




































Highlights in BioScience
ISSN:2682-4043
DOI:10.36462/H.BioSci.202402

Research Article

Open Access

1 Department of Chemistry, Faculty of Mathemat-

ics and Natural Sciences, Universitas Negeri

Malang, Jl. Semarang No. 5 Malang, Malang,

East Java 65145, Indonesia.
2 Biotechnology Program, Department of Applied

Science, Faculty of Mathematics and Natural

Sciences, Universitas Negeri Malang, Jl. Se-

marang No. 5 Malang, Malang, East Java 65145,

Indonesia.
3 Research Group, Department of Chemistry, Fac-

ulty of Science, Universiti Teknologi Malaysia,

Johor, Malaysia
4 Enzyme Technology and Green Synthesis,

Department of Chemistry, Faculty of Science,

Universiti Teknologi Malaysia, Johor, Malaysia

* To whom correspondence should be
addressed: evi.susanti.fmipa@um.ac.id

Editor: Morad M. Mokhtar, Agricultural Genetic
Engineering Research Institute (AGERI), Giza,
Egypt.

Reviewer(s):
Suleiman Aminu, Department of Biochemistry,
Ahmadu Bello University, Zaria, Nigeria.

AbdulAziz Ascandari, Chemical and Biochemical
Sciences-Green Process Engineering, University
Mohammed VI Polytechnic, BenGuerir, Morocco.

Received: October 13, 2023

Accepted: February 17, 2024

Published: May 21, 2024

Citation: Wijayanti C, Sanjaya EH, Wahab RA,
Susanti E. Exploration of the polypropylene
degrading bacteria candidates from the passive
zone of the Supit Urang landfill in Malang city
by using the next generation sequencing (NGS)
method. 2024 May 21;7:bs202402

Copyright: © 2024 Wijayanti et al.. This is an
open access article distributed under the terms of
the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduc-
tion in any medium, provided the original author and
source are credited.
Data Availability Statement: All relevant data are
within the paper and supplementary materials.
Funding: The authors have no support or funding to
report.
Competing interests: The authors declare that they
have no competing interests.

Exploration of the polypropylene degrading bacteria candidates from
the passive zone of the Supit Urang landfill in Malang city by using the
next generation sequencing (NGS) method

Chandra Wijayanti1
>< ,Eli Hendrik Sanjaya1,2

>< �, Roswanira Abdul Wahab2,3,4
>< �,

Evi Susanti*,1,2
><�

Abstract

The buildup of plastic waste in the passive zone of the Supit Urang landfill located in
Malang City has been ongoing since 2018. Currently, plastic waste in this area appears
brittle and cracked, potentially providing a habitat for PP-degrading bacteria. This
research aims to explore the potential of polypropylene-degrading bacteria using
Next Generation Sequencing (NGS) techniques in the passive zone of the Supit
Urang landfill, Malang City. Our study was conducted in four steps: 1) sampling and
sample collection, 2) DNA sequencing, 3) bioinformatics analysis, and 4) bibliometric
analysis for identification of polypropylene-degrading bacteria. Based on the results
of full-length sequencing using Oxford Nanopore Technologies with whole amplicon
sequencing techniques, a total of 2,496 sequences were read, and 1,713 sequences
were identified as species in the passive zone of Supit Urang landfill. The most
abundant bacterial phyla in this region were Proteobacteria (51%), Firmicutes (21%),
Acidobacteria (7%), Bacteroidetes (6%), Planctomycetes (4%), Actinobacteria (3%),
Gemmatimonadetes (2%), Nitrospirae (2%), and Chloroflexi (2%). These results
indicate that Proteobacteria and Firmicutes are abundant in the passive zone of TPA
Supit Urang and could potentially biodegrade microplastics such as polypropylene. The
narrative review’s research showed that numerous bacterial species, including Bacillus
thuringiensis, B. cereus, and Bacillus sp., were identified by NGS analysis as possible
polypropylene-degrading bacteria.

Keywords: Bacterial diversity, Next generation sequencing, Polypropylene degradation

Introduction
Reports of synthetic waste contamination began in the 1970s [1]. Since then, the number

has continued to become a serious problem and global concern. This is caused by the natural

biodegradation of plastic waste that takes a very long time, i.e., 50 to more 100 years [2]. One type

of plastic waste that often becomes problematic is polypropylene (PP) waste. PP is an extremely

versatile polymer that is preferred for its ease of processing, barrier properties, gloss, and dimensional

stability, but as its use grows, so does the problem of managing its waste. PP is utilized in diverse

commercial applications, including packaging, labelling, and fibre production for indoor and outdoor

carpeting. More than 79.01 million tons of plastic waste is estimated to have reached our oceans.

According to data Making Oceans Plastic Free (2017), the use of the PP in Indonesia reaches 1,278

million metric tons. Important to note, many types of PP plastic are scattered in the passive zone of

the landfills [3; 4]. Synthetic polymers such as PP have the potential to become microplastics that

are harmful to the environment and human health. Important proteins such as albumin, globulin and

fibrinogen are altered by microplastics when they enter the body and become dysfunctional due to

their interaction with blood particles [4]. Changes in chromosome structure caused by microplastics

entering cells and tissues have been linked to cancer, obesity and infertility [5]. As a result, the

presence of mountains of plastic waste will endanger the environment and human health [6].

Highlights in BioScience Page 1 of 10 May 2024|Volume 7

https://doi.org/10.36462/H.BioSci.202402
https://creativecommons.org/licenses/by/4.0/
mailto:chandrawijayanti1811@gmail.com
mailto:eli.hendrik.fmipa@um.ac.id
https://orcid.org/0000-0002-6513-5198
mailto:roswanira@utm.my
https://orcid.org/0000-0002-9982-6587
mailto:evi.susanti.fmipa@um.ac.id
https://orcid.org/0000-0002-0201-7303
http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

In recent years, natural degradation has become critical as
social pressures increase for safe plastic waste management. One
alternative step is biodegradation by utilizing indigenous degrad-
ing bacteria. Several studies show that using indigenous bacteria
can increase the degradation rate and support ecosystem recovery
without causing adverse impacts and is environmentally friendly
[7]. This leads to the need of an alternative solution in form of
the development of biological degradation [8].

