more effective exploitation of biomass Innovative enzyme technology - - PDF document

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Innovative enzyme technology for more effective exploitation of biomass

Vincent Eijsink Norwegian University of Life Sciences (UMB)

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

The biobased economy

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

The biobased economy:

Using renewable materials as source for energy, chemical and materials. Some key R&D challenges: We need to understand our raw materials (plants, seaweeds, by-products) We need chemical or enzymatic conversion technologies

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Key technology: enzymatic processing

Enzymes are great! Mild conditions Precise (no by-products) Catalyze almost any reaction Can be engineered Non-toxic, natural, degradable BUT: (most) enzymes are expensive

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Enzymology – applied projects UMB

Chitin/ Chitosan Production of CHOS Protein hydrolysates Protein and more Proteins, Carbohydrates Prebiotics, Protein hydrolysates Cellulose Hemicellulose Sugar, ethanol, gas

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Key process: biomass deconstruction (saccharification)

From biomass to monosugars - the central starting point for in the ”biorefinery”.

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

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. Enzymes are also crucial in other steps of the biorefining process. Nutrient recycling is crucial.

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Enzymatic conversion of recalcitrant polysaccharides:

Classic “dogma”: “Endo” and processive “Exo” enzymes do the job.

Does this make

sense?

Could there be a

missing link?

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Tibtech 26:228-235 (2008)

Through long-term research on chitin and chitinases we have gained novel insights in biomass-converting enzymes which is now applied to lignocellulosic biomass.

Innovative enzyme technology

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Unravelling the role of processivity

Picture: NREL Picture: NREL

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

The discovery of accessory proteins

Gustav Vaaje-Kolstad et al., Journal of Biological Chemistry 280: 11313 - 11319 & 280:28492-28497 (2005) + Patent application

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

12

UMN-UMB, Team 4 project

Designer proteins for efficient enzymatic degradation of recalcitrant cellulose Goals:

• Use of directed evolution 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

www.umb.no

13

A screening assay has been developed (a major time investment) (UMN). Mutant libraries have been constructed and novel ones are under construction (UMN). Library screening has yielded the first interesting mutants:

• ther binding preferences, changed activities (UMN).

UMN-UMB, Team 4 – Status

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

14

Mutants are being tested in detail; researcher exchanges (UMN & UMB). More mutants are being selected (UMN). Biophysical characterization and structural work using solid state NMR (UMN & UMB). Novel mechanistic insights (UMB). Novel wild-type accessory proteins are being produced and characterized (UMB)

UMN-UMB, Team 4 – Status

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Novel developments – GH61

NB! GH61 proteins are abundant in cellulose degrading fungi

CBP21 GH61

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Novel developments – unravelling of the mechanism and boosting of activity

Science, Oct 8, 2010, Vol 330, pp 219-222

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

A missing link in cellulose degradation

Chain cleavage in a fully crystalline context!

CBP21 GH61

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Concluding remarks

• The UMB-UMN project is very ”hot”; the partners

have world-leading, complementary expertises.

• Completely new paradigms for enzymatic biomass

conversion may emerge.

• Major challenges in the directed evolution part &

lack of labour power have been delaying the joint project; current potential is huge and progress is good.

• Short line from fundamental research to application.

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

Applied bioenergy projects at Ås (Nb. new projects; since 2009)

• Conversion of wood, straw, bagasse, seaweed to

sugar or ethanol.

• Conversion of any biomass, household waste,

manure, by-products from food production to methane.

• Combined, integrated processes.

Industry is heavily involved (Cambi, Borregaard, Statoil, Xynergo, Hafslund, Weyland, SES) Approximately 7 persons (+ more at partners) Institutional collaborations with UMN (Minneapolis) and SLU (Uppsala)

NORWEGIAN UNIVERSITY OF LIFE SCIENCES

www.umb.no

Department of Chemistry, Biotechnology and Food Science

How can we get better?

UMN – UMB:

• More labour power
• More intense (regular) contacts, more exchange
• Larger network ?

Norway / Nordic:

• Much more funding for long-term researcher-driven

science in Norway (and Europe)

• More funding for equipment
• Nordic bioenergy initiative ?