Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports
2016
Authors
Mihailović, MladenTrbojević-Ivić, Jovana
Banjanac, Katarina
Milosavić, Nenad
Veličković, Dušan
Simović, Milica
Bezbradica, Dejan
Article (Published version)
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In this study, two commercial supports (Eupergit (R) C and Purolite (R) A109) were chemically modified in order to introduce thiosulfonate groups, which could subsequently exclusively react with the cysteine residues on the surface of enzymes. Thereafter, the maltase from Saccharomyces cerevisiae was immobilized onto the obtained thiosulfonate-activated supports, resulting in high expressed enzymatic activities (around 50 %), while on the other hand, immobilization on unmodified supports yielded expressed activities less than 5 %. Moreover, protein loadings up to 12.3 mg g(-1) and immobilized activities up to 3580 IU g(-1) were achieved by employment of these thiosulfonate supports. Desorption experiments, performed on samples taken during immobilization, proved that immobilization on the thiosulfonate supports was the first step of fast adsorption onto the supports and the formation of covalent bonds between the thiosulfonate groups and the thiol groups of cysteine represented a secon...d slower step. More importantly, although enzyme coupling occurred via covalent bond formation, the performed immobilization proved to be reversible, since it was shown that 95 % of the immobilized activity could be detached from the support after treatment with a thiol reagent (beta-mercaptoethanol). Thus, the support could be reused after enzyme inactivation.
Keywords:
alpha-glucosidase / covalent disulfide bridges / Eupergit (R) C / Purolite (R) A109Source:
Journal of the Serbian Chemical Society, 2016, 81, 12, 1371-1382Publisher:
- Srpsko hemijsko društvo, Beograd
Funding / projects:
- Ministry of Education, Science and Technological Development of the Republic of Serbia
DOI: 10.2298/JSC160730099M
ISSN: 0352-5139
WoS: 000392946100004
Scopus: 2-s2.0-85009798661
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Tehnološko-metalurški fakultetTY - JOUR AU - Mihailović, Mladen AU - Trbojević-Ivić, Jovana AU - Banjanac, Katarina AU - Milosavić, Nenad AU - Veličković, Dušan AU - Simović, Milica AU - Bezbradica, Dejan PY - 2016 UR - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3422 AB - In this study, two commercial supports (Eupergit (R) C and Purolite (R) A109) were chemically modified in order to introduce thiosulfonate groups, which could subsequently exclusively react with the cysteine residues on the surface of enzymes. Thereafter, the maltase from Saccharomyces cerevisiae was immobilized onto the obtained thiosulfonate-activated supports, resulting in high expressed enzymatic activities (around 50 %), while on the other hand, immobilization on unmodified supports yielded expressed activities less than 5 %. Moreover, protein loadings up to 12.3 mg g(-1) and immobilized activities up to 3580 IU g(-1) were achieved by employment of these thiosulfonate supports. Desorption experiments, performed on samples taken during immobilization, proved that immobilization on the thiosulfonate supports was the first step of fast adsorption onto the supports and the formation of covalent bonds between the thiosulfonate groups and the thiol groups of cysteine represented a second slower step. More importantly, although enzyme coupling occurred via covalent bond formation, the performed immobilization proved to be reversible, since it was shown that 95 % of the immobilized activity could be detached from the support after treatment with a thiol reagent (beta-mercaptoethanol). Thus, the support could be reused after enzyme inactivation. PB - Srpsko hemijsko društvo, Beograd T2 - Journal of the Serbian Chemical Society T1 - Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports EP - 1382 IS - 12 SP - 1371 VL - 81 DO - 10.2298/JSC160730099M ER -
@article{ author = "Mihailović, Mladen and Trbojević-Ivić, Jovana and Banjanac, Katarina and Milosavić, Nenad and Veličković, Dušan and Simović, Milica and Bezbradica, Dejan", year = "2016", abstract = "In this study, two commercial supports (Eupergit (R) C and Purolite (R) A109) were chemically modified in order to introduce thiosulfonate groups, which could subsequently exclusively react with the cysteine residues on the surface of enzymes. Thereafter, the maltase from Saccharomyces cerevisiae was immobilized onto the obtained thiosulfonate-activated supports, resulting in high expressed enzymatic activities (around 50 %), while on the other hand, immobilization on unmodified supports yielded expressed activities less than 5 %. Moreover, protein loadings up to 12.3 mg g(-1) and immobilized activities up to 3580 IU g(-1) were achieved by employment of these thiosulfonate supports. Desorption experiments, performed on samples taken during immobilization, proved that immobilization on the thiosulfonate supports was the first step of fast adsorption onto the supports and the formation of covalent bonds between the thiosulfonate groups and the thiol groups of cysteine represented a second slower step. More importantly, although enzyme coupling occurred via covalent bond formation, the performed immobilization proved to be reversible, since it was shown that 95 % of the immobilized activity could be detached from the support after treatment with a thiol reagent (beta-mercaptoethanol). Thus, the support could be reused after enzyme inactivation.", publisher = "Srpsko hemijsko društvo, Beograd", journal = "Journal of the Serbian Chemical Society", title = "Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports", pages = "1382-1371", number = "12", volume = "81", doi = "10.2298/JSC160730099M" }
Mihailović, M., Trbojević-Ivić, J., Banjanac, K., Milosavić, N., Veličković, D., Simović, M.,& Bezbradica, D.. (2016). Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports. in Journal of the Serbian Chemical Society Srpsko hemijsko društvo, Beograd., 81(12), 1371-1382. https://doi.org/10.2298/JSC160730099M
Mihailović M, Trbojević-Ivić J, Banjanac K, Milosavić N, Veličković D, Simović M, Bezbradica D. Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports. in Journal of the Serbian Chemical Society. 2016;81(12):1371-1382. doi:10.2298/JSC160730099M .
Mihailović, Mladen, Trbojević-Ivić, Jovana, Banjanac, Katarina, Milosavić, Nenad, Veličković, Dušan, Simović, Milica, Bezbradica, Dejan, "Immobilization of maltase from Saccharomyces cerevisiae on thiosulfonate supports" in Journal of the Serbian Chemical Society, 81, no. 12 (2016):1371-1382, https://doi.org/10.2298/JSC160730099M . .