Reversible Photoswitchable Inhibitors Enable Wavelength‐Selective Regulation of Out‐of‐Equilibrium Bi‐enzymatic Systems

Abstract

The construction of synthetic enzymatic reaction networks can provide new insights into the design principles of living systems. However, the programmable connection of enzymes into a wide range of network topologies has been challenging due to the lack of a general strategy enabling a reversible activity regulation of individual network enzymes. Here, we exploit a general and modular strategy based on the external regulation of enzymes using light and photoswitchable inhibitors (PIs) that enables the bottom-up construction and control of enzymatic systems studied under out-of-equilibrium conditions. Upon synthesis and incorporation of potent photoswitchable trypsin inhibitors (Tr-PIs), the output of several functional enzymatic systems could be photoregulated using 390/460 nm light as a trigger signal. In addition, the wavelength-selective control over the activity of two enzymes within a functional bi-enzymatic system was achieved using a suitable combination of two PIs.

Exploring Emergent Properties in Enzymatic Reaction Networks: Design and Control of Dynamic Functional Systems

“…The warhead, a functional group that is known to inhibit the enzyme, can be covalently attached to a chromophore that can change their 3D structure upon light irradiation . Using this strategy, Teders et al recently designed an azobenzene-based inhibitor for Tr and Cr. , The activity of these enzymes could be reversibly adjusted under flow conditions upon different light intensity, duration, and wavelength of light, see Figure b. These systems exhibited an ultrasensitive response, illustrating potential for using light as an input for controlling complex dynamics in ERNs.…”