Impact of donor binding on polymerization catalyzed by KfoC by regulating the affinity of enzyme for acceptor
| Autor: | Xinke Zhang, Lan Jin, Peixue Ling, Fengshan Wang, Jiajun Xue, Juzheng Sheng |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Stereochemistry Biophysics Biochemistry Catalysis Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Biosynthesis Glycosyltransferase Escherichia coli Chondroitin Chondroitin sulfate Molecular Biology Polymerase chemistry.chemical_classification 030102 biochemistry & molecular biology biology Chondroitin Sulfates Substrate (chemistry) Directed evolution carbohydrates (lipids) 030104 developmental biology Enzyme Hexosyltransferases chemistry biology.protein Directed Molecular Evolution |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - General Subjects. 1860:844-855 |
| ISSN: | 0304-4165 |
| DOI: | 10.1016/j.bbagen.2016.01.018 |
| Popis: | Background Currently marketed chondroitin sulfate isolated from animal sources and structurally quite heterogeneous. Synthesis of structurally defined chondroitin sulfate is highly desired. The capsular polysaccharide from Escherichia coli strain K4 is similar to chondroitin, and its biosynthesis requires a chondroitin polymerase (KfoC). The essential step toward de novo enzymatic synthesis of chondroitin sulfate, synthesis of chondroitin, could be achieved by employing this enzyme. Methods Structurally defined acceptors and donor-sugars were prepared by chemoenzymatic approaches. In addition, surface plasmon resonance was employed to determine the binding affinities of individual substrates and donor–acceptor pairs for KfoC. Results KfoC has broad donor substrate specificity and acceptor promiscuity, making it an attractive tool enzyme for use in structurally-defined chimeric glycosaminoglycan oligosaccharide synthesis in vitro. In addition, the binding of donor substrate molecules regulated the affinity of KfoC for acceptors, then influenced the glycosyl transferase reaction catalyzed by this chondroitin polymerase. Conclusion and general significance These results assist in the development of enzymatic synthesis approaches toward chimeric glycosaminoglycan oligosaccharides and designing future strategies for directed evolution of KfoC in order to create mutants toward user-defined goals. |
| Databáze: | OpenAIRE |
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