Chapter 2. Biochemistry of Anaerobic Digestion. Anaerobic Digestion
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1 Chapter Biochemistry of Anaerobic Digestion Anaerobic Digestion Complex Organics (Carbohydrates, proteins, lipids) Mono and Oligomers (sugars, aminoacids, longchained fatty acids) Intermediates 3 3 (Propionate, butyrate, alcohols). Hydrolysis. Fermentation 3. Acetogenesis 4. Methanogenesis 3 Acetate 4 CH 4 4
2 Hydrolysis Solubilization of insoluble particles and biological decomposition of organic polymers to monomers or dimers, which can pass through cell membrane. Usually carried out by extracellular enzymes. It is not necessarily an enzymatic process catalyzed by biologically produced enzymes but could take place due to physico-chemical reactions as well. Hydrolysis of a complex, insoluble substrate depends on different parameters such as; (i) Particle size (ii) ph (iii) production of enzymes and (iv) diffusion and adsorption of enzymes to particles Hydrolysis Substrate Carbohydrates Cellulose Proteins Lipids Hydrolysis rate, d Ref. Christ O,Wilderer PA,Angerhofer R,Faulstich M (000) Water Sci Technol 4:6 Gujer W, Zehnder AJB (983) Water Sci Technol 5:7
3 Fermentation (Acidogenesis) Dissolved organic matter is biodegraded mainly to volatile fatty acids (VFAs) and alcohols by a heterogeneous microbial population. Dominant species in anaerobic digesters are bacteria while small populations of protozoa, fungi and yeasts have also been reported. It is mainly the obligatory and facultative anaerobic bacteria that carry out the fermentation. Most important factors that influence the fermentation are; (i) interspecies hydrogen transfer (ii) ph (iii) hydraulic retention time (iv) previous acclimation of the anaerobic culture Acetogenesis Oxidation of fermentation products into a substrate (acetate, and CO ) appropriate for methanogens. Homoacetogenesis: Production of acetate as a sole end product from CO and. Thermodynamically, it is less favorable than methanogenesis and sulfate reduction. Synthrophic Acetogenesis: Anaerobic oxidation of propionate and butyrate to acetate and. Propionate and butyrate oxiditation are inhibited by even low partial pressures, therefore can occur only if is consuming by methanogens, SRB and homoacetogens. 3
4 Acetogenesis Syntrophic Acetogenesis Propionate + 3 O Acetate + HCO 3 + H G 0 * = +76. kj/mol substrate Butyrate + O Acetate + H + + Consumed G 0 * = kj/mol Homoacetogenesis 4 + HCO 3 + H + Acetate + 4 O G 0 * = kj/mol * Thauer RK, Jungermann K, Decker K (977) Bacteriol Rev 4:00 Syntrophic Acetogenesis Interspecies transfer CH 4 VFA oxidizing bacteria d utilizing methanogen propionate, butyrate, etc. Distance d is critical The lower the concentration the better are the thermodynamics of the VFA degradation 4
5 Methanogenesis A limited number of organic compounds are used as carbon and energy sources in methanogenesis. They are; CO, CO, formic and acetic acid, methanol, methylamines and dimethyl sulfide. Almost 65-70% of CH 4 produced in anaerobic digesters comes from acetate. Methanogenesis from CO and has a significant role as well by keeping a low hydrogen pressure and thus supporting the anaerobic oxidation VFAs to acetate &. Methanogenesis is extremely sensitive to temperature, loading rate and ph fluctuations and inhibited by a number of organic & inorganic compounds. Methanogenesis 4 + HCO 3 + H + CH O G 0 = kj/mol substrate Acetate + O CH 4 + HCO 3 G 0 = -3.3 kj/mol Methanol ¾ CH 4 + ¼ HCO 3 + ¼ H + + ¼ O G 0 = kj/mol Formate - + H + ¼ CH 4 + ¾ CO + ½ O G 0 = -36. kj/mol * Thauer RK, Jungermann K, Decker K (977) Bacteriol Rev 4:00 5
6 AD with Sulfate Reduction Complex Organics (Carbohydrates, proteins, lipids) Monosaccharides, Amino acids Intermediates 3 3 (Propionate, butyrate, alcohols) 3 Higher Fatty Acids, Alcohols Acetate 5. Hydrolysis. Fermentation 3. Acetogenesis 4. Methanogenesis 5. Sulfate reduction 4 CH S Sulfate Reduction Sulfate (SO 4 - ) or sulfite (SO 3 - ) can be used by SRB as acceptor of electrons released during the oxidation of organic materials under anaerobic conditions. The end product is hydrogen sulfide ( S). VFAs, several aromatic acids,, methanol, ethanol, glycerol, sugars, amino acids and some phenol compounds are the substrates used in sulfate reduction. There is a competition for the substrate available to be used in sulfate reduction instead of fermentation (sugars), acetogenesis (VFAs) and methanogenesis (acetate, ). SO 4 - /COD ratio is the critical parameter. 6
7 Sulfate Reduction 4 + SO 4 + H + HS + 4 O G 0 = -5.9 kj/mol substrate Acetate + SO 4 HCO 3 + HS G 0 = kj/mol Propionate + ¾ SO 4 Acetate + HCO 3 + ¾ HS + ¼ H + G 0 = kj/mol Butyrate + ½ SO 4 Acetate + ½ HS + ½ H + G 0 = -7.8 kj/mol Lactate + ½ SO 4 Acetate + HCO 3 + ½ HS + ½ H + G 0 = kj/mol Ethanol + ½ SO 4 Acetate + ½ HS + ½ H + + O G 0 = kj/mol * Thauer RK, Jungermann K, Decker K (977) Bacteriol Rev 4:00 Anaerobic Digestion Complex Organic Materials 9% 30% 5% % Intermediates (propionate, butyrate, etc.) 9% Acetate 30% 70% CH 4 Carbon flow in anaerobic environments with active methanogens 7
8 Anaerobic Digestion Complex Organic Materials 0-30% 50-70% 0-30% Intermediates (propionate, butyrate, etc.) Acetate X: Inhibited because of high partial pressure Carbon flow in anaerobic environments without active methanogens Syntrophic Acetate Conversion Complex Organics (Carbohydrates, proteins, lipids) Mono and Oligomers (sugars, aminoacids, longchained fatty acids) Intermediates 3 3 (Propionate, butyrate, alcohols). Hydrolysis. Fermentation 3. Acetogenesis 4. Methanogenesis 3 Acetate 4 CH 4 8
9 Syntrophic Acetate Conversion When acetate-utilizing methanogens are inhibited by high concentrations of ammonia or sulfite, other groups of microorganisms replace them to obtain energy from oxidation of acetate to & CO. Due to thermodynamic constrains this reaction proceeds much better at temperatures higher than 60 C (upper limit of thermophilic acetate-utilizing methanogens) and is the way of acetate transformation. Both syntrophic acetate oxidation and methanogenesis from acetate can be simultaneously occur in an AD. Biochemical Pathways Lipids Particulate Proteins Particulate Carbohydrates Long Chain Fatty Acids Soluble Proteins Simple Sugars Acetate Valerate Butyrate Acetate Lactate Propionate Acetate CH 4 9
10 AD of Sewage Sludge AD of Lipids & Proteins 0
11 AD with Sulfate Reduction
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