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<p>This study extracted, purified, and characterized a β-glucosidase produced by the thermophilic fungus <i>Rasamsonia composticola</i> (β-GluRc) and compared the performance of the enzyme with that of a commercial enzyme cocktail in sugarcane bagasse saccharification. The enzyme was purified 19-fold, with a 14% yield. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) revealed a molecular mass of 42&#xa0;kDa. The optimal temperature and pH for enzyme activity were 70&#xa0;°C and 5.0, respectively. The enzyme showed increased activity in the presence of Triton X-100 (14%), saponin (21%), tetramethylethylenediamine (TEMED) (45%), and mercaptoethanol (56%). Furthermore, the enzyme exhibited activity toward <i>p</i>-nitrophenyl-β-<span>d</span>-glucopyranoside (<i>p</i>NP-glc) (112 U mg<sup>− 1</sup>) and <i>p</i>-nitrophenyl-β-<span>d</span>-xylopyranoside (<i>p</i>NP-xyl) (13,0 (U mg<sup>− 1</sup>) and was highly tolerant to glucose. Enzyme activity was enhanced by xylose concentrations ranging from 1 to 100 mM. <i>K</i><sub>m</sub> and <i>V</i><sub>max</sub> values for <i>p</i>NP-glc hydrolysis were 2.35 ± 0.11 mM and 112.2 ± 2.4 µmol min<sup>− 1</sup>, respectively, with a <i>K</i><sub>cat</sub>/<i>K</i><sub>m</sub> of 145.6 × 10<sup>− 10</sup>. Saccharification experiments using pretreated sugarcane bagasse revealed synergism between a commercial enzymatic cocktail (Celluclast<sup>®</sup>) and purified β-GluRc (80:1), resulting in increased release of total reducing sugars (1.58&#xa0;g L<sup>− 1</sup>) and total sugars (15.87&#xa0;g L<sup>− 1</sup>) compared with the control (Celluclast only). The findings demonstrate that <i>R. composticola</i> holds promise as a source of β-glucosidase for biotechnological applications, particularly in the hydrolysis of lignocellulosic biomass.</p>

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Biotechnological Potential of a Glucose-Tolerant β-Glucosidase from Rasamsonia composticola (β-GluRc) in Sugarcane Bagasse Saccharification

  • 期刊论文

  • 发表日期：

    2025年11月5日

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  • Isabela Pavão Vargas,
  • Rodrigo Mattos Silva Galeano,
  • Aline Pereira de Almeida,
  • Ana Lorena de Oliveira Simas,
  • João Victor Souza Ribeiro,
  • Maria de Lourdes Teixeira de Moraes Polizeli,
  • Douglas Chodi Masui,
  • Giovana Cristina Giannesi,
  • Roberto Ruller,
  • Fabiana Fonseca Zanoelo

摘要

This study extracted, purified, and characterized a β-glucosidase produced by the thermophilic fungus Rasamsonia composticola (β-GluRc) and compared the performance of the enzyme with that of a commercial enzyme cocktail in sugarcane bagasse saccharification. The enzyme was purified 19-fold, with a 14% yield. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) revealed a molecular mass of 42 kDa. The optimal temperature and pH for enzyme activity were 70 °C and 5.0, respectively. The enzyme showed increased activity in the presence of Triton X-100 (14%), saponin (21%), tetramethylethylenediamine (TEMED) (45%), and mercaptoethanol (56%). Furthermore, the enzyme exhibited activity toward p-nitrophenyl-β-d-glucopyranoside (pNP-glc) (112 U mg− 1) and p-nitrophenyl-β-d-xylopyranoside (pNP-xyl) (13,0 (U mg− 1) and was highly tolerant to glucose. Enzyme activity was enhanced by xylose concentrations ranging from 1 to 100 mM. Km and Vmax values for pNP-glc hydrolysis were 2.35 ± 0.11 mM and 112.2 ± 2.4 µmol min− 1, respectively, with a Kcat/Km of 145.6 × 10− 10. Saccharification experiments using pretreated sugarcane bagasse revealed synergism between a commercial enzymatic cocktail (Celluclast®) and purified β-GluRc (80:1), resulting in increased release of total reducing sugars (1.58 g L− 1) and total sugars (15.87 g L− 1) compared with the control (Celluclast only). The findings demonstrate that R. composticola holds promise as a source of β-glucosidase for biotechnological applications, particularly in the hydrolysis of lignocellulosic biomass.