NOTES BY Nimisha J Chennakadan : environmental biotechnology

Unit 4: Biofuels and Metal biotechnology: Biofuels and biological control of air pollution, plant derived fuels,
biogas, landfill gas, bioethanol, biohydrogen; use of biological techniques in controlling air pollution;
Removal of chlorinated hydrocarbons from air. Biotechnology application to hazardous waste management -
examples of biotechnological applications to hazardous waste management – cyanide detoxification -
detoxification of oxalate, urea etc. - toxic organics - phenols. Mining –heavy metals. Microbial transformation,
accumulation and concentration of metals, metal leaching, extraction and future prospects.

What are Biofuels? hydrocarbon fuel that is produced from an organic matter (living or once living
material) in a short period of time (days, weeks, or even months) is considered a biofuel.
▪ Biofuels may be solid, liquid or gaseous in nature.
o Solid: Wood, dried plant material, and manure
o Liquid: Bioethanol and Biodiesel
o Gaseous: Biogas
These can be used to replace or can be used in addition to diesel, petrol or other fossil fuels for transport,
stationary, portable and other applications. Also, they can be used to generate heat and electricity.
Some of the main reasons for shifting to biofuels are the rising prices of oil, emission of greenhouse
gases from fossil fuels and the interest in obtaining fuel from agricultural crops for the benefit of farmers.

• Biofuel, any fuel that is derived from biomass-that is, plant or algae material or animal waste.
The common commercially used biofuels are bioethanol, biodiesel and biomethane.
✓ Bioethanol is made from sugar, algae, wheat and sugar beet
✓ Biodiesel is made from vegetable oil, algal lipids, animal fats
✓ Biomethane can be produced from waste organic material, sewage, agriculture waste and domestic
wastes.
What are the Different Categories of Biofuels?
1. First Generation Biofuels: First-generation biofuels are made from food crops such as sugar, starch,
vegetable oil, or animal fats.
Common types include bio-alcohols (e.g., ethanol, butanol), biodiesel, vegetable oil, bioethers, and biogas.
Production Process: These biofuels are produced using conventional technology. For example, ethanol can be
produced through the fermentation of sugars from crops like corn and sugarcane, while biodiesel is made from
vegetable oils or animal fats through a process called transesterification.
Advantages:
• The production process is relatively simple and well-established.
• These biofuels can be used directly or blended with fossil fuels in existing engines and infrastructure.
Disadvantages:
• Food vs. Fuel Debate: Using food crops for fuel production can lead to increased food prices and food
shortages, exacerbating hunger and malnutrition.
• Environmental Impact: The cultivation of biofuel crops can lead to deforestation, loss of biodiversity, and
increased use of fertilizers and pesticides.
2. Second Generation Biofuels: Second-generation biofuels are produced from non-food crops or the non-
edible parts of food crops, such as stems, husks, wood chips, and fruit peels.
Examples: Cellulosic ethanol, biodiesel from lignocellulosic biomass.
Production Process: These biofuels are produced through more complex processes like thermochemical
reactions (e.g., gasification, pyrolysis) or biochemical conversion (e.g., enzymatic hydrolysis, fermentation).
Advantages:
• They do not compete with food crops, thus not affecting the food economy.
• Second-generation biofuels generally emit fewer greenhouse gases compared to first-generation biofuels.
Disadvantages:
• The production processes are technologically complex and often more expensive.
• There can be challenges in efficiently breaking down lignocellulosic biomass into fermentable sugars.
3. Third Generation Biofuels Third-generation biofuels are derived from microorganisms like algae.
Example: Butanol produced from algae.
Production Process: Algae can be grown in various environments, including land and water unsuitable for
traditional agriculture. Algae are then processed to extract oils, which can be converted into biofuels.
Advantages:
• Algae can be cultivated on non-arable land and use wastewater or saltwater, reducing the strain on
freshwater resources.
• High yield per unit area compared to traditional crops.
Disadvantages:
• The use of fertilizers and other inputs can lead to environmental pollution.
• The current technology for large-scale production of algae-based biofuels is still under development and
can be costly.
Fourth Generation Biofuels : Fourth-generation biofuels involve genetically engineered crops designed to
absorb high amounts of carbon dioxide. These crops are converted into fuel using techniques similar to second-
generation biofuels.
Production Process: The crops are grown and harvested as biomass, then converted into biofuel. The fuel
undergoes pre-combustion, and the carbon dioxide emitted is captured and sequestered in geological
formations, such as depleted oil or gas fields or unmineable coal seams.
Advantages:
• Carbon Negative: These fuels can be carbon-negative because the crops absorb CO2 during growth, and
the captured CO2 is stored, reducing the overall carbon footprint.
• They offer a sustainable way to produce energy while addressing climate change by removing CO2 from
the atmosphere.
Disadvantages:
• The technology is still in the research and development stage and is not yet widely implemented.
• There are concerns about the ecological and ethical implications of using genetically modified organisms
(GMOs).
Biofuel feedstock

