SLIDE 1
Yves Verhertbruggen
October 2014
Agreenskills Annual Meeting - Toulouse
The biochemical and physiological function of mannan and CELLULOSE - - PowerPoint PPT Presentation
The biochemical and physiological function of mannan and CELLULOSE - - PowerPoint PPT Presentation
The biochemical and physiological function of mannan and CELLULOSE SYNTHASE LIKE D2 protein in Triticum endosperm Yves Verhertbruggen October 2014 Agreenskills Annual Meeting - Toulouse Who? Where? How? What? What? What? Context Albumen
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SLIDE 3
Context
Outer layers 14% Albumen 83% Embryo 3%
65-70 % feruloylated arabinoxylan (AX) 20-25 % (1-3)(1-4)-β-D Glucan (MLG)
- ca. 7 % Mannan
2-4 % Cellulose Trace of pectin (HG, RGI, RGII)
???
Trace of Xyloglucan (XG)
SLIDE 4
Defining:
- 1. The structure
- 2. The biological function
- 3. The biosynthesis of mannan
in wheat endosperm Aims of the project
Which one(s) ???
SLIDE 5
Defining:
- 1. The structure
- 2. The biological function
- 3. The biosynthesis of mannan
in wheat endosperm Aims of the project
150°J 350°J
aleurone Albumen amylacé
Why ???
Mannan occurrence
SLIDE 6
Defining:
- 1. The structure
- 2. The biological function
- 3. The biosynthesis of mannan
in wheat endosperm Aims of the project
GDP-D-mannose Man-(Man)n-Man
mannan β-(1→4)- mannosyltransferase
GDP Man-Man-(Man)n-Man
How ???
SLIDE 7
- 1. Biochemistry
Methodology
HPLC/GC MS
Enzymatic fingerprint
- 100000
100000 200000 300000 400000 500000 5 10 15 20 25 30 35 Tim e ( m in) Xylanase (eau) betaglucanase (eau)
X1 X2 DP4 DP5 DP6 DP9 DP7
G3 G4
- 100000
100000 200000 300000 400000 500000 5 10 15 20 25 30 35 Tim e ( m in) Xylanase (eau) betaglucanase (eau)
X1 X2 DP4 DP5 DP6 DP9 DP7 X1 X2 DP4 DP4 DP5 DP5 DP6 DP6 DP9 DP9 DP7 DP7
G3 G4 G3 G4 NaOAc 1M: 0.3% Flow: 0.4 ml/min Absorbance (nC)
Mono mannose Mannobiose Mannotriose
Time of retention (min)
Wheat sample treated with a mannanase Mannan extract treated with a mannanase
NaOAc 1M: 0.3% Flow: 0.4 ml/min Time of retention (min) Absorbance (nC)
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- 2. Molecular biology
Methodology
Collaboration with R. Mitchell (Rothamsted Research, United Kingdom)
TaCSLD2 RNAi transformant
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- 2. Molecular biology
Methodology
Test of enzymatic activity
Transient expression in Nicotiana benthamiana
Activity (% of control)
400 300 200 100
Endogenous acceptors
- 3. Preparation of
microsomes and affinity purification of GT
- 4. Incubation with donors
and acceptors and detection of potential products
SLIDE 10
- 1. Milling and food industry
- 2. Biofuel industry
- 3. « How to feed a growing global population? »
Impact
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Come and ask me
For more information
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Cell wall synthesis
T h a n k y
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