Improving the cellobiose hydrolysis activity of glucose-stimulating β-glucosidase Bgl2A
Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved..
β-Glucosidases with high catalytic activity and glucose tolerant properties possess promising applications in lignocellulose-based industries. To obtain enzymes possessing these properties, a semi-rational strategy was employed to engineer the glucose-stimulating β-glucosidase Bgl2A for high cellobiose hydrolysis activity. A total of 18 mutants were constructed. A22S, V224D, and A22S/V224D exhibited high specific activities of 272.06, 237.60, and 239.29 U/mg toward cellobiose, which were 2.5- to 2.8-fold of Bgl2A. A22S, V224D, and A22S/V224D exhibited increased kcat values, which were 2.7- to 3.1-fold of Bgl2A. A22S and V224D maintained glucose-stimulating property, whereas A22S/V224D lost it. Using 150 g/L cellobiose as the substrate, the amount of glucose produced by A22S was the highest, yielding 129.70 g/L glucose after 3 h reaction at 35 °C. The synergistic effects of the engineered enzymes with commercial cellulase on hydrolyzing cellulose were investigated. Supplemented with the commercial cellulase and A22S, the highest glucose amount of 23.30 g/L was yielded from cellulose with hydrolysis rate of 21.02 %. Given its high cellobiose hydrolysis activity and glucose-stimulating properties, A22S can be used as a component of enzyme cocktail to match mesophilic cellulases for efficient cellulose hydrolysis.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:169 |
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| Enthalten in: |
Enzyme and microbial technology - 169(2023) vom: 05. Sept., Seite 110289 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Liu, Shuaifeng [VerfasserIn] |
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| Links: |
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| Themen: |
β-glucosidase |
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| Anmerkungen: |
Date Completed 27.07.2023 Date Revised 27.07.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.enzmictec.2023.110289 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM359741339 |
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| 245 | 1 | 0 | |a Improving the cellobiose hydrolysis activity of glucose-stimulating β-glucosidase Bgl2A |
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| 520 | |a Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a β-Glucosidases with high catalytic activity and glucose tolerant properties possess promising applications in lignocellulose-based industries. To obtain enzymes possessing these properties, a semi-rational strategy was employed to engineer the glucose-stimulating β-glucosidase Bgl2A for high cellobiose hydrolysis activity. A total of 18 mutants were constructed. A22S, V224D, and A22S/V224D exhibited high specific activities of 272.06, 237.60, and 239.29 U/mg toward cellobiose, which were 2.5- to 2.8-fold of Bgl2A. A22S, V224D, and A22S/V224D exhibited increased kcat values, which were 2.7- to 3.1-fold of Bgl2A. A22S and V224D maintained glucose-stimulating property, whereas A22S/V224D lost it. Using 150 g/L cellobiose as the substrate, the amount of glucose produced by A22S was the highest, yielding 129.70 g/L glucose after 3 h reaction at 35 °C. The synergistic effects of the engineered enzymes with commercial cellulase on hydrolyzing cellulose were investigated. Supplemented with the commercial cellulase and A22S, the highest glucose amount of 23.30 g/L was yielded from cellulose with hydrolysis rate of 21.02 %. Given its high cellobiose hydrolysis activity and glucose-stimulating properties, A22S can be used as a component of enzyme cocktail to match mesophilic cellulases for efficient cellulose hydrolysis | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Cellobiose | |
| 650 | 4 | |a Glucose tolerance | |
| 650 | 4 | |a Specific activity | |
| 650 | 4 | |a β-glucosidase | |
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| 700 | 1 | |a Zhang, Meng |e verfasserin |4 aut | |
| 700 | 1 | |a Hong, Dengwang |e verfasserin |4 aut | |
| 700 | 1 | |a Fang, Zemin |e verfasserin |4 aut | |
| 700 | 1 | |a Xiao, Yazhong |e verfasserin |4 aut | |
| 700 | 1 | |a Fang, Wei |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Xuecheng |e verfasserin |4 aut | |
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