Production and capture of β-glucosidase from Thermoascus aurantiacus using a tailor made anionic cryogel

• Autor(a) principal: Mól, Paula Chequer Gouveia [UNESP]
• Data de Publicação: 2019
• Outros Autores: Veríssimo, Lizzy Ayra Alcântara, Minim, Luis Antonio, Boscolo, Maurício [UNESP], Gomes, Eleni [UNESP], da Silva, Roberto [UNESP]
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://dx.doi.org/10.1016/j.procbio.2019.03.029; http://hdl.handle.net/11449/190248
• topic: Adsorption; Cryogel; DMAEMA; Ion-exchange; β-Glucosidase

In this study, an anion exchange cryogel was prepared and characterized using morphological, hydrodynamic and thermal techniques and it was used to capture β-glucosidase from Thermoascus aurantiacus. The enzyme was produced by solid state fermentation. The effect of the stirring time (15, 30 and 60 min)on the β-glucosidase extraction was assessed and was not significant (p > 0.05). The adsorption behavior of the β-glucosidase produced was studied as a function of pH (3.0, 5.0 and 7.0), and the results of yield (%)and purification factor were calculated and submitted to ANOVA at a significance level of 95%. The efficiency was considerably affected (p < 0.05)by the pH and the best result was achieved at pH 5.0 (82%). Purification factors did not vary and the results were low since isocratic elution was performed (1.25–1.33). SDS-PAGE was also realized to investigate purity. The point of zero charge of the adsorbent was found at pH 8.0. The evaluation of functional groups confirmed the incorporation of the ion exchanger onto the cryogel surface. Thermal degradation of the cryogel started at 220 °C. These results indicated that the cryogel exhibited good properties to be used as a chromatographic media to purify biomolecules such as β-glucosidase.