Microbial degradation of plastic and

application of nanoparticles : A Review

-Diya Ganapathy K
190950002
 Contents

1. Abstract 3
2. Introduction 3-4
3. Literature Reviews 4-13
4. Conclusion 13
5. References 14-15

2
Microbial degradation of plastic
with application of nanoparticles:
A review

1.Abstract
With the increased global demand for plastic, the manufacturing and disposal
has been increasing day by day. Plastic are corrosion resistant, inexpensive
and easily available. Though plastic has unfortunately become a convenient
material in the daily uses, it’s effects on the environment and our ecosystem
cannot be avoided. Plastic accounts to almost 70% of the total solid wastes
produced and it’s reckless disposal has lead to severe pollution that has been
taking a toll on humans, animals and the surrounding environment. With time,
the world has recognized this issue and the degradation of this highly calcitrant
material has become a major goal to be accomplished in order to reduce it’s
negative effects on our environment. Microbial degradation has been one of the
most sought after solutions to tackle this problem. This review paper discusses
the role of microbes and nanoparticles in the biodegradation of these polymers.

2.Introduction
The word plastic originated from the Greek word ‘plastikos' which means ‘ability
to be modified into desired shapes'. Plastics can be generally defined as a
polymer containing carbon, nitrogen, hydrogen, oxygen, silicon,chlorine and
bromine that has the ability to be reshaped under the effect of heat and
pressure. Plastics are made by linking polymer chains made up of various
monomers linked together by chemical bonds. Polythene comprises of 64% of
total plastic and is a linear hydrocarbon polymer consisting of long chains of
ethylene monomers.

Plastic can be considered to be the most widely used manmade substance . For
the past two decades plastic has gained widespread use due to it’s use in
3
packaging, transportation, industries, medical materials etc. Plastics are
composed of resins that are petroleum based and resistant to biodegradation.
Though plastics have been commercially acceptable on a very broad spectrum,
it cause severe problems the major loophole being it’s resistant to biodegrade.
The reckless disposal of plastic in open landfills and bins have been known to
cause blockage of pipes and sewer lines, site for fly contaminations, choking of
animals etc. Discarded plastic, besides being aesthetically unpleasant are a
rapidly increasing percentage of solid wastes that are harmful to the natural
environment.

The efficient degradation of plastic takes about a 1000 years. Burning of plastics
releases dioxins and carbon dioxide and the commonly used methods for plastic
waste management have proved to be inadequate. This calls for an alternative
solution that’s environment friendly and economical at the same time. Hence,
there is a growing demand in the use of effective microorganisms capable of
degrading these synthetic polymers. Another alternative accepted widely
besides recycle and reuse of plastic, is the manufacturing of biodegradable
plastics. These biodegradable plastics are designed to be degraded by
microbes at a faster rate, because they are made up of organic materials like
lignin, cellulose, starch and hemicellulose.

3.Literature Reviews
3.1 Properties of plastics and their basic classification

Based on the thermal properties, plastics are classified into thermoplastics and
thermoset plastics. Thermoplastics can be hardened and softened by repeated
heating and cooling methods. These plastics are produced from the breakage
of the double bond and are linear solids. Examples are polyethylene,
polypropylene, PVC, polystyrene etc. Thermoset plastics have highly cross
linked structures and the chemical changes are irreversible but cannot be
recycled. An example is phenol formaldehyde polyurethanes.

Plastics can also be classified based on properties like electrical conductivity,

tensile strength, degradability, durability and thermal stability. Plastics can be
biodegradable or non-biodegradable. Non biodegradable plastics are made up

4
of small monomers and have higher molecular weights. Biodegradable plastics
are made of starch and have lower molecular weight.

Physical or chemical changes in polymers due to factors like light,

heat ,moisture, chemical and biological conditions is termed as degradation.
The deterioration of these polymers due to the effects of microbial activity is
known as biodegradation. [1]

3.2 Factors affecting plastic degradation

Factors like light, moisture content, pH, skylight, presence and absence of
oxygen, temperature etc not only affect the polymer degradation, but also have
a role to play in the microbial activity and their enzymatic processes. The
physical and chemical properties of the polymers also affects it’s degradation.

3.2.1 Molecular weights of polymers are a deciding factor in the extent to which
they can be degradable. Clearly, low density polymers are easily degraded
when compared to high density polymers.
3.2.2 The melting point the polymer is another parameter that decides the effect
on enzymatic degradation. Polymers with higher melting points usually
have a lower degradation potential.
3.2.3 Enzymatic degradation efficiency also lowers with increasing time.
3.2.4Additive anti-oxidants, stabilizers used in the manufacturing of the polymer
is tougher to be microbially degraded and also might be toxic to the microbes.[2]

3.3 Microbial degradation of plastic

Chemical and physical processes of plastic degradation though effective, pose
other threats to the environment, therefore microbial degradation has been
and active methodology accepted by a larger scale for the plastic degradation.
Different microbes have different degrading potentials on different materials.
Microbes like actenomycets, bacteria and fungi are known to degrade both
natural and synthetic plastics.

5
Microbes have been known to play an effective role in the degradation of
plastic, as per several studies. Fungi have been analyzed to degrade plastic
by the action of degrading enzymes and extracellular polymers like
polysaccharide , along with the ability of the fungal hyphae to penetrate.
Commonly, A.niger, A.niger, Lysinibacillus spp. have been known to have
degrading potentials upto 0.86% per week at an average. Microorganisms are
involved in the degradation of both synthetic and natural polymers thus
forming biofilms. Different microbes are known to degrade different types of
plastics.

The mode of degradation can be of two ways

3.3.1 Direct action- deterioration of plastics which serves as food for the
microbes
3.3.2 Indirect action- influence of enzymes or metabolic products of microbes
causing discoloration and thus deterioration. [3]

Aerobic biodegradation occurs in the presence of oxygen and carbon dioxide is

produced. In anaerobic degradation methane is produced instead of carbon
dioxide. Mineralisation is the conversion of biomass or biodegradable materials
to gases, water,salts,minerals etc. The chemical structure is the most important
factor affecting the biodegradablity of polymers. Other factors include elasticity,
morphology, porosity etc.[4]

Microorganisms attach themselves to the surface of the large molecular

substrate and cleave it into smaller segments, by extracellular or intracellular
enzyme secretions. Microorganisms use polymers as a source of nutrition and
organic compounds.[1] Microorganisms have the natural ability to transform
compounds like hydrocarbons, metals and pharmaceutical compounds as a
part of their secondary metabolism. The polymer chains are broken down into
lower molecular weight monomers ,thus making it easier for the accumulation
of the microbial colonies and the plastic is hydrolysed by the microbial enzymes
to create functional groups that improve the hydrophilicity. The polymers can
also be broken down to organic molecules like alcohol, acids and ketones
6
depending on the type of enzymatic activity. The microbes then absorb these
water soluble cleaved products . Aerobic metabolisms are reported to result in
carbon dioxide and water formation, whereas anaerobic metabolisms result in
carbon dioxide, water and methane as the end products.[2]

The plastic degradation is typically a surface erosion process and the enzymatic
degradation is usually a two step process. First, the enzyme binds to the
polymer surface and eventually catalyses the hydrolytic cleavage. Many fungi
like Aspergillus spp, Fusarium, Mucor , Rhizopus, Penicillium etc have been
studied to analyse their potential in degrading plastics. The fungal hyphae
penetrate the polymer solids, thus causing swelling and bursting of the surface.
Bacterial species like Pseudomonas spp, Xylosys spp, Azotobacter spp and
several actinomycetes have been used in the microbial degradation of
polymeric solids. [2]

3.4 Biodegradation of plastic by Aspergillus spp isolated from

the polluted cities in Chennai, Tamil Nadu ( Raaman et al 2012)

Approximately 60 percent of the entire plastic production is reported has been

reported to comprise of low density polyethylene. In India the wide use of
polyethylene bags
Has become an environmental nuisance , contributing to the littering of a large
percentage of highly Calcitrant solid waste materials. This study by (Raman et
al 2012) was to estimate heterotrophic fungi diversity and a broad study of their
polythene degrading ability.
The samples for these studies were polythene materials collected from highly
polluted cities in Chennai , Tamil Nadu. To isolate the fungal strains the samples
were subjected to serial dilution and pour plated onto PD Agar under sterile
conditions and incubated at 37 C for 48 Hours. The colonies were selected on
morphological basis and different colonies were transferred onto different PDA
slates for the pure cultures and further characterisation. Macroscopic
observation and straining techniques were applied for the fungal
characterisation.

7
Screening and identification of polyethylene degrading fungus: substrates like
1% starch , 1% gelatine and 1%tween 80 mixed with agar were used as suitable
substrate mediums to analyse the enzyme action of the fungal extracts. The
media are poured on to plates under sterile conditions and once set deep wells
are cut to load the isolated fungal filtrates aseptically. After incubations under
standard conditions the plates were observed for opacity around the loaded well
surface, which indicates a positive result for the presence of enzymatic activity.

Microbial degradation of polyethylene: 1 cm diameter of LDPE strips were

weighed and transferred on to 50 ml of rose Bengal broth medium and
inoculated with the selected fungal strains. LDPE strips in a microbial medium
was used as a control medium. The flasks are left in a beaker incubator for a
month after which the strips are washed dried and weighed
The weight loss of the LDPE strips were calculated and SEM were used to
analyse the surface of the strips to observe the morphological degradation
change.
Several fungal strains that were isolated from the polluted sites of Chennai were
found to have around 6-12 percent of efficiency in degrading the LDPE around
a time span of 1 month.
SEM analysis were performed at different magnifications to confirm the
degradation activity in each LDPE strip by observing for surface corrosion
folding and cracks which supposedly might be due to the metabolites or
enzymes.[5]

3.5 A comparative study of commercially available plastic carry

bag biodegradation by micro organisms isolated by
hydrocarbon effluent enriched soil (Mukherjee et al 2014)

Plastics are traditionally composed of petroleum base resins like polythene and
polypropylene which are very stable and do not degrade in the very ambient
environment (Raaman et al, 2012), leading to its accumulation and as a result
as pollution. Under the study strains like E.colli, A.niger etc were isolated from
the soil enriched with hydrocarbons effluents for further comparative studies non
the efficiencies on plastic degradation. A.niger has been to biodegrade
polyethylene and plastics, the sampling soils were collected from the petroleum
refinery at West Bengal, Purba Mednipur and the plastics were collected from
the local stores.
8
 In this context Apergillus niger , Baciflus weihenstephanensis , Burkhoideria
cepecia, Escherichia coli were isolated from hydrocarbon effluent enriched soil
and pursued further for a comparative study on their efficiency in plastic
degradation. It was observed that the microbes partially degraded the plastic
strips by utilising them as sole carbon source. A.niger exhibited the formation of
fungal mats and plastic lumps with time, which resulted in it’s deterioration. Cell
surface hydrophobicity of these microbes was the leading factor for the
formation of these biofilms, consequently enhancing the plastic degradation.
These properties of the microbes can help the microbiologists to have a better
insight into production of commercial biodegradable and eco-friendly plastic
carry bags.[6]

3.6 Nanoparticles and their characteristics

Nanotechnology has made several revolutionary developments in the field of

medicine, biotechnology, physics etc. Nanoparticles are materials that lie in the
size range of less than 100nm at least. Nanoparticles cab be broadly classified
into carbon NP's, metal NP's, ceramic NP's, semiconductor NP's, polymeric and
lipid based NP's. Nanoparticles can be manufactured by physical, mechanical,
chemical and microbial synthesis methods. This review is based on the metal
nanoparticles that are synthesized with the help of bacteria. Following are the
general characteristics of nanoparticles that explains it’s wide applications in
several domains of science and technology:

3.6.1 Nanoparticles are comprised of three layers, the surface, shell and the
core.
3.6.2 Nanoparticles can be encountered as emulsions, aerosols or
suspensions.
3.6.3 Nanoparticles have high surface area to volume ratio and therefore make
excellent catalysts.
3.6.4 Nanoparticles can be mixed with other materials to form composite matter
with enhanced properties and better efficiencies.[12]

9
A new advancement in the study of microbial degradation of waste is the
application of nanoparticles. Through several studies nanoparticles have been
proved to enhance the microbial degradation process.

3.7 Nanoparticles as an enhancer for biodegradation

Several nanoparticles used in the microbial degradation are as follows[3]

3.7.1 SPION (super magnetic iron oxide nanoparticles) 10-37.8 nm. These
nanoparticles along with microbial consortiums of Microbacterium
spp,Pseudomonas putida were monitored in minimal broth Davis medium
without iron and dextrose. The study reported that the nanoparticles
accelerated the bacterial growth. SPION of size 10.6 nm were found to
significantly increase the efficiency of biodegradation. The results were
analysed by FTIR and simultaneous TG-DTG-DTA.
3.7.2 NBT ( nano barium titanate)- NBT was supplied in the minimal broth to
study it’s influence on the growth cycle of the LDPE degrading microbial
consortia. It was reported to influence the lag phase, exponential phase
and stationary phase of the cycle by reducing the duration of the lag phase
and reducing the latter phases. For P.putida, Microbacterium spp the
favoured nanoparticle size is 38nm. UV- spectrophotometer was used to
monitor the bacterial growth and the residues were analyzed by FTIR and
TG-DTG-DTA where the LDPE was taken as the control. A shift in lambda
max was observed from 209 – 225.3 nm in 4 days in the absence of NBT
as the polymeric structure changed. In the presence of NBT the spectrum
was seen to shift from 209 – 224.11 nm in 2days , therefore showing an
improved biodegradation.

3.7.3 Fullerene 60 nanoparticles – fullerene 60 nanoparticles were studied in a

minimal broth Davis without dextrose. Fullerene 60 are detrimental to
bacterial growth at concentrations more than 0.01% (w/v). UV
spectrophotometer was done to analyse the bacterial growth curve, to
study the effect of fullerene 60 np on individual bacterial strains and on
consortia . After treatment with fullerene 60 nanoparticles and the
microbial consortia, the plastic samples are evaporated dried and
analysed using FTIR and TG-DTG-DTA. The shift in lambda max due to
10
 the change in the polymeric structure was observed from 209 to 223 nm
after 4 days without the fullerene 60 nanoparticles. Where as, the samples
with fullerene 60 np, lambda max was observed to shift from 209 – 224.97
nm on the first day itself, therefore showing an improved rate of
degradation.

3.8 Application of nanoparticles as an enhancer in the

degradation process of plastic using A.niger

A study conducted by ( Patwari, Sabat 2017) reported the role of silver

nanoparticles as an enhancer in the biodegradation of plastic. A.niger was
isolated from spoilt vegetables like onion and was characterized by microscopic
and staining techniques. The fungus was inoculated in CPZ Himedia enriched
with 1% peptone, 1% dextrose and 1% starch to increase the longevity of the
fungus. The plastic strips were treated with 1% SDS and 10% NaOCl and
washed and surface sterilised with 90% ethyl alcohol . The strips were placed
on the agar and observed for zone of clearance.

For the fullerene reactions mixture synthesis, 6% clay solution with 15mM silver
nitrate were microwaved till a colour change to a constant yellowish brown
colour was observed. The nanoparticles were characterised by SEM analysis
and later immobilized using 2% sodium alginate and 1M calcium chloride with
1microl of the reaction mixture. This was introduced the inoculated broth and
inoculated for 3 weeks. The plastic degradation was observed by FTIR analysis.
The agar plates with the treated plastic strips showed a clearance zone and
wrinkling of the surface that suggests deterioration of the plastic strip. The effect
of silver nanoparticles as an enhancer in the plastic degradation by Aspergillus
niger was promptly observed by the FTIR analysis. The clay particles helped in
the surface adsorption property that facilitated the cell adhesion off the fungus
that facilitated further enzyme reactions to promote the degradation.[7]

11
3.9 Studying the effect of biosilver nano particles on
polyethylene degradation 2018

Polyethylene has a lower tendency for degradation when compared to other

plastics. The process for this biodegradation includes photocatalysis and
microbial degradation . It is known from various studies that nanoparticles
increases the biodegradablity of a polymer. In a study by ( Jayaprakash et al
2018) the polyethylene samples were buried for 6 months and later the soil
adhering to the polyethylene was scraped and serially diluted to isolate the
fungal strains (Aspergillus oryzae). The fungal filtrate from the inoculated and
incubated media was treated with silver nitrate for the synthesis of silver
nanoparticles. During the incubation of this mixture a colour change was
observed which confirmed the reaction between the microbial enzyme
reductase and silver nitrate. The enzyme reductase reduced the size of the
silver ions to nanoparticle size. These silver nanoparticles were characterized
by UV-VIS spectral analysis , particle size analysis and zeta potential. TEM
analysis was performed to study the size range of the silver nanoparticles
present in the mixture.

To study the degradation, polyethylene strips (LDPE and HDPE) were added to
the culture broth with 1% AgNP and incubated for 5 weeks. FTIR analysis,
weight loss determination and GC-MS analysis were performed to observe any
possible signs of degradation. The FTIR analysis showed significant peaks
which represented the bending of the functional groups or alkane. It was
observed that the phytotoxicity of AgNP degraded the polyethylene and were
capable of degrading 64% of LDPE and 44% of HDPE. 8]

4.0 Biodegradation of oil based plastics

The production of oil based synthetic plastics has led to a severe accumulation
of plastic waste in the environment. It has caused drastic changes in the marine
system as well. Micro plastics have been a growing concern in the
environmental platform, and several studies have been made to study the bio-
remediation of these plastic wastes.[9]

12
Some specific studies have described that certain bacteria inhabiting in the
gastrointestinal duct of some insect larvae have the ability to degrade the
petroleum derived synthetic plastics like PE and PS.( Yang et al 2014) reported
that waxworms were capable of chewing and digesting LLDPE. Two 9 bacterial
strains, Enterobacter asburiae YT1 and Bacillus spp YP1 were isolated from
the guts of these wax worms and were reported of degrading untreated LLDPE
and formed biofilms to decrease the hydrophobic nature and caused surficial
damage to the LLDPE films. A gravimetric weight loss up to 13% was observed
after 28 days of incubation with the bacterial isolates.[10]

In another such study (Yang et al 2015), gut associated microbes from

mealworms were reported to play a role in the polystyrene mineralisation.
Exiguobacterium sp Yt2 was isolated and 28 days after incubation the polymer
strips were observed to have changes in surface topography, reduction in
hydrophobicity and molecular weight upto11%.[11]

4.0 Conclusion
Plastic waste management has become a global concern and various platforms
are open to tackle this issue effectively and economically. Landfills aren’t a wise
option to discard these calcitrant materials as it’s unsightly and would take
thousands of years to degrade. Methods like incineration release toxic gases
into the atmosphere which again poses various threats. Biodegradation has
become a popular phenomenon in waste management. Microbial cells can be
genetically modified and the degradation processes can be optimised, as
several studies have shown the high capability of certain microbes in degrading
polymers under standard conditions. Nanotechnology has been a revolutionary
subject that has wide applications in various domains. The addition of
nanoparticles with microbes to degrade the polymers have been reported to
enhance the process and achieve better percentage of degradation. Therefore,
this field has the potential to be explored so that we can have better solutions
to treat plastic wastes and reduce it’s risks. Another wise option is the use of
biodegradable polymers synthesized from plant materials which might
eventually lead to a complete reduction in the usage of plastic materials like
carry bags that are the most widely used plastic products.

13
5.0 References
1. Raziyafathima M, Praseetha PK, Rimal Isaac R S. Microbial Degradation of
Plastic Waste: A Review. Journal of Pharmaceutical, Chemical and Biological
Sciences. June-August 2016;4(2): 231-242

2. Swapnil K.Kale, Amit G.Deshmukh, Mahendra S.Dudhare, Vikram B.Patil.

Microbial degradation of plastic a review. J Biochem Tech 2015;6(2): 952-961

3. Poornima Pandey, P.Swati, Harshita, Manimita, Shraddha, Mahavir Yadav,

Archana Tiwari. Nanoparticles accelerated in-vitro biodegradation of LDPE :A
review.Advances in Applied Science Research, 2015, 6(4):17-22

4. Katarzyna Leja, Grażyna Lewandowiez. Polymer Biodegradation and

Biodegradable Polymers- a Review.Polish J. of Environ. Stud. Vol. 19, No.2
(2010), 225-266

5. Suman Mukherjee, Shamba Chatterjee. A comparative study of commercially

available plastic carry bag biodegradation by microorganisms isolated from
hydrocarbon effluent enriched soil. Intl. J. Curr. Microbial. App. Sci. 2014 3(5):
pp, 318-325

6. N Raaman, N. Rajitha, A.Jayashree, R. Jagadeesh. Biodegradation of plastic

by Aspergillus spp isolated from polythene polluted sites around Chennai. J.
Acad. Res, 1(6): pp, 313-316

7. Shreya Patwari, Sasmita Sabat. Application of nanoparticles as an enhancer

in the degradation process of plastics using A.niger. Scire Science and
MultidisciplinaryJournal.2017.1(1). https://doi.org/10.25129/SSMJSC2017.153

8. Vijayashree Jayaprakash, Uma Maheswari Devi Palempalli. Studying the

effect of biosilver nanoparticles on polyethylene degradation. Applied
Nanoscience. 2018. https://doi.org/10.1007/s13204-018-0922-6.

9. Noura Raddadi, Fabio Fava. Biodegradation of oil-based plastics in the

environment: Existing knowledge and needs of research and innovation.
Science of the Total Environment 679 (2019) 148-158.
14
10. Yang Y, Yang J, Wu, W.M, Zhao, J, Jiang,L, 2014. Evidence of polyethylene
biodegradation by bacterial strains from the guts of plastic-eating waxworms :
part 2. Role of gut microorganisms. Environ. Sci. Technol. 49, 13776-13784.

11. Yang,Y., Yang,J., Wu, W.M., Zhao,J., Song, Y., Gao,L., Yang., Jiang, L,
2015. Biodegradation and mineralization of polystyrene plastic-eating
mealworms: part 2. Role of gut microorganisms. Environ. Sci. Technol. 49,
12087-12093.

12. Ibrahim Khan, Khalid Saeed, Idrees Khan. Nanoparticles: Properties,

applications and toxicities. Arabian Journal of Chemistry (2017).
www.ksu.edu.sa , www.sciencedirect.com .

15