Patent Number: 6,168,931

Title: Enhanced in vitro synthesis of biological macromolecules using a novel ATP regeneration system

Abstract: Compositions and methods are provided for the enhanced in vitro synthesis of biological molecules where ATP is required for synthesis. Of particular interest is the synthesis of polymers, e.g. nucleic acids, polypeptides, and complex carbohydrates. A homeostatic system is used for production of ATP, where the required high energy phosphate bonds are generated in situ, e.g. through coupling with an oxidation reaction. The homeostatic energy source will typically lack high energy phosphate bonds itself, and will therefore utilize free phosphate in the reaction mix during generation of ATP. Since inorganic phosphate can be an inhibitory by-product of synthesis, the period of time when synthesis is maintained in vitro can be extended. The homeostatic energy source is provided in combination with an enzyme that catalyzes the creation of high energy phosphate bonds and with an enzyme that can use that high energy phosphate bond to regenerate ATP.

Inventors: Swartz; James R. (Menlo Park, CA), Kim; Dong-Myung (Palo Alto, CA)

Assignee: The Board of Trustees of the Leland Stanford Junior University

International Classification: C07K 14/435 (20060101); C07K 14/525 (20060101); C12P 21/00 (20060101); C12N 9/10 (20060101); C12P 021/06 ()

Expiration Date: 01/02/2018