Bioethanol:

Bioethanol is produced by the fermentation of carbohydrate-rich sources. Common feedstocks include

sugarcane, sugar beet, and corn.
The basic steps involved in the production of bioethanol are:

1. Feedstock Preparation: The raw materials (sugar or starch-based crops) are prepared by milling or
crushing.
2. Saccharification: Starches are converted into fermentable sugars using enzymes or acids.
3. Fermentation: The sugars are fermented by yeast or other microorganisms to produce ethanol and carbon
dioxide.
4. Distillation: The ethanol is separated from the fermentation mixture through distillation to achieve the
desired purity.
5. Dehydration: Further purification removes water to produce anhydrous ethanol suitable for fuel use.

Properties:

• Bioethanol is a colorless and clear liquid.

• It has a mild characteristic odor.
• It is highly flammable and miscible with water in all proportions.

Uses: Bioethanol is one of the most widely used alternative automotive fuels globally. It can be used in its
pure form (E100) or blended with gasoline in various proportions (e.g., E10, E15, E85). In addition to its use
as a fuel, bioethanol is also used as an industrial solvent and in the production of alcoholic beverages.

Advantages:

• Environmental Friendly: Bioethanol burns more cleanly than gasoline, producing fewer greenhouse
gases and pollutants. It contributes to a reduction in carbon dioxide emissions.
• Renewable: As it is derived from biomass, bioethanol is a renewable resource.
• Lower Toxicity Levels: It is less toxic compared to traditional fossil fuels and can reduce air and water
pollution
.
Biodiesel:

Biodiesel is an alternative diesel fuel made from renewable biological sources such as vegetable oils, animal
fats, and animal waste.

Advantages:

• Cost-Effective: Biodiesel can be produced from waste materials, reducing costs and providing an
environmentally friendly way to utilize waste.
• Renewable: It is derived from renewable resources, making it a sustainable alternative to fossil
fuels.
• Compatibility: Biodiesel can be used in diesel engines with little or no modification.

Production Process:

o 1. Transesterification: Transesterification is a chemical process that converts fats and oils

(triglycerides) into biodiesel (methyl or ethyl esters) and glycerol. This process involves reacting
triglycerides with an alcohol (usually methanol or ethanol) in the presence of a catalyst.

Steps in the Transesterification Process:

1. Feedstock Preparation:
o Sources such as vegetable oils (soybean, canola, sunflower) or animal fats (tallow, lard) are
collected and filtered to remove impurities.
2. Catalyst Preparation:
o Common catalysts used are sodium hydroxide (NaOH) or potassium hydroxide (KOH).
3. Reaction:
o The triglycerides (TAG) from the feedstock react with an alcohol (ethanol or methanol) in the
presence of the catalyst.
o The general reaction can be represented as:

4. Separation:
o The reaction mixture is allowed to settle, and the biodiesel is separated from the glycerol.
Biodiesel forms the upper layer, and glycerol, the lower layer.
5. Purification:
o The crude biodiesel is washed to remove residual catalysts, soaps, and impurities. It is then
dried to remove water.

