ENERGY PRODUCTION FROM

PLASTIC WITH MICROBIAL FUEL

CELL BASED TECHNOLOGY

GUIDE: Dr. PRANAV VYAS

SHIVANGI MISHRA
M.Sc. (Previous)
 INTRODUCTION
• Plastics are synthetic polymers and are non metallic moldable compound which
can be pushed into any desirable shape and size on heating.

• Term plastic is derived from the Latin word “plasticus”, which is derived from the
Greek “plastikos” that was used to describe something able to be molded or fit for
molding. (PlasticsEurope2018)

• Plastics are composed of organic and inorganic raw materials comprising

hydrogen, carbon, oxygen, silicon, nickel and chloride that are often obtained
from oil, coal and natural gas.

• Due to its better physical and chemical properties, such as strength, lightness,
resistance to water and most waterborne microorganisms they are used worldwide
in packaging of food, detergents, chemicals, drugs, etc.

• These properties make plastic superior to other materials in many applications and
leads to many fold increase in its production since 1964, exceeding the world
plastic production to 360 million tonnes in 2018.
(PlasticsEurope2019)
 Contd…
• Synthetic plastics accumulate at a rate of 25 million tons per year in the
terrestrial and marine coastal environment.
(Balasubramanian et al., 2010)
• Municipal Solid Waste consists of waste fractions including organic wastes,
such as fruits, vegetables, food waste and inorganic wastes, such as plastics,
glass, metal pieces among others. Plastic waste fractions constitute a major
part of inorganic fractions of MSW.

(Moharir et al.,2019)
Contd…
❑Polyethylene
(PE):

• Thermosetting
plastics
• Non • Dangerous to
• Hydrophobic
biodegradable
• High Environment
molecular (naturally)
weight

• Polyethylene and other Plastics being strong and highly durable, takes up to
1000 of years for natural degradation in both terrestrial and aquatic
environment.
(Sivan, 2011)
• The presence of plastics in the terrestrial and aquatic environments lead to :
 soil infertility, release of toxic substances, ingestion by aquatic
organisms, entanglement of marine organisms, etc.
Contd…
Q. What is degradation?

• Heat
• Light
Physical • Moisture
and Change
• Chemicals
chemical • Biological
conditions

UV treatment, Heat
Physical treatment Toxic
treatment, Photo-oxidative
by-product
treatment
and
Mode of disposal
treatment Chemical treatment Use of chemicals to issue
of plastic break the polymer chain

Bacterial and Environmental friendly,

Biological treatment no harmful by-product
fungal species
(Moharir et al.,2019)
Contd…
❖Bio degradation:
• Biodegradation is the process in which living organisms are responsible for
breaking the organic substances.
➢ It can be achieved by following ways: (Gu J. D, et al., 2000a)
i. Aerobic
ii. Anaerobic Aerobic Anaerobic
iii. Semi-aerobic condition: condition:
CO2, H2O CO2, H2O, CH4

• Microorganisms such as bacteria

and fungi are involved in
degradation of both natural and
synthetic plastics.

General mechanism of plastic biodegradation

(Mueller, 2006)
Contd…
Main interest of this research is to check whether the plastic degrading micro-
organism isolated from various samples are able to generate power/electricity
along with their degradation ability through microbial fuel cell.
❖Microbial fuel cell:
• M.C. Potter was first to use MFC, in
1911.

(Rabaey et al., 2005)

Contd…
Electron transfer (ANODE)

Mediated
Direct
electron transfer

Secondary
Nanowires Outer membrane metabolites
(Pili) Cytochromes

Anode (A.J. Slate at el., 2019)

(Schroder. 2007)
Contd…
Types of MFC and its Applications

1. Submersible microbial electrolysis cell

• Biohydrogen production.

2. Microbial Desalination cell.

• Waste water and Water desalination, and
production of renewable energy.

3. Biosensor
• Monitoring organic matter, microbial
activity and toxicity.

4. Sediment MFC
• Nitrate and nitrite removal and electricity
generation from eutrophic lakes
(Y. Zhang 2012)
 RATIONALE

Uncontrolled use and non-biodegradable nature of plastics lead to large number of

plastic waste production and accumulation globally.

Plastic biodegradation does not occur efficiently in natural environment.

Many researches has been conducted to mitigate plastic waste production and
plastic degradation by using various microorganism such as bacteria and fungi.

In this project ability of plastic degrading bacteria will be checked, whether they
can generate electricity simultaneously with plastic degradation
 OBJECTIVES

1.Isolation of plastic degrading bacteria from different locations.

2. Identification and characterization of plastic degrading bacteria.

3.Screening plastic degrading activity of isolated bacteria.

4. Using most efficient plastic degrading bacteria in Microbial Fuel Cell.

 Plastic sample to be used

• < 50 micron
thickness

Polyethylene
bags

• 20-100 micron
thickness
WORK PLAN
OBJECTIVE 1: Isolation of plastic degrading bacteria from
different location.
1) SAMPLE COLLECTION PLASTIC
DUMPING
SITE WASTE
WATER
POLLUTED SAMPLE
RIVER BEFORE
PRIMARY
TREATMENT

WASTE
WATER
OIL SAMPLE SAMPLE
REFINERY SITES AFTER
PRIMARY
TREATMENT

POLLUTED INDUSTRIAL
LAKE EFFLUENT

ROOT OF
OLD TREE
 2) ENRICHMENT OF SAMPLE

❖Soil and water sample collected from different sites were used as source of
microbial inoculum for enrichment culture.

