Frontispiz: Vektorielle Katalyse mit oberflächenverankerten nano‐metallorganischen Gerüsten in mikrofluidischen Reaktoren

Designing an enzyme assembly line for green cascade processes using bio-orthogonal chemistry

“…63,64 Thus, in the aligned assembly of AKR and ADH in the -X-Y-X-Ymanner alignment impedes diffusion of the intermediates into the bulk solution 65,66 and reduces the transient time, 10 resulting in a higher reaction rate through substrate chanelling. 46,67 This is analogous to that observed in enzymatic cascades in vivo, such as the natural polyketide synthase module, 68,69 In addition, intensified artificial vectorial catalysis in the close alignment of AKR and ADH can also cause macromolecular crowding 70,71 and induce conformational changes that directly affect the structure of active sites in favour of increasing affinity for substrates. 72 In these crowded environments the conformational flexibility of enzymes and protein dynamics critical to enzyme catalysis 73 can be altered.…”

“…In the present work, we report the incorporation of ncAAs containing azide and alkyne groups into an aldo-keto reductase (AKR) and an alcohol dehydrogenase (ADH), respectively, at preselected sites. Each mutation site is kept away from the catalytic triad but close to the catalytic pocket in order to achieve vectorial biocatalysis, 45,46 Positioning of the ncAAs on the protein surface facilitates subsequent cross-linking and the overall enhanced efficiency is an important stimulus for operation in continuous flow. 47 The engineered bacterium used for the expression of the enzyme proteins containing ncAAs was E. coli C321.ΔA.…”