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Biomass Energy I Biology of methanogenesis

Knowledge of the fundamental processes

Anaerobic digestion (AD) provides an ef-
involved in methane fermentation is nec-
fective method for turning residues from
essary for planning, building and operat-
PPRE / EUREC-REMA livestock farming, agri-residues and food
ing biogas plants.
Winter term 2009/10 processing industries into:

Anaerobic digestion involves the activities

of three different bacterial communities:

• Biogas (rich in methane) which can • Fermentative bacteria (hydrolysis),
be used to generate heat and/or elec-
• Acetogenic bacteria (acidification &
• Fibre which can be used as a nutrient- acetogenesis),
• What is ANAEROBIC DIGESTION? rich soil conditioner, and
• Role of Microrganisms
• Parameters that influence Methana- • Liquor which can be used as liquid fer- • Methanogenic bacteria (methanogen-
tion tiliser esis).
• Basic Concepts of Biogas Plants

Biogas and the global carbon cycle

The process of biogas-production depends
• 590-880 million tons of methane per on various parameters.
year are released worldwide into the at-
mosphere through uncontrolled microbial For example, changes in ambient temper-
activity. ature or pH can have a negative effect on
bacterial activity.
• About 90% of the emitted methane de-
rives from biogenic sources, i.e. from the
decomposition of biomass.

Anaerobic digestion −→a complex process:

• The rest is of fossil origin (e.g. petro-

chemical processes, but also release of
reservoirs through climatic change).
• conditions that encourage the
natural breakdown of organic The tropospheric methane concentration
matter amounts to about 1.65 ppm (methane is
• bacteria working in the absence a very effective greenhouse gas).
of air (oxygen).

Anaerobic digestion is a series of processes Hydrolysis (Stage I)

The introduction of Anaerobic Digestion
in which microorganisms break down bio-
as a technology can thus
degradable material in the absence of oxy- Organic matter is dissolved externally by
gen. extracellular enzymes (cellulase, amylase,
• help farmers to reduce fossil fuels com- protease and lipase).
The digestion process begins with bacte- sumption, and
rial hydrolysis of the input materials break- Bacteria decompose the long chains of
ing down insoluble organic polymers such the complex carbohydrates, proteins and
• the slurry reduces demand for synthetic
as carbohydrates and make them avail- lipids
fertilisers and other soil conditioners
able for other bacteria. into smaller molecules.

Acidogenic bacteria then convert the sug-

ars and amino acids into carbon dioxide,
• help avoid methane emissions into the
hydrogen, ammonia, and organic acids. For example, polysaccharides are converted
into monosaccharides.
Acetogenic bacteria then convert these
Proteins are split into peptides and amino
resulting organic acids into acetic acid, • in large, centralised installations it may acids.
along with additional ammonia, hydrogen, contribute to national energy supply.
and carbon dioxide.

Methanogens convert these products to

methane and carbon dioxide.
Under natural conditions, methane pro-
ducing microorganisms occur under anaer-
obic condition, e.g. under water ( sedi-
ments) and in animal stomaches.

They are obligatory anaerobic and very

sensitive to environmental changes.

Acidification (Stage II) Parameters which influence the biometha-

In contrast to the acidogenic and aceto- nation
Conversion of products of hydrolysis into genic bacteria, the methanogenic bacte-
acetic acid (CH3COOH), hydrogen (H2) ria belong to the archaebacteria. The metabolic activity involved in micro-
and carbon dioxide (CO2) by acid-producing biological methanation is dependent on
bacteria. They are a group of bacteria with a very the following factors:
heterogeneous morphology and a num- • Substrate temperature
These bacteria are facultatively anaero- ber of common properties that distinguish
them from all other bacterial in general. • Available nutrients
bic and can grow under acidic (low pH)
conditions. • Retention time (flow-through time)

Symbiosis of bacteria
To produce acetic acid, they need oxygen
and carbon. • pH level
Methane- and acid-producing bacteria act
in a symbiotical way. • Nitrogen inhibition and C/N ratio
By using the oxygen solved in the solution
or bounded-oxygen, the acid-producing bac- • Substrat solid content and agitation
Acid-producing bacteria create an envi-
teria create an anaerobic condition which • Inhibitory factors
ronment with ideal conditions for methane-
is essential for the methane producing mi-
producing bacteria (anaerobic conditions,
croorganisms (next stage).
compounds with a low molecular weight).

