Photosensitizers combine the inherent reactivity of reactive oxygen species with the sophisticated reaction control of light. Through selective targeting these light-active molecules have the potential to overcome certain limitations in drug disco...
Engineering Glucose Dehydrogenase to Favor Totally Synthetic Biomimetic containing carboxyl group
Von Wiley-VCH zur Verfügung gestellt
The utilization of unnatural nicotinamide cofactors for reactions catalyzed by oxidoreductases has gained increasing interest. Totally synthetic nicotinamide cofactor biomimetics (NCBs) are cost-effective and convenient to synthesize. Thus, it has becoming increasingly important to develop enzymes that accept NCBs. Here, we have engineered SsGDH to favor a newly synthesized unnatural cofactor 3-carbamoyl-1-(4-carboxybenzyl) pyridin-1-ium (BANA+). Using in situ ligand minimization tool, sites 44 and 114 were identified as hotspots for mutagenesis. All the double mutants demonstrated 2.7–7.7-fold improvements in catalytic activity, and the best double mutant E44D/E114L exhibited 10.6-fold increased catalytic efficiency toward BANA+. These results provide valuable information for the rational engineering of oxidoreductases with versatile NCBs-dependency, as well as the design of novel biomimetic cofactors.Zum Volltext
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