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Table of contents
Chapter 1. Literature review
1.1 ANAEROBIC DIGESTION
1.1.1 A four steps biological reaction
1.1.2 Anaerobic digestion of lignocellulosic crop residues
1.1.2.1 Crop residues: an important resource
1.1.2.2 Main constituents of lignocellulosic biomass
1.1.2.3 Other possibilities for energy recovery from crop residues
1.1.2.4 A solid substrate adapted to Solid-State anerobic digestion
1.1.3 Solid-State Anaerobic Digestion: definition, advantages and limits
1.1.4 Main parameters influencing anaerobic digestion
1.1.4.1 Temperature
1.1.4.2 Mixing
1.1.4.3 pH and alkalinity
1.1.4.4 Carbon/Nitrogen (C/N) ratio
1.1.4.5 Nutrient supplementation
1.1.4.6 Inhibitors
1.1.4.7 Particle size of the substrate
1.1.4.8 Inoculum
1.1.5 Anaerobic digestion processes
1.1.5.1 Biochemical Methane potential (BMP)
1.1.5.2 SSAD processes
1.2 PRETREATMENTS FOR ANAEROBIC DIGESTION
1.2.1 Non-biological pretreatments
1.2.2 Biological treatments
1.3 ENZYMATIC MECHANISMS OF WRF AND CULTURE PARAMETERS
1.3.1 Enzymatic degradation by WRF and Solid-State Fermentation (SSF)
1.3.2 Cultivation parameters for Solid-State Fermentation (SSF)
1.4 QUANTIFICATION OF ANAEROBIC DIGESTIBILITY IMPROVEMENT AFTER WRF RETREATMENT
1.4.1 Anaerobic digestion studies with WRF pretreatment
1.4.1.1 Nitrogen metabolism for anaerobic digestion of fungal pretreated substrates
1.4.1.2 Slight acidification of pretreated substrates
1.4.2 In Vitro Dry Matter Digestibility (IVDMD) studies
1.5 PARAMETERS OF LIGNOCELLULOSIC BIOMASS INFLUENCING ITS ANAEROBIC DEGRADABILITY
1.5.1 Polymers composition and enzymatic accessibility
1.5.2 The lignin-carbohydrates complex (LCC): linkages and derived compounds
1.5.2.1 Major lignin linkages in cell-wall: ferulic acid (FA) and p-coumaric acid (pCA)
1.5.2.2 Anaerobic degradability of FA and pCA
1.5.2.3 Other cell-wall linkages in anaerobic digestion
1.5.2.4 Lignin-derived compounds
1.5.3 Influence of histology
1.5.3.1 Epidermis role and composition
1.5.3.2 Anaerobic digestion at cell scale
1.6 QUALITATIVE EFFECTS OF FUNGAL PRETREATMENT ON ANAEROBIC DIGESTIBILITY
1.6.1 Modifications of lignin: G/S (guaiacyl/synapyl) ratio
1.6.2 Cellulose digestibility improvement
1.6.3 Changes in hemicelluloses X/A (xylose/arabinose) ratio and acetate
1.6.4 Increase of porosity
1.6.5 Actions of WRF on lignin-carbohydrates complex (LCC) linkages
1.6.6 Release of lignocellulosic polymers main constituents
1.6.6.1 Lignin-derived compounds
1.6.6.2 Hemicelluloses and cellulose derived compounds
1.6.7 Influence of histology: fungal degradation at cell scale
1.7 PYROLYSIS-GAZ CHROMATOGRAPHY-MASS SPECTROPHOTOMETRY (PY-GC-MS): A POWERFUL TOOL FOR STRUCTURE ANALYSIS
1.7.1 Definition and interest
1.7.2 Pyrolysis products (pyrolysates)
1.7.2.1 Formation mechanisms
1.7.2.2 Parameters influencing pyrolysates yield and nature
1.7.3 Various existing technologies for Py-GC-MS
1.7.4 Py-GC-MS compared to chemical methods to study lignin structure
