SBIR-STTR Award

Hypotheses about the stabilization of enzymes by covalent attachmcnt to soluble polymers are to be tested in detail to isolate the effect of multi-point attachment from other factors known to affect enzyme stability including chemical modification, reduced proteolysis, diffusions] effects, and subunit dissociation. Two model enzyme systems have been selected based on criteria which will allow measurement of the effect of multi-point attachment alone. Each enzyme will be covalently attached through its lysine residues to two different activated, soluble polymers. Stability to heat, pH change, and denaturation by urea will be studied as a function ol'the number of points of attachment to the soluble polvnier. The ultimate goal or the research is to develop a geineral procedure for producing exceptionally stable enzymes for a variety of applications.The potential commercial application as described by the awardee: Manufacture of chemicals and pharmaceuticals, food processing, and diagnostics are all industries that will be significantly benefited through the development ot'nicthods for stabilizing enzymes.

The purpose of the proposed work is directed toward the investigation of stabilization of enzymes by attachment to soluble polymers. Methods for the stabilization of enzymes by multi-point attachment to soluable polymers will be developed, with the emphasis on improving the scope of the technique with respect to the types of chemistry employed, the types of stabilizing molecules used, and the number of different enzymes to which the methods can be applied. The tresyl chloride method developed in Phase I of Small Business Innovation Research (SBIR) will be further examined, using several different hydroxylic polymers and also lower molecular weight diols and polyethylene glycols. Other chemistries to be explored include reactions to produce multi-point attachment through chemically modified lysine residues and through carboxyl groups. Once refined, the methods will be applied to a number of different enzymes. Finally, the techniques will be developed for the production of stabilized, immobilized enzymes. This work is to be funded under Small Business Innovation Research (SBIR)Phase II program. The funds to be granted here will permit Genetics Institute to undertake further research to fully demonstrate the technical feasibility for commercialization under Phase II of the SBIR Program. The technical staff of Genetics Institute has an outstanding track record on enzyme technology, and the success of this project could benefit the food, pharamaceutical and feed industries.