Beruflich Dokumente
Kultur Dokumente
$$$$$
What fibre ?
Why Hemp ?
In 03/1998, the Industrial Hemp Regulations enabled commercial cultivation of industrial hemp which produces little or no THC. Grower associations in almost all provinces (Quinte, ON) Interest in hemp production increasing (28% increase in licensed area in 2004) Converting 5X more CO2 to biomass vs. forestry, without herbicide or pesticide Strongest and most durable among natural fibres Advanced products: priced hemp garments, automobile panels, windmill blade Renewed interest for medical, pulp and fibre purposes Feasibility of hemp uniforms for Canadas 2010 Olympic team (Hemptown, Van)
Ash
55 44
16 18
18 4
4 28
1 1
2 3
4 2
! Present chemical process (NaOH) dirty and fibre-damaging ! Glycosidases (pectinases & others) potentially capable to clean fibre without damaging durability ! The green advantage of enzymes environmental saving (less waste, reduced energy consumption)
From one end of Canada to the other, bio-composite projects are emerging; and the ability to thoroughly clean and yet preserve the strength and durability of natural fibres will be critical. John Baker
Unprocessed fibre
Endo-polygalacturonase Exo-polygalacturonase Pectate lyase Pectin (methyl) esterase Pectin lyase polygalacturonate (pectin main chain)
www.rcsb.org/pdb www.ncbe.reading.ac.uk
Biocatalyst Business
The name of the game is to find the right catalyst. You can do this by mutation, or you can do it by finding the perfect enzyme in nature. But you have to do it quickly and efficiently.
Frances H. Arnold, Professor of Chemical Engineering and Biochemistry, Caltech. In Innovations, Chemistry & Biology, 12:503-505, 2005.
Enzymes Made-to-Measure
Changes in catalytic properties Increase in Vmax Decrease in Km Change in pH optimum Alteration of substrate specificity Changes in structural properties Thermostability Stability in organic solvents
Erwinia endopolygalacturonase
Genome Mining of Potential Pectinase-encoding Sequences Mining of microbial genome databases an economical approach to new biocatalyst discovery Bait a known bacterial pectinase sequence Catches Multiple; selected 9 representing the three subclasses of pectinases for cloning and expression (5 putative lyases, 2
polygalacturonase and 2 methylesterases from Xanthomonas campestris and Streptomyces coelicolor)
Phylogram Tree
Control Sc 626147 47.7 kDa Sc 627050 53.3 kDa Xc 638163 37.4 kDa
Activity Assay
Polygalacturonic acid assay Three good candidates - two lyases (from Sc and Xc) and one hydrolase (from Xc)
Thermostability
Effect of 1hr Incubation at Temperature
120 % remaining activity 100 80 60 40 20 0 20 30 40 50 60 70 temperature (C) pectate lyase from S. coelicolor polygalacturonase from X. campestris
Characterization of Pectinases
Ongoing Investigation
! ! ! ! Further characterization of cloned pectinases Engineering of pectinases with optimal activity at elevated temperature and alkaline pH Recombination of individual pectinase enzymes for an optimal efficiency of fibre extraction. Preparation of pectinase combination without other enzyme contaminants (cellulase, etc.) which may attack the integrity of the fibre (A winning synergistic cocktail)
Scourzyme Application
Chemical Process 5450 382 1100 3480 331 Enzyme Process 3810 281 792 1200 238 Change
Impact Category
Energy consumption (MJ LHV) Global warming (kg CO2 eqv.) Acidification (g SO2 eqv.) Nutrient enrichment (g PO4 eqv.) Summer smog
Source: Novozyme
(g ethylene eqv.)
73 % 40 % 70 %
O W N
Bound FA
BRI
Vinyl guaiacol
Characterization
(SEM NIR NMR)
Biocomposites
IMI
Biochemicals
Thank You
Stay tuned (Whats hempening ?)*