1.8 CONCLUSION
Chapter 2. Main experimental strategy
2.1 MATERIAL AND METHOD COMMON TO ALL RESULTS CHAPTERS: MATERIAL ORIGIN AND BMP TESTS
2.1.1 Fungal strains
2.1.2 Wheat straw
2.1.3 Cultivation of Polyporus brumalis BRFM 985 in liquid medium
2.1.4 Total Solids and Volatile Solids determination
2.1.5 Biochemical Methane Potential (BMP) measurements: general case
2.2 OVERVIEW OF ANALYSED SAMPLES
2.3 MATERIAL AND METHODS SPECIFIC TO RESULTS OVERVIEW (CHAPTER 7)
2.3.1 Contamination problem
2.3.1.1 Pilot-reactor cleaning verification
2.3.1.2 Wheat straw microflora
2.3.1.3 BMP-tests
2.3.2 Enzyme assays
2.3.2.1 Glycoside Hydrolases (GH) activities
2.3.2.2 Ligninolytic activities
2.3.3 Wheat straw compositional analyses
2.3.3.1 Data obtained for ethanol production study and PCA
2.3.3.2 Fourier Transform InfraRed (FTIR) spectroscopy
2.3.3.3 Crystallinity measurement with X-Ray Diffractometry (XRD)
Chapter 3. White-rot Fungi selection for pretreatment of lignocellulosic biomass for anaerobic digestion and impact of glucose supplementation
3.1 INTRODUCTION
3.2 MATERIAL AND METHODS SPECIFIC TO SELECTION STEP
3.2.1 Fungal pretreatment
3.2.1.1 SSF in 24-well plates
3.2.1.2 SSF in columns
3.2.2 Anaerobic digestion: samples characterization and normalization of BMP results
3.2.2.1 Total Carbon (TC) measurements for pretreated samples in columns
3.2.2.2 BMP-tests
3.2.2.3 Normalization of BMP results for samples pretreated in deep-well
3.2.3 Impact of nutrient solution amount
3.2.3.1 SSF in deep well
3.2.3.2 Acid hydrolysis (Klason lignin, cellulose and hemicelluloses)
3.2.3.3 Determination of residual glucose
3.3 BMP IMPROVEMENT
3.3.1 Straw pretreated in 24-well plates
3.3.2 Straw pretreated in SSF columns
3.3.3 Pretreatments comparison
3.4 INFLUENCE OF STARTER SOLUTION ON PRETREATMENT
3.4.1 Fate of the nutrient solution during the pretreatment
3.4.2 Nutrient solution and wheat straw composition
3.4.2.1 Limited delignification
3.4.2.2 Carbohydrates consumption
3.5 CONCLUSION
Chapter 4. Influence of Polyporus brumalis culture parameters for the fungal pretreatment: optimization step
4.1 INTRODUCTION
4.2 MATERIAL AND METHODS SPECIFIC TO OPTIMIZATION STEP
4.2.1 Fungal pretreatment for anaerobic digestion
4.2.1.1 SSF in columns
4.2.1.2 BMP-tests
4.2.2 Experimental design
4.2.2.1 Building of the experimental design
4.2.2.2 Analyze of the experimental design and response variables
4.2.3 Substrate characterization
4.2.3.1 Dry matter losses after pretreatment
4.2.3.2 Klason lignin, cellulose and hemicelluloses determination by acid hydrolysis
4.2.3.3 Lignin losses in pretreated samples compared to Non inoculated Controls (NIC)
4.2.3.4 Soluble sugars
4.2.3.5 Enzymatic hydrolysis of cellulose: glucose yield
4.2.3.6 Real Time Quantitative Polymerase Chain Reaction (qPCR)
4.3 RELATIONSHIPS BETWEEN ANAEROBIC DIGESTION AND SUBSTRATE CHARACTERISTICS
4.3.1 Methane production results during BMP tests
4.3.2 Parameters influencing anaerobic degradability
4.3.3 Negative impact of lignin amount on methane production
