1Department of Chemistry/Mass Spectrometry Laboratory – University of Liège, B-4000 Liège (Sart Tilman), Belgium
2Department of Chemistry/Organic and Biological Analytical Chemistry - University of Liège, B-4000 Liège (Sart Tilman), Belgium 3Department of Functional and Evolutionary Entomology - Gembloux Agro-Bio Tech, University of Liège, B-5030 Gembloux, Belgium 4Department of Bio-Industries - Gembloux Agro-Bio Tech, University of Liège, B-5030 Gembloux, Belgium
5Department of Microbiology and Genomics - Gembloux Agro-Bio Tech, University of Liège, B-5030 Gembloux, Belgium
TERMITES AS A TOOL TO IMPROVE LIGNOCELLULOSE BIOMASS
VALORIZATION: ANALYSIS OF INTERMEDIATE OLIGOSACCHARIDES
PRODUCED BY RETICULITERMES SANTONENSIS
Use of termites as a tool to improve lignocellulose biomass valorization:
study of enzymatic complex in termites and its common symbionts by
proteomic, genomic and metabolomic approaches.
TERMITOFUEL PROJECT :
PARTNERS :
- Bio-Industries Unit (BIO) MICROBIOLOGY
- Functional and Evolutionary Entomology Unit (EFE) PROTEOMIC
- Microbiology and Genomics Unit (BAM) GENOMIC
Mass Spectrometry Unit (LSM) METABOLOMIC
WHY TERMITES?
Efficient System from :
- Termites?
- Symbionts?
SYNERGY?
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 N u m b e r o f s u b s t it u e n t s Degree of polymerization Trichoderma reesei T-gut extract TFA 2MMass spectra of CM-oligomers obtained from acid or enzymatic hydrolysis of carboxymethylcellullose (CMC) :
1199.5681419.626 979.522 1639.673 1535.638 1303.598 1755.701 1801.714 1859.733 2079.786 2021.763 2299.833 0.0 0.2 0.4 0.6 0.8 1.0 4 x10 Int en s. [ a.u .] 1000 1250 1500 1750 2000 2250 2500 2750 m /z 0 2000 4000 6000 8000 Inte ns. [a. u.] 1000 1500 2000 2500 3000 3500 m /z Acid hydrolysis Enzymatic hydrolysis
The hydrolysis products are classified according to their degree of polymerization and number of substituents. Specific enzymatic or acid hydrolysis patterns are reported and compared. CM-oligomers are produced when testing gut extracts of
Reticulitermes santonensis
but higher selectivity is observed compared to the pattern obtained with a commercial cellulase of
Trichoderma reesei, used as
reference.
MALDI-TOFMS ANALYSIS:
Hydrolysis of crystalline cellulose substrates were tested with termite gut extracts and no cellodextrins were detected by MALDI-TOFMS although hydrolysis profiles were obtained with CMC in the same conditions. CMC is a derivative soluble substrate that has been preferably used for termite studies, mostly because of its high solubility. However, CMC is not well representative of naturally occurring cellulose. The use of derivative cellulose substrates is appropriate to recognize an enzymatic activity and evaluate its selectivity which should be of interest for specific applications
Cello3-Na+ B A x105 x105 5.0 1.0 0.5 0.1 I ( a. u.) 5.0 0.1 1.0 3.0 2.0 4.0 527.0 527.0 689.1 689.1 851.2 851.2 1013.2 1175.3 1337.3 1499.3 Cello3-Na+ Cello4-Na+ Cello4-Na+ Cello5-Na+ Cello5-Na+ Cello6-Na+ Cello7-Na+ Cello8-Na+ Cello9-Na+ m/z 569.0* 701.1* * 833.2 965.2* α- Cellulose Avicel® Mass spectra of cellodextrins obtained from T. reesei cellulase hydrolysis of cellulose powders:
x103 I (a. u .) 1.5 0.5 1.0 m/z 701.1 833.2 569.0 965.2 1097.3 1229.3 1361.4 X y lo4 -Na + X y lo5 -Na + X y lo6 -Na + X y lo7 -Na + 61 71 81 31 41 51 91 101 111 X y lo8 -Na + X y lo9 -Na + X y lo10 -Na +
Xylon-Na+ MeGlcA
Mass spectrum of xylo-oligosaccharides obtained from Reticulitermes santonensis hydrolysis of Xylan:
(DeBoy et al. 2008)
Enzymatic activity on xylan seems to be the predominant source of oligosaccharides from termites. Xylooligosaccharides production is of interest for industrial applications. Moreover, analysis of intermediate degradation products is important when studying a cellulolytic activity as some compounds can be involved in retro-inhibition mechanisms, undergo post cleavage modification or induce enzyme gene expression. This study is of interest for comprehension of the mechanisms involved and identification of specific enzymes.
