Structural and functional characterisation of CBP21, a non

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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
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