EAGER: Rapid Development of the "Rosettazyme" - A Novel Enzyme Sequestration Platform for Cellulosic Substrate Deconstruction

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R. Michael CeballosProposal#0929484EAGER: Rapid Development of the 'Rosettazyme' - A Novel Enzyme Sequestration Platform for Cellulosic Substrate DeconstructionCurrent technology in bioethanol production is limited by the ability to efficiently breakdown cellulosic biomass into fermentable sugars. In addition, many pre-treatment regimens for cellulosic substrate require acidic washes and high temperature processes that inhibit the catalytic efficiency of cellulose-degrading enzymes used in bioethanol production processes. Using proteins derived from hyperthermoacidophilic microbes from volcanic hot springs, the investigators on this project have developed a temperature- and acid-resistant enzyme sequestration platform capable of binding multiple cellulose-deconstruction enzymes to form a 'super-enzyme', called a 'rosettazyme'. In preliminary studies, it has been shown that these rosettazymes can improve simple sugar production from cellulosic substrates by more than two-fold. The purpose of this project is to further develop this novel technology using various combinations of bound lignocellulosic deconstruction enzymes from several species in order to generate a 5 to 10-fold increase in cellulose breakdown/conversion efficiency. This level of improvement in substrate deconstruction efficiency has been determined to be the threshold for making this technology an economically viable alternative to current bioethanol production methods. The proposed work will be a joint effort between several academic institutions and government laboratories throughout the United States. This project will also serve to provide science students from underrepresented groups and economically underserved backgrounds with research experiences in protein biochemistry, microbiology, genetic engineering, and alternative energy sciences. Within the two years of this project, the investigators will develop several rosettazyme subtypes using bacterial, archaeal, and/or fungal enzymes to increase the efficiency of lignocelluose conversion to simple fermentable sugars.