The soil microbial community may contain indigenous bacte-
ria candidates that are more adaptive. Observation results found
that waste in the zone that has been passive for a long time in
Malang City, namely the Supit Urang Landfill, since 2018 con-
tains a lot of plastic that has experienced degradation, became
cracked and brittle. Since the condition and nutritional character-
istics of each landfill soil are likely to be different, resulting in
diversity, different isolates of PP degrading bacteria can therefore
be obtained from different landfills. From contaminated sites,
including landfills, bacterial species capable of biodegrading
plastics have previously been isolated [9].

The exploration of PP-degrading bacteria isolates can be done
using bacterial culture techniques or a genomic approach. The
bacterial culture technique takes a relatively long period of time
because of some stages as sampling, isolate culturing, screening
and isolate identification are influenced by many parameters such
as nutrition in growth medium and environmental conditions.
Meanwhile, identifying the presence of PP-degrading bacteria
with the fastest culture technique takes 40 days [4]. Thereby,
isolate culturing is slightly abandoned. On the other hands, the
genomic technique using Next Generation Sequencing (NGS)
methods takes only a few days. NGS uses a parallel approach
to sequencing, in which millions of DNA fragments can be se-
quenced at the same time in a single reaction, allowing for high
throughput analysis [10]. NGS automation is very high because
almost all steps in the sequencing process, including sample
preparation, amplification, library preparation, sequencing, and
data analysis, can be done simultaneously. After sequencing,
the data is obtained and subjected to bioinformatics analysis to
identify colonizing bacterial species.

This research aims to explore candidate polypropylene de-
grading bacteria isolates in the passive zone of Supit Urang Land-
fill, Malang City using NGS methods. It is hoped that the findings
of this study could serve as evidence for the search of polypropy-
lene degrading bacteria from landfills.

Experimental section
This research uses an explorative descriptive method to assess

the biodiversity of bacterial communities in the passive zone of
TPA Supit Urang Malang City, consisting of: 1) sampling site and
sample colection, 2) DNA sequencing, 3) bioinformatics analysis,
and 4) bibliometric analysis for identification of polypropylene
degrading bacteria. The analysis of the passive zone samples
begins with the isolation of genomic DNA from the samples from
passive zone of TPA Supit Urang Landfill using Zymo Research’s

ZymoBIOMICS DNA MiniPrep Kit, followed by amplification
of the isolated DNA with 16S 27F - 1429R primers using (NGS)
using Oxford Nanopore Technologies technology, sequencing of
the DNA amplification results using NGS, downstream analysis
of the results using Pavian Krona software, instrumentation and
library preparation using Oxford Nanopore Technology kits, and
review of blibliometric analysis to identify native polypropylene-
degrading bacterial candidates.

Sampling and Sample Collection
Soils were collected from the passive zone of the TPA Supit

Urang in Mulyorejo Village, Sukun District, Malang City, East
Java, with the coordinates of 7o59’20.04" latitude in the South
and 112o38’17.52" longitude in the East. The sampling technique
refers to Helen et al. (2017) [4] and Anah et al., (2020) [11]. Soil
sediment samples were collected with a shovel from 30 to 60
cm above the soil surface at 5 different locations. Samples were
taken at each end as much as 200 g to obtain a total piece of 1
kg sediment. Then, the samples from the five points were stirred
evenly and sifted to get a homogeneous soil. Homogeneous
samples were subjected to preliminary physico-chemical tests
to determine the temperature, moisture, pH, organic carbon and
total N content of the soil. The soil samples were then placed
in dark container and containing ice gel to maintain a consistent
temperature during transport.

DNA from the soil samples (Sample Code: CA20) were ex-
tracted using the ZymoBIOMICS DNA MiniPrep Kit from Zymo
Research with minimum sample amount of 250 mg. The basic
principle of using the ZymBIOMICS DNA MiniPrep Kit is to
lyse and disrupt all microbes in samples using lysis buffer and
beads. Then, the purification step is repeated several times to en-
sure better separation and more efficient protein precipitation, and
the final step uses affinity chromatography to obtain pure DNA
extracts. DNA concentration measurements were performed us-
ing a Qubit fluorometer. Qubit provides DNA concentration
information using fluorogenic dyes that bind selectively to DNA
or RNA. The dye emits a signal only when bound to the target.
It can then be used to determine the purity of DNA using the
NaNoDrop spectrophotometer, providing a direct measurement
of the A260/280 purity ratio. Using the two together can help
ensure that the DNA sample is of sufficient concentration and
purity for sequencing applications.

DNA Sequencing
All obtained amplicons were sequenced using Full-Length

next-generation sequencing (NGS) techniques. Full-Length Se-
quencing using Oxford Nanopore Technologies technology. Ox-
ford Nanopore Technologies’ technology is a nanopore-based
DNA sequencing technology in which DNA or RNA molecules
can be passed through nanopores from end to end, causing elec-
trical changes that produce a complete sequence of the molecule.

Highlights in BioScience Page 2 of 10 May 2024|Volume 7

http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

Bioinformatics analysis
DNA amplification results were sequenced with the special-

ized software for Oxford Nanopore Technologies’ technology,
namely GRIDION instrument using MinKNOW software ver-
sion 20.06.9. Base calling was carried out using Guppy software
to visualize the data from genetic sequence data resulting in
±2400 sequences. Quality control was performed using nano
plots and classified as bacteria and archaea indices downloaded
from the website (https://ccb.jhu.edu/software/centrifuge). The
analysis was carried out downstream using the software Pavian
Krona Tools: to visualize microbial diversity data using krona
visualization.

