Thermostability Improvement of the Chitinase from <i>Bacillus circulans</i> for Efficient Chitin Oligosaccharide Production via Computational Design
The chitinase A1 from <i>Bacillus circulans</i> WL-12 (BcChiA1) exhibits promising potential for producing chitin oligosaccharides (CHOs), while its application is limited by its poor thermal stability. In this study, a set of thermostable variants were obtained by modifying BcChiA1 usin...
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MDPI AG
2025-02-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/15/3/330 |
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| author | Jingwei Liu Jie Xie Si Wang Hong Feng Ganggang Wang |
| author_facet | Jingwei Liu Jie Xie Si Wang Hong Feng Ganggang Wang |
| author_sort | Jingwei Liu |
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| description | The chitinase A1 from <i>Bacillus circulans</i> WL-12 (BcChiA1) exhibits promising potential for producing chitin oligosaccharides (CHOs), while its application is limited by its poor thermal stability. In this study, a set of thermostable variants were obtained by modifying BcChiA1 using a comprehensive strategy based on a computer-aided design. A combination of five beneficial single-point mutations (S67G/K177R/A220V/N257Y/N271E) to BcChiA1 generated a markedly improved variant, Mu5. Mu5 exhibited a half-life of 295 min at 60 °C, which was 59 times higher than that of BcChiA1. Furthermore, Mu5 was reused for chitin conversion, releasing 86.14 ± 3.73 mM of CHOs after five reaction cycles. Molecular dynamics simulation and structural analysis revealed that these enhancements were driven by increased structural rigidity and compactness, resulting in a protein conformation that was less prone to thermal denaturation. This combined approach through computational design yielded a thermostable BcChiA1 variant, potentially facilitating CHOs production in economical way. |
| format | Article |
| id | doaj-art-eb2a9c5dc88c479582c73e794115bdfb |
| institution | Kabale University |
| issn | 2218-273X |
| language | English |
| publishDate | 2025-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj-art-eb2a9c5dc88c479582c73e794115bdfb2025-08-20T03:43:26ZengMDPI AGBiomolecules2218-273X2025-02-0115333010.3390/biom15030330Thermostability Improvement of the Chitinase from <i>Bacillus circulans</i> for Efficient Chitin Oligosaccharide Production via Computational DesignJingwei Liu0Jie Xie1Si Wang2Hong Feng3Ganggang Wang4Key Laboratory of Environmental Microbiology of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, ChinaKey Laboratory of Environmental Microbiology of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, ChinaKey Laboratory of Environmental Microbiology of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, ChinaCollege of Life Sciences, Sichuan University, Chengdu 610064, ChinaKey Laboratory of Environmental Microbiology of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, ChinaThe chitinase A1 from <i>Bacillus circulans</i> WL-12 (BcChiA1) exhibits promising potential for producing chitin oligosaccharides (CHOs), while its application is limited by its poor thermal stability. In this study, a set of thermostable variants were obtained by modifying BcChiA1 using a comprehensive strategy based on a computer-aided design. A combination of five beneficial single-point mutations (S67G/K177R/A220V/N257Y/N271E) to BcChiA1 generated a markedly improved variant, Mu5. Mu5 exhibited a half-life of 295 min at 60 °C, which was 59 times higher than that of BcChiA1. Furthermore, Mu5 was reused for chitin conversion, releasing 86.14 ± 3.73 mM of CHOs after five reaction cycles. Molecular dynamics simulation and structural analysis revealed that these enhancements were driven by increased structural rigidity and compactness, resulting in a protein conformation that was less prone to thermal denaturation. This combined approach through computational design yielded a thermostable BcChiA1 variant, potentially facilitating CHOs production in economical way.https://www.mdpi.com/2218-273X/15/3/330chitinasethermostabilitycomputational designchitin oligosaccharidesprotein engineering |
| spellingShingle | Jingwei Liu Jie Xie Si Wang Hong Feng Ganggang Wang Thermostability Improvement of the Chitinase from <i>Bacillus circulans</i> for Efficient Chitin Oligosaccharide Production via Computational Design Biomolecules chitinase thermostability computational design chitin oligosaccharides protein engineering |
| title | Thermostability Improvement of the Chitinase from <i>Bacillus circulans</i> for Efficient Chitin Oligosaccharide Production via Computational Design |
| title_full | Thermostability Improvement of the Chitinase from <i>Bacillus circulans</i> for Efficient Chitin Oligosaccharide Production via Computational Design |
| title_fullStr | Thermostability Improvement of the Chitinase from <i>Bacillus circulans</i> for Efficient Chitin Oligosaccharide Production via Computational Design |
| title_full_unstemmed | Thermostability Improvement of the Chitinase from <i>Bacillus circulans</i> for Efficient Chitin Oligosaccharide Production via Computational Design |
| title_short | Thermostability Improvement of the Chitinase from <i>Bacillus circulans</i> for Efficient Chitin Oligosaccharide Production via Computational Design |
| title_sort | thermostability improvement of the chitinase from i bacillus circulans i for efficient chitin oligosaccharide production via computational design |
| topic | chitinase thermostability computational design chitin oligosaccharides protein engineering |
| url | https://www.mdpi.com/2218-273X/15/3/330 |
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