Application of Immobilized β-Glucosidase from Candida boidinii in the Hydrolysis of Delignified Sugarcane Bagasse

利用博伊丁假丝酵母来源的固定化β-葡萄糖苷酶水解脱木素甘蔗渣

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Abstract

Candida boidinii is a methylotrophic yeast with wide geographical distribution. In the present study, the microorganism was isolated from the Bahian semiarid and the enzymatic extract containing β-glucosidase was obtained through submerged fermentation. Response surface methodology was employed to optimize of fermentation medium. The higher production of β-glucosidase was obtained after 71 h of fermentation in an optimized medium composed of 3.35% glucose, 1.78% yeast extract and 1% peptone. The optimum pH and temperature of enzymatic activity were 6.8 (citrate-phosphate buffer) and 71.7 °C, respectively. Salts tested (10 mM) CaCl(2), Na(2)SO(4) and ZnSO(4) promotes the increase of 91%, 45% and 80% of activity, respectively. The enzyme retained 64% ± 2.3 of its initial activity after 1 h heating at 90 °C. The production of reducing sugars was 95.94% after 24 h of hydrolysis and, with the addition of metal ions, this value increased more than 2 times. Among the supports analyzed for immobilization, chitosan showed higher residual activity during reuse. The immobilized enzyme showed higher activity at 60 °C with pH 6 and preserved almost 100% of the initial activity after 30 min at 70 °C.

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