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BIOSCIENCES BIOTECHNOLOGY RESEARCH ASIA, June 2021. Vol. 18(2), p. 347-355
Published by Oriental Scientific Publishing Company © 2021
This is an Open Access article licensed under a Creative Commons license: Attribution 4.0 International (CC-BY).
*Corresponding author E-mail:
Biodegradation of Ultra-violet Irradiated Waste Polyethylene
Bags by Bacterial Community from Soil around Coal-fired
Thermal Power Plant
Arun Kalia and M.S. Dhanya*
Department of Environmental Science and Technology,
Central University of Punjab, Bathinda – 151401, India.
http://dx.doi.org/10.13005/bbra/2921
(Received: 17 March 2021; accepted: 04 June 2021)
The current study focused on biotic degradation of waste polyethylene bags using
bacterial community from hydrocarbon contaminated soil near coal fired thermal power
plant and also the effect of UV irradiation on its biodegradation.The predominant groups
in the bacterial community in the hydrocarbon contaminated soil near coal fired thermal
power plant were identified by 16s DNA sequencing were Steroidobacter, Flavisolibacter,
Planctomyces, Balneimonas, Gemmata, Alicyclobacillus, Lactobacillus, Mycobacterium,
Geodermatophilus, Prevotella, Virgisporangium and Adhaeribacter. The native bacterial
community from hydrocarbon contaminated soil was capable of polyethylene degradation.
The bacterial community in the hydrocarbon contaminated soil metabolized 12.85± 0.16
percent of polyethylene (10 g/L) as sole carbon source in mineral salt media within 30 days.The
UV irradiation of polyethylene enhanced weight loss of 22.80 percent higher than untreated
polyethylene. The improvement in bacterial degradation by UV exposure of waste polyethylene
in-vitro for 144 hresulted 15.78± 0.32 percent weight loss in 30 days. The photo-oxidation by
UV irradiation of polyethylene had surface disruption and was confirmed by Field Emission
Scanning Electron Microscopy (FE-SEM) and Fourier Transform Infrared Spectroscopy (FTIR).
The photochemical reaction induced by UV irradiation of polyethylene resulted in formation of
carbonyl peaks on polymer surface and addition as well as shifting of peaks. The morphological
changes of polyethylene by UV exposure enhanced colonization, metabolism by and synergistic
effect on polyethylene biodegradation by bacterial community from hydrocarbon contaminated
soil.
Keywords: Bacterial Community; Carbonyl Index; Waste Polyethylene Bags;
Weight Loss,Synergism; UV-Irradiation.
Polyethylene is one of the polymers with
versatile applications in packaging, insulation,
agricultural mulch lms, domestic and industrial
uses. The polyethylene is categorized based
on branching into low-density polyethylene,
high-density polyethylene, linear low-density
polyethylene and cross-linked polyethylene1. The
low-density polyethylene (LDPE) is the prominent
component of plastic waste which accounts for
60% of the total plastic production2. About 94
percent of plastic waste is thermoset plastic and
of great concern to the environment3.
348 KALIA & DHANYA, Biosci., Biotech. Res. Asia, Vol. 18(2), 347-355 (2021)
The conventional methods of disposal of
polyethylene waste include recycling, incineration
and landlling. The biodegradation of polyethylene
waste is environment friendly and sustainable than
the conventional physicochemical breakdown4.
The biodegradation has been aected by inert and
persistent nature of polyethylene for longer time
and interference of hydrophobic nature in polymer
availability to microorganisms5. The microbes
with high polyethylene degradation potential were
isolated from plastic contaminated sites such as
garbage soil, crude oil spilled sites, plastic dumping
site and soil form volcano crater6, 7.
The most important step in microbial
degradation of polyethylene is the surface
attachment of bacterial cells and biolm formation
on the polymer surface8. The alterations in
polyethylene properties make easy availability of
polyethylene for microbial biodegradation. The
researchers have studied the additive eect of
prior abiotic pretreatments of the polymer such as
thermal oxidation, UV irradiation and chemical
disintegration on microbial biodegradation of
polyethylene9. The UV treatment of polyethylene
resulted from photo-oxidative reactions by the
absorption of ultraviolet radiation10. The enzymes
secreted by the microbes initiate the biological
degradation of petro-based polymers and break the
polymer chain into oligomers and to monomers.
These smaller monomeric products are metabolized
in the microbial cells as carbon source11. In the
present study, the bacterial community from
hydrocarbon contaminated soil around coal red
thermal power plant was evaluated for their
ability to degrade polyethylene in liquid media
in-vitro and also the eect of UV-radiation on its
biodegradation.
MATERIAL AND METHODS
Collection of Waste polyethylene bags
The waste polyethylene bags were
collected from the local waste dumping site in
Bathinda, Punjab, India. The plastic bags were
translucent and of 20 microns thickness. The plastic
bags were thoroughly washed with deionized water
followed by ethanol sterilization (70% v/v), drying
overnight and stored aseptically for further use.
UV irradiation of polyethylene bags
The plastic bags were cut into small strips
of 1.5× 1.5 cm, sterilized with 70% v/v ethanol for
30 minutes. The polyethylene strips were irradiated
under UV lamps (16W) placing at 5 cm distance
for 144 h in UV protected glass chamber.
Soil sample for polyethylene degrading bacterial
community
Soil was collected from coal fired
Thermal Power Plant, Bathinda, Punjab (30Ú13’
59.57" N and 74Ú55’ 48.92" E) from the depth of
approximately 0–10 cm in sterile zip lock plastic
bags. The bacterial community used in the present
study was prepared by soil enrichment method
using waste polyethylene as sole carbon source
in Mineral Salt media (MSM)12. The bacterial
community with mixed culture of bacteria was
preserved at 4°C and used in further degradation
studies. The dominant bacterial groups in the
hydrocarbon contaminated soil were identied by
16s rDNA sequencing.
Biodegradation of waste polyethylene by
bacterial populations
The degradation study was performed with
untreated and UV irradiated waste polyethylene
(1% w/v) added to mineral salt media as a sole
source of carbon and energy at 30°C and continuous
agitation of 150 rpm for an incubation period of
1 month. The16 h old bacterial populations were
grown in Luria-Bertani broth was used as the
inoculum for degradation studies. The bacterial
growth and biodegradation study was investigated
by culturing polyethylene enriched medium
containing NH4 NO3 (1.0 gL-1); K2HPO4 (1.0
gL-1); KCl (0·15 gL-1); MgSO4·7H2O (0.2 gL-1);
CaCl22H2O (0·1 gL-1) and yeast extract (0·1 (gL-
1) along with 1·0 mgL”1 of ZnSO4 7H2O, FeSO4
6H2O, and MnSO4 13. The samples were collected
at 5 days intervals for estimation of biomass
growth of bacterial consortium. The weight loss
of polyethylene samples from the culture media
was estimated14 after washing with sodium dodecyl
sulfate (2 % (v/v) followed by distilled water15. All
the experiments were conducted in triplicates and
the results were the mean values with standard
deviations. The results were statistically analyzed
by single factor Analysis of Variance followed by
Post- hoc Tukey test.
Quantication of bacterial adherence on the
polyethylene
The bacterial populations on the
polyethylene surface were measured indirectly
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KALIA & DHANYA, Biosci., Biotech. Res. Asia, Vol. 18(2), 347-355 (2021)
by determining the concentration of extractable
protein. The protein estimation by Lowry’s method
was followed for the supernatant obtained from
boiling polyethylene samples in 0.5 N NaOH for
30 min16.
Structural changes on polyethylene surface
The Fourier Transform Infrared
Spectroscopy (FTIR) and Scanning Electron
Microscope (SEM) analysis of untreated
polyethylene, UV irradiated polyethylene
and samples of polyethylene lms after biotic
degradation for 30 days were studied for
understanding structural changes. The change in the
polyethylene surface and generation of new peaks
were obtained by FTIR spectroscopy (FTIR Bruker,
Model: Tensor 27) from 400 to 4000 cm”1. Using
the FTIR spectrum, carbonyl residue as carbonyl
index and double bond index were measured on the
basis of ratio of relative intensities of the carbonyl
band (1715 cm”1) and double bond band (1650
cm”1) to that of methylene scissoring band at 1460
cm”1. The bacterial growth and structural changes
on polyethylene was checked by SEM analysis as
per the method given by Tribedi and Dey17.
RESULTS AND DISCUSSION
The hydrocarbon contaminated soil from
the coal red Thermal Power Plant, Bathinda used
for screening polyethylene degrading bacterial
community was slightly alkaline with pH 8.10 ±
0.08 and electric conductivity of 556 ± 1.8 µS/cm.
The total petroleum hydrocarbon content present
in contaminated soil aected the physicochemical
properties18. The soil sample had bacterial
population of 3.11× 109 CFU/g (Figure 1).
The 16s rDNA sequencing of native
bacterial community from hydrocarbon
contaminated soil near coal red thermal power
plant identied predominant bacterial genera as
Steroidobacter, Flavisolibacter, Planctomyces,
Balneimonas, Gemmata, Alicyclobacillus,
Lactobacillus, Mycobacterium, Geodermatophilus,
Prevotella, Virgisporangium and Adhaeribacter.
The bacterial populations in the hydrocarbon
contaminated soil were able to grow using
polyethylene as sole carbon and energy source.
It was clear from the protein concentration at 5
days with an increase of 3.69 times than 0 h. The
continuous increase in protein concentration over
time from the colonization of microbial populations
on the polyethylene surface. The increase of
protein concentration was 12.93 times and 25.98
times at 15 days and 30 days respectively than the
inoculation. But the increase in biomass growth
was declined to 44 percent in 10th day than 5th day
(Graph 1). The increase was 2.44 times in the next 5
days which was again reduced to 59 percent in 20th
day of biodegradation study. Further the percent
increase was reduced to 9.69 percent in 30th day
of incubation.
The biomass growth using polyethylene
as carbon source and biodegradation was
conrmed from the 12.85± 0.16 percent weight
loss of waste polyethylene bags in 30 days.
The range of weight loss in polyethylene after
Fig. 1. (a) Hydrocarbon contaminated soil from Thermal power plant for polyethylene degrading bacteria (b)
Viable bacteria from hydrocarbon contaminated soil on nutrient agar plates.
350 KALIA & DHANYA, Biosci., Biotech. Res. Asia, Vol. 18(2), 347-355 (2021)
Fig. 2. SEM images of (a) Untreated polyethylene (b) UV irradiated polyethylene for 144 h
Fig. 3. SEM images of bacterial growth in 30 days on (a) untreated polyethylene (b) UV irradiated polyethylene.
biodegradation of 30 days was from 12.67 to
13.05 percent. This weight loss of polyethylene
by bacterial community from hydrocarbon
contaminated soil from coal red thermal power
plant was higher than LDPE biodegradation in
60 days by Pseudomonas aeruginosa (ISJ14)
from waste dumping site19. The polyethylene
degradation was increased by dierent microbes
than pure culture20. The microbial populations
present in the hydrocarbon contaminated sites
had high biosurfactant production capability
and presence of alkane hydroxylase gene for
competent biodegradation21. The degradation
of polyethylene was due to interaction between
dierent polyethylene degrading microorganisms22.
The UV irradiation increased the bacterial
biomass (protein concentration) to 61.89 percent
than untreated polyethylene lms in 5 days and
33.82 percent in 30 days of degradation. The
protein concentration of bacterial community from
utilizing UV treated polyethylene as carbon source
was 24.30 and 38.82 times higher in 15th day and
30th day of degradation respectively than the day of
inoculation. The initial growth was also higher in
UV treated polyethylene with 6.67 times in 5 days
of inoculation. The rate of growth in UV treated
polyethylene was doubled in 10 days which was
reduced by 65.56, 24.45, 15.41 and 11.2 percent
respectively for 15, 20, 25 and 30 -days incubation
period. The UV exposure of waste polyethylene
351
KALIA & DHANYA, Biosci., Biotech. Res. Asia, Vol. 18(2), 347-355 (2021)
bags improved initial growth and more bacterial
colonization on polymer surface.
The increase in biomass growth of bacterial
community from hydrocarbon contaminated soil
utilizing UV irradiated polyethylene as carbon and
energy source was conrmed with biodegradation
of 15.81± 0.32 percent weight loss of waste
polyethylene bags in 30 days. The polyethylene
after biodegradation had a weight loss ranged
from 15.43 to 16.22 percent in 30 days. The
UV irradiation on bacterial biodegradation of
polyethylene bags was statistically found highly
signicant at P value <0.01 by one way ANOVA
with Post- hoc Tukey test. The biomass growth was
also found signicant (P value <0.01) with the UV
treatment of polyethylene bags.
The UV irradiation increased the
carbonyl index and caused signicant alteration
on the polymer structure that favored the enhanced
microbial attachment on polymer surface. The
Graph 1. Protein concentration of bacterial population as biomass indicator on olyethylene surface using
nontreated and UV irradiated polyethylene lms.
Graph 2. FTIR spectra of non-treated and UV irradiated polyethylene lms.
352 KALIA & DHANYA, Biosci., Biotech. Res. Asia, Vol. 18(2), 347-355 (2021)
increased degradation of polyethylene was also
reported by Jeon and Kim23 and Montazer and
coworkers 24.
The structural modifications of UV
treated polyethylene was clear from FTIR spectra.
FTIR spectra of nontreated polyethylene lms
and UV irradiated lms is shown in Graph 2.
The infrared spectrum of the waste polyethylene
bags after photooxidation (UV irradiation for 144
h) showed a peak at 1716 cm”1 and 1055 cm”1
representing carbonyl groups (-C=O) and ether
groups (-C-O-C-) respectively. The generation of
additional peaks at 3807 cm-1, 3420 cm-1, 2362
cm-1, 2121 cm-1 and 1917 cm-1 after 144 h of UV
irradiation also conrmed the structural changes
in the polymer. The functional groups identied in
the FTIR spectra were indicator precursors in the
photochemical reactions of the polyethylene25. The
prior UV pretreatments of polyethylene increased
surface hydrophilicity by formation of additional
carbonyl groups. The radical was formed initially
from absorption of UV-radiation on polyethylene
and further to hydro-peroxide formation and then
terminal carbonyl groups 26.
The carbonyl index showed the oxidation
of polyethylene by UV radiation from the carbonyl
species in the FTIR spectra. The UV pretreatment
of the polyethylene for 144 h enhanced the ester
carbonyl index and keto carbonyl index by 7.97 and
9.49 percent respectively. The increase in carbonyl
index from UV treatment was due to photo-
oxidative reaction of plasticizer and stabilizers
present in polyethylene that later leads to chain
scission27.
The UV irradiation of waste
polyethylene bags also increased the brittleness
of the polyethylene, surface roughness, cracks
and depressions on the polymer surface as clear
from scanning electron microscopy (SEM)
images (gure 2). The structural changes on the
UV irradiated polyethylene enhanced bacterial
consortium from hydrocarbon contaminated soil
to improve degradation by 22.80 percent higher
than nontreated waste polyethylene lms in vitro.
The SEM images after 30 days
biodegradation study was also confirmed the
bacterial biodegradation of polyethylene (gure 3).
The adherence of microorganisms on the surface of
plastics followed by the colonization of the exposed
surface are the major mechanisms involved in the
microbial degradation of plastics28. The microbes
adhered to the surface increased biodegradation
and cause cracks and cavities29.
The decrease in pH in the growth media
with polyethylene evidenced the metabolism
and the biodegradation process in four weeks
period by bacterial populations from hydrocarbon
contaminated soil. This was also in accordance with
reports of Das and Kumar 30.
The weight loss of untreated polyethylene
without pro-oxidant additives and oxidative
pretreatment by UV irradiation of polyethylene
conrmed the biodegradation potential of bacterial
community from hydrocarbon contaminated soil.
The bacterial populations in the community from
hydrocarbon contaminated soil had the polyethylene
degrading ability from surface adhesion, bacterial
proliferation and exopolysaccharide production 31.
The interactions among bacterial populations in
the community with dierent oxidative enzymes
resulted in higher polyethylene biodegradation 32,33.
The improvement in weight loss of the
waste polyethylene by UV treatment and biotic
degradation conrmed the synergistic eect from
photooxidation by UV radiation and biodegradation
by bacterial community from hydrocarbon
contaminated soil24. The similar results were also
reported by Albertsson et al. 26 and Esmaeili et al.
34.
The UV light exposure induced the
photooxidation of the polymer and generated the
free radicals that facilitate the microbial attack26,
35,36. The photooxidation by UV irradiation
increased carbonyl bond and terminal double-bond
index which in turn increased microbial utilization
of carbonyl residues17,37,38. The biodegradation was
initiated with adherence of microbes on surface
of polyethylene39. The biodegradation of these
recalcitrant pollutants was also enhanced by the
surfactants produced by the microbes which
increase bioavailability40. The enzymes present in
the polyethylene degrading microorganisms from
hydrocarbon contaminated soil cleave the polymer
chain into monomers and smaller fragments that
can be easily used up by the microorganism31,41.
This depolymerization process with intracellular
and extracellular depolymerases helped microbes
from hydrocarbon contaminated soil to utilize
polyethylene as carbon and energy sources 23,
42,43. The products were easily metabolized by
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KALIA & DHANYA, Biosci., Biotech. Res. Asia, Vol. 18(2), 347-355 (2021)
microbes via the â-oxidation pathway and citric
acid cycle44. . The biodegradation eciency was
improved in UV irradiated polyethylene and
with microbial communities resulted in more
microbial colonization and synergistic effect
of photooxidation and biodegradation 26,45,46.
Microbial biodiversity inhabiting contaminated
sites have already proved high eciency to degrade
polyethylene. The nature and type of polyethylene,
types of additive present in the polymer and extent
of prior photooxidation pretreatment play a crucial
role in biodegradation47.
CONCLUSION
The hydrocarbon contaminated soil
from coal red thermal power plant was a good
habitat for dierent ecient bacterial populations
capable of degrading polyethylene wastes.
The prior UV irradiation resulted in increased
bacterial adherence, colonization and synergism in
polyethylene biodegradation. The combination of
photo-oxidation and biodegradation helps in easy
metabolism of polyethylene and in plastic waste
management without any ecological impacts.
ACKNOWLEDGMENTS
The authors are grateful to Central
University of Punjab, Bathinda (India) for providing
facilities to undertake the present work. The authors
specially thank Central Instrumentation Facility,
Central University of Punjab for FTIR and SEM
analysis. The rst author thanks University Grants
Commission for the Junior Research Fellowship
(UGC-JRF).
Conict of interest
The authors declare that there is no
conict of interest.
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