Department of Chemistry, Biotechnology and Food Science NORWEGIAN UNIVERSITY OF LIFE SCIENCES Team 4 – Lignocellulose to biofuels Topic: enzyme technology for conversion of lignocellulosic biomass www.umb.no Department of Chemistry, Biotechnology and Food Science NORWEGIAN UNIVERSITY OF LIFE SCIENCES The biorefinery, 2nd generation biofuels and enzymes – Notes Enzymatic deconstruction is the method of choice. Enzymes are a major cost. Biofuel is only one of many possible products. www.umb.no Department of Chemistry, Biotechnology and Food Science NORWEGIAN UNIVERSITY OF LIFE SCIENCES UMN-UMB, Team 4 specific project: CBP21, a helper protein from the CBM33 family Gustav Vaaje-Kolstad et al., Journal of Biological Chemistry 280: 11313 11319 & 280:28492-28497 (2005)www.umb.no + Patent application 4 NORWEGIAN UNIVERSITY OF LIFE SCIENCES UMN-UMB, Team 4 specific project – starting point Goals: Use of directed evolution (mutagenesis) to produce accessory proteins that act on cellulose. Generation of fundamental knowledge about how helper proteins such as CBP21 work. People: Claudia Schmidt-Dannert group with post-doc Jake Vick. Eijsink group with post-doc Gustav Vaaje-Kolstad and (since 2010) Ph.D. student Zarah Forsberg. www.umb.no 5 NORWEGIAN UNIVERSITY OF LIFE SCIENCES UMN-UMB, Team 4 specific project – Developments since 2007 GH61 proteins act synergistically with cellulose and show structural similarity with CBM33 (CBP21) CBM33 proteins are enzymes that break down chitin chains GH61 proteins are enzymes that break down cellulose chains Identification of natural CBM33 proteins that break down cellulose Interesting ideas and findings at UMN concerning mechanism GH61 CBM33 www.umb.no Department of Chemistry, Biotechnology and Food Science NORWEGIAN UNIVERSITY OF LIFE SCIENCES A new paradigm for degradation of crystalline polysaccharides Endo Exo-processive + ”Oxidohydrolase” (”CBM33” or ”GH61”), catalyzing chain cleavage in a fully crystalline context Vaaje-Kolstad et al., 2010, Science 330:219-222 www.umb.no 7 NORWEGIAN UNIVERSITY OF LIFE SCIENCES UMN-UMB, Team 4 – Status June 2011 Very “hot” project. IP issues. Lots of mutagenesis work on CBP21 has been done and mutant characterization is in progress. Several new, active CBM33s available. NMR structure of CBP21 (a CBM33) has been solved. Shift of focus from ”enzyme development” to enzyme ”understanding”. Several joint papers, including potential ”breakthrough” papers on mechanism, are on their way. Funding is running out (?) www.umb.no Department of Chemistry, Biotechnology and Food Science NORWEGIAN UNIVERSITY OF LIFE SCIENCES Notes about the future Funding ? (currently only one PhD student at UMB) Many applications sent in Norway (but……) MSc student exchange: Sophanit Mengesha PhD student exchange: Zarah Forsberg (?) Nb. Current potential is huge and progress is good, but very high complexity (protein production, analytical tools, theoretical biochemistry, many partners). www.umb.no