WWW.NESUNGRANT.CORNELL.EDU
U.S. Department of
Transportation
Northeast Region
HIGH-THROUGHPUT ENGINEERING OF CELLULASE CONSORTIA
USING A GEL THAT CAN PRODUCE PROTEINS
Dr. Dan Luo, Cornell University
With the advent of directed molecular evolution techniques, a whole new
potential for enzyme engineering (instead of microbial engineering) and
improvement has opened. The greatest advantage of directed evolution is
that it is independent of knowledge of enzyme structure and of the interactions between enzyme and substrate. Recently, we have developed a
novel hydrogel (termed P-gel) which can efficiently produce proteins
without any living cells. The unique advantage of the P-gel system is that
it is totally cell-free, thus it is ideally suited to express a consortium of
proteins without worrying about the toxic effect to or low-yield problems
PI: Dr. Dan Luo
in living cells.
Cornell University,
Enzymatic conversion of natural cellulose, in particular, non-food cellu- Biological and Environlose, to bioethanol will have great impact on alternative and renewable
mental Engineering
energy development, local economy, and environment protection. However, economical production of bioethanol from cellulosic biomass is currently impeded by the high cost and low efficiency of cellulase comFunded: $150,000
plexes.
Start Date: 07/01/2009
Three goals for this project: 1) Using P-gel to establish cellulase mutant End Date: 06/30/2011
selection methods based on insoluble substrates; 2) Using microfluidics
to generate P-gel microdroplets in order to establish a high-throughput
method for screening the high activity cellulase mutants; and 3) Highthroughput molecular engineering of cellulase consortia. We will combine the molecular evolution approach with our cell-free P-gel and engineer cellulase consortia in a high-throughput fashion via the microfluidic
format.
We expect that we will establish a robust, P-gelbased, high-throughput protein engineering platform that is able to molecularly evolve not just
single proteins, but also protein consortia. We
also expect to achieve a five- to ten-fold reduction in the cost of cellulase enzymes by dramatically improving the efficiency of these enzymes.
Source of Sun Grant Funding:
United States Department of Transportation
Other Sources of Funding:
Cornell University