Methane-producing microorganisms use the

Moreover, they reduce the compounds with intermediates of the acid-producing bac-
a low molecular weight into alcohols, or- teria. EFFECT OF CONDITIONS
ganic acids, amino acids, carbon dioxide,
hydrogen sulphide and traces of methane. Without consuming them, toxic conditions
• Each of the types of bacteria responsi-
for the acid-producing microorganisms would
ble for the three stages of the anaero-
This process is only possible with energy bic digestion is affected differently by the
input. above parameters.
In fermentation processes the metabolic
actions of various bacteria all act in

• Since interactive effects between the

various determining factors exist, no pre-
cise quantitative data on gas production
as a function of the above factors are
No single bacteria is able to produce fer- available.
mentation products alone.
• Discussion of the various factors is lim-
ited to their qualitative effects on the pro-
cess of fermentation.

Methane formation (Stage III)

Temperature range of anaerobic fermen-
Methane-producing bacteria feed on com-
pounds with low molecular weight.
• Anaerobic fermentation is in principle
possible between 3°C and approximately
They utilize hydrogen, carbon dioxide and
70°C. Differentiation is generally made be-
acetic acid to form methane.
tween three temperature ranges:
1. The psychrophilic temperature range
Biomass Energy I • Depending on the digestor geometry,
the means of mixing, etc., the effective
lies below 20°C, retention time may vary widely for the in-
2. the mesophilic temperature range be- dividual substrate constituents.
tween 20°C and 40°C, and
3. the thermophilic temperature range • Selection of a suitable retention time
above 45°C. thus depends not only on the process tem-
... Five minutes break ... perature, but also on the type of sub-
strate used.

• Mesophilic digestion tends to be robust Role of Nutrients for Bacterial Growth

and more tolerant than the thermophilic For liquid manure undergoing fermenta-
process, but gas production is slower i.e. • Bacteria need more than just a supply of tion in the mesophilic temperature range,
retention time is longer. organic substances as a source of carbon the following approximate values for re-
and energy. tention time apply:
• Larger digestion tanks are required for
• They also require mineral nutrients.
the mesophilic process compared to the
thermophilic one. • liquid cow manure: 20-30 days
• In addition to carbon, oxygen and hydro-
gen, the generation of bio-mass requires • liquid pig manure: 15-25 days
• The speed of methane production in- nitrogen, sulfur, phosphorous, potassium, • liquid chicken manure: 20-40 days
creases with temperature. Since the amount calcium, magnesium.

of free ammonia also increases with tem-

perature, the performance is reduced as • A number of trace elements such as
a result. • animal manure mixed with plant ma-
iron, manganese, molybdenum, zinc, cobalt,
• Unheated biogas plants only work where selenium,tungsten, nickel etc.
50-80 days
mean annual temperatures are around 20°C are essential for bacterial growth.
or above or where the average daily tem- • addition of (sterilised) slaugtherhouse
perature is at least 18°C. • Agricultural residues or municipal sewage wastes may reduce retention time to
usually contain adequate amounts of the 10-15 days.
• Within the range of 20-28°C mean tem-
perature, gas production increases over- all above mentioned elements.

• If the temperature of the biomass is be-

low 15°C, gas production will be so low
that the biogas plant is no longer eco-
• Higher concentration of any individual
nomically feasible because of the required If the retention time is too short, the
huge digester volume. substance usually has an inhibitory effect,
bacteria in the digester are "washed
• Thermophilic digestion systems on the out" faster than they can reproduce,
• Analyses on case-to-case basis to de-
other hand offer higher methane produc- so that the fermentation practically
termine which amount of which nutrients
tion, faster throughput, better pathogen comes to a standstill.
needs to be added.
and virus ’kill’, but require more expensive
technology, greater energy input (heat-
ing) and a high degree of process control.

