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

被引:14
作者
Gouveia Mol, Paula Chequer [1 ]
Alcantara Verissimo, Lizzy Ayra [2 ]
Minim, Luis Antonio [3 ]
Boscolo, Mauricio [1 ]
Gomes, Eleni [1 ]
da Silva, Roberto [1 ]
机构
[1] Sao Paulo State Univ, UNESP, Lab Biochem & Appl Microbiol, BR-15054000 Sao Jose Do Rio Preto, SP, Brazil
[2] Univ Fed Lavras, Dept Food Sci, BR-37200000 Lavras, MG, Brazil
[3] Univ Fed Vicosa, Dept Food Technol, BR-36570000 Vicosa, MG, Brazil
基金
巴西圣保罗研究基金会;
关键词
beta-Glucosidase; Adsorption; Cryogel; DMAEMA; Ion-exchange; AUREOBASIDIUM-PULLULANS; EXCHANGE CHROMATOGRAPHY; PURIFICATION; IMMOBILIZATION; FERMENTATION; ENHANCEMENT; HYDROLYSIS; LYSOZYME; LECTINS;
D O I
10.1016/j.procbio.2019.03.029
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In this study, an anion exchange cryogel was prepared and characterized using morphological, hydrodynamic and thermal techniques and it was used to capture beta-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 beta-glucosidase extraction was assessed and was not significant (p > 0.05). The adsorption behavior of the beta-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 degrees C. These results indicated that the cryogel exhibited good properties to be used as a chromatographic media to purify biomolecules such as beta-glucosidase.
引用
收藏
页码:75 / 83
页数:9
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