Brazilian Journal of Microbiology (2024) 55:3449–3463

[Link]

ENVIRONMENTAL MICROBIOLOGY - RESEARCH PAPER

A comparative study on biodegradation of low density polyethylene

bags by a Rhizopus arrhizus SLNEA1 strain in batch and continuous
cultures
Randa Harrat1 · Ghania Bourzama2 · Nouari Sadrati3 · Amina Zerroug3 · Gaëtan Burgaud4,5 ·
Houria Ouled-Haddar6 · Boudjema Soumati1

Received: 22 November 2023 / Accepted: 15 July 2024 / Published online: 30 September 2024
© The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia 2024

Abstract
Biodegradation poses a challenge for environmentalists and scientific community, offering a potential solution to the
plastic waste problem. This study aims to investigate the biological degradation of low-density polyethylene (LDPE)
bags by a fungus in both batch and continuous cultures, with the goal of identifying an eco-friendly and cost-effective
waste management strategy. The fungal strain Rhizopus arrhizus SLNEA1, isolated from a landfill located in northeastern
Algeria, was tested for its capability to degrade LDPE films and utilize them as a sole carbon source in batch (α-LDPE)
and continuous (γ-LDPE) cultures. The results indicated a higher rate of weight loss for γ-LDPE (29.74%) compared to
α-LDPE (23.77%). The biodegradation effect was examined using scanning electron microscopy (SEM), Energy Disper-
sive X-ray Spectroscopy (EDS) and Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) to evaluate
morphological and chemical changes in LDPE samples, highlighting alterations of LDPE films through cracks, veins
and holes under SEM and chemical transformation and appearance of new functional groups in the FTIR data. Rhizopus
arrhizus SLNEA1 demonstrated the ability to break down and utilize LDPE films as a carbon source. This isolate shows
promise for LDPE biodegradation applications, which may be leveraged for the development of future plastic degradation
systems involving fungi.

Keywords LDPE films · Rhizopus arrhizus · Batch culture · Continuous culture · . Biodegradation

Introduction
Responsible Editor: Luis Henrique Souza Guimaraes.

Randa Harrat Conventional plastic materials are typically derived from

harratranda@[Link]; [Link]@[Link] petrochemicals extracted from coal and natural gas, exist-
1
ing in various forms as synthetic or semi-synthetic organic
Laboratory of Biochemistry and Environmental Toxicology,
Badji Mokhtar-Annaba University, [Link] 12, Annaba. polymers [1]. Plