Immobilization of Lipase B from Candida antarctica in Octyl-Vinyl Sulfone Agarose: Effect of the Enzyme-Support Interactions on Enzyme Activity, Specificity, Structure and Inactivation Pathway

Lipase B from was immobilized on heterofunctional support octyl agarose activated with vinyl sulfone to prevent enzyme release under drastic conditions. Covalent attachment was established, but the blocking step using hexylamine, ethylenediamine or the amino acids glycine (Gly) and aspartic acid (As...

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Bibliographic Details
Published in:International journal of molecular sciences 2022-11, Vol.23 (22), p.14268
Main Authors: Souza, Priscila M P, Carballares, Diego, Gonçalves, Luciana R B, Fernandez-Lafuente, Roberto, Rodrigues, Sueli
Format: Article
Language:English
Subjects:
Aspartic acid
Biocatalysts
Butyrates
Candida antarctica
Enzymatic activity
Enzyme activity
Enzymes
Enzymes, Immobilized - chemistry
Ethylenediamine
Ethylenediamines
Fluorescence
Glycine
Hydrophobic surfaces
Immobilization
Lipase
Lipase - metabolism
Nitrophenol
p-Nitrophenol
Physiology
Reagents
Sepharose - chemistry
Spectra
Triacetin
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Summary:Lipase B from was immobilized on heterofunctional support octyl agarose activated with vinyl sulfone to prevent enzyme release under drastic conditions. Covalent attachment was established, but the blocking step using hexylamine, ethylenediamine or the amino acids glycine (Gly) and aspartic acid (Asp) altered the results. The activities were lower than those observed using the octyl biocatalyst, except when using ethylenediamine as blocking reagent and -nitrophenol butyrate ( ) as substrate. The enzyme stability increased using these new biocatalysts at pH 7 and 9 using all blocking agents (much more significantly at pH 9), while it decreased at pH 5 except when using Gly as blocking agent. The stress inactivation of the biocatalysts decreased the enzyme activity versus three different substrates ( NPB, S-methyl mandelate and triacetin) in a relatively similar fashion. The tryptophane (Trp) fluorescence spectra were different for the biocatalysts, suggesting different enzyme conformations. However, the fluorescence spectra changes during the inactivation were not too different except for the biocatalyst blocked with Asp, suggesting that, except for this biocatalyst, the inactivation pathways may not be so different.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms232214268