Bibliometric Analysis for Identification of
Polypropylene-Degrading Bacteria

The data sources used were scientific articles in the 1960-2021
period, sourced from the Scholarly Database (https://scopus.com).
Search for scientific articles was made by using Publish or Per-
ish (PoP) Version 7 software with the keywords degradation of
polypropylene. The software collects and processes data from
sources such as Scopus to provide statistics about one’s scholarly
work, such as number of citations, h-index, g-index, and other in-
dicators. Data obtained from Publish or Perish was filtered based
on the suitability with research, then stored in CSV (Comma
Separated Values) and RIS (Research Information System) files.
The results obtained were analysed using VOSviewer version
1.6.20 software for bibliometric map analysis to determine the
development of the topic of polypropylene-degrading bacteria.
VOSviewer is software used to visualize and analyze bibliometric
networks and overall concept maps emerging from the scientific
literature.

Results and Discussion
Extracted DNA from the sample with the ZymBIOMICS

DNA MiniPrep Kit were 50 µL with a concentration of 57.4
ng/µL, which means there is 2.86 µg of DNA. Amplification of
the 16S rRNA gene by polymerase chain reaction (PCR) using the
primers resulted in an amplicon size of around 1,500 bp (Figure
1) and 2,496 sequences. According to Sacchi et al. (2002), the
PCR band sizes of about 1,500 bp 1,600 bp are those of the
bacterial 16S rRNA sequences as they can distinguish between
specific taxa or strains and are indispensable for describing new
species [12].

A total of 2,496 sequences were successfully identified. All
sequences were then subjected to quality control using NanoPlot
to ensure that the sequences obtained were Bacteria or Archaea,
and classified using the source data downloaded from the website
(https://ccb.jhu.edu/software/centrifuge). Based on this classifi-
cation, 2,496 sequences were bacterial sequences [12]. A total
of 2,496 bacterial sequences were identified from the full-length
16S rRNA sequence results as Kingdom (1), Phylum (25), Class
(10), Order (10), Family (16), Genus (19), Phylum (659), Species
(1,713), and Subspecies (43). In addition, the data obtained was
further analyzed using Pavian Krona Tools software to visualize

Figure 1. Electrophoregram of amplified bacterial DNAs from CA20 (passive

zone sample of Supit Urang Landfill, Malang City).

microbial diversity data using Krona visualization.
Based on the results of the Krona graph, the diversity of

bacterial phyla abundant in the passive zone of Supit Urang land-
fill are Proteobacteria (51%), Firmicutes (21%), Acidobacteria
(7%), Bacteroidetes (6%), Planctomycetes (4%), Actinobacteria
(3%), Gemmatimonadetes (2%), Nitrospirae (2%), and Chlo-
roflexi (2%). Meanwhile, relative abundance less than 0.6% is
categorized into other categories. Proteobacteria and Firmicutes
exhibited the highest bacterial abundance among the analyzed
samples. The overall structure of the present bacteria in the pas-
sive zone of Supit Urang Landfill, Malang, is visualized in a
Krona Graph (Figure 2), showing a broader taxonomic hierarchy.
The style of reading the Krona Graph from inside to outside
showed the abundance of the bacterial domain consisting of 9
phyla (Proteobacteria, Firmicutes, Acidobacteria, Bacteroidetes,
Planctomycetes, Actinobacteria, Gemmatimonadetes, Nitrospi-
rae, Chloroflexi) (Figure 3). Each consisted of classes, orders,
families, genera, and species. The advantage of the Krona Graph
visualization is that it has a more extensive or highly detailed
taxonomic hierarchy [13].

Another similar study with the same conditions and research
methods but different sampling sites [14] from Bestari Landfill,
Probolinggo, resulted in different phyla identified (Table 1). This

Highlights in BioScience Page 3 of 10 May 2024|Volume 7

http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

Figure 2. The Krona graph represents bacterial diversity distribution in the Passive Zone of Supit Urang Landfill Malang, East Java.

proves the initial assumption of this study that each landfill has its
own characteristics resulting in different microbial diversity. The
characteristic properties of each landfill mainly depend on fac-
tors of different environmental conditions, including pH, oxygen
availability, temperature, humidity, and landfill age which will
influence differences in microbial diversity [13]. Differences in
environmental conditions at the Supit Urang Landfill and Bestari
Landfill are shown in Table 2. The Bestari Landfill location is
in the lowlands compared to the Supit Urang Landfill location,
which is in the highlands. Thus, Bestari, Probolinggo Landfill
is used as a comparison because it has different environmental
conditions (physicochemical conditions, types of waste disposed,
topographical differences) will affect the diversity of native bac-
teria that are adaptive to the environment.

Based on our preliminary investigations, the differences in the

Table 1. Comparison of the diversity of identified bacterial phyla with those in
the reference [14].

Phylum Supit Urang Landfill, Malang Presence Reference Percentage (%)
Proteobacteria identified 51 Identified 70
Firmicutes identified 21 Identified 15
Acidobacteria identified 7 not identified
Bacteroidetes identified 6 not identified
Planctomycetes identified 4 Identified 2
Actinobacteria identified 3 not identified
Gemmatimonadetes identified 2 not identified
Nitrospirae identified 2 not identified
Chloroflexi identified 0.6 not identified
Cyanobacteria not identified identified 0.3

abundance of some phyla in the Supit Urang Landfill, Malang,
and in the Bestari Landfill, Probolinggo, could be due to the
differences in some of the environmental parameters in the differ-
ent soil profiles. Based on research [15], several Pseudomonas
strains have been reported to aid in PP degradation [3; 4]. In

Highlights in BioScience Page 4 of 10 May 2024|Volume 7

http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

Table 2. Comparison of identified environmental conditions with reference [14].