Uses:

• Biodiesel can be used in diesel engines, either as a pure fuel (B100) or blended with petroleum diesel
in various proportions (e.g., B20, which is 20% biodiesel and 80% petroleum diesel).

Environmental Benefits:

• Lower Emissions: Biodiesel produces fewer greenhouse gases and pollutants compared to
conventional diesel. It reduces emissions of carbon monoxide, particulate matter, unburned
hydrocarbons, and sulfur oxides.
• Biodegradable: Biodiesel is non-toxic and biodegradable, making it a safer alternative for the
environment in case of spills.
Biobutanol: Biobutanol is a type of butanol (an alcohol) made from renewable resources such as grains or
cornstalks through a fermentation process.

Biobutanol Production Process

Biobutanol is produced through a biological fermentation process using renewable feedstocks and specific
microorganisms. Here’s a detailed breakdown of the process:

1. Feedstock Preparation

• Feedstocks: Renewable materials such as grains (corn, wheat), agricultural residues (cornstalks, straw),
and energy crops.
• Pretreatment: The biomass is often pretreated to break down complex carbohydrates into simpler
sugars. This may involve mechanical, chemical, or enzymatic methods to increase the accessibility of
fermentable sugars.

2. Hydrolysis
• Enzymatic Hydrolysis: Enzymes are added to the pretreated biomass to convert complex carbohydrates
(cellulose and hemicellulose) into fermentable sugars (glucose, xylose).
• Saccharification: This process releases sugars from the biomass, making them available for
fermentation.

3. Fermentation

• Microorganisms: Solventogenic Clostridia bacteria (e.g., Clostridium acetobutylicum, Clostridium

beijerinckii) are used to ferment the sugars.
• Fermentation Process:
o Inoculation: The prepared feedstock (sugar solution) is inoculated with Clostridia bacteria.
o Fermentation: The bacteria convert the sugars into biobutanol and other by-products through
anaerobic fermentation. The key metabolic pathways involved are:
▪ Glycolysis: Conversion of glucose into pyruvate.
▪ Butanol Production Pathway: Pyruvate is converted into butyryl-CoA and then reduced
to butanol.
o By-products: Along with biobutanol, the process also produces acetone and ethanol.

4. Product Recovery

• Distillation: The fermentation broth contains butanol, water, and other fermentation products.
Distillation is used to separate and purify biobutanol from the mixture.
• Solvent Extraction: In some processes, solvent extraction is used to enhance the recovery of butanol.
• Dehydration: Any remaining water in the biobutanol is removed through dehydration processes to
achieve the desired purity.

5. Refining and Blending

• Purification: Further refining steps may be employed to ensure that the biobutanol meets fuel-grade
specifications.
• Blending: Biobutanol can be blended with gasoline or used directly as a biofuel.

Detailed Fermentation Pathway

1. Inoculation and Culture Growth:

o The bacteria are introduced into a nutrient-rich medium containing the hydrolyzed sugars.
o The culture is grown under anaerobic conditions (absence of oxygen).
2. Acidogenesis:
o Initial stage of fermentation where sugars are converted into organic acids (e.g., acetic acid,
butyric acid).
o Glycolysis breaks down glucose into pyruvate, producing ATP and NADH.
3. Solventogenesis:
o The metabolic shift from acid production to solvent (butanol) production.
o Organic acids are converted into solvents (butanol, acetone, and ethanol).
o Key enzymes: Butyryl-CoA dehydrogenase, butanol dehydrogenase.
4. Product Recovery and Purification:
o Distillation: Separates butanol from water and other components based on boiling points.
o Solvent Extraction: Uses organic solvents to selectively extract butanol.
o Dehydration: Removes residual water to purify the butanol.