4.3.4 Influence of fungal mycelium on CH4 production
4.3.5 Contribution of cellulose degradation to methane production
4.4 RESPONSE SURFACES FOR METHANE PRODUCTION
4.4.1 Methane production after 6 days
4.4.2 Methane production after 57 days
4.4.2.1 Samples pretreated with metals addition
4.4.2.2 Samples pretreated without metals addition
4.4.3 BMP
4.5 CONCLUSION
Chapter 5. Pyrolysis-GC-MS study of White-Rot Fungi pretreated wheat straws for anaerobic digestion
5.1 INTRODUCTION
5.2 MATERIAL AND METHODS SPECIFIC TO PY-GC-MS STUDY OF PRETREATED STRAWS
5.2.1 Fungal pretreated samples
5.2.2 Pyrolysis-GC-MS
5.2.2.1 Spectra obtaining
5.2.2.2 Spectrum analysis and interpretation
5.2.3 Characterization of fungal pretreated straws for anaerobic digestion
5.2.3.1 Acid hydrolysis (Klason lignin, cellulose and hemicelluloses) with NREL method
5.2.3.2 Qpcr
5.2.3.3 BMP-tests
5.2.4 Statistical analysis
5.2.4.1 Analysis of Variances (ANOVA)
5.2.4.2 Principal Component Analysis (PCA)
5.3 ORIGIN OF THE MAIN COMPOUNDS FOUND IN PYROLYSATES
5.3.1 Fungus characterization
5.3.1.1 N-containing (N) and unspecific compounds (UN)
5.3.1.2 Polysaccharide-derived products (PS)
5.3.1.3 Possible presence of small lignin amount
5.3.2 Straws characterization
5.3.2.1 Lignin derived pyrolysis products
5.3.2.2 Polysaccharide-derived products
5.3.2.3 N-containing compounds
5.4 COMPARISON BETWEEN PY-GC-MS DATA, OTHER CHARACTERIZATION TECHNICS AND ANAEROBIC DIGESTIBILITY
5.4.1 Variations of the S/G ratio
5.4.2 PCA to study relationships between samples characteristics
5.4.2.1 qPCR variable opposed to straw composition variables in the plan 1-2
5.4.2.2 Correlation between BMP and PS/LIG-pyr or PS/LIG NREL in the plan 1-3
5.4.3 Fungal biomass (qPCR variable)
5.4.4 Polysaccharides (PS)/Lignin (LIG) ratio determined by py-GC-MS
5.5 CONCLUSION
Chapter 6. Solid-State Anaerobic Digestion of wheat straw: impact of substrate/inoculum ratio and of fungal pretreatment
6.1 INTRODUCTION
6.2 MATERIAL AND METHODS SPECIFIC TO SSAD IN BATCH REACTORS (CHAPTER 6)
6.2.1 Wheat straw pretreatment in a pilot-reactor
6.2.2 Solid state Anerobic Digestion in batch
6.2.2.1 SSAD reactors design
6.2.2.2 Experiment I: tests of several S/I
6.2.3 Experiment II: fungal pretreatment for SSAD
6.2.3.1 Substrates and inoculum
6.2.3.2 Experimental configuration
6.2.4 Analytical methods
6.2.4.1 SSAD monitoring parameters: leachate pH, VFA, alkalinity and biogas composition
6.2.4.2 BMP-tests
6.2.4.3 Total Kjeldahl Nitrogen (TKN)
6.2.4.4 Ammonium concentration in final leachate
6.2.4.5 Final Total Carbon (TC) in digestate
6.2.4.6 Analysis of Variances (ANOVA)
6.3 EXPERIMENT I: TESTS OF SEVERAL S/I
6.3.1 Anaerobic digestion of wheat straw: start-up phase progress
6.3.2 TVFA/alkalinity as process stability indicator
6.4 EXPERIMENT II: FUNGAL PRETREATMENT FOR SSAD
6.4.1 Substrates anaerobic digestibility
6.4.2 SSAD start-up phase
6.4.3 Methane production of fungal pretreated straw in SSAD
6.5 CONCLUSION
