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Biochemical Characterization of Thermostable Carboxymethyl Cellulase and β-Glucosidase from Aspergillus fumigatus JCM 10253

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Abstract

Second-generation biofuel production has emerged as a prominent sustainable and alternative energy. The biochemical properties of cellulolytic enzymes are imperative for cellulosic biomass conversion into fermentable sugars. In the present study, thermostable CMCase and β-glucosidase were purified and characterized from Aspergillus fumigatus JCM 10253. The enzymes were purified through 80% ammonium sulfate precipitation, followed by dialysis and DEAE-cellulose ion-exchange chromatography. The molecular masses of the purified CMCase and β-glucosidase were estimated to be 125 kDa and 90 kDa, respectively. The CMCase and β-glucosidase demonstrated optimum activities at pH 6.0 and 5.0, respectively. Their respective maximum temperatures were 50 and 60 °C. The cellulase activities were stimulated by 10 mM concentration of Ca2+, Ni2+, Fe2+, Mg2+, Cu2+, Mn2+, Zn2+, and Pb2+ ions. The CMCase activity was enhanced by surfactant Triton X-100 but marginally influenced by most inhibitors. The β-glucosidase retained its activity in the presence of organic solvents (30%) isoamyl alcohol, heptane, toluene, and ethyl acetate, while CMCase was retained with acetone during a prolonged incubation of 168 h. The Km and Vmax values of the two cellulases were studied. The properties of high thermostability and good tolerance against organic solvents could signify its potential use in biofuel production and other value-added products.

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Acknowledgements

The authors acknowledge the support provided by the Department of Biotechnology, National Institute of Technology Warangal, Telangana, India.

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  1. Department of Biotechnology, National Institute of Technology Warangal, Warangal, 506004, Telangana, India

    Paramjeet Saroj & Korrapati Narasimhulu

  2. Department of Biotechnology, National Institute of Technology Andhra Pradesh, Tadepalligudem, 534101, India

    Manasa P

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Paramjeet Saroj performed the research experiments, data analysis, and writing of the manuscript. Manasa P contributed to the data collection and manuscript writing. Korrapati Narasimhulu supervised during research experiments and manuscript preparation. All authors contributed to the study conception and design. All authors read and approved the final manuscript.

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Saroj, P., P, M. & Narasimhulu, K. Biochemical Characterization of Thermostable Carboxymethyl Cellulase and β-Glucosidase from Aspergillus fumigatus JCM 10253. Appl Biochem Biotechnol 194, 2503–2527 (2022). https://doi.org/10.1007/s12010-022-03839-2

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