Advantages:

• Renewable: Produced from biomass, making it a sustainable alternative to fossil fuels.

• Compatibility: Can be used in gasoline engines without major modifications and blended with
gasoline.
 • Energy Density: Biobutanol has a higher energy content compared to ethanol, making it more
efficient as a fuel.

Environmental Benefits:

• Lower Emissions: Reduces greenhouse gas emissions compared to conventional fossil fuels.
• Biodegradable: Non-toxic and biodegradable, posing less risk to the environment.

Biogas

Biogas is a renewable energy source produced through the anaerobic digestion of organic matter by
microorganisms. The primary components of biogas are methane (CH4) and carbon dioxide (CO2), with trace
amounts of other gases like hydrogen sulfide (H2S) and ammonia (NH3). The process occurs in an oxygen-
free environment, typically within a biogas digester or anaerobic digester.

Composition of Biogas

• Methane (CH4): 50-70%

• Carbon Dioxide (CO2): 30-50%
• Other Gases: Hydrogen sulfide (H2S), water vapor, ammonia (NH3), and trace gases.

Production Process

Biogas is a renewable energy source produced through the anaerobic digestion of organic matter by
microorganisms. Here’s a detailed overview of the biogas production process:
1. Feedstock Collection and Preparation

Feedstocks: Organic materials such as agricultural residues, animal manure, food waste, and sewage sludge.

Pre-treatment: The feedstock may be shredded or mixed to ensure uniformity and enhance the digestion
process. This can involve mechanical, thermal, or chemical pre-treatment to break down complex organic
matter.

2. Anaerobic Digestion

• Process Stages:
o Hydrolysis: Complex organic polymers (carbohydrates, proteins, fats) are broken down into simpler
monomers (sugars, amino acids, fatty acids) by hydrolytic bacteria.
o Acidogenesis: The monomers are further degraded into volatile fatty acids (VFAs), alcohols,
hydrogen, and carbon dioxide by acidogenic bacteria.
o Acetogenesis: VFAs and alcohols are converted into acetic acid, hydrogen, and carbon dioxide by
acetogenic bacteria.
o Methanogenesis: Methanogenic archaea convert acetic acid, hydrogen, and carbon dioxide into
methane (CH4) and carbon dioxide (CO2), forming biogas.

3. Biogas Collection and Storage

• Biogas Composition: Biogas typically contains about 50-70% methane, 30-50% carbon dioxide, and
trace amounts of other gases such as hydrogen sulfide (H2S) and ammonia (NH3).
• Collection: Biogas is collected from the top of the digester and transferred to storage tanks.
• Storage: Biogas is stored in gas holders or tanks under pressure until it is used or further processed.

4. Biogas Purification

• Removal of Impurities: The raw biogas may contain impurities like H2S, water vapor, and siloxanes
that need to be removed.
o Desulfurization: Removal of hydrogen sulfide using iron oxide filters, activated carbon, or
biological filters.
o Dehydration: Removal of water vapor using drying agents or refrigeration.
o Siloxane Removal: Activated carbon or other specialized filters are used to remove siloxanes.

5. Utilization

• Electricity Generation: Biogas can be used in combined heat and power (CHP) systems to generate
electricity and heat.
• Heating: Biogas can be burned directly in boilers for space heating or industrial processes.
• Transportation Fuel: After upgrading to biomethane (removal of CO2), biogas can be used as a
vehicle fuel.
• Injection into Natural Gas Grid: Upgraded biomethane can be injected into the natural gas grid.

