In vitro expression of a cellulase library on a microfluidics chip
Prof. Hermann Gaub, Department of Physics, Ludwig-Maximilians-University Munich (LMU)
Core of this project will be spotting the genes of a large number of the known cellulosome enzymes on a microfluidics chip and to create by in vitro expression a chip with a large variety of the known cellulases and other supporting proteins like cellulose binding proteins. These proteins will all have the same dockerin domain as anchor and an additional peptide as handle. An AFM cantilever tip functionalized with an antibody against this peptide will pick up individual cellulases and place them on an area, which is functionalized with a dense layer of cohesins, the modular binding partners of the dockerins. By placing one enzyme after the other we will assemble a large number of different designer cellulosomes, which differ both in composition and geometry. With the same technique we will complement the cellulase assemblies with nanocatalysts. Once a large number of different designer cellulosomes are assembled, we will measure their cellobiose production by quantitative fluorescence microscopy of thousands of different designer cellulosomes in parallel. We will go in iterative cycles through this process with the goal not only to identify the most productive arrangement but to derive a profound understanding of the basic functional mechanisms. This assay will also provide the technology platform for a very general Synthetic Biology approach for the high throughput investigation of enzymatic networks.
See also http://www.biophysik.physik.uni-muenchen.de/research/designer_cellulosomes/index.html
More information about the group:
Prof. Gaub's website