Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde
Abstract
Immobilization of enzymes on glutaraldehyde-activated supports has been largely used on supports previously activated with amine groups. Therefore, the supports are positively charged hence usually the immobilization is promoted through a two step mechanism: in a first step the enzyme is adsorbed on the support via an anionic exchange mechanism and then, the covalent immobilization occurs. In this paper a new glutaraldehyde activated support without a net charge is presented and characterized in immobilizations of trypsin, penicillin acylase G, lipase and E. coli BL21 cell extract. Immobilization mechanism was studied and this was produced without an adsorption step. This support promoted initially a reversible immobilization, converting into irreversible after incubation of the enzyme-support for several days or after a reduction step. In addition the stability of glutaraldehyde groups was studied retaining around 50 and 25% of its immobilization capacity for 24 h at pH 7 and 10 respe...ctively. This fact allows the incubation of the enzyme with the support even at alkaline pH promoting an extra stabilization factor for trypsin on this support.
Keywords:
Enzyme immobilization / Glutaraldehyde / Cysteine / Agarose / StabilitySource:
Journal of Molecular Catalysis B-Enzymatic, 2014, 102, 218-224Publisher:
- Elsevier, Amsterdam
Funding / projects:
- 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)
DOI: 10.1016/j.molcatb.2014.02.021
ISSN: 1381-1177
WoS: 000335872900031
Scopus: 2-s2.0-84896334037
Institution/Community
Tehnološko-metalurški fakultetTY - JOUR AU - Bezbradica, Dejan AU - Mateo, Cesar AU - Guisan, Jose M. PY - 2014 UR - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/2813 AB - Immobilization of enzymes on glutaraldehyde-activated supports has been largely used on supports previously activated with amine groups. Therefore, the supports are positively charged hence usually the immobilization is promoted through a two step mechanism: in a first step the enzyme is adsorbed on the support via an anionic exchange mechanism and then, the covalent immobilization occurs. In this paper a new glutaraldehyde activated support without a net charge is presented and characterized in immobilizations of trypsin, penicillin acylase G, lipase and E. coli BL21 cell extract. Immobilization mechanism was studied and this was produced without an adsorption step. This support promoted initially a reversible immobilization, converting into irreversible after incubation of the enzyme-support for several days or after a reduction step. In addition the stability of glutaraldehyde groups was studied retaining around 50 and 25% of its immobilization capacity for 24 h at pH 7 and 10 respectively. This fact allows the incubation of the enzyme with the support even at alkaline pH promoting an extra stabilization factor for trypsin on this support. PB - Elsevier, Amsterdam T2 - Journal of Molecular Catalysis B-Enzymatic T1 - Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde EP - 224 SP - 218 VL - 102 DO - 10.1016/j.molcatb.2014.02.021 ER -
@article{ author = "Bezbradica, Dejan and Mateo, Cesar and Guisan, Jose M.", year = "2014", abstract = "Immobilization of enzymes on glutaraldehyde-activated supports has been largely used on supports previously activated with amine groups. Therefore, the supports are positively charged hence usually the immobilization is promoted through a two step mechanism: in a first step the enzyme is adsorbed on the support via an anionic exchange mechanism and then, the covalent immobilization occurs. In this paper a new glutaraldehyde activated support without a net charge is presented and characterized in immobilizations of trypsin, penicillin acylase G, lipase and E. coli BL21 cell extract. Immobilization mechanism was studied and this was produced without an adsorption step. This support promoted initially a reversible immobilization, converting into irreversible after incubation of the enzyme-support for several days or after a reduction step. In addition the stability of glutaraldehyde groups was studied retaining around 50 and 25% of its immobilization capacity for 24 h at pH 7 and 10 respectively. This fact allows the incubation of the enzyme with the support even at alkaline pH promoting an extra stabilization factor for trypsin on this support.", publisher = "Elsevier, Amsterdam", journal = "Journal of Molecular Catalysis B-Enzymatic", title = "Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde", pages = "224-218", volume = "102", doi = "10.1016/j.molcatb.2014.02.021" }
Bezbradica, D., Mateo, C.,& Guisan, J. M.. (2014). Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde. in Journal of Molecular Catalysis B-Enzymatic Elsevier, Amsterdam., 102, 218-224. https://doi.org/10.1016/j.molcatb.2014.02.021
Bezbradica D, Mateo C, Guisan JM. Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde. in Journal of Molecular Catalysis B-Enzymatic. 2014;102:218-224. doi:10.1016/j.molcatb.2014.02.021 .
Bezbradica, Dejan, Mateo, Cesar, Guisan, Jose M., "Novel support for enzyme immobilization prepared by chemical activation with cysteine and glutaraldehyde" in Journal of Molecular Catalysis B-Enzymatic, 102 (2014):218-224, https://doi.org/10.1016/j.molcatb.2014.02.021 . .