Patent ID: 8688427
Filing Date: 2014-04-01
Classification: C12N,C12Y

Abstract:
1. A method for making an enzyme that catalyzes a Diels-Alder reaction, the method comprising: creating a Diels-Alder active site model comprising a diene component, a dienophile component, and hydrogen bond donor and hydrogen bond acceptor amino acid functional groups positioned to support catalysis; placing the Diels-Alder active site model in a Diels-Alder scaffold selected from introducing mutations in the scaffold to prepare an active site having the protein functional groups in position to stabilize the Diels-Alder transition state, and having a surface complementary to the Diels-Alder transition state; introducing mutations in the scaffold as guided by computational design and Diels-Alder assay; and expressing the Diels-Alder enzyme and testing for the Diels-Alder enzyme activity.