Parameter Supit Urang Landfill, Malang Bestari Landfill, Probolinggo
Temperature 33oC 36oC
Humidity 46% 52%
pH 7 7
Carbon Organic 5.71% 2.85%
Nitrogen Total 0.36% 0.15%

both aquatic and terrestrial environments, Pseudomonas has a
pH range of 4.5 to 9.5 [16]. Unfortunately, in the abundance of
microbial diversity of the NGS results did not show the presence
of Pseudomonas (Figure 3). It is possible that the pH range of
the Supit Urang Landfill habitat is within the growth range of
Pseudomonas, but there are other nutrients that do not support
Pseudomonas growth. Therefore, Pseudomonas has not been
identified in the Supit Urang Landfill.

Most members of the phylum Firmicutes are Gram-positive
bacteria with the ability to form endospores. They are primarily
found in soil habitats [17]. The most frequently mentioned organ-
isms are the genus Bacillus members [18]. Bacteria in this genus
can produce oval or cylindrical endospores and can function as
aerobes or facultative aerobes. Certain strains can generate extra-
cellular hydrolytic enzymes that break down complex polymers,
such as lipids, to be used as a carbon source and electron donor.
Moreover, several of these bacteria synthesize antibiotics and
insecticides [17]. Growth took place between pH values of 5.4
and 8.5, with the most favorable growth at 7.0 [19]. Bacillus is
known to be able to degrade PP [20; 21; 22; 23].

Pseudomonas was suggested by 21.0% of the studies, Bacil-
lus by 15%, and a combination of the two by 17% of the studies
as the bacteria that effectively initiated the biodegradation of syn-
thetic polymers. Thus, the environmental conditions of the waste
piles are severe because of the existence of bio toxic compounds
and various synthetic substrates [24].

Based on Table 2, the temperature in the passive zone is 33oC;
mesophilic bacteria grow well at that temperature. The optimal
temperature for plastic-degrading bacteria to grow optimally is
20oC-40oC [25]. Humidity in the sampling zone is 46% (moder-
ate), which is not suitable for bacterial growth [26]. The pH at the
side locations ranged from 6.97.2, indicating that the sampling
location was neutral. Furthermore, in the analysis, the organic
C content was found to be 5.71%, which was classified as very
high (> 5%), and the organic N content was 0.36% (classified
as moderate). Soil total nitrogen is used as an essential index of
soil fertility. Based on the physicochemical characteristics of the
soil, dealing with Proteobacteria or Firmicutes bacteria requires
selective media to inhibit the growth of other unwanted species.

Identity of Polypropylene-Degrading Bacteria Based on
Bibliometric Analysis

The data source used was scientific articles based on the Sco-
pus.com database, with the keyword "degradation of polypropy-
lene" in the keywords. Article searches are based on the Scopus
database, because Scopus is one of the databases whose scope
of reputable international journals can be recognized or well

received by all researchers around the world [27]. As shown
in Table 3, 200 articles were published by scopus.com, filtered
according to research on polypropylene degradation from 1960-
2021 in the scopus.com database, which has increased signifi-
cantly yearly. The bibliometric analysis was performed at the
end of 2022, but the articles that could be filtered by PoP only
reached 2021. This is because Scopus-indexed research on bacte-
rial degradation lasts until 2021.

Based on the graphical data of the number of publications
from 1960-2021, it can be seen that the number of publications
on polypropylene degradation has an unstable trend from 2001-
2022. The beginning of the research topic on polypropylene
degradation started in 1960 with one study and was significant
until 1992. However, since 2001, research on polypropylene
degradation has experienced ups and downs in the number of
publications. Based on the graph of the number of publications
(Figure 4), there were 12 publications in 2006 and 2018. In
addition, only 1 publication was recorded in 2022, and even no
publication records were found in 2023.

In terms of document type (Figure 5), most publications were
research articles (166; 83%), followed by reviews (29; 14.5%),
conferences (3; 1.5%), and short surveys (2; 1%). Information
from the Scopus database showed that most of the publications
were all open access.

Table 3. Analysis of PoP Metrics of Relevant Articles

Results Metrics
Publication years 1960-2021
Citation years 62
Papers 200
Citations 58412
Cites/year 942.13
Cites/paper 292.06
Authors/paper 1.00
h-index 156
g-index 200
hI, norm 156
hI, annual 2.42
hA-index 38

The terms in the title, abstract, index keywords, and year of
publication were identified for each publication. A circle repre-
sented each time. The size of the process reflected the number
of publications. Meanwhile, the distance between the two terms
offered an approximate indication of the relatedness of the terms.
On the other hand, colors represent groups of words that are
strongly related to each other [28]. The development map of
the publication of polypropylene-degrading bacteria showed that
there were 4 clusters (Figure 6 & Table 4). Keyword analysis
provides a comprehensive overview of research trajectories and
research topics (Donthu et al., 2021). Based on the keyword
analysis (Table 4), cluster 1 has the highest number of selected
keywords, which means that it contains keywords that are fre-
quently used in the research context, which is "ability". In terms
of main keywords, "ability" has the highest occurrence, followed

Highlights in BioScience Page 5 of 10 May 2024|Volume 7

http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

Figure 3. Relative abundance (%) at the phylum (A) and species (B) levels obtained from the Passive Zone of the Supit Urang Landfill Malang City, East Java,

Indonesia.

Figure 4. Global publication trend by document types.

Highlights in BioScience Page 6 of 10 May 2024|Volume 7

http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

Figure 5. Publication types used for bibliometrics of 200 published articles.

by "biodegradability". This refers to the ability of a substance to
be decomposed or broken down by microorganisms into simpler
and less harmful components in the environment. The process of
biodegradability is part of the natural cycle of plastic degradation,
in which microorganisms such as bacteria and fungi break down
complex compounds into simpler forms.

