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Enhancement of Alpha-amylase’s Stability and Catalytic Efficiency After Modifying Enzyme Structure Using Calcium and Ultrasound

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Abstract

High thermostability and catalytic efficiency of α-amylase are required for the industrial applications. To improve catalytic parameters including Michaelis constant, maximum velocity, turnover number, enzyme half-life, and the catalytic efficiency of α-amylase, it was treated alone and/or simultaneously with Ca2+ ions (12.5 and 25 mM) and/or ultrasound (USN) (64.5 W, 25 + 40 kHz). The structural analysis of the enzyme (the enzyme’s secondary structure and conformation) was carried out using Fourier transform infrared (FTIR) spectroscopy and circular dichroism (CD) techniques. Moreover, the thermal properties of the enzyme were evaluated using differential scanning calorimetry (DSC). USN + 12.5 mM Ca2+ and 25 mM Ca2+, changed the maximum velocity (+ 6.5% and + 37.4%), turnover number (+ 6.6% and + 38.2%), enzyme half-life (+ 51.6% and + 113.7%), and catalytic efficiency (+ 18.1% and -3.3%) of α-amylase. The energy barriers of thermo-inactivation and inactivation enthalpy of α-amylase exposed to USN + 25 mM Ca2+ were increased by 25% and 5.8%, respectively, without affecting entropy of inactivation, showing how enzyme’s thermal stability was improved after the treatments. The increase in midpoint temperature (35%) and α-helix (14%) values as well as the decrease in the β-sheet structure (39%) of the α-amylase after applying USN + Ca2+ (25 mM) confirmed the thermal stability enhancement of α-amylase after the treatment compared to control.

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Abbreviations

K m :

Michaelis constant

V max :

Maximum velocity

K cat :

Turnover number

t 1/2 :

Enzyme half-life

K cat/K m :

Catalytic efficiency

T m :

Midpoint temperature

ΔH:

Enthalpy

Ea(in):

Energy barriers of thermo-inactivation

∆G*:

Entropy of inactivation

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Authors and Affiliations

  1. Department of Food Science and Technology, Faculty of Agriculture, Fasa University, Fasa, Iran

    Elahe Abedi

  2. Department of Chemical Technologies, Food Science and Technology Group, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran

    Homa Torabizadeh

  3. Centro de Investigación E Innovación Agroalimentaria y Agroambiental (CIAGRO-UMH), GIAAMA Research Group, Universidad Miguel Hernández, Orihuela, Spain

    Luciano Orden

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  1. Elahe Abedi
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  2. Homa Torabizadeh
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  3. Luciano Orden
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Contributions

Elahe Abedi wrote the manuscript conceived and designed the research, analyzed data d Writing – review & editing. Homa Torabizadeh and conceived and designed the research and analyzed the data. Luciano Orden: Data curation, Writing – review & editing. All the authors read and approved the manuscript.

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Correspondence to Elahe Abedi or Homa Torabizadeh.

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Abedi, E., Torabizadeh, H. & Orden, L. Enhancement of Alpha-amylase’s Stability and Catalytic Efficiency After Modifying Enzyme Structure Using Calcium and Ultrasound. Food Bioprocess Technol 17, 1546–1562 (2024). https://doi.org/10.1007/s11947-023-03213-y

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