Detailed Anaerobic Digestion Pathway

1. Hydrolysis:
o Enzymes break down complex organic matter into simpler molecules.
o Proteins → Amino acids
o Carbohydrates → Simple sugars
o Fats → Fatty acids and glycerol
2. Acidogenesis:
o Acidogenic bacteria convert the products of hydrolysis into volatile fatty acids (VFAs),
alcohols, and gases.
 oSimple sugars and amino acids are transformed into acetic acid, propionic acid, butyric acid,
hydrogen, and carbon dioxide.
3. Acetogenesis:
o Acetogenic bacteria convert VFAs and alcohols into acetic acid, hydrogen, and carbon dioxide.
o Propionic acid, butyric acid, and other intermediates are broken down into acetic acid.
4. Methanogenesis:
o Methanogenic archaea produce methane and carbon dioxide from acetic acid, hydrogen, and
carbon dioxide.
o Acetic acid → Methane + Carbon dioxide
o Hydrogen + Carbon dioxide → Methane + Water

Removal of Chlorinated Hydrocarbons from Air

Chlorinated hydrocarbons, such as trichloroethylene (TCE) and dichloromethane (DCM), are volatile organic
compounds that pose significant environmental and health risks. Biological methods for removing these
pollutants from the air include biofiltration, biotrickling filters, and bioscrubbers. These systems use
microorganisms to degrade chlorinated hydrocarbons into less harmful substances. In biofilters, contaminated
air passes through a medium like compost or wood chips, where microbes break down pollutants. Biotrickling
filters use a liquid phase to enhance contact between microbes and contaminants, while bioscrubbers employ
a scrubbing liquid to capture pollutants before microbial degradation. These biological approaches are
effective, sustainable, and can be integrated into industrial air pollution control strategies.

Biological methods such as biofiltration, biotrickling filters, and bioscrubbers are effective in removing
chlorinated hydrocarbons like trichloroethylene (TCE) and dichloromethane (DCM) from the air. Here's a
detailed explanation of how each method works:

1. Biofiltration:
o Description: Biofilters are porous beds filled with a medium such as compost, wood chips, or
synthetic materials. These materials provide a habitat for microorganisms that can degrade
pollutants.
o Operation: Contaminated air is passed through the biofilter bed. As the air moves through
the medium, microorganisms attached to the surface or living within the medium break down
the chlorinated hydrocarbons into simpler, less harmful substances through biological
oxidation.
 o Process: The pollutants are adsorbed onto the surface of the biofilter medium, where
microbial activity (biodegradation) occurs. Microbes utilize the pollutants as a carbon source
and transform them into carbon dioxide, water, and other harmless by-products.
2. Biotrickling Filters:
o Description: Biotrickling filters are similar to biofilters but operate with a liquid phase. They
typically consist of a packed bed where the packing material is irrigated with a nutrient
solution that promotes microbial growth.
o Operation: Contaminated air is introduced into the biotrickling filter, and the air pollutants
dissolve into the liquid phase. Microorganisms in the nutrient-rich liquid degrade the
dissolved pollutants.
o Process: The liquid phase enhances contact between the contaminants and the microbial
biofilm, speeding up the biodegradation process compared to biofilters. The nutrients
provided in the liquid phase support microbial metabolism and facilitate the breakdown of
chlorinated hydrocarbons.
3. Bioscrubbers:
o Description: Bioscrubbers are air pollution control devices that use a liquid scrubbing
solution to capture pollutants from the air before microbial degradation occurs.
o Operation: Polluted air is passed through the bioscrubber, where it comes into contact with
the scrubbing solution. The solution captures the chlorinated hydrocarbons through
absorption or chemical reaction.
o Process: After capturing the pollutants, the scrubbing solution is directed to a bioreactor
where microorganisms degrade the pollutants dissolved in the liquid phase. The bioreactor
ensures that the contaminants are fully degraded before the scrubbing solution is recycled
back into the bioscrubber.

Advantages and Applications:

• Effectiveness: These biological methods are highly effective in removing chlorinated hydrocarbons
from air streams, often achieving high removal efficiencies.
• Sustainability: They are environmentally friendly and sustainable compared to traditional chemical
methods.
• Integration: Biofiltration, biotrickling filters, and bioscrubbers can be integrated into existing
industrial processes for air pollution control, offering a cost-effective and efficient solution.