Temperature Changes (during operation) Batch-type and continuous plants pH value

• The process of biomethanation is very • The methane-producing bacteria live best

sensitive to changes in temperature. under neutral to slightly alkaline condi-
• The degree of sensitivity, in turn, is de- tions.
pendent on the temperature range.
• Brief fluctuations not exceeding the fol- • Once the process of fermentation has
lowing limits may be regarded as still un- stabilized under anaerobic conditions, the
inhibitory with respect to the process of pH will normally take on a value of be-
fermentation: tween 7 and 8.5.

The hydraulic retention time can only be pH value is no indication of energy con-
accurately defined in batch-type facilities. tents
• psychrophilic range: ± 2°C/h
For continuous systems, the mean reten-
• mesophilic range: ± 1°C/h tion time is approximated by dividing the • Due to the buffer effect of carbon dioxide-
• thermophilic range: ± 0,5°C/h digester volume by the daily influent rate. bicarbonate (OH−+CO2
HCO− 3 ) and
ammonia-ammonium (H++NH3
NH− 4 ),
V the pH level is no sign of substrate acids
The daily temperature changes are no prob- RT = and/or potential biogas yield.
Vin f l
lem for underground plants, since the tem-
• A digester containing a high volatile-
perature of the earth quite is constant. RT : rentention time [day]; V : digester vol- acid concentration requires a somewhat
ume [m3]; Vin f : daily influent rate [m3/day] higher-than-normal pH value.
• Since the results of mixing depend on
• No generally valid guidelines can be of- the substrate in use, it is not possible
fered with regard to specific biogas pro- to achieve a general evaluation of various
If the pH value drops below 6.2, the
duction for any particular solids percent- mixing systems.
medium will have a toxic effect on the
methanogenic bacteria.
Thus, such system will be designed on the
basis of empirical data.

Nitrogen inhibition
Inhibitory factors
• All substrates contain nitrogen. The
• The presence of heavy metals, antibi-
table below lists the nitrogen content of
Many substrates and various modes of otics and detergents used in livestock hus-
some organic substances and the C/N ra-
fermentation require substrate agitation bandry has an inhibitory effect on the pro-
or mixing in order to maintain process sta- cess of anaerobic digestion.
bility within the digester. The objectives
• For higher pH values, even a relatively
of agitation are: The following table lists the limit concen-
low nitrogen concentration may inhibit the
trations (mg/l) for some inhibitors.
process of fermentation.

• removal of the produced gas inhibitor limit [g/m3]

• mixing of fresh substrate and bacterial Copper 10-250
• Noticeable inhibition occurs above a ni-
population (inoculation) Calcium 8000
trogen concentration of roughly 1700 mg
• preclusion of scum formation and sed- Sodium 8000
ammonium-nitrogen (NH4-N) per liter sub- imentation Magnesium 3000
strate. • no big temperature gradients within Nickel 100
the digester Zinc 350
• BUT – Microorganisms need both nitro- • provision of a uniform bacterial popu- Chromium 200
gen and carbon for assimilation into their lation density Sulfide (as Sulfur) 200
• prevention of the formation of dead Cyanide 2
cell structures.

• Experiments have shown that the metabolic
activity of methanogenic bacteria is best The digestion process takes place in a
at a C/N ratio of 8-20, For designing of agitation “tools”, the warm (thermally insulated or heated), sealed
following has to be considered: airless volume (the digester) which cre-
• Optimum value depending on the nature ates the ideal conditions for the bacteria
of the substrate. to ferment the organic material in oxygen-
free conditions.

material N in [%] C/N

Cow dung 1.8 19.9 1. The Biogas fermentation process in-
The digestion tank needs to be warmed
Chicken manure 6.3 7.3 volves a symbiotic relationship between
and mixed thoroughly to create the ideal
Pig manure 2.8 13.7 various strains of bacteria,
Kitchen waste 1.9 28.6 conditions for the bacteria to convert or-
consequently, too frequent mixing is
Corn stalks 1.2 56.6 ganic matter into biogas (a mixture of
bad for the process. Slow stirring is
Rice straw 0.7 51.0 carbon dioxide, methane and small amounts
better than fast. ∗
Bagasse 0.4 31.0 of other gases).
Soybean stalks 1.3 33.0
Peanut hulls 1.7 31.0 ∗
Please contact Mr Kulschewski for a deeper in-
sight into the mixing processes in a biogas plant!

