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Purification and characterisation of Saccharomyces cerevisiae external invertase isoforms

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2010
Authors
Anđelković, Uroš
Picuric, Srdjan
Vujčić, Zoran
Article (Published version)
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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 / Imobillization
Source:
Food Chemistry, 2010, 120, 3, 799-804
Publisher:
  • Elsevier Sci Ltd, Oxford
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
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URI
http://cer.ihtm.bg.ac.rs/handle/123456789/742
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