Engineering of Cellobiose Dehydrogenases for Improved Glucose Sensitivity and Reduced Maltose Affinity
| Autor: | Miguel D. Toscano, Maria Silow, Lo Gorton, Mahbubur Rahman, Roland Ludwig, Christoph Sygmund, Roberto Ortiz, Beatrice Zangrilli, Pernille Ollendorff Micheelsen |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
chemistry.chemical_classification
Cellobiose dehydrogenase Stereochemistry Glucose Measurement Substrate (chemistry) 02 engineering and technology Maltose Cellobiose 010402 general chemistry 021001 nanoscience & nanotechnology 01 natural sciences Catalysis 0104 chemical sciences chemistry.chemical_compound Enzyme chemistry Biochemistry Electrochemistry Thiol 0210 nano-technology Cysteine |
| Zdroj: | ChemElectroChem. 4:846-855 |
| ISSN: | 2196-0216 |
| DOI: | 10.1002/celc.201600781 |
| Popis: | Cellobiose dehydrogenase (CDH) is a fungal extracellular flavocytochrome capable of direct electron transfer (DET). Unlike other CDHs, the pH optimum for CDHs from Corynascus thermophilus (CtCDH) and Humicola insolens (HiCDH) is close to the human physiological pH in blood (7.4). These are, therefore, interesting candidates for glucose measurements in human blood and the application in enzymatic fuel cells is, however, limited by their relatively low activity with this substrate. In this work, the substrate specificities of CtCDH and HiCDH have been altered by a single cysteine to tyrosine substitution in the active sites of CtCDH (position 291) and HiCDH (position 285), which resulted in improved kinetic constants with glucose while decreasing the activity with several disaccharides, including maltose. The DET properties of the generated CDH variants were tested in the absence and in the presence of substrates, on graphite electrodes and thiolic self-assembled monolayer (SAM)-modified Au electrodes. Seven different thiols with different spacer lengths were used, containing -COOH, -OH, and -NH2 end groups. The length and head functionality of the thiol govern the efficiency of the DET reaction and indicate different DET properties of CtCDH and HiCDH (Less) |
| Databáze: | OpenAIRE |
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