Substrate solids content

Different Types of Biogas Plants
2. A thin layer of scum on top must not
• Mobility of the methanogenic bacteria necessarily have an adverse effect on
within the substrate is restricted by in- The three main types of simple biogas
the process.
creasing solids content, and the biogas plants are shown in the next figure :
yield may suffer as a result.
3. Some types of biogas systems can func-
• balloon plants
• However, reports of relatively high bio- tion well without any mechanical agi-
gas yields from landfill material with a tation at all. • fixed-dome plants
high solids content are found in the lit-
• floating-drum plants
The gas drum is prevented from tilting by
a guiding frame. If the drum floats in a
water jacket, it cannot get stuck, even in
substrate with high solid content.

Balloon plants Advantages are the relatively low construc-

Advantages are the simple, easily under-
tion costs, the absence of moving parts
Balloon plants consists of a digester bag stood operation - the volume of stored
(e.g. PVC) in the upper part of which the and corroding steel parts. If well con-
gas is directly visible. The gas pressure
gas is stored. structed, fixed dome plants have a long
is constant, determined by the weight of
life span. The underground construction
Inlet and outlet are attached directly to the gas holder. The construction is rela-
saves space and protects the digester from
the plastic skin of the balloon. tively easy, construction mistakes do not
temperature changes. The construction
lead to major problems in operation and
provides opportunities for skilled local em-
Sufficient gas pressure is achieved through gas yield.
the elasticity of the balloon and by added ployment.
weights placed on the balloon.

Advantages are low cost, ease of trans- Disadvantages are mainly the frequent prob-
portation, low construction sophistication, lems with the gas-tightness of the brick-
high digester temperatures, uncomplicated work gas holder (a small crack in the up- Disadvantages are high material costs of
cleaning, emptying and maintenance. per brickwork can cause heavy losses of
the steel drum, the susceptibility of steel
Disadvantages can be the relatively short biogas).
parts to corrosion. Floating drum plants
life span, high susceptibility to damage, Fixed-dome plants are, therefore, recom-
have a shorter life span than fixed-dome
little creation of local employment and, mended only where construction can be
plants and regular maintenance costs for
therefore, limited self-help potential. supervised by experienced biogas techni-
A variation of the balloon plant is the cians. The gas pressure fluctuates sub- the painting of the drum.
channel-type digester, which is usually cov- stantially depending on the volume of the
ered with plastic sheeting and a sunshade. stored gas.

High-tec Biogas Plants

Even though the underground construc-

To contrast these simple biogas plants,
tion buffers temperature extremes, digester
the next figure gives an impression about
temperatures are generally low.
dimensions of ‘industrial plants’ which are
being constructed in developed countries.

Fixed-dome plants Floating-drum plants

The fixed-dome plant consists of a di-

gester with a fixed, non-movable gas holder,
which sits on top of the digester.

When gas production starts, the slurry is

displaced into the compensation tank.

Floating-drum plants consist of an under-

ground digester and a moving gas-holder.
Gas pressure increases with the volume
of gas stored and the height difference Sources of Info for Self-Study
The gas-holder floats either directly on
between the slurry level in the digester
the fermentation slurry or in a water jacket
and the slurry level in the compensation
of its own.

The gas is collected in the gas drum, which

rises or moves down, according to the
amount of gas stored.
PROBLEMS Some Resources
1. Please find short explanations for the
following key-words:

(a) hydrolysis
(b) acidogenation / acetogenation There are useful links and documents linked
(c) methanogenation to the weekly plan in physik-multimedial
(d) anaerobic digestion

2. Please give chemical formulas or struc-

tural explanations of the following sub-

(a) polysaccharide <–> monosaccha-


(b) proteins, lipids, fatty acids

(c) enzymes