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Título: | Dramatic hyperactivation of lipase of Thermomyces lanuginosa by a cationic surfactant: Fixation of the hyperactivated form by adsorption on sulfopropyl-sepharose |
Autor: | Moreno-Pérez, Sonia CSIC ORCID; Ghattas, Nesrine; Filice, Marco CSIC ORCID; Guisán, José Manuel CSIC ORCID ; Fernández-Lorente, Gloria CSIC ORCID | Palabras clave: | Novel hyperactivated lipase derivatives Hydrolysis of small synthetic substrates Hydrolysis of fish oil Immobilization of hyperactivated TLL |
Fecha de publicación: | 2015 | Editor: | Elsevier | Citación: | Journal of Molecular Catalysis B: Enzymatic 122: 199-203 (2015) | Resumen: | Soluble lipase from Thermomyces lanuginosa (TLL) was dramatically hyper-activated by a cationic surfactant, cetyltrimethylammonium bromide (CTAB). The greatest hyperactivation (above 340-fold) was observed with 0.005% CTAB. In addition to that, very high hyperactivation was also observed with much higher concentrations of surfactant (100-fold in the presence of 0.3% CTAB). Without surfactant or with very low surfactant concentrations, TLL was not adsorbed to the cationic exchangers. However, in the presence of high concentrations of surfactant (0.3%), the lipase was completely and strongly adsorbed on sulfopropyl-sepharose. The adsorbed enzyme remained hyperactivated (80-fold more active than the soluble enzyme) after elimination of the excess of surfactant. Complete desorption of the hyperactivated TLL from the cationic exchanger is only achieved at 2 M NaCl. The same level of hyperactivation was observed for the hydrolysis of a large substrate: fish oil. The release of EPA (eicosapentaenoic acid) was 80-fold more rapid with hyperactivated TLL derivatives than with TLL very mildly immobilized on CNBr activated agarose. Hyperactivated derivatives were very stable at 25 °C and 37 °C. Full activity was preserved after 1 week. | URI: | http://hdl.handle.net/10261/149817 | DOI: | 10.1016/j.molcatb.2015.08.015 | Identificadores: | doi: 10.1016/j.molcatb.2015.08.015 issn: 1381-1177 e-issn: 1873-3158 |
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