Design of an enzyme-chaperone chimera as a new approach to enzyme immobilization and self-renaturation

BIOT 30

Lisa M Bergeron, LBergero@berkeley.edu1, Talar Tokatlian, ttokatli@uclink.berkeley.edu1, and Douglas S Clark, clark@cchem.berkeley.edu2. (1) Department of Chemical Engineering, University of California at Berkeley, 473 Tan Hall, Berkeley, CA 94720, (2) Department of Chemical Engineering, University of California, Berkeley, 497 Tan Hall, Berkeley, CA 94720
The use of enzymes in the presence of organic solvents is often limited by enzyme stability, particularly in solutions containing high concentrations of water-miscible organic co-solvents. Immobilization has been used to overcome this challenge by providing the enzyme with a stabilizing support; however, this can result in a reduction of enzyme activity. We have shown that some molecular chaperones can function to maintain enzyme activity under solvent-denaturing conditions. A single subunit isolated from the thermosome of the hyperthermophile Methanocaldococcus jannaschii functions in the presence of several water-miscible organic co-solvents. We have designed a chimera in which enzyme is fused to this chaperone and immobilized. This allows for continuous use of the enzyme in a flow-through reactor and provides it with the protection of a molecular chaperone to resist loss of activity in the presence of organic co-solvents.
 

Upstream Processing: Advances in Biocatalysis
8:00 AM-11:00 AM, Sunday, August 19, 2007 BCEC -- 107C, Oral

Division of Biochemical Technology

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007