The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from <i>Marinimicrobium</i> sp. H1
Alginase lyase is an important enzyme for the preparation of alginate oligosaccharides (AOS), that possess special biological activities and is widely used in various fields, such as medicine, food, and chemical industry. In this study, a novel bifunctional alginate lyase (AlgH) belonging to the PL7...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-09-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/17/10/545 |
| _version_ | 1811302317803175936 |
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| author | Junjun Yan Peng Chen Yan Zeng Yan Men Shicheng Mu Yueming Zhu Yefu Chen Yuanxia Sun |
| author_facet | Junjun Yan Peng Chen Yan Zeng Yan Men Shicheng Mu Yueming Zhu Yefu Chen Yuanxia Sun |
| author_sort | Junjun Yan |
| collection | DOAJ |
| description | Alginase lyase is an important enzyme for the preparation of alginate oligosaccharides (AOS), that possess special biological activities and is widely used in various fields, such as medicine, food, and chemical industry. In this study, a novel bifunctional alginate lyase (AlgH) belonging to the PL7 family was screened and characterized. The AlgH exhibited the highest activity at 45 °C and pH 10.0, and was an alkaline enzyme that was stable at pH 6.0−10.0. The enzyme showed no significant dependence on metal ions, and exhibited unchanged activity at high concentration of NaCl. To determine the function of non-catalytic domains in the multi-domain enzyme, the recombinant AlgH-I containing only the catalysis domain and AlgH-II containing the catalysis domain and the carbohydrate binding module (CBM) domain were constructed and characterized. The results showed that the activity and thermostability of the reconstructed enzymes were significantly improved by deletion of the F5/8 type C domain. On the other hand, the substrate specificity and the mode of action of the reconstructed enzymes showed no change. Alginate could be completely degraded by the full-length and modified enzymes, and the main end-products were alginate disaccharide, trisaccharide, and tetrasaccharide. Due to the thermo and pH-stability, salt-tolerance, and bifunctionality, the modified alginate lyase was a robust enzyme which could be applied in industrial production of AOS. |
| first_indexed | 2024-04-13T07:25:51Z |
| format | Article |
| id | doaj.art-9ff3a1edce2741da94914c3160e1f614 |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-04-13T07:25:51Z |
| publishDate | 2019-09-01 |
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| series | Marine Drugs |
| spelling | doaj.art-9ff3a1edce2741da94914c3160e1f6142022-12-22T02:56:29ZengMDPI AGMarine Drugs1660-33972019-09-01171054510.3390/md17100545md17100545The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from <i>Marinimicrobium</i> sp. H1Junjun Yan0Peng Chen1Yan Zeng2Yan Men3Shicheng Mu4Yueming Zhu5Yefu Chen6Yuanxia Sun7College of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, ChinaNational Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, ChinaNational Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, ChinaNational Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, ChinaCollege of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, ChinaNational Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, ChinaCollege of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, ChinaNational Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, ChinaAlginase lyase is an important enzyme for the preparation of alginate oligosaccharides (AOS), that possess special biological activities and is widely used in various fields, such as medicine, food, and chemical industry. In this study, a novel bifunctional alginate lyase (AlgH) belonging to the PL7 family was screened and characterized. The AlgH exhibited the highest activity at 45 °C and pH 10.0, and was an alkaline enzyme that was stable at pH 6.0−10.0. The enzyme showed no significant dependence on metal ions, and exhibited unchanged activity at high concentration of NaCl. To determine the function of non-catalytic domains in the multi-domain enzyme, the recombinant AlgH-I containing only the catalysis domain and AlgH-II containing the catalysis domain and the carbohydrate binding module (CBM) domain were constructed and characterized. The results showed that the activity and thermostability of the reconstructed enzymes were significantly improved by deletion of the F5/8 type C domain. On the other hand, the substrate specificity and the mode of action of the reconstructed enzymes showed no change. Alginate could be completely degraded by the full-length and modified enzymes, and the main end-products were alginate disaccharide, trisaccharide, and tetrasaccharide. Due to the thermo and pH-stability, salt-tolerance, and bifunctionality, the modified alginate lyase was a robust enzyme which could be applied in industrial production of AOS.https://www.mdpi.com/1660-3397/17/10/545alginate lyasePL7 familyalginate oligosaccharidesbifunctionalrobust enzyme |
| spellingShingle | Junjun Yan Peng Chen Yan Zeng Yan Men Shicheng Mu Yueming Zhu Yefu Chen Yuanxia Sun The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from <i>Marinimicrobium</i> sp. H1 Marine Drugs alginate lyase PL7 family alginate oligosaccharides bifunctional robust enzyme |
| title | The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from <i>Marinimicrobium</i> sp. H1 |
| title_full | The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from <i>Marinimicrobium</i> sp. H1 |
| title_fullStr | The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from <i>Marinimicrobium</i> sp. H1 |
| title_full_unstemmed | The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from <i>Marinimicrobium</i> sp. H1 |
| title_short | The Characterization and Modification of a Novel Bifunctional and Robust Alginate Lyase Derived from <i>Marinimicrobium</i> sp. H1 |
| title_sort | characterization and modification of a novel bifunctional and robust alginate lyase derived from i marinimicrobium i sp h1 |
| topic | alginate lyase PL7 family alginate oligosaccharides bifunctional robust enzyme |
| url | https://www.mdpi.com/1660-3397/17/10/545 |
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