From Waste to Energy – The Bio-Chemical Process

Biogas Compact Workshop

Project Planning
Postgraduate for
Programme
Renewable
Biodigesters in Developing
Energy (PPRE) and
Industrialized Countries
26 – 28 April, 2011
University of Oldenburg, Germany

Henri Spanjers
LeAF Lettinga Associates Foundation

Content

• History
• Definitions
• Biochemical processes
– Hydrolysis
– Acidogenesis
– Acetogenesis
– Methanogenesis
• Reactors

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 Natural Anaerobic Environments

20% O2 AIR 20% O2

Interface
8 ppm O2

8 ppm O2
Water

0 ppm O2

- organic matter + organic matter

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Carbon cycle
(CH2O)n
Photosynthesis: Respiration:
Algae Organic compounds plants, animals,
Green -plants microorganisms
Cyanobacteria

Methane-oxidizing
bacteria
Aerobic
CH4 (CO2)

Anaerobic
Methanogenic
bacteria
sedimentation
Anaerobic respiration
(Methyl Phototrophic
compounds) bacteria Fermentation

Organic compounds

(CH2O)n

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 Some history

First to report about natural methane production

Hey, there is a flammable

gas coming out of rotting
marshes and swamps!!!

ALESSANDRO VOLTA
Italy
1770

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Figure 1 Natural gas and the chemist. John Dalton (1766–1844) collecting marsh gas by poking a
stick into pond sediments (Picture by Ford Madox Brown). Marsh gas (methane, CH4, the
simplest alkane) is the main component of natural gas. As shown by the work of Zengler et al.
new aspects of this economically important bacterial process are still being revealed.

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 Fermentation

Fermentation: process of deriving energy from the oxidation

of organic compounds, such as carbohydrates, and using
an endogenous electron acceptor, usually an organic
compound
German chemist and zymologist, Eduard Buchner, winner of
the 1907 Nobel Prize in chemistry, determined that
fermentation is actually caused by a yeast secretion that
he termed zymase.

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Fermentation Products

• Microbial cells or biomass (single cell protein, bakers

yeast, lactobacillus, E. coli, etc.)
• Microbial enzymes: catalase, amylase, protease,
pectinase, glucose isomerase, cellulase, hemicellulase,
lipase, lactase, streptokinase, etc.
• Microbial metabolites:
– Primary metabolites – ethanol, citric acid, glutamic acid, lysine,
vitamins, polysaccharides etc.
– Secondary metabolites: all antibiotic fermentation
• Recombinant products: insulin, HBV, interferon, GCSF,
streptokinase
• Biotransformations: phenyl acetyl carbinol, steroid
biotransformation, etc.

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 Fermentation Products (cont’d)

• Alcoholic fermentation: sugars (glucose, fructose,

sucrose) are converted into Ethanol and carbon dioxide
• Dark fermentation is the fermentative conversion of
organic substrate to H2

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Anaerobic biodegradation

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 Anaerobic biodegradation

• Anaerobic biodegradation or anaerobic digestion: very

complex biological and biochemical process performed by
various different species of bacteria working together
• End products of anaerobic digestion are “biogas” and
more bacteria that grow out of the consumed organic
matter

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living organisms

Archaebacteria were the

first living organisms on
eukaryotes prokaryotes earth. Before there was
oxygen in the atmosphere!

archaebacteria eubacteria

Methanogenic
bacteria
extreme halophiles thermoacidophiles

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 Comparison Aerobic - Anaerobic

Characteristic Aerobic Anaerobic

Reaction C6H12O6 + 6O2 → 6CO2 + 6H2O C6H12O6 → 3CO2 + 3CH4

Energy release ∆G°’ = -2840 kJ/mol glucose ∆G°’ = -393 kJ/mol glucose

Carbon balance 50% → CO2 95% → CH4 + CO2 (= biogas)

50% → biomassa 5% → biomassa
Energy balance 60% → biomassa 90% retained in CH4
40% → heat production 5% → biomassa
5% → heat production
Biomass Fast growth of biomass, Slow growth of biomass
production Resulting in a sewage sludge
problem

