Biochemical characteristics and potential application of a novel ethanol and glucose-tolerant β-glucosidase secreted by Pichia guilliermondii G1.2

• Published in: Journal of biotechnology 2019-03, Vol.294, p.73-80
• Main Authors: da Silva, Ronivaldo Rodrigues, da Conceição, Pâmela Joyce Previdelli, de Menezes, Cíntia Lionela Ambrosio, de Oliveira Nascimento, Carlos Eduardo, Machado Bertelli, Maryane, Pessoa Júnior, Adalberto, de Souza, Gisele Monteiro, da Silva, Roberto, Gomes, Eleni
• Format: Article
• Language: English
• Subjects: beta-Glucosidase - antagonists & inhibitors; beta-Glucosidase - chemistry; beta-Glucosidase - isolation & purification; Cellulose - chemistry; Ethanol - chemistry; Fermentation; Glucose - chemistry; Hydrogen-Ion Concentration; Hydrolysis; Microbial glycosidase; Pichia - enzymology; Temperature; Wine; Winemaking; Yeast enzyme
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2019.02.001

•A novel ethanol and glucose-tolerant β-glucosidase.•Potential application for cellulosic ethanol production and in winemaking.•β-glucosidase was no able to hydrolyze other substrates, such as pNPA, pNPX, beechwood xylan, CMC, and avicel. β-glucosidases are glycoside hydrolases that—particularly those from filamentous fungi—have been extensively explored in cellulose fiber saccharification and wine quality improvement. However, these enzymes from yeast have been poorly studied. In this study, an ethanol-glucose tolerant β-glucosidase that is secreted by Pichia guilliermondii (current name Meyerozyma guilliermondii) was purified and characterized. This enzyme exhibited an estimated molecular mass of 97 kDa and the highest activity between pH 3.5–5.5 and 55 °C. The β-glucosidase was also tolerant to acetone, ethanol, isopropanol, and methanol up to 30% and glucose at 1 M. It was also stable up to 55 °C for 80 min, maintaining 70% of its initial activity and in a wide pH range (pH 3–10). The enzyme exhibited 90–100% of its initial activity for 72 h at 20, 25, and 30 °C in presence of 10% ethanol at pH 3.5, which is a similar condition to winemaking. Studies that identify new enzymes and describe their purification are required for oenology applications. The β-glucosidase described herein is a promising candidate for use in the preparation of wine. Additionally, its tolerance to glucose is an important biochemical property that adds value to this enzyme and enables it to be used during the final saccharification process.