Multi-Functional Alginate Lyase <i>AlgVR7</i> from <i>Vibrio rumoiensis</i>: Structural Insights and Catalytic Mechanisms
In this study, we identified <i>AlgVR7</i>, a novel bifunctional alginate lyase from <i>Vibrio rumoiensis</i> and characterized its biochemical properties and substrate specificity. Sequence alignment analysis inferred the key residues K267, H162, N86, E189, and T244 for <...
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2025-03-01
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| Series: | Marine Drugs |
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| author | Zhe Huang Shuai Liang Wulong Jiang Li Wang Yuan Wang Hua Wang Lianshun Wang Yuting Cong Yanan Lu Guojun Yang |
| author_facet | Zhe Huang Shuai Liang Wulong Jiang Li Wang Yuan Wang Hua Wang Lianshun Wang Yuting Cong Yanan Lu Guojun Yang |
| author_sort | Zhe Huang |
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| description | In this study, we identified <i>AlgVR7</i>, a novel bifunctional alginate lyase from <i>Vibrio rumoiensis</i> and characterized its biochemical properties and substrate specificity. Sequence alignment analysis inferred the key residues K267, H162, N86, E189, and T244 for <i>AlgVR7</i> catalysis, and it is derived from the PL7 family; exhibited high activity towards sodium alginate, polyM (PM), and polyG (PG); and can also degrade polygalacturonic acid (PGA) efficiently, with the highest affinity and catalytic efficiency for the MG block of the substrate. The optimal temperature and pH for <i>AlgVR7</i> were determined to be 40 °C and pH 8, respectively. The enzyme activity of <i>AlgVR7</i> was maximum at 40 °C, 40% of the enzyme activity was retained after incubation at 60 °C for 60 min, and enzyme activity was still present after 60 min incubation. <i>AlgVR7</i> activity was stimulated by 100 Mm NaCl, indicating a halophilic nature and suitability for marine environments. Degradation products analyzed using ESI-MS revealed that the enzyme primarily produced trisaccharides and tetrasaccharides. At 40 °C and pH 8.0, its <i>K</i><sub>m</sub> values for sodium alginate, PM, and PG were 16.67 μmol, 13.12 μmol, and 22.86 μmol, respectively. Structural analysis and molecular docking studies unveiled the key catalytic residues involved in substrate recognition and interaction. Glu167 was identified as a critical residue for the PL7_5 subfamily, uniquely playing an essential role in alginate decomposition. Overall, <i>AlgVR7</i> exhibits great potential as a powerful bifunctional enzyme for the efficient preparation of alginate oligosaccharides, with promising applications in biotechnology and industrial fields. |
| format | Article |
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| institution | OA Journals |
| issn | 1660-3397 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj-art-4e717a459f964a42bc30da907f5733382025-08-20T01:48:52ZengMDPI AGMarine Drugs1660-33972025-03-0123312410.3390/md23030124Multi-Functional Alginate Lyase <i>AlgVR7</i> from <i>Vibrio rumoiensis</i>: Structural Insights and Catalytic MechanismsZhe Huang0Shuai Liang1Wulong Jiang2Li Wang3Yuan Wang4Hua Wang5Lianshun Wang6Yuting Cong7Yanan Lu8Guojun Yang9College of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaCollege of Fisheries and Life Science, National Demonstration Center for Experimental Aquaculture Education, Dalian Ocean University, Ministry of Education, Dalian 116023, ChinaIn this study, we identified <i>AlgVR7</i>, a novel bifunctional alginate lyase from <i>Vibrio rumoiensis</i> and characterized its biochemical properties and substrate specificity. Sequence alignment analysis inferred the key residues K267, H162, N86, E189, and T244 for <i>AlgVR7</i> catalysis, and it is derived from the PL7 family; exhibited high activity towards sodium alginate, polyM (PM), and polyG (PG); and can also degrade polygalacturonic acid (PGA) efficiently, with the highest affinity and catalytic efficiency for the MG block of the substrate. The optimal temperature and pH for <i>AlgVR7</i> were determined to be 40 °C and pH 8, respectively. The enzyme activity of <i>AlgVR7</i> was maximum at 40 °C, 40% of the enzyme activity was retained after incubation at 60 °C for 60 min, and enzyme activity was still present after 60 min incubation. <i>AlgVR7</i> activity was stimulated by 100 Mm NaCl, indicating a halophilic nature and suitability for marine environments. Degradation products analyzed using ESI-MS revealed that the enzyme primarily produced trisaccharides and tetrasaccharides. At 40 °C and pH 8.0, its <i>K</i><sub>m</sub> values for sodium alginate, PM, and PG were 16.67 μmol, 13.12 μmol, and 22.86 μmol, respectively. Structural analysis and molecular docking studies unveiled the key catalytic residues involved in substrate recognition and interaction. Glu167 was identified as a critical residue for the PL7_5 subfamily, uniquely playing an essential role in alginate decomposition. Overall, <i>AlgVR7</i> exhibits great potential as a powerful bifunctional enzyme for the efficient preparation of alginate oligosaccharides, with promising applications in biotechnology and industrial fields.https://www.mdpi.com/1660-3397/23/3/124multifunctional alginate lyasealginate oligosaccharidesstructural insightscatalytic mechanisms |
| spellingShingle | Zhe Huang Shuai Liang Wulong Jiang Li Wang Yuan Wang Hua Wang Lianshun Wang Yuting Cong Yanan Lu Guojun Yang Multi-Functional Alginate Lyase <i>AlgVR7</i> from <i>Vibrio rumoiensis</i>: Structural Insights and Catalytic Mechanisms Marine Drugs multifunctional alginate lyase alginate oligosaccharides structural insights catalytic mechanisms |
| title | Multi-Functional Alginate Lyase <i>AlgVR7</i> from <i>Vibrio rumoiensis</i>: Structural Insights and Catalytic Mechanisms |
| title_full | Multi-Functional Alginate Lyase <i>AlgVR7</i> from <i>Vibrio rumoiensis</i>: Structural Insights and Catalytic Mechanisms |
| title_fullStr | Multi-Functional Alginate Lyase <i>AlgVR7</i> from <i>Vibrio rumoiensis</i>: Structural Insights and Catalytic Mechanisms |
| title_full_unstemmed | Multi-Functional Alginate Lyase <i>AlgVR7</i> from <i>Vibrio rumoiensis</i>: Structural Insights and Catalytic Mechanisms |
| title_short | Multi-Functional Alginate Lyase <i>AlgVR7</i> from <i>Vibrio rumoiensis</i>: Structural Insights and Catalytic Mechanisms |
| title_sort | multi functional alginate lyase i algvr7 i from i vibrio rumoiensis i structural insights and catalytic mechanisms |
| topic | multifunctional alginate lyase alginate oligosaccharides structural insights catalytic mechanisms |
| url | https://www.mdpi.com/1660-3397/23/3/124 |
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