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Anaerobic digestion

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 Scheme anaerobic biodegradation
Polymers
(proteins, polysaccharides, lipids)

h
Monomers
(sugars, amino acids, peptides)

1
Methanogenic 1 Propionate 1
butyrate
Consortium 2 2
H2 + CO2 acetate
3
4 4
CH4 + CO2

h Hydrolytic enzymes 3 Homoacetogenic bacteria

1 Fermentative bacteria 4 Methanogens
2 Syntrophic acetogenic bacteria
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From www.uasb.org
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ADM1 Model Structure Overview

Complex particulate waste and Inactive
biomass

Inert particulate

Carbohydr. Proteins Lipids

Inert soluble

Sugars Amino acids LCFA

1 2 3

Propionate HVa, HBu

5
4

Acetate H
2
Death
6 7

CH
4
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 Anaerobic Conversion of Organic Matter
Organic Polymers
proteins carbohydrates lipids
Hydrolysis
Hydrolytic enzymes
Mono- and oligomers
amino acids, sugars, fatty acids

Acidogenesis
Fermentative bacteria
Volatile Fatty Acids
Lactate
Ethanol
Acetogenesis
Syntrophic acetogenic bacteria

H2 / CO2 Acetate
Homoacetogenic bacteria

Methanogenesis
Methanogens
CH4 / CO2

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Hydrolysis
Organic Polymers
proteins carbohydrates lipids

Mono- and oligomers

amino acids, sugars, fatty acids

Volatile Fatty Acids

Lactate
Ethanol

H2 / CO2 Acetate
Homoacetogenic bacteria

CH4 / CO2

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 Hydrolysis: Characteristics

• Polymeric compounds  monomer or dimeric

components
• By extra-cellular enzymes
• Slow process (rate limiting): dS/dt = -Kh•S
• Retention time and particle size rate determining
• Optimum pH = 6
• Cellulose/hemicellulose degradation depends on lignin
fraction
• Hydrolysis of fats hardly proceeds <15-20°C (rate limit ing)
• (Product) inhibition by: LCFA. NH3, amino acids, H2?

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Hydrolysis: Enzymes

• Hydrolysis of suspended solids is surface-related process

• The more specific surface, the faster the process

• Individual enzymes work at constant rate (constant T, pH)

• Number of enzymes determines the total rate

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 Hydrolysis: Surface related

Hydrolysis as a surface-related process

More enzymes
“attack” the
substrate

Rate
increases

Particle
breakdown
or “lysis”

From: Wendy Sanders

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Hydrolytic enzymes

Long Chain Fatty Acids (LCFA)

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 Hydrolysis: Carbohydrates

• Cellulose is hydrolysed by cellulase (mixture of exo-

glucanases, endo-glucanases and cellobiases)
• The hydrolysis of starch is performed by a mixture of
amylases that is able to hydrolyse the α-1,4 bonds and α-
1,6 bonds of the amylose and amylopectin.

From Dr. Wendy Sanders

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Hydrolysis: Proteins

• Proteinases (Peptidases + Proteases)

• Protein  polypeptides  peptides  amino acids

From Dr. Wendy Sanders

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 Hydrolysis: Lipids

most lipids in waste(water) are present as triacylglycerides

From Dr. Wendy Sanders

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Bio-degradation of cellulitic matter versus lignin content

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 Digestible part of wood

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Acidogenesis
Organic Polymers
proteins carbohydrates lipids

Mono- and oligomers

amino acids, sugars, fatty acids

Volatile Fatty Acids

Lactate
Ethanol

H2 / CO2 Acetate

CH4 / CO2

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 Acidogenesis: Sugars
• Release of protons (H+) and reaction products (proton acceptors)

• H2 formation (catalyzed by the enzyme hydrogenase)

• Performed by a very large group of bacteria (about 1% of all bacteria

facultative fermenters)

C12H22O11 + 9 H2O → 4 CH3COO- + 4 HCO3- + 8 H+ + 8 H2

C12H22O11 + 5 H2O → 2 CH3CH2CH2COO- + 4 HCO3- + 6 H+ + 4 H2

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Acidogenesis: Sugars

End products depend on circumstances, e.g.:

• Glucose fermentation in a two-step system

– more reduced products like ethanol, lactate, propionate, butyrate,
CO2 and H2

• Glucose fermentation in a one-step system

– acetate, H2 and CO2

• Production of acids proceeds up to pH = 4

(product inhibition)

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 Acidogenesis of sugars: most rapid step!

