NORWEGIAN UNIVERSITY OF LIFE SCIENCES Department of Chemistry, Biotechnology and Food Science Team 4 – Lignocellulose to biofuels Topic: enzyme technology for conversion of lignocellulosic biomass

NORWEGIAN UNIVERSITY OF LIFE SCIENCES Department of Chemistry, Biotechnology and Food Science 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.

NORWEGIAN UNIVERSITY OF LIFE SCIENCES Department of Chemistry, Biotechnology and Food Science UMN-UMB, Team 4 specific project: CBP21, a helper protein from the CBM33 family Gustav Vaaje-Kolstad et al., Journal of Biological Chemistry 280: & 280: (2005) + Patent application

NORWEGIAN UNIVERSITY OF LIFE SCIENCES 4 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.

NORWEGIAN UNIVERSITY OF LIFE SCIENCES 5 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 CBM33 GH61

NORWEGIAN UNIVERSITY OF LIFE SCIENCES Department of Chemistry, Biotechnology and Food Science A new paradigm for degradation of crystalline polysaccharides ” Oxidohydrolase” (”CBM33” or ”GH61”), catalyzing chain cleavage in a fully crystalline context + Endo Exo-processive Vaaje-Kolstad et al., 2010, Science 330:

NORWEGIAN UNIVERSITY OF LIFE SCIENCES 7  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 (?) UMN-UMB, Team 4 – Status June 2011

NORWEGIAN UNIVERSITY OF LIFE SCIENCES Department of Chemistry, Biotechnology and Food Science 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).