Purification and characterisation of Saccharomyces cerevisiae external invertase isoforms
Abstract
Four external invertase isoforms (EINV1, EINV2, EINV3 and EINV4) from Saccharomyces cerevisiae were highly purified by isoelectric precipitation, ethanol precipitation, ion-exchange on QAE-Sephadex and gel filtration using Sephacryl S-200. Unlike previously published procedures for external invertase purification, a specially designed step elution was applied on QAE-Sephadex which enabled the separation of four isoforms. The isoforms have the same molecular mass and catalytic properties: K-m for sucrose (25.6 mM), pH optimum (3.5-5.0) and temperature optimum (60 degrees C), but they exhibit significant difference in pl values, thermal stability and chemical reactivity. Deglycosylation Studies showed that the observed differences between isoforms arise from posttranslational modifications. Results showed that external invertase is a mixture of at least four isoforms, but in order to improve the efficiency of food industry processes, only the most stable isoform (EINV1) should be purifie...d and utilised. Substantially different chemical reactivity of the isoforms could be used to improve the yield of covalent immobilization procedures.
Keywords:
Invertase / Isoforms / Saccharomyces cerevisiae / Enzyme stability / Deglycosylation / ImobillizationSource:
Food Chemistry, 2010, 120, 3, 799-804Publisher:
- Elsevier Sci Ltd, Oxford
Funding / projects:
- Interakcije prirodnih proizvoda i njihovih analoga sa proteinima i nukleinskim kiselinama (RS-142026)
DOI: 10.1016/j.foodchem.2009.11.013
ISSN: 0308-8146
WoS: 000275014200024
Scopus: 2-s2.0-74149088237
Collections
Institution/Community
IHTMTY - JOUR AU - Anđelković, Uroš AU - Picuric, Srdjan AU - Vujčić, Zoran PY - 2010 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/742 AB - Four external invertase isoforms (EINV1, EINV2, EINV3 and EINV4) from Saccharomyces cerevisiae were highly purified by isoelectric precipitation, ethanol precipitation, ion-exchange on QAE-Sephadex and gel filtration using Sephacryl S-200. Unlike previously published procedures for external invertase purification, a specially designed step elution was applied on QAE-Sephadex which enabled the separation of four isoforms. The isoforms have the same molecular mass and catalytic properties: K-m for sucrose (25.6 mM), pH optimum (3.5-5.0) and temperature optimum (60 degrees C), but they exhibit significant difference in pl values, thermal stability and chemical reactivity. Deglycosylation Studies showed that the observed differences between isoforms arise from posttranslational modifications. Results showed that external invertase is a mixture of at least four isoforms, but in order to improve the efficiency of food industry processes, only the most stable isoform (EINV1) should be purified and utilised. Substantially different chemical reactivity of the isoforms could be used to improve the yield of covalent immobilization procedures. PB - Elsevier Sci Ltd, Oxford T2 - Food Chemistry T1 - Purification and characterisation of Saccharomyces cerevisiae external invertase isoforms VL - 120 IS - 3 SP - 799 EP - 804 DO - 10.1016/j.foodchem.2009.11.013 ER -
@article{ author = "Anđelković, Uroš and Picuric, Srdjan and Vujčić, Zoran", year = "2010", abstract = "Four external invertase isoforms (EINV1, EINV2, EINV3 and EINV4) from Saccharomyces cerevisiae were highly purified by isoelectric precipitation, ethanol precipitation, ion-exchange on QAE-Sephadex and gel filtration using Sephacryl S-200. Unlike previously published procedures for external invertase purification, a specially designed step elution was applied on QAE-Sephadex which enabled the separation of four isoforms. The isoforms have the same molecular mass and catalytic properties: K-m for sucrose (25.6 mM), pH optimum (3.5-5.0) and temperature optimum (60 degrees C), but they exhibit significant difference in pl values, thermal stability and chemical reactivity. Deglycosylation Studies showed that the observed differences between isoforms arise from posttranslational modifications. Results showed that external invertase is a mixture of at least four isoforms, but in order to improve the efficiency of food industry processes, only the most stable isoform (EINV1) should be purified and utilised. Substantially different chemical reactivity of the isoforms could be used to improve the yield of covalent immobilization procedures.", publisher = "Elsevier Sci Ltd, Oxford", journal = "Food Chemistry", title = "Purification and characterisation of Saccharomyces cerevisiae external invertase isoforms", volume = "120", number = "3", pages = "799-804", doi = "10.1016/j.foodchem.2009.11.013" }
Anđelković, U., Picuric, S.,& Vujčić, Z.. (2010). Purification and characterisation of Saccharomyces cerevisiae external invertase isoforms. in Food Chemistry Elsevier Sci Ltd, Oxford., 120(3), 799-804. https://doi.org/10.1016/j.foodchem.2009.11.013
Anđelković U, Picuric S, Vujčić Z. Purification and characterisation of Saccharomyces cerevisiae external invertase isoforms. in Food Chemistry. 2010;120(3):799-804. doi:10.1016/j.foodchem.2009.11.013 .
Anđelković, Uroš, Picuric, Srdjan, Vujčić, Zoran, "Purification and characterisation of Saccharomyces cerevisiae external invertase isoforms" in Food Chemistry, 120, no. 3 (2010):799-804, https://doi.org/10.1016/j.foodchem.2009.11.013 . .