Kinetic Properties Acidifiers / Methanogens

Process Rx Y Ks µ-max Td
gCOD/gVSS/d g VSS/g COD mg COD/l day-1 days

Acidogenesis 13 0.15 200 2.0 0.35

Methanogenesis 3 0.03 30 0.12 5.8

Overall 2 0.03 –0.18 - 0.12 5.8

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Substrate availability and bacterial growth

Monod’s equation:

S
µ = µmax ⋅ µ
Ks + S µmax

At S = Ks → µ = ½ µmax

Ks S

dX
= µ⋅X
dt
Where: dS/dt = substrate utilisation rate
dX dS
= −Y ⋅ Y = yield coefficient
dt dt

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 Acidogenesis: Acidification
Methane Poor
Capacity Buffering
Exceeded Capacity

Methanogenic Toxicity
VFA Increasing
increases

pH Unionized VFA
decreases increasing

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Acidogenesis: Proteins
• Organically bound N (amino acids) is released as NH4+
(Stickland reaction: oxidation-reduction)

Alanine: CH3CHNH2COO- + 3 H2O → CH3COO- + HCO3- + NH4+ + 2 H2

Glycine: 2 CH2NH2COO- + 2 H2 → 2 CH3COO- + 2 NH3

alanine + glycine + 3 H2O → 3 acetate + 2 NH3 + NH4+ + HCO3-

(2 NH3 + 2 H2O + 2 CO2 → 2 NH4+ + 2 HCO3-)

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 Stickland reaction

Alanine Glycine

4 e-
ATP
(ATP)

acetate, CO2, NH4 2 acetate, 2 NH4

Oxidative branch Reductive branch

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Acidogenesis: Long Chain Fatty Acids

• Anaerobic degradation of LCFA proceeds via β-oxidation

CH3-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-CH2-COO-

• Palmitic acid:
CH3-(CH2)14-COO- + 14 H2O → 8 CH3COO- + 7 H+ + 14 H2

• With uneven numbers: acetate + propionate are formed:

CH3-(CH2)14-CH2COO- + 14 H2O → 7 CH3COO- + CH3CH2COO- 7 H+
+ 14 H2

• Unsaturated LCFA are firstly hydrogenated before degradation

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 Acetogenesis
Organic Polymers
proteins carbohydrates lipids

Mono- and oligomers

amino acids, sugars, fatty acids

Volatile Fatty Acids

Lactate
Ethanol

H2 / CO2 Acetate
Homoacetogenic bacteria

CH4 / CO2

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Acetogenesis (Acetate formation)

• Conversion of fermentation products into acetic acid, CO2, and H2

• Mainly from propionic acid, butyric acid and ethanol

propionate- + 3H2O → acetate- + HCO3- + H+ + 3H2 ∆ G0’ = + 76.1 kJ/mole

butyrate- + 2H2O → 2 acetate- + H+ + 2H2 ∆ G0’ = + 48.1 kJ/mole

ethanol + 2H2O → acetate- + H+ + 2H2 ∆ G0’ = + 9.6 kJ/mole

_______________________________________________________________________________________________

4 H2 + CO2 → CH4 + 2H2O ∆ G0’ = -138.9 kJ/mole

Need for syntrophic associations !!!

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 Impact of pH2 on thermodynamics
[C ]c ⋅ [ D]d
∆G ' = ∆G '0 + RT ln
∆G’ (kJ/mole) [ A]a ⋅ [ B ]b
-100
propionate- + 3H2O → acetate- + HCO3- + H+ + 3H2

-50
Reaction butyrate- + 2H2O → 2 acetate- + H+ + 2H2
possible
Methanogenic niche
0
Reaction
impossible 4 H2 + CO2 → CH4 + 2H2O
50

2 4 6 8
pH2=-log (H2)
High H2 Low H2
pressure pressure
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Methanogenesis
Organic Polymers
proteins carbohydrates lipids

