The co-immobilization of P450-type nitric oxide reductase and glucose dehydrogenase for the continuous reduction of nitric oxide via cofactor recycling
| Autor: | Jan A. Verschoor, Seike Garny, Isak B. Gerber, Justin Jordaan, Natasha Beeton-Kempen |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Immobilized enzyme Nitric-oxide reductase Coenzymes Bioengineering Nitric Oxide 01 natural sciences Applied Microbiology and Biotechnology Biochemistry Cofactor Nitric oxide Fungal Proteins 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins Cytochrome P-450 Enzyme System Glucose dehydrogenase Multienzyme Complexes Enzyme Stability chemistry.chemical_classification biology 010405 organic chemistry Cytochrome P450 reductase Glucose 1-Dehydrogenase Enzymes Immobilized Enzyme assay Microspheres 0104 chemical sciences 030104 developmental biology Enzyme chemistry biology.protein Oxidoreductases Oxidation-Reduction Biotechnology |
| Zdroj: | Enzyme and microbial technology. 85 |
| ISSN: | 1879-0909 |
| Popis: | The co-immobilization of enzymes on target surfaces facilitates the development of self-contained, multi-enzyme biocatalytic platforms. This generally entails the co-immobilization of an enzyme with catalytic value in combination with another enzyme that performs a complementary function, such as the recycling of a critical cofactor. In this study, we co-immobilized two enzymes from different biological sources for the continuous reduction of nitric oxide, using epoxide- and carboxyl-functionalized hyper-porous microspheres. Successful co-immobilization of a fungal nitric oxide reductase (a member of the cytochrome P450 enzyme family) and a bacterial glucose dehydrogenase was obtained with the carboxyl-functionalized microspheres, with enzyme activity maintenance of 158% for nitric oxide reductase and 104% for glucose dehydrogenase. The optimal stoichiometric ratio of these two enzymes was subsequently determined to enable the two independent chemical reactions to be catalyzed concomitantly, allowing for near-synchronous cofactor conversion rates. This dual-enzyme system provides a novel research tool with potential for in vitro investigations of nitric oxide, and further demonstrates the successful immobilization of a P450 enzyme with potential application towards the immobilization of other cytochrome P450 enzymes. |
| Databáze: | OpenAIRE |
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