250ml of Erlenmeyer flask containing 100ml / 90ml

of sterilized synthetic medium (SM) Synthetic
medium
10g soil sample / 10ml water sample • NH4NO3 = 1g/l
• MgSO4.7H2O =0.2 g/l
• K2HPO4 =1 g/l
• CaCl2.2H2O = 0.1 g/l
300mg small pieces of polyethylene • KCl = 0.15g/l
• Yeast extract = 0.1g/l
• Micronutrients =
1.0mg/l (FeSO4.6H2O,
Incubation at 30C for 3 months ZnSO4.7H2O, MnSO4)

Isolation of culture (V. Balasubramanian

et al.,2010)
 3) ISOLATION OF BACTERIA
After enrichment of each samples for 3 months, 5ml of sample from each flask will
be taken into sterilized testube under aseptic conditions.

1ml from each samples will be serially diluted in 9ml sterile distill water up to 105
dilution.

0.1ml from each 103, 104 and 105 dilution tubes of each sample will be pipette out
onto each nutrient agar plate aseptically.

With the help of “Glass spreader” sterilized with ethanol, spread the inoculum on the
surface of agar.

Plates will be incubated at 37C for 48 hours.

After incubation, number and types of colonies will be counted . Pure bacterial
colonies from plates will be isolated and streak plate method will be performed on
Nutrient agar plates. Pure culture is maintained on nutrient agar slant.
OBJECTIVE 2: Identification and Characterization of plastic
degrading bacteria.

Identification

Macroscopic Microscopic
examination examination

• Colony morphology,
• surface pigment,
• shape and size on Staining Motility
agar plate
• Gram staining
• Endospore staining
• Capsule staining
Contd…
Characterization

Antibiotic
Biochemical tests Enzyme assay
susceptibility test
1. Catalase test
• Kirby-Bauer disk • Esterase activity
2. Triple sugar test
diffusion method • Protease activity
3. Indole test
4. Methyl red and
Vogesproskauer
test
5. Oxidase test
6. Starch
7. Simmons Citrate
test
8. Urease test
OBJECTIVE 3: SCREENING PLASTIC DEGRADING
ACTIVITY OF ISOLATED BACTERIA.

➢ Plastic degrading activity of all bacterial species isolated from collected

samples will be checked individually using Luria Bertani broth.

100ml Luria Bertani broth will be taken in

Erlenmeyer flask and is autoclaved.

LB broth will be inoculated with bacterial

culture and sterile plastic pieces of appropriate
dimensions aseptically

The mixture will be incubated for 1 month at

37C in rotatory shaker

After 1 month, plastic pieces will be removed,

washed and weighed
Contd…

Plastic degrading
screening test

Visual analysis Determination of dry weight

OBJECTIVE 4: Using most efficient plastic degrading
bacteria in Microbial Fuel Cell.
REQUIREMENTS:
• Omega glass bottle • Resistor
• Omega glass rod • Voltmeter
• Graphite electrodes • Copper wire
• Small bulb • Paraffin wax

Omega glass bottle Glass rod

(Animesh Devala et al., 2013)

Contd…
The bacteria which can efficiently degrade plastic will be used for MFC.

ANODE CATHODE

Luria Bertain broth Salt bridge 100mM potassium

+ ferricyanide
powder polyethylene Contains electrolytes like KCl, KNO3, +
+ K2SO4, etc. 100mM phosphate
Bacterial culture buffer (pH 7).

Electricity production
activity will checked
 SUMMARY
• MICROSCOPIC
Identification • BIOCHEMICAL
Samples Enrichment
TESTS

Isolation

Most efficient
plastic degrading
L.B medium
Polyethylene

Plastic degrading activity

Visual Dry
analysis weight

MFC
 REFERENCES
• Balasubramanian, V., Natarajan, K., Hemambika, B., Ramesh, N., Sumathi, C.
S., Kottaimuthu, R., & Rajesh Kannan, V. (2010). High‐density polyethylene
(HDPE)‐degrading potential bacteria from marine ecosystem of Gulf of Mannar,
India. Letters in applied microbiology, 51(2), 205-211.

• Deval, A., & Dikshit, A. K. (2013). Construction, working and standardization

of microbial fuel cell. APCBEE procedia, 5, 59-63.

• Gu, J. D., Ford, T. E., Mitton, D. B., & Mitchell, R. (2000). Microbial corrosion
of metals. The Uhlig Corrosion Handbook. 2nd Edition. New York: Wiley, 915-
27.

• Moharir, R. V., & Kumar, S. (2019). Challenges associated with plastic waste
disposal and allied microbial routes for its effective degradation: a
comprehensive review. Journal of cleaner production, 208, 65-76.

• Mueller, R. J. (2006). Biological degradation of synthetic polyesters—enzymes

as potential catalysts for polyester recycling. Process Biochemistry, 41(10),
2124-2128.
Contd…
• Rabaey, K., & Verstraete, W. (2005). Microbial fuel cells: novel biotechnology
for energy generation. TRENDS in Biotechnology, 23(6), 291-298.

• Schröder, U. (2007). Anodic electron transfer mechanisms in microbial fuel

cells and their energy efficiency. Physical Chemistry Chemical Physics, 9(21),
2619-2629.

• Sivan, A. (2011). New perspectives in plastic biodegradation. Current opinion

in biotechnology, 22(3), 422-426.

• Slate, A. J., Whitehead, K. A., Brownson, D. A., & Banks, C. E. (2019).

Microbial fuel cells: An overview of current technology. Renewable and
sustainable energy reviews, 101, 60-81.

• Zhang, Y. (2012). Energy recovery from waste streams with microbial fuel cell
(MFC)-based technologies.