Mono- and oligomers

amino acids, sugars, fatty acids

Volatile Fatty Acids

Lactate
Ethanol

H2 / CO2 Acetate

CH4 / CO2

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 Methanogenesis

• Aceticlastic methanogenesis (70%):

CH3COOH → CH4 + CO2

• Hydrogenotrophic methanogenesis (30%):

CO2 + H2 → CH4 + CO2

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Methanogenesis: Substrates
∆G0 (kJ/mole CH4)
4H2 + CO2 => CH4 + 2H2O -130.4
4HCOOH => CH4 + 3CO2 + 2H2O -119.5
4CO + 2H2O => CH4 + 3CO2 -185.5
4CH3OH => 3CH4 + CO2 + 2H2O -103.0
CH3OH + H2 => CH4 + H2O -112.5
+
4CH3NH3 + 2 H2O => 3CH4 + CO2 + 4NH4 - 74.0
+
2(CH3) 2NH2 + 2H2O => 9CH4 + 3CO2 + 4NH4 - 74.0
CH3COOH => CH4 + CO2 - 32.5
Most important substrates: hydrogen and acetate
Furthermore: formate, carbon monoxyde, methanol and methylamines

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 Maximum Production of Biogas

1,40
production of biogas
production of methane
1,20

1,00

0,80
[Nm³/kg]

0,60

0,86
0,40

0,50
0,20 0,40

0,00
Lipids Carbohydrates Proteins (ATV-DVWK M 363)

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Animal manure Biogas Practice Area

Inorganic Organic

Phosphorous Nitrogenous Carbonaceous Sulphurous

Proteines Lipids Carbohydrates

Inositiol
phosphate,
phospholipids, Peptides
nucleic acids, Glycerol Sugars
ATP Fibers
Amino acids
Alcohols
Fatty acids
Sulphites

Compounds Volatile acids

of Cu, P, K,
Zn, Mn, Co, Inorganic Cellulose
Ca, Fe, H, O phosphates lignin
N, NH4 H2 O CH4 CO2 Hs S

Biochemical processes
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 Take-home message

• Anaerobic microbial conversion differs from aerobic

• Anaerobic digestion is a complex process

• Ultimate COD removal via production of CH4

• Anaerobic bacteria have a narrow substrate spectrum:

complex consortia are needed for complete COD removal

• Environmental factors affect the process

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Reactors

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 Different Technologies of Process Management

Water content in the wet (TS <= 10%)

reactor dry (TS = 30-35%)
Process Continuous Stirred
management Plug-Flow
Mesophilic (35-37ºC)
Temperature
Thermophilic (55-60ºC)
Single-stage
Steps of process
Multi-Stage
Semi-continuous
Operational mode
Discontinuous (Batch)
vertical (conventional, oval, etc.)
Form of reactor
horizontal
Agitation
Mixing process Circulation
Percolation

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Fixed Dome Domestic Digester

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 Digester: Schematic
Digester with rubber membrane cover > 50 % of
all digesters

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Biogas plant in the UK

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 Development of “high-rate” anaerobic treatment systems

Completely mixed Immobilised Enhanced

Physical retention
biomass contact
(Bio)gas

influent effluent

Relative
Relative
capacity: 1 Relative
capacity: 5
capacity: 25

Relative
capacity: 75

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UASB and EGSB

Auto immobilization / granulation EGSB

UASB

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 Reactor Technologies for Liquids
gas gas
effluent
effluent
gas

effluent
second stage sludge bed

first stage influent

influent influent
UASB-Reactor IC-Reactor Biobed-Reactor
gas
gas gas
effluent effluent
effluent
recirculation

recirculation

recirculation
(loop)

(loop)

influent influent
influent

Anaerobic Contact Reactor Fixed bed Reactor Fluidized bed Reactor

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UASB Reactor: Sewage

Bucaramanga, Colombia, 12000 m3/d

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 UASB reactors: Sewage

Mirzapur, India, 14 m3/d plant

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UASB: Sewage

Accra, Ghana

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 Anaerobic Industrial Wastewater Treatment
Anaerobic UASB-Reactor CSM

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IC-Reactor: Distillery Hanover

(Kraul & Wilkening u. Stelling)

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