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.

Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE

Original Publication: Basel, Switzerland : MDPI, [2000-

Lipase*/metabolism ; Enzymes, Immobilized*/chemistry; Sepharose/chemistry ; Butyrates ; Ethylenediamines

Lipase B from Candida antarctica 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 p -nitrophenol butyrate ( pNPB ) 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 ( p 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.

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01 Coordenação de Aperfeicoamento de Pessoal de Nível Superior; PR2-0101-00012.01.00/15 National Council for Scientific and Technological Development

Keywords: enzyme inactivation; heterofunctional supports; tuning enzyme specificity

EC 3.1.1.3 (Lipase)
9012-36-6 (Sepharose)
5PFN71LP8M (divinyl sulfone)
0 (Enzymes, Immobilized)
0 (Butyrates)
60V9STC53F (ethylenediamine)
0 (Ethylenediamines)

Moesziomyces antarcticus

Date Created: 20221126 Date Completed: 20221129 Latest Revision: 20221213

20231215

PMC9697615

10.3390/ijms232214268

36430745