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Carnegie Mellon U. develops microgel to recover enzymes for manufacturing, research assays

aging, and we are excited about extending this concept to other enzymes," noted Armitage. In addition to the food and dairy industries, enzymes are used in a wide variety of applications, including DNA testing in forensic labs, clinical tests for diagnosing diseases and synthesis of new pharmaceutical agents.

Armitage and Patterson relied on biomolecular recognition to create their microgel particles. For each particle, single DNA strands were used to create a three-way junction (TWJ) that looks like spokes of a wheel. To the end of each spoke, they tethered a strand of peptide nucleic acid (PNA), a synthetic material that recognizes and binds to DNA. Finally, enzyme complexes were then attached irreversibly to the tips of up to four PNA strands in a way that led to cross-linking of different TWJs, resulting in the microgel. At room temperature, individual microgel particles are suspended in solution, but lowering the temperature causes the microgel particles to cluster together and precipitate from solution.

To test these enzymatic microgel particles, the Carnegie Mellon chemists used an analogue of lactose that changes color when broken down by the microgel enzyme. When the investigators added this substrate to the microgel, a yellow color instantly appeared, indicating the reaction's success. Once the reaction was complete, the temperature was lowered, causing the microgel particles to precipitate. The product remained in solution and was separated simply by pouring the liquid into a separate container. The microgel particles could be subjected to several cycles of reaction, precipitation and reconstitution with no loss of activity, meaning that the enzyme was not leaching from the microgel, nor was it becoming inactivated.

The project demonstrates how effectively chemistry can bridge the biological and materials sciences, according to Armitage. "This is becoming a dominant theme in interdisciplinary research, particularly relating to nanotechno
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Contact: Lauren Ward
wardle@andrew.cmu.edu
412-268-7761
Carnegie Mellon University
8-Apr-2004


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