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Biomass Conversion and Biorefinery

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12-03-2020 | Original Article

Kinetics, thermodynamics, and physicochemical properties of 1, 4-α-d-glucan glucohydrolase from Aspergillus oryzae NRRL1560

Authors: Bilqees Fatima, Muhammad Mohsin Javed

Published in: Biomass Conversion and Biorefinery | Issue 6/2021

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Abstract

The aim of the study was to produce Ca²⁺ and Mg²⁺ independent raw starch hydrolyzing 1,4-α-d-glucan glucohydrolase (GGH) for glucose and fructose syrup production. A thermophilic GGH was produced from Aspergillus oryzae NRRL 1560. Two iso-forms of the enzyme, i.e., GGH1 and GGH2, were purified to homogeneity up to 4.93- and 8.82-fold that showed broad bands of 40 and 48 kDa on SDS-PAGE, respectively. K_m and V_max values of GGH2 were 1.315 mg mL⁻¹ and 70.864 μmol mL⁻¹ min⁻¹, respectively. Enzyme showed optimum activity at 60 °C, pH 5.0. It displayed a broad range of pH (3.0–9.0) and temperature (30–60 °C) stability. Inactivation kinetics parameters were calculated as K_d = 0.014 min⁻¹, t_1/2 = 49.5 min, D = 164.5 min, z = 22.23 °C, T_m = 77 °C, and E_a(d) = 337.481 kJ/mol. Thermodynamics parameters for thermal unfolding were ΔG*_(d) = 264.323, ΔH*_(d) = 334.713 kJ/mol, and ΔS*_(d) = 0.211 kJ/mol K. GGH2 was activated by 1 mM Na⁺ (18%), K⁺ (15%), Mg²⁺ (6%), Ca²⁺ (1%) and inhibited by Cu²⁺ (21%), Zn²⁺ (10%), Mn²⁺ (6%) at the same concentrations. The enzyme was also inhibited by 5 mM SDS (95%), DTT (8%), β-mercaptoethanol (4%), and EDTA (2%). GGH2-hydrolyzed raw starches were in the following order rice > wheat > corn > sweet potato > potato. This is the first report on raw starch digesting enzyme from A. oryzae. All these features make this enzyme a potential candidate for industrial production of glucose/fructose syrups and ethanol from raw starch via biological conversion.

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Title: Kinetics, thermodynamics, and physicochemical properties of 1, 4-α-d-glucan glucohydrolase from Aspergillus oryzae NRRL1560
Authors: Bilqees Fatima
Muhammad Mohsin Javed
Publication date: 12-03-2020
Publisher: Springer Berlin Heidelberg
Published in: Biomass Conversion and Biorefinery / Issue 6/2021
Print ISSN: 2190-6815
Electronic ISSN: 2190-6823
DOI: https://doi.org/10.1007/s13399-020-00663-8

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