Figure 6. Knowledge map based on index keywords from Elsevier Scopus from

1960-2021.

The resulting clusters of keywords provide an overview of
the research themes. For example, cluster 1 describes "ability"
or "biodegradability", while the term in cluster 2 is the bacterial
species Bacillus, which is most often found in research as a
potential bacterium that can degrade plastics commonly isolated
from sediments. Clusters 3 and 4, on the other hand, show
interest in exploring bacterial species and sample locations to
obtain more candidate bacterial species with biodegradation and
bioremediation capabilities.

Table 5 shows that only some studies of PP biodegradation
by bacteria have been reported. Bacillus thuringiensis has been
reported to degrade PP by 12% for 40 days [3]. The study was
conducted using PP pre-treatment, which involved irradiation
or thermal treatment [29; 30] and was shown to reduce the hy-
drophobicity of the polymer. B. flexus has also been shown to

Table 4. Keyword clusters in degradation of polypropylene research

Clusters No. of keywords Selected keywords Color

1 37 Ability, B. cereus, bacteria isolate, bacteria

isolated, bacterial strain, biodegradability,

commercial application, day, environment,

environmental pollution, extent, important

role, incubation, isolate, mangrove

ecosystems, medium, microorganism,

microplastic material, mineral salt medium,

polymer reduction, polypropylene

degradation, polypropylene granule,

polypropylene polymer, positive potential,

the present study, rate, recalcitrance,

reduction, remediation, Sglobispora, sole

carbon source, Sporasarcina globispora,

term, versatile polymer, vitro, weight loss

Red

2 13 Bacillus, Bacillus biofilm, biodeterioration,

degradation potential, growth, growth

kinetic, mangrove sediment, polypropylene

microplastic, pretreated polypropylene,

Pseudomonas, Rhodococcus, synergistic

growth

Green

3 11 Bacillus paralicheniformis, biodegradation,

low-density polyethylene, Lysinibacillus

fusiformi, Madurai, microbe, municipal

solid waste, polypropylene, polypropylene

film, soil, Vaigai river

Yellow

4 11 aquatic ecosystem, Bacillus strain,

bacterium, compost, degradation,

mangrove ecosystem, microplastic

degradation, peninsular Malaysia,

polypropylene l lactated, screening, soil

consortia

Blue

degrade PP by UV treatment [23]. Bacillus sp. can degrade
polypropylene plastic during the 40-day incubation period [31].
B. cereus showed 12% in degrading polypropylene for 40 days
[4]. The B. flexus + B. subtilis consortium for one year showed
a degradation of 1.45% [32]. This confirmed that the bacterial
diversity, namely B. thuringiensis, B. cereus, and Bacillus sp.,
identified from the passive zone of the Supit Urang Landfill,
could potentially degrade polypropylene.

Highlights in BioScience Page 7 of 10 May 2024|Volume 7

http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

Table 5. List of bibliography studies on the topic Degradation of Polypropylene

No Publisher Title Bacteria Comment References
1 Elsevier Degradation of unpretreated and thermally pretreated

polypropylene by soil consortia

Bacillus flexus Able to degrade polypropylene film (PP-TT), which has

not been processed for 12 months, by 10.7%

[22]

2 Elsevier Growth of Pseudomonas and Bacillus biofilms on the

pretreated polypropylene surface

Bacillus and Pseudomonas Pseudomonas azotoformans, Pseudomonas stutzeri,

Bacillus subtilis and Bacillus flexus separately for 12

months. P. azotoformans and B. subtilis are relatively

hydrophobic, produce biosurfactants, and form biofilms

on polymers with higher carbohydrates and proteins than

the other two organisms.

[23]

3 Taylor and Francis Synergistic growth of Bacillus and Pseudomonas and its

degradation potential on pretreated polypropylene

Consortium of B. flexus + P. azotoformans,

Consortium B. flexus + B. subtilis

Consortium B. flexus + P. azotoformans for one year

showed maximum degradation (22.7%). Consortium B.

flexus + B. subtilis showed a 1.45% degradation for one

year.

[32]

4 Elsevier Screening of Bacillus strains isolated from mangrove

ecosystems in Peninsular Malaysia for microplastic

degradation

B. gottheilii B. gottheilii was able to degrade PP for 40 days by 3.6% [20]

5 Biochemistry and Bioinformatics Screening for Polypropylene Degradation Potential

of Bacteria Isolated from Mangrove Ecosystems in

Peninsular Malaysia

Bacillus cereus, Sporosarcina globispora Bacillus cereus showed 12%, and Sporosarcina

globispora showed an 11% decrease in body weight in 40

days.

[4]

6 Taylor and Francis Degradation of polypropylenepoly-L-lactide blend by

bacteria isolated from compost

Bacillus licheniformis (isolate P6),

Bacillus thuringiensis (isolate P8)

Isolate P8 was 12%, and P6 was 10%, capable of

degrading PP for 40 days.

[3]

7 Elsevier Growth kinetics and biodeterioration of polypropylene

microplastics by Bacillus sp. and Rhodococcus sp.

isolated from mangrove sediment

Rhodococcus, Bacillus sp. Able to degrade PP by 6.4% Rhodococcus sp. strain 36

and lessen PP by 4.0% by Bacillus sp. strain 27 after 40

days of incubation.

