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- ChitoBioEngineering “Metabolic and Enzyme Engineering
for the Biotechnological Production
of Partially Acetylated Chitosans”
Pre-Final Report
Bruno Moerschbacher . [email protected]
Westfälische Wilhelms-Universität . Münster . Germany
A collaborative research project supported by the ERA-NET Scheme of the 6th EU Framework Programme
partially acetylated chitosans
plant protection
GlcN
Partner:
GlcNAc
scar-free
wound healing
Bruno Moerschbacher . WWU-IBBP
structure-function relationships of chitosans
Partner:
chitosan matrix
GlcNAc
GlcN
Partner:
chemical generation of chitosans
GlcNAc
1000
GlcN
100
DP
random
10
DP
1
DA
0
25
50
75
100
DA
Partner:
enzymatic modification of chitosans
GlcNAc
1000
DP
GlcN
100
DP
random
10
1
0
25
50
75
blockwise
PA
100
regular
patterned
0
25
50
DA
75
100
DA
Partner:
knowledge-based and un-biased discovery programs
endophytic fungi
from tropical trees
metagenomics
from soil samples
screening assays
enzymatic
fingerprinting
novel chitosan
modifying enzymes
HNO2
mass-spectrometric
sequencing
bioinformatic
modeling
optimisation by
protein
engineering
Partner:
chitosans
with non-random PA
plant disease
protection
scar-free
wound healing
nanoparticles for
drug delivery
knowledge-based chitosans
CuChiBCA
disease
resistance
&
stress
tolerance
&
yield
increase
seed coating
(CaraSeed)
&
foliar spray
(CaraPlant)
Partner:
next generation chitosans
CuChiBCA
agriculture
Partner:
next generation chitosans
biotechnological
monoclonal
chitosans
CuChiBCA
agriculture
Partner:
cosmetics
veterinary medicine
ChitoBioEngineering
project aim
to biotechnologically produce partially acetylated
chitosan oligomers with fully defined architecture,
i.e. - known degree of polymerisation (DP)
- known degree of acetylation (DA)
- known pattern of acetylation (PA)
for detailed analysis of structure-function relationships
and, hence, reliable bio-activities and applications
general project approach
by expressing and optimising known and novel chitin
synthase and chitin deacetylase genes in suitable host
strains, metabolically engineered for maximum yield
Partner:
role of academic partners
genetic engineering of suitable host
strains over-expressing known and novel
chitin synthases and chitin deacetylases
protein engineering of chitin synthases
and chitin deacetylases to produce fully
defined chitosan oligomers
metabolic engineering of host strains
for maximum yield of fully defined
chitosan oligomers
Partner:
role of industrial partners
technology transfer and up-scaling
of fermentation from the lab scale
to the pilot scale
technology transfer and co-ordination of
trial applications with partners in the
cosmetics & veterinary medicine industry
Partner:
state of the art
chitin
chitin
deaceteylase
synthase
nodB chitosan
UDP- nodC
chitin
GlcNAc
pentamer
pentamer
GlcN
GlcNAc
Partner:
ChitoBioEngineering strategies
strain
enzyme
pathway
optimisation
UDPGlcNAc
CS
chitin
oligomers
chitin
synthases
CDA
chitosan
oligomers
chitin
deacetylases
GlcN
GlcNAc
Partner:
ChitoBioEngineering targets
different DPs
different DAs
UDPGlcNAc
CS
CDA
chitin
oligomers
chitosan
oligomers
different PAs
GlcN
GlcNAc
Partner:
targets reached at mid-term (in vivo)
GlcN
GlcNAc
Partner:
targets reached at mid-term (in vivo & in vitro)
GlcN
GlcNAc
Partner:
targets reached pre-final (in vivo & in vitro)
dimers: 4 of 4
trimers: 5 of 8
GlcN
GlcNAc
Partner:
tetramers: 12 of 16
GlcN
GlcNAc
Partner:
pentamers
GlcN
GlcNAc
Partner:
yields reached pre-final (in vivo & in vitro)
pentamers
100 L fermenter
ca. 1 kg
ca. 99 % pure
!
Megazyme >ca. 95 % pure
20 mg = 200 €
in vitro deacetylation
ca. 50 mg each
ca. 99 % pure
GlcN
GlcNAc
Partner:
bio-assays for plant protection
„pathogen !“
(= „stopp all activities,
and defend yourself“)
„pathogen ?“
(= „keep going, but
prepare for defense“)
Partner:
resistance reaction [H2O2] / µM
addition of COS
as a priming inducer
bio-assays for cellular priming
+
2 µM priming active COS
+
2 µM chitin hexamer
200 µM priming inactive COS
-3h
time after elicitation / min
addition of chitin hexamer as an elicitor
Partner:
ChitoBioEngineering progress
X 11
VII 11
IV 11
II 14
Belgium
Spain
Germany
tasks for the cost-neutral extension
Belgium
down-stream processing of partially acetylated oligomers
Spain
more oligomer diversity from deacetylase chimeras
Germany more bio-assays and application tests
Partner:
CuChiBCA
nano & micro
nano & CCME
PA
&
CCME
DP & DA
monoclonal
polymers
Partner:
monoclonal
oligomers
thank you
University Ghent
Prof Dr. Wim Soetaert
Prof. Dr. Marjan De Mey
Dr. Jo Maertens
Dr. Joeri Beauprez
Pieter Coussement
Wouter Van Bellegem
University Münster
Prof Dr. Bruno Moerschbacher
Dr. Stephan Kolkenbrock
Shoa Naqvi
Stefanie Hamer
Stefan Cord-Landwehr
Heppe Medical Chitosan
Katja Richter
Torsten Richter
University Ramon Llull Barcelona
Prof. Dr. Antoni Planas
Dr. Eduardo Andrés
Dr. Xevi Biarnés
Xavi Pézez
www.chitobio
Partner:
BioBase Europe Pilot Plant
Prof. Dr. Wim Soetaert
Dr. Hendrik Waegeman
Kristof Callemin
Diedrik Snoeck
Georges Moureau
An Vermeulen
Jeroen De Smet
engineering.eu

Results of ChitoBioEngineering Presentation (click here)

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