Immobilization and Characterization of a Processive Endoglucanase EG5C-1 from <i>Bacillus subtilis</i> on Melamine–Glutaraldehyde Dendrimer-Functionalized Magnetic Nanoparticles
Exploring an appropriate immobilization approach to enhance catalytic activity and reusability of cellulase is of great importance to reduce the price of enzymes and promote the industrialization of cellulose-derived biochemicals. In this study, Fe<sub>3</sub>O<sub>4</sub> ma...
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| Iaith: | English |
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MDPI AG
2024-02-01
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| Cyfres: | Nanomaterials |
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| Mynediad Ar-lein: | https://www.mdpi.com/2079-4991/14/4/340 |
| _version_ | 1827343024763437056 |
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| author | Xiaozhou Li Jie Chen Bin Wu Zhen Gao Bingfang He |
| author_facet | Xiaozhou Li Jie Chen Bin Wu Zhen Gao Bingfang He |
| author_sort | Xiaozhou Li |
| collection | DOAJ |
| description | Exploring an appropriate immobilization approach to enhance catalytic activity and reusability of cellulase is of great importance to reduce the price of enzymes and promote the industrialization of cellulose-derived biochemicals. In this study, Fe<sub>3</sub>O<sub>4</sub> magnetic nanoparticles (MNPs) were functionalized with meso-2,3-dimercaptosuccinic acid to introduce carboxyl groups on the surface (DMNPs). Then, melamine–glutaraldehyde dendrimer-like polymers were grafted on DMNPs to increase protein binding sites for the immobilization of processive endoglucanase EG5C-1. Moreover, this dendrimer-like structure was beneficial to protect the conformation of EG5C-1 and facilitate the interaction between substrate and active center. The loading capacity of the functionalized copolymers (MG-DMNPs) for EG5C-1 was about 195 mg/g, where more than 90% of the activity was recovered. Immobilized EG5C-1 exhibited improved thermal stability and increased tolerability over a broad pH range compared with the free one. Additionally, MG-DMNP/EG5C-1 biocomposite maintained approximately 80% of its initial hydrolysis productivity after five cycles of usage using filter paper as the substrate. Our results provided a promising approach for the functionalization of MNPs, enabling the immobilization of cellulases with a high loading capacity and excellent activity recovery. |
| first_indexed | 2024-03-07T22:19:17Z |
| format | Article |
| id | doaj.art-a71eb851e61f4644be825fcf8b72a4ae |
| institution | Directory Open Access Journal |
| issn | 2079-4991 |
| language | English |
| last_indexed | 2024-03-07T22:19:17Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Nanomaterials |
| spelling | doaj.art-a71eb851e61f4644be825fcf8b72a4ae2024-02-23T15:29:23ZengMDPI AGNanomaterials2079-49912024-02-0114434010.3390/nano14040340Immobilization and Characterization of a Processive Endoglucanase EG5C-1 from <i>Bacillus subtilis</i> on Melamine–Glutaraldehyde Dendrimer-Functionalized Magnetic NanoparticlesXiaozhou Li0Jie Chen1Bin Wu2Zhen Gao3Bingfang He4College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, ChinaCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, ChinaSchool of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 211800, ChinaExploring an appropriate immobilization approach to enhance catalytic activity and reusability of cellulase is of great importance to reduce the price of enzymes and promote the industrialization of cellulose-derived biochemicals. In this study, Fe<sub>3</sub>O<sub>4</sub> magnetic nanoparticles (MNPs) were functionalized with meso-2,3-dimercaptosuccinic acid to introduce carboxyl groups on the surface (DMNPs). Then, melamine–glutaraldehyde dendrimer-like polymers were grafted on DMNPs to increase protein binding sites for the immobilization of processive endoglucanase EG5C-1. Moreover, this dendrimer-like structure was beneficial to protect the conformation of EG5C-1 and facilitate the interaction between substrate and active center. The loading capacity of the functionalized copolymers (MG-DMNPs) for EG5C-1 was about 195 mg/g, where more than 90% of the activity was recovered. Immobilized EG5C-1 exhibited improved thermal stability and increased tolerability over a broad pH range compared with the free one. Additionally, MG-DMNP/EG5C-1 biocomposite maintained approximately 80% of its initial hydrolysis productivity after five cycles of usage using filter paper as the substrate. Our results provided a promising approach for the functionalization of MNPs, enabling the immobilization of cellulases with a high loading capacity and excellent activity recovery.https://www.mdpi.com/2079-4991/14/4/340processive endoglucanaseimmobilizationmagnetic nanoparticlesmelamine–glutaraldehyde dendrimer |
| spellingShingle | Xiaozhou Li Jie Chen Bin Wu Zhen Gao Bingfang He Immobilization and Characterization of a Processive Endoglucanase EG5C-1 from <i>Bacillus subtilis</i> on Melamine–Glutaraldehyde Dendrimer-Functionalized Magnetic Nanoparticles Nanomaterials processive endoglucanase immobilization magnetic nanoparticles melamine–glutaraldehyde dendrimer |
| title | Immobilization and Characterization of a Processive Endoglucanase EG5C-1 from <i>Bacillus subtilis</i> on Melamine–Glutaraldehyde Dendrimer-Functionalized Magnetic Nanoparticles |
| title_full | Immobilization and Characterization of a Processive Endoglucanase EG5C-1 from <i>Bacillus subtilis</i> on Melamine–Glutaraldehyde Dendrimer-Functionalized Magnetic Nanoparticles |
| title_fullStr | Immobilization and Characterization of a Processive Endoglucanase EG5C-1 from <i>Bacillus subtilis</i> on Melamine–Glutaraldehyde Dendrimer-Functionalized Magnetic Nanoparticles |
| title_full_unstemmed | Immobilization and Characterization of a Processive Endoglucanase EG5C-1 from <i>Bacillus subtilis</i> on Melamine–Glutaraldehyde Dendrimer-Functionalized Magnetic Nanoparticles |
| title_short | Immobilization and Characterization of a Processive Endoglucanase EG5C-1 from <i>Bacillus subtilis</i> on Melamine–Glutaraldehyde Dendrimer-Functionalized Magnetic Nanoparticles |
| title_sort | immobilization and characterization of a processive endoglucanase eg5c 1 from i bacillus subtilis i on melamine glutaraldehyde dendrimer functionalized magnetic nanoparticles |
| topic | processive endoglucanase immobilization magnetic nanoparticles melamine–glutaraldehyde dendrimer |
| url | https://www.mdpi.com/2079-4991/14/4/340 |
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