[21]

8 Elsevier Complete genome sequence of marine Bacillus sp. Y-01,

isolated from the plastics contamination in the Yellow

Sea

Bacillus sp. Bacillus sp. isolated from plastic contamination in the

Yellow Sea

[28]

9 Elsevier Microplastic degradation by bacteria in aquatic

ecosystem

Bacillus sp. BCBT21, Bacillus amyloliq-

uefaciens BSM-1, B. amyloliquefaciens

BSM-2, Pseudomonas putida, Bacillus

subtilis, Bacillus cereus, Brevibaccil-

lus borstelensis, Bacillus vallismortis

bt-dsce 01, P. protegens bt-dsce 02,

Stenotrophomonas sp. bt-dsce03, and

Paenibacillus sp.bt-dsce04

Extracellular hydrolytic enzymes such as CMCase,

lipase, xylanase, keratinase, chitinase, and protease

secreted by these bacteria play a charismatic role in

plastic degradation. Polyurethanes depolymerize urethane

and ester bonds due to the hydrolytic properties of urease,

esterase, and protease enzymes. Papain and urease

can degrade medical polyester due to their proteolytic

properties.

[33]

10 PubMed Microbial and Enzymatic Degradation of Synthetic

Plastics

- Outlining the progress made in the microbial degradation

of synthetic plastics and an overview of the enzymes

involved in biodegradation.

[34]

11 Elsevier Plastic biodegradation: Frontline microbes and their

enzymes

- Literature study on plastic degradation by bacteria. [35]

12 Springer Biodegradation of low-density polyethylene and

polypropylene by microbes isolated from Vaigai River,

Madurai, India

Bacillus paramycoides (BP) and Bacillus

cereus (BC)

The highest degradation of PP and PE was observed in

BP (78.99 ± 0.005%) and BC (63.08 ± 0.009%) in the

single approach, whereas in the combined system, BC &

BP recorded the highest degradation in both PP (78.62 ±

2.16%) and PE (72.50 ± 20.53%).

[36]

13 Elsevier Biodegradation of polypropylene films by Bacillus

paralicheniformis and Lysinibacillus fusiformis isolated

from municipality solid waste contaminated soil

Bacillus paralicheniformis and Lysinibacil-

lus fusiformis

Growth of Bacillus paralicheniformis and Lysinibacillus

fusiformis has shown OD values at 600nm after a 4-week

degradation period increased from 0.131 to 0.334 and

0.148 to 0.213, respectively.

[37]

14 Elsevier Microplastics spatiotemporal distribution and plastic-

degrading bacteria identification in the sanitary and

non-sanitary municipal solid waste landfills

- The microplastic (MP) surface gradually fades, becomes

rough, and even produces cracks and holes with landfill

depth and age. Small-size MPs (<100 m) were the most

abundant, and their numbers increased significantly from

28.14% to 49.13% in SL and from 24.54% to 59.51% in

NSL, while prominent-size MPs decreased significantly.

[38]

Conclusion
NGS employing 16S rRNA analysis at the Supit Urang Land-

fill showed 1,713 bacterial species from 2,496 sequences. Bac-
terial species with high abundance, namely Bacillus thuringien-
sis (15%), B. cereus (13%), B. velezensis (10%), and Bacillus
sp. (7%) were identified. Based on the bibliometric analysis
results, B. thuringiensis, B. cereus, and Bacillus sp. showed po-
tential in the degradation of polypropylene. These indigenous
PP-degrading bacteria are expected to be cultured and used to
develop PP biological degradation using suitable selective media.
The limitation of the study is that it only provides the profile
of candidate bacteria that can degrade plastics. To improve the

results of this research in the future, confirmation can be carried
out on a wet lab scale by performing proteomic analysis aimed
at obtaining information on cellular protein expression, so that it
can find specific proteins that can only be found in plastic degrad-
ing bacterial candidates, and not in bacteria that do not degrade
plastic. From the specific proteins obtained, it can then proceed
to bioinformatics analysis through molecular docking to deter-
mine the specific binding to the active site of the target protein.
Bioinformatics analysis can specifically predict the target protein
that causes the microbe to have the ability to degrade xenobiotic
plastics, and it can be combined with molecular docking to see
its ability to different types of xenobiotic plastics.

Highlights in BioScience Page 8 of 10 May 2024|Volume 7

http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

Acknowledgment
The author would like to thank Lembaga Penelitian dan

Pengabdian Masyarakat (LP2M) Universitas Negeri Malang (UM),
which funded this research through the UM internal funding the-
sis research grant scheme in 2022.

Reference
1. Carpenter E, Smith K. Plastics on the Sargasso sea surface.

Science. 1972;175(4027):1240-1.

2. Chamas A, Moon H, Zheng J, Qiu Y, Tabassum T, Jang J,
et al. Degradation Rates of Plastics in the Environment. ACS
Sustain Chem Eng. 2020;8(9):3494-511.

3. Jain K, Bhunia H, Sudhakara Reddy M. Degrada-
tion of polypropylenepoly-L-lactide blend by bacteria iso-
lated from compost. Bioremediat J. 2018;22(3–4):73-90.
Available from: https://doi.org/10.1080/10889868.
2018.1516620.

4. Helen A, Uche E, Hamid F. Screening for Polypropylene
Degradation Potential of Bacteria Isolated from Mangrove
Ecosystems in Peninsular Malaysia. Int J Biosci Biochem
Bioinforma. 2017;7(4):245-51.

5. Moulia E. Analysis of acid sulfate soil bacterial communi-
ties from two types of swamps in Kalimantan using Next-
Generation Sequencing (NGS) approach; 2019. Not pub-
lished.

6. Malek M, Jackowski M, Lasica W, Kadela M. Characteristics
of recycled polypropylene fibers as an addition to concrete
fabrication based on portland cement. Materials (Basel).
2020;13(8).

7. Vincent A, Derome N, Boyle B, Culley A, Charette S. Next-
generation sequencing (NGS) in the microbiological world:
How to make the most of your money. J Microbiol Methods.
2017;138:60-71. Available from: http://dx.doi.org/
10.1016/j.mimet.2016.02.016.

