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Kinetic Study of a Glucose Tolerant β-Glucosidase from Aspergillus fumigatus ABK9 Entrapped into Alginate Beads

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Abstract

A β-glucosidase from Aspergillus fumigatus ABK9 was purified from a pre-optimized solid state fermentation medium. The molecular weight of the purified enzyme (62.78 kDa) was determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, zymogram analysis and confirmed by MALDI-TOF mass spectrometry. The purified enzyme was entrapped in 4 % alginate beads and some physico-chemical properties of free and immobilized β-glucosidase were analyzed. The immobilized enzyme displayed higher Km and Vmax values but lower Kcat/Km value in comparison to its free counterpart. The pH and temperature stability of the enzyme were enhanced after immobilization. Increased thermostability of the immobilized enzyme was evidenced by the high activation energy (48.80 kJ mol−1) for thermal denaturation, longer half-life (T1/2) (1,037 min at 50 °C), higher melting temperature (Tm) (85 °C), and temperature coefficient (Q10) values (1.0). Besides superior thermodynamic properties, increased storage stability (80 % after 30 days), glucose tolerance (Ki = 430 mM) and reusability of the immobilized enzyme (nine cycles until E1/2) confirmed its promising industrial applicability.

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Acknowledgments

Authors are thankful to the University Grant Commission [Sanction No: F-3/2006 (BSR)/11-114/2008 (BSR)], Govt. of India, for the financial contribution in this study.

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Authors and Affiliations

  1. Department of Microbiology, Vidyasagar University, Midnapore, 721102, West Bengal, India

    Arpan Das, Tanmay Paul, Priyanka Ghosh, Suman Kumar Halder, Pradeep Kumar Das Mohapatra, Bikas Ranjan Pati & Keshab Chandra Mondal

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  1. Arpan Das
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  2. Tanmay Paul
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  3. Priyanka Ghosh
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  7. Keshab Chandra Mondal
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Correspondence to Keshab Chandra Mondal.

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Das, A., Paul, T., Ghosh, P. et al. Kinetic Study of a Glucose Tolerant β-Glucosidase from Aspergillus fumigatus ABK9 Entrapped into Alginate Beads. Waste Biomass Valor 6, 53–61 (2015). https://doi.org/10.1007/s12649-014-9329-0

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