Industrial Biotechnology

(11102301)
By
Dr-Hemanga Kumar Das
Dept. Of Biotechnology
Parul Institute of Applied Sciences
 Unit-2

Bioremediation
 Introduction:

➢ Bioremediation is the process of reduction, elimination, alteration, and transformation

of contaminants present in the natural environment like soil, sediments, air, and water
through the application of microorganisms, fungi, green plants, or their enzymes.
➢ It is a waste management technique that uses naturally occurring biological organisms to
break down hazardous substances into less toxic or non-toxic forms.
➢ Bioremediation includes a series of redox reactions for the production of energy within
microbial cells for cell maintenance and reproduction.
➢ Bioremediation is a global, regional and local application for removing pollutants from the
environment restoring the contaminated sites.
 Factors affecting bioremediation:

➢ The bioremediation process depends upon the different factors for the removal of the
contaminants, some of them are:
1. Concentration of the contaminant
✓ The concentration of the contaminants directly affects microbial activity.
✓ Lower the concentration of the contaminants there will be decreasing rate of
degrading enzymes produced by bacteria in the soil.
✓ Toxic effects are observed in presence of higher concentrations of contaminants.
✓ The decomposition rate of catabolic enzymes can be increased by the synergistic
interactions between different components of the contaminants.
 Factors affecting bioremediation:

➢ The bioremediation process depends upon the different factors for the removal of the
contaminants, some of them are:
2. Nutrient availability
✓ Carbon, nitrogen, phosphorus, potassium, and calcium are the basic requirement for
the growth of microorganisms, the concentration of the nutrient availability directly
affects the degradation of the contaminants.
✓ The excessive presence of nitrogen, potassium, and phosphorus shows a negative
impact on the degradation of hydrocarbons.
✓ The rate of bioremediation can also be determined by knowing the accessibility of
organic matters towards microorganisms; which is known as bioavailability.
 Factors affecting bioremediation:

➢ The bioremediation process depends upon the different factors for the removal of the
contaminants, some of them are:
3.Characteristics of the contaminated soil
The bioremediation process is significantly affected by the different parameters of the
contaminated soil such as pH, texture, permeability, water holding capacity, temperature,
and oxygen availability.
a. pH
Optimum pH is required for the bioremediation process which ranges from 6-8. Neutral pH is
suitable for the degradation of petroleum hydrocarbons whereas some fungi and acidophilic
microbes degrade contaminants in an acidic environment.
 Factors affecting bioremediation:

➢ The bioremediation process depends upon the different factors for the removal of the contaminants, some
of them are:
3.Characteristics of the contaminated soil
b. Temperature
The degradation of the contaminants is also affected by temperature especially in the case of hydrocarbons
under both in situ and ex-situ conditions. It has been found that a higher temperature of 30°C-40°c increases
the bioremediation in the soil as well as in the marine environment.
c. Oxygen availability
Oxygen is a very important factor to determine the extent and rate of biodegradation of contaminants. Aerobic
biodegradation is much faster than anaerobic biodegradation. For the aerobic respiratory breakdown of
organic contaminants, oxygen availability plays a significant role. In the majority of cases, the addition of
hydrogen peroxide is used to introduce oxygen. Hydrogen peroxide is about seven times more soluble in water
than oxygen.
 Types of bioremediation:

➢ On the basis of removal and transportation of waste for treatment, bioremediation is of

two different types.
✓ In Situ Bioremediation
✓ Ex Situ Bioremediation
 In situ bioremediation:

➢ In situ remediation is the in-site treatment of contaminants using biological agents. It is a

cleanup approach between microbes and the contaminants directly for
biotransformation.
➢ There are two major types of in situ bioremediation.
1.Intrinsic Bioremediation
•Intrinsic bioremediation or natural attenuation is a passive process of treatment of polluted
sites without any human intervention through naturally occurring microbial population.
•It is the natural remediation of environmental pollutants into non-toxic or less toxic forms using
the inherent capacity of microorganisms without taking any engineering steps to enhance the
process.
•The major application of intrinsic bioremediation is for control of contaminants at the waste
sites
 In situ bioremediation:

2.Engineered Bioremediation
•Engineered bioremediation or accelerated bioremediation is the advanced process of
application of engineered systems to stimulate microbial activity for remediation of
environmental pollutants.
•When intrinsic bioremediation isn’t feasible, accelerated bioremediation is proceeded where,
either substrate or nutrients are added to the environment to degrade the toxicity of
contaminants making the microorganism grow more rapidly.
•This process accelerates the biodegradation process through optimization of physical and
chemical conditions vitalizing the growth of microorganisms.
•Usually the microorganism is indigenous, but occasionally microorganism that are very efficient
at degrading a certain contaminant are additionally added.
 In situ bioremediation techniques:

1.Bioaugmentation
2.Biostimulation
3.Bioslurping
4.Bioventing
5.Biosparging
 Bioaugmentation:

➢ Bio augmentation is the process of addition of culture microbial population which have
the ability to degrade specific soil and groundwater contaminants.
➢ A technique of bioremediation in which strains of natural or genetically engineered
bacteria with unique metabolic profiles are added to the contaminated site in order to
supplement indigenous microflora and speed up biodegradation.
➢ Common application involves bio augmentation for chlorinated contaminants, petroleum
hydrocarbon etc.
➢ Microorganisms are isolated either from contaminated sites, historical sites or genetically
modified to support remediation process of contaminated sites.
 Biostimulation:

➢ Microbes cannot use pollutants as the only source of energy thus they need to be
accessed with supplied nutrients.
➢ Bio-stimulation is the process of environmental modification via addition of limiting
nutrients and electron acceptors like phosphorous, nitrogen, oxygen or carbon in order to
stimulate the existing microbial population which are involved in bioremediation.
➢ It is most common remediation approach against petroleum pollutants in soil.
 Bioslurping:

➢ Bioslurping, also called as multi-phase extraction is the process of in situ aerobic

bioremediation of contaminated soils using bioventing and vacuum enhanced free-
product recovery that extracts light, no aqueous-phase liquids (LNAPLs) from the capillary
fringe.
➢ Bioslurping is limited to 25 feet below ground surface as contaminants cannot be lifted
more than 25 feet by this method and effective for removing free product that is floating
on the water table.
 Biosparging:

➢ Biosparging the injection of a gas and gas-phase nutrients pressure into the saturated
zone applying pressure to promote aerobic biodegradation.
➢ It is the most recommended approach for aerobic degredation of sites affected with
lighter to heavier petroleum contaminants such as oils, diesel, gasoline, jetfuels etc.
➢ Lighter ones are removed easily but heavier ones due to minimum level of microbial
bioavaibility requires longer process of treatment.
➢ In it the cost can be reduce by reducing the diameter of injection point.
 Bioventing:

➢ The most common in situ treatment and involves supplying air and nutrients through
wells to contaminated soil to stimulate the indigenous microorganism.
➢ Bioventing is applied for remediation of petroleum hydrocarbons contaminants in soil
through air supply to an unsaturated soil zone using a combination of pumps and blowers
for continuous injection of low volumes of air.
➢ It can be categorized as either aerobic, anaerobic or co-metabolic depending on the
amendments used.
➢ The slow removal of air and maintaining 5% oxygen in subsurface is generally practice for
bioventing.
 Ex-Situ Bioremediation:

➢ Ex-situ bioremediation or off-site bioremediation is the removal/excavation of

contaminants and pollutants by subsequent transportation of contaminants from one site
to another. Similar to in situ techniques, remediation occurs with the role of
microorganisms. These techniques are based on the type of contaminants, site of
pollution, degree of pollution, and cost of treatment.
➢ Techniques in ex situ bioremediation are:
1.Biofiltration
2.Biopile
3.Bioreactor
4.Composting
5.Land Farming
 Biofiltration:

➢ Biofiltration is the biological treatment process of biodegradable waste which relies on

biodegrading microbial populations.
➢ It is the process of purification of contaminated air evolved from volatile organic
compounds.
➢ The treatment of contaminants is done using various materials like bio-filters, bio-trickling
filters, bio-scrubbers, conventional bio-filters etc.
➢ Biofilter is treatment bed consisting of compost, soil or peat media inside which the
pollutants come into contact with microorganisms and eventually get biodegraded.
 Biopile:

➢ Biopile is type of remediation process that involves enhancement techniques via above-
ground piling of excavated polluted soil, nutrient amendment, and sometimes aeration to
increase the microbial population and their activity.
➢ This technique involves aeration, irrigation, nutrient and a treatment bed.
➢ Biopile can reduce and limit volatilization of low molecular weight (LMW) pollutants, also
help in effective remediation of extreme polluted environments
➢ It is a cost effective approach which ensures effective biodegradation.
 Bioreactor:

➢ Bioreactor is an engineered system involving series of biological reactions in which

pollutants are fed into the bioreactor vessel for their degradation that facilitate the
growth of biological mass.
➢ There 5 different operating mechanisms in bio reactor which includes: batch, fed-batch,
sequencing batch, continuous and multistage.
➢ Bioreactors maintains suitable controlled environment for the optimum growth
conditions that leads to the proliferation of microbial populations.
➢ Pollutants are fed either in the dry or slurry form.
 Composting:

➢ Composting is the process of degradation and decaying or organic waste under favorable
controlled conditions with the action waste degrading microorganisms.
➢ Composting is a self-heating, substrate-dense and solid phase treatment process.
➢ Microbial population metabolize the organic waste and degrade it to volume by 50%
reduction forming the end product called compost or humus.
➢ Compost is a nutrient rich soil which is very useful in application to the crops and plants
for their effective growth.
➢ The steps involved in the process include sorting and separating, size reduction, and
digestion of the refuse.
 Landfarming:

➢ One of the simplest bioremediation process of excavation of polluted soil transported to

above the ground surface allowing aerobic biodegradation of pollutant by autochthonous
microorganisms.
➢ The autochthonous microorganisms are stimulated by tilling process which involves
nutrients amendments (nitrogen, phosphorous etc.), aeration process and irrigation.
➢ If the in-site treatment is the process can also fall under in-situ remediation.
➢ It is also a cost effective approach which requires minimal environment and energy for
treatment of large volume of polluted soils.
 Advantages:

✓ Complete remediation of harmful contaminants presents in the environment instead to

transferring contaminants from one site to another.
✓ Cost effective method with minimal requirements of complex tools and equipment
✓ Environment friendly approach with use of microorganism instead of harmful chemicals
✓ In majority cases, can be carried out on site reducing transportation cost
✓ Minimum site destruction and disruption
✓ Lower liability level
✓ Low energy consumption
✓ Being a very effective reliable and easy approach, is therefore accepted publicly and by
the regulatory authorities.
 Disdvantages:

✓ Only limited to biodegradable waste and contaminants

✓ Requires extensive monitoring
✓ Being a biological process, specificity is a major drawback in terms factors like type of
environmental growth conditions, types of microorganisms, type of nutrient requirements
and type of contaminants.
✓ Possibility of production unknown and potentially toxic byproducts
✓ Comparatively a time consuming process
 Applications:

✓ Bioremediation is highly effective on petroleum products.

✓ Sites where production-related chemicals are leaked or released as wastewater. Heavy
metals such as lead and chromium are difficult to remediate, while many minor pollutants
can be eliminated biologically.
✓ Bioremediation is ideally suited for landfills that are overfilled, leaking, or closed.
Methane gas is a frequent byproduct that can be regulated by air stripping and scrubbing.
✓ Farms that have been over-fertilized are ideal candidates for bioremediation. This
comprises both synthetic fertilisers and animal waste.
 Applications:

✓ Wood preservatives often pollute lumber processing yards. They typically seep into the
soil and groundwater, however bioremediation efforts can remove them.
✓ When septic tanks and disposal fields fail, onsite sewage systems contaminate the soil and
groundwater. These sewage system overflows respond exceptionally well to biological
treatment.
✓ Tailings from mines can be exceedingly toxic. Efforts to decontaminate old mine quarries
and pits by bioremediation have shown to be quite successful.
✓ Alongside traffic routes, accidental chemical leaks have been remedied via biological
therapy. This includes even road salts and petroleum spills.