The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite Hybrid
Enzyme immobilization is a powerful method to improve the stability, reuse, and enzymatic properties of enzymes. The immobilization of the α-amylase enzyme from Aspergillus fumigatus on a chitin-bentonite (CB) hybrid has been studied to improve its stability. Therefore, this study aims to obtain the...
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| Format: | Article |
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Wiley
2022-01-01
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| Series: | Biochemistry Research International |
| Online Access: | http://dx.doi.org/10.1155/2022/5692438 |
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| author | Ezra Rheinsky Tiarsa Yandri Yandri Tati Suhartati Heri Satria Bambang Irawan Sutopo Hadi |
| author_facet | Ezra Rheinsky Tiarsa Yandri Yandri Tati Suhartati Heri Satria Bambang Irawan Sutopo Hadi |
| author_sort | Ezra Rheinsky Tiarsa |
| collection | DOAJ |
| description | Enzyme immobilization is a powerful method to improve the stability, reuse, and enzymatic properties of enzymes. The immobilization of the α-amylase enzyme from Aspergillus fumigatus on a chitin-bentonite (CB) hybrid has been studied to improve its stability. Therefore, this study aims to obtain the higher stability of α-amylase enzyme to reduce industrial costs. The procedures were performed as follows: production, isolation, partial purification, immobilization, and characterization of the free and immobilized enzymes. The CB hybrid was synthesized by bentonite, chitin, and glutaraldehyde as a cross-linker. The free enzyme was immobilized onto CB hybrid using 0.1 M phosphate buffer pH 7.5. The free and immobilized enzymes were characterized by optimum temperature, Michaelis constant (KM), maximum velocity Vmax, thermal inactivation rate constant (ki), half-life (t1/2), and transformation of free energy because of denaturation (ΔGi). The free enzyme has optimum temperature of 55°C, KM = 3.04 mg mL−1 substrate, Vmax=10.90 μmolemL−1min−1, ki = 0.0171 min−1, t1/2 = 40.53 min, and ΔGi = 104.47 kJ mole−1. Meanwhile, the immobilized enzyme has optimum temperature of 60°C, KM = 11.57 mg mL−1 substrate, Vmax=3.37 μmolemL−1min−1, ki = 0.0045 min−1, t1/2 = 154.00 min, and ΔGi = 108.17 kJ mole−1. After sixth cycle of reuse, the residual activity of the immobilized enzyme was 38%. The improvement in the stability of α-amylase immobilized on the CB hybrid based on the increase in half-life was four times of the free enzyme. |
| format | Article |
| id | doaj-art-bac01950b9c64fcd9068edf518d3611b |
| institution | Kabale University |
| issn | 2090-2255 |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Wiley |
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| series | Biochemistry Research International |
| spelling | doaj-art-bac01950b9c64fcd9068edf518d3611b2025-02-03T05:58:10ZengWileyBiochemistry Research International2090-22552022-01-01202210.1155/2022/5692438The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite HybridEzra Rheinsky Tiarsa0Yandri Yandri1Tati Suhartati2Heri Satria3Bambang Irawan4Sutopo Hadi5Department of ChemistryDepartment of ChemistryDepartment of ChemistryDepartment of ChemistryDepartment of BiologyDepartment of ChemistryEnzyme immobilization is a powerful method to improve the stability, reuse, and enzymatic properties of enzymes. The immobilization of the α-amylase enzyme from Aspergillus fumigatus on a chitin-bentonite (CB) hybrid has been studied to improve its stability. Therefore, this study aims to obtain the higher stability of α-amylase enzyme to reduce industrial costs. The procedures were performed as follows: production, isolation, partial purification, immobilization, and characterization of the free and immobilized enzymes. The CB hybrid was synthesized by bentonite, chitin, and glutaraldehyde as a cross-linker. The free enzyme was immobilized onto CB hybrid using 0.1 M phosphate buffer pH 7.5. The free and immobilized enzymes were characterized by optimum temperature, Michaelis constant (KM), maximum velocity Vmax, thermal inactivation rate constant (ki), half-life (t1/2), and transformation of free energy because of denaturation (ΔGi). The free enzyme has optimum temperature of 55°C, KM = 3.04 mg mL−1 substrate, Vmax=10.90 μmolemL−1min−1, ki = 0.0171 min−1, t1/2 = 40.53 min, and ΔGi = 104.47 kJ mole−1. Meanwhile, the immobilized enzyme has optimum temperature of 60°C, KM = 11.57 mg mL−1 substrate, Vmax=3.37 μmolemL−1min−1, ki = 0.0045 min−1, t1/2 = 154.00 min, and ΔGi = 108.17 kJ mole−1. After sixth cycle of reuse, the residual activity of the immobilized enzyme was 38%. The improvement in the stability of α-amylase immobilized on the CB hybrid based on the increase in half-life was four times of the free enzyme.http://dx.doi.org/10.1155/2022/5692438 |
| spellingShingle | Ezra Rheinsky Tiarsa Yandri Yandri Tati Suhartati Heri Satria Bambang Irawan Sutopo Hadi The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite Hybrid Biochemistry Research International |
| title | The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite Hybrid |
| title_full | The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite Hybrid |
| title_fullStr | The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite Hybrid |
| title_full_unstemmed | The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite Hybrid |
| title_short | The Stability Improvement of Aspergillus fumigatus α-Amylase by Immobilization onto Chitin-Bentonite Hybrid |
| title_sort | stability improvement of aspergillus fumigatus α amylase by immobilization onto chitin bentonite hybrid |
| url | http://dx.doi.org/10.1155/2022/5692438 |
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