8. Kumar Sen S, Raut S. Microbial degradation of low den-
sity polyethylene (LDPE): A review. J Environ Chem Eng.
2015;3(1):462-73. Available from: http://dx.doi.org/
10.1016/j.jece.2015.01.003.

9. Matjai T, Simi T, Medveek N, Bajt O, Dreo T, Mori N. Criti-
cal evaluation of biodegradation studies on synthetic plastics
through a systematic literature review. Sci Total Environ.
2021;752.

10. Aminu S, Ascandari A, Laamarti M, Safdi N, El Allali A,
Daoud R. Exploring microbial worlds: a review of whole
genome sequencing and its application in characterizing the
microbial communities. Crit Rev Microbiol. 2023 Nov:1-25.

11. Anah I, Aniriani G, Sulistiono E. Biodegradation of
LDPE (Low Density Polyethlene) Plastic using Winogradsky
Columns. J Enviscience. 2020;4(2):96.

12. Ghosh S, Das A. Metagenomic insights into the micro-
bial diversity in manganese-contaminated mine tailings and
their role in biogeochemical cycling of manganese. Sci Rep.
2018;8(1):1-12. Available from: http://dx.doi.org/10.
1038/s41598-018-26311-w.

13. Naveen B, Mahapatra D, Sitharam T, Sivapullaiah P, Ra-
machandra T. Physico-chemical and biological characteri-
zation of urban municipal landfill leachate. Environ Pollut.
2017;220:1-12. Available from: http://dx.doi.org/10.
1016/j.envpol.2016.09.002.

14. Qodriyah N, Sanjaya E, Jatmiko Y, Santoso A, Susanti
E. Biodiversity of Polypropylene Degrading Bacteria from
Communal Waste Landfill of Probolinggo City and its Poten-
tial for Biodegradation of Medical Mask Waste; 2022. Not
published.

15. Jacquin J, Cheng J, Odobel C, Pandin C, Conan P, Pujo-
Pay M, et al. Microbial ecotoxicology of marine plastic
debris: A review on colonization and biodegradation by the
plastisphere. Front Microbiol. 2019;10(APR):1-16.

16. Klein S, Lorenzo C, Hoffmann S, Walther J, Storbeck S,
Piekarski T, et al. Adaptation of Pseudomonas aerugi-
nosa to various conditions includes tRNA-dependent for-
mation of alanyl-phosphatidylglycerol. Mol Microbiol.
2009;71(3):551-65.

17. Madigan M, Martinko J, Parker J. Brock biology of microor-
ganisms. 11th ed. Upper Saddle River, NJ: Prentice hall;
1997.

18. Parte A. LPSN - List of prokaryotic names with standing
in nomenclature (Bacterio.net), 20 years on. Int J Syst Evol
Microbiol. 2018;68(6):1825-9.

19. Combet-Blanc Y, Kalamba K, Kergoat P. Effect of pH on
Bacillus thermoamylovorans growth and glucose fermenta-
tion. Appl Environ Microbiol. 1995;61(2):656-9.

20. Auta H, Emenike C, Fauziah S. Screening of Bacillus
strains isolated from mangrove ecosystems in Peninsular
Malaysia for microplastic degradation. Environ Pollut.
2017;231:1552-9. Available from: https://doi.org/10.
1016/j.envpol.2017.09.043.

21. Auta H, Emenike C, Jayanthi B, Fauziah S. Growth kinet-
ics and biodeterioration of polypropylene microplastics by
Bacillus sp. and Rhodococcus sp. isolated from mangrove
sediment. Mar Pollut Bull. 2018;127:15-21.

Highlights in BioScience Page 9 of 10 May 2024|Volume 7

https://doi.org/10.1080/10889868.2018.1516620
https://doi.org/10.1080/10889868.2018.1516620
http://dx.doi.org/10.1016/j.mimet.2016.02.016
http://dx.doi.org/10.1016/j.mimet.2016.02.016
http://dx.doi.org/10.1016/j.jece.2015.01.003
http://dx.doi.org/10.1016/j.jece.2015.01.003
http://dx.doi.org/10.1038/s41598-018-26311-w
http://dx.doi.org/10.1038/s41598-018-26311-w
http://dx.doi.org/10.1016/j.envpol.2016.09.002
http://dx.doi.org/10.1016/j.envpol.2016.09.002
https://doi.org/10.1016/j.envpol.2017.09.043
https://doi.org/10.1016/j.envpol.2017.09.043
http://bioscience.highlightsin.org/


Wijayanti et al., 2024 Identification of polypropylene-degrading bacteria in Supit Urang landfill using NGS

22. Arkatkar A, Arutchelvi J, Bhaduri S, Uppara P, Doble
M. Degradation of unpretreated and thermally pretreated
polypropylene by soil consortia. Int Biodeterior Biodegrad.
2009;63(1):106-11. Available from: http://dx.doi.org/
10.1016/j.ibiod.2008.06.005.

23. Arkatkar A, Juwarkar A, Bhaduri S, Uppara P, Doble M.
Growth of Pseudomonas and Bacillus biofilms on pre-
treated polypropylene surface. Int Biodeterior Biodegrad.
2010;64(6):530-6. Available from: http://dx.doi.org/
10.1016/j.ibiod.2010.06.002.

24. Bakke T, Klungsøyr J, Sanni S. Environmental impacts
of produced water and drilling waste discharges from the
Norwegian offshore petroleum industry. Mar Environ Res.
2013;92:154-69. Available from: http://dx.doi.org/
10.1016/j.marenvres.2013.09.012.

25. Thiel T, May V, Kalk M, Alters S, Alters B, OBrian J. Sci-
ence in the real world: Microbes in action. Introd to Bact.
1999:1-8.

26. Rudiyansyah A, Wahyuningsih N, Kusumanti E. Effect of
Temperature, Humidity, and Sanitation on the Presence of
Eschericia Coli and Salmonella Bacteria in Chicken Cages
at Broiler Farms in Karanggeneng Village, Semarang City. J
Kesehat Masy. 2015;3(2):196-201.

27. Ascandari A, Aminu S, Safdi N, El Allali A, Daoud R. A
bibliometric analysis of the global impact of metaproteomics
research. Front Microbiol. 2023;14:1217727.

28. Wang X, Qu C, Wang W, Zheng Z, Liu F, An M, et al.
Complete genome sequence of marine Bacillus sp. Y-01,
isolated from the plastics contamination in the Yellow Sea.
Mar Genomics. 2019;43(6):72-4. Available from: http:
//dx.doi.org/10.1016/j.margen.2018.05.002.

29. Alariqi S, Pratheep Kumar A, Rao B, Singh R. Biodegra-
dation of -sterilised biomedical polyolefins under compost-
ing and fungal culture environments. Polym Degrad Stab.
2006;91(5):1105-16.

30. Iwamoto A, Tokiwa Y. Enzymatic degradation of
plastics containing polycaprolactone. Polym De-
grad Stab. 1994;45(2):205-13. Available from:
https://www.sciencedirect.com/science/
article/pii/0141391094901384.

31. Auta H, Emenike C, Jayanthi B, Fauziah S. Growth ki-
netics and biodeterioration of polypropylene microplastics
by Bacillus sp. and Rhodococcus sp. isolated from man-
grove sediment. Mar Pollut Bull. 2018;127:15-21. Available
from: https://doi.org/10.1016/j.marpolbul.2017.
11.036.

32. Aravinthan A, Arkatkar A, Juwarkar A, Doble M. Syner-
gistic growth of Bacillus and Pseudomonas and its degrada-
tion potential on pretreated polypropylene. Prep Biochem
Biotechnol. 2016;46(2):109-15.

33. Chandra P, Enespa, Singh D. Microplastic degradation
by bacteria in aquatic ecosystem. In: Microorganisms
for Sustainable Environment and Health. INC; 2020. p.
431-67. Available from: http://dx.doi.org/10.1016/
B978-0-12-819001-2.00022-X.

34. Mohanan N, Montazer Z, Sharma P, Levin D. Microbial
and Enzymatic Degradation of Synthetic Plastics. Front
Microbiol. 2020 November;11.

35. Amobonye A, Bhagwat P, Singh S, Pillai S. Plastic biodegra-
dation: Frontline microbes and their enzymes. Sci To-
tal Environ. 2021;759:143536. Available from: https:
//doi.org/10.1016/j.scitotenv.2020.143536.

36. Nanthini Devi K, Raju P, Santhanam P, Dinesh Kumar S,
Krishnaveni N, Roopavathy J, et al. Biodegradation of
low-density polyethylene and polypropylene by microbes
isolated from Vaigai River, Madurai, India. Arch Mi-
crobiol. 2021;203(10):6253-65. Available from: https:
//doi.org/10.1007/s00203-021-02592-0.

37. Kaviraj R, Mridul U, Kathirvel P. Biodegradation
of polypropylene films by Bacillus paralicheni-
formis and Lysinibacillus fusiformis isolated from
municipality solid waste contaminated soil. Res J
Chem Environ. 2021;25(7):71-8. Available from:
https://api.elsevier.com/content/abstract/
scopus_id/85109159500.

38. Li N, Han Z, Guo N, Zhou Z, Liu Y, Tang Q. Microplas-
tics spatiotemporal distribution and plastic-degrading bacte-
ria identification in the sanitary and non-sanitary municipal
solid waste landfills. J Hazard Mater. 2022;438:129452.
Available from: https://www.sciencedirect.com/
science/article/pii/S0304389422012456.

Highlights in BioScience Page 10 of 10 May 2024|Volume 7

http://dx.doi.org/10.1016/j.ibiod.2008.06.005
http://dx.doi.org/10.1016/j.ibiod.2008.06.005
http://dx.doi.org/10.1016/j.ibiod.2010.06.002
http://dx.doi.org/10.1016/j.ibiod.2010.06.002
http://dx.doi.org/10.1016/j.marenvres.2013.09.012
http://dx.doi.org/10.1016/j.marenvres.2013.09.012
http://dx.doi.org/10.1016/j.margen.2018.05.002
http://dx.doi.org/10.1016/j.margen.2018.05.002
https://www.sciencedirect.com/science/article/pii/0141391094901384
https://www.sciencedirect.com/science/article/pii/0141391094901384
https://doi.org/10.1016/j.marpolbul.2017.11.036
https://doi.org/10.1016/j.marpolbul.2017.11.036
http://dx.doi.org/10.1016/B978-0-12-819001-2.00022-X
http://dx.doi.org/10.1016/B978-0-12-819001-2.00022-X
https://doi.org/10.1016/j.scitotenv.2020.143536
https://doi.org/10.1016/j.scitotenv.2020.143536
https://doi.org/10.1007/s00203-021-02592-0
https://doi.org/10.1007/s00203-021-02592-0
https://api.elsevier.com/content/abstract/scopus_id/85109159500
https://api.elsevier.com/content/abstract/scopus_id/85109159500
https://www.sciencedirect.com/science/article/pii/S0304389422012456
https://www.sciencedirect.com/science/article/pii/S0304389422012456
http://bioscience.highlightsin.org/

	Abstract
	Introduction
	Experimental section
	Sampling and Sample Collection
	DNA Sequencing
	Bioinformatics analysis
	Bibliometric Analysis for Identification of Polypropylene-Degrading Bacteria

	Results and Discussion
	Identity of Polypropylene-Degrading Bacteria Based on Bibliometric Analysis

	Conclusion
	Acknowledgment

