Published version)
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Our investigation of the catalytic properties of Saccharomyces cerevisiae alpha-glucosidase (AGL) using hydroxybenzyl alcohol (HBA) isomers as transglucosylation substrates and their glucosides in hydrolytic reactions demonstrated interesting findings pertaining to the aglycon specificity of this important enzyme. AGL specificity increased from the para(p)- to the ortho(o)-HBA isomer in transglucosylation, whereas such AGL aglycon specificity was not seen in hydrolysis, thus indicating that the second step of the reaction (i.e., binding of the glucosyl acceptor) is rate-determining. To study the influence of substitution pattern on AGL kinetics, we compared AGL specificity, inferred from kinetic constants, for HBA isomers and other aglycon substrates. The demonstrated inhibitory effects of HBA isomers and their corresponding glucosides on AGL-catalyzed hydrolysis of p-nitrophenyl a-glucoside (PNPG) suggest that HBA glucosides act as competitive, whereas HBA isomers are noncompetitive, ...inhibitors. As such, we postulate that aromatic moieties cannot bind to an active site unless an enzyme-glucosyl complex has already formed, but they can interact with other regions of the enzyme molecule resulting in inhibition.
Keywords:
Aglycon specificity / alpha-glucosidase / Transglucosylation kinetics / Hydrolysis kinetics / Substrate inhibition / Hydroxybenzyl alcoholSource:
Applied Microbiology and Biotechnology, 2014, 98, 14, 6317-6328Publisher:
- Springer, New York
Funding / projects:
- Allergens, antibodies, enzymes and small physiologically important molecules: design, structure, function and relevance (RS-MESTD-Basic Research (BR or ON)-172049)
- Novel encapsulation and enzyme technologies for designing of new biocatalysts and biologically active compounds targeting enhancement of food quality, safety and competitiveness (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-46010)
- 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-FP7-256716)
DOI: 10.1007/s00253-014-5587-9
ISSN: 0175-7598
PubMed: 24682477
WoS: 000338237400013
Scopus: 2-s2.0-84903612924
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Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Dusan, Velickovic AU - Nenad, Milosavic AU - Dejan, Bezbradica AU - Bihelović, Filip AU - Segal, Ann Marie AU - Šegan, Dejan M. AU - Trbojević-Ivić, Jovana AU - Aleksandra, Dimitrijevic PY - 2014 UR - https://cherry.chem.bg.ac.rs/handle/123456789/1797 AB - Our investigation of the catalytic properties of Saccharomyces cerevisiae alpha-glucosidase (AGL) using hydroxybenzyl alcohol (HBA) isomers as transglucosylation substrates and their glucosides in hydrolytic reactions demonstrated interesting findings pertaining to the aglycon specificity of this important enzyme. AGL specificity increased from the para(p)- to the ortho(o)-HBA isomer in transglucosylation, whereas such AGL aglycon specificity was not seen in hydrolysis, thus indicating that the second step of the reaction (i.e., binding of the glucosyl acceptor) is rate-determining. To study the influence of substitution pattern on AGL kinetics, we compared AGL specificity, inferred from kinetic constants, for HBA isomers and other aglycon substrates. The demonstrated inhibitory effects of HBA isomers and their corresponding glucosides on AGL-catalyzed hydrolysis of p-nitrophenyl a-glucoside (PNPG) suggest that HBA glucosides act as competitive, whereas HBA isomers are noncompetitive, inhibitors. As such, we postulate that aromatic moieties cannot bind to an active site unless an enzyme-glucosyl complex has already formed, but they can interact with other regions of the enzyme molecule resulting in inhibition. PB - Springer, New York T2 - Applied Microbiology and Biotechnology T1 - The specificity of alpha-glucosidase from Saccharomyces cerevisiae differs depending on the type of reaction: hydrolysis versus transglucosylation VL - 98 IS - 14 SP - 6317 EP - 6328 DO - 10.1007/s00253-014-5587-9 ER -
@article{ author = "Dusan, Velickovic and Nenad, Milosavic and Dejan, Bezbradica and Bihelović, Filip and Segal, Ann Marie and Šegan, Dejan M. and Trbojević-Ivić, Jovana and Aleksandra, Dimitrijevic", year = "2014", abstract = "Our investigation of the catalytic properties of Saccharomyces cerevisiae alpha-glucosidase (AGL) using hydroxybenzyl alcohol (HBA) isomers as transglucosylation substrates and their glucosides in hydrolytic reactions demonstrated interesting findings pertaining to the aglycon specificity of this important enzyme. AGL specificity increased from the para(p)- to the ortho(o)-HBA isomer in transglucosylation, whereas such AGL aglycon specificity was not seen in hydrolysis, thus indicating that the second step of the reaction (i.e., binding of the glucosyl acceptor) is rate-determining. To study the influence of substitution pattern on AGL kinetics, we compared AGL specificity, inferred from kinetic constants, for HBA isomers and other aglycon substrates. The demonstrated inhibitory effects of HBA isomers and their corresponding glucosides on AGL-catalyzed hydrolysis of p-nitrophenyl a-glucoside (PNPG) suggest that HBA glucosides act as competitive, whereas HBA isomers are noncompetitive, inhibitors. As such, we postulate that aromatic moieties cannot bind to an active site unless an enzyme-glucosyl complex has already formed, but they can interact with other regions of the enzyme molecule resulting in inhibition.", publisher = "Springer, New York", journal = "Applied Microbiology and Biotechnology", title = "The specificity of alpha-glucosidase from Saccharomyces cerevisiae differs depending on the type of reaction: hydrolysis versus transglucosylation", volume = "98", number = "14", pages = "6317-6328", doi = "10.1007/s00253-014-5587-9" }
Dusan, V., Nenad, M., Dejan, B., Bihelović, F., Segal, A. M., Šegan, D. M., Trbojević-Ivić, J.,& Aleksandra, D.. (2014). The specificity of alpha-glucosidase from Saccharomyces cerevisiae differs depending on the type of reaction: hydrolysis versus transglucosylation. in Applied Microbiology and Biotechnology Springer, New York., 98(14), 6317-6328. https://doi.org/10.1007/s00253-014-5587-9
Dusan V, Nenad M, Dejan B, Bihelović F, Segal AM, Šegan DM, Trbojević-Ivić J, Aleksandra D. The specificity of alpha-glucosidase from Saccharomyces cerevisiae differs depending on the type of reaction: hydrolysis versus transglucosylation. in Applied Microbiology and Biotechnology. 2014;98(14):6317-6328. doi:10.1007/s00253-014-5587-9 .
Dusan, Velickovic, Nenad, Milosavic, Dejan, Bezbradica, Bihelović, Filip, Segal, Ann Marie, Šegan, Dejan M., Trbojević-Ivić, Jovana, Aleksandra, Dimitrijevic, "The specificity of alpha-glucosidase from Saccharomyces cerevisiae differs depending on the type of reaction: hydrolysis versus transglucosylation" in Applied Microbiology and Biotechnology, 98, no. 14 (2014):6317-6328, https://doi.org/10.1007/s00253-014-5587-9 . .