Rational design of Shewanella sp. l-arabinose isomerase for d-galactose isomerase activity under mesophilic conditions
| Autor: | Dilip Venkataraman, Thirukumaran Kandasamy, Arun Baskaran Jayaraman, Sankaranarayanan Meenakshisundaram |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Shewanella Isomerase activity Stereochemistry Bioconversion Bioengineering L-arabinose isomerase Isomerase 01 natural sciences Applied Microbiology and Biotechnology Biochemistry 03 medical and health sciences chemistry.chemical_compound 010608 biotechnology Escherichia coli Cloning Molecular Site-directed mutagenesis Aldose-Ketose Isomerases Hexoses Rational design Substrate (chemistry) Galactose Molecular Docking Simulation 030104 developmental biology chemistry Biotechnology |
| Zdroj: | Enzyme and microbial technology. 147 |
| ISSN: | 1879-0909 |
| Popis: | d-Tagatose, a potential low calorific substitute for sucrose, can be produced by bioconversion of d-galactose catalysed by l-arabinose isomerase. l-Arabinose isomerase from Shewanella sp. ANA-3 is unique for its ability to catalyse bioconversion reactions under mesophilic conditions. However, d-galactose not being a natural substrate for l-arabinose isomerase is catalysed at a slower rate. We attempted to increase the biocatalytic efficiency of Shewanella sp. l-arabinose isomerase by rational design to enhance galactose isomerisation activity. In silico molecular docking, analysis has revealed that F279 is sterically hindering the binding of d-galactose at the C6 position. Substitution of bulky Phe residue with smaller hydrophilic residues such as Asn and Thr increased the galactose isomerase activity by 86 % and 12 % respectively. At mesophilic conditions, F279N mutant catalysed the bioconversion of d-galactose more efficiently than l-arabinose, indicating a shift in substrate preference. |
| Databáze: | OpenAIRE |
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