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Photoresponsive polymer-enzyme switches
Tsuyoshi Shimoboji1, Edmund Larenas, Tim Fowler
1Department of Bioengineering, University of Washington, Seattle, WA 98195; and Genencor International, Inc., Palo Alto, CA 94304, USA.
Summary
Enzyme activity can be controlled with light using smart polymers. This photo-regulation enables on/off switching for applications in diagnostics and bioprocessing, even in microfluidic devices.
Area of Science:
- Bioconjugation Chemistry
- Polymer Science
- Enzyme Engineering
Background:
- Controlling enzyme activity is crucial for advanced biotechnological applications.
- Developing methods for reversible enzyme activity modulation is an ongoing challenge.
- Photoresponsive materials offer potential for non-invasive control mechanisms.
Purpose of the Study:
- To demonstrate the photoregulatory control of enzyme activity using conjugated "smart" polymers.
- To investigate the mechanism of light-induced substrate access modulation.
- To assess the functionality of photoresponsive polymer-enzyme conjugates in solution and immobilized states.
Main Methods:
- Conjugation of photoresponsive polymers to enzymes.
- Utilizing changes in polymer size and hydration upon light exposure.
- Assessing enzyme activity via substrate conversion measurements.
- Employing magnetic beads for immobilization studies.
Main Results:
- Photoinduced changes in polymer coil size and hydration reversibly modulated enzyme activity.
- The polymer-enzyme conjugates exhibited light-dependent "on" and "off" states.
- Activity regulation was observed for both free and immobilized conjugates.
- The system demonstrated potential for sequential bioprocessing and diagnostic assays.
Conclusions:
- Smart polymer conjugation provides an effective strategy for photoregulating enzyme activity.
- This approach offers precise, reversible control over enzyme function using optical signals.
- The developed system is versatile, applicable in solution and on solid supports, including microfluidics.