Characterization of Thermostable and Chimeric Enzymes via Isopeptide Bond-Mediated Molecular Cyclization
| Autor: | De-Ying Gao, Hui-En Zhang, Yan-Ni Liu, Jiakun Wang, Xin-Lei Shi, Qian Wang, Xiao-Bao Sun, Ming-qi Liu, Yang-Nan Li, Shang-Jun Yin |
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| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
Hot Temperature Stereochemistry Bacillus subtilis Ring (chemistry) Protein Engineering 01 natural sciences Bacterial Proteins Enzyme Stability Phosphofructokinase 2 Thermostability chemistry.chemical_classification Isopeptide bond biology Chemistry 010401 analytical chemistry beta-Mannosidase Substrate (chemistry) General Chemistry Hydrogen-Ion Concentration biology.organism_classification 0104 chemical sciences Enzyme Biocatalysis Cyclization General Agricultural and Biological Sciences 010606 plant biology & botany |
| Zdroj: | Journal of agricultural and food chemistry. 67(24) |
| ISSN: | 1520-5118 |
| Popis: | Mannooligosaccharides are released by mannan-degrading endo-β-1,4-mannanase and are known as functional additives in human and animal diets. To satisfy demands for biocatalysis and bioprocessing in crowed environments, in this study, we employed a recently developed enzyme-engineering system, isopeptide bond-mediated molecular cyclization, to modify a mesophilic mannanase from Bacillus subtilis. The results revealed that the cyclized enzymes showed enhanced thermostability and ion stability and resilience to aggregation and freeze-thaw treatment by maintaining their conformational structures. Additionally, by using the SpyTag/SpyCatcher system, we generated a mannanase-xylanase bifunctional enzyme that exhibited a synergistic activity in substrate deconstruction without compromising substrate affinity. Interestingly, the dual-enzyme ring conformation was observed to be more robust than the linear enzyme but inferior to the single-enzyme ring conformation. Taken together, these findings provided new insights into the mechanisms of molecular cyclization on stability improvement and will be useful in the production of new functional oligosaccharides and feed additives. |
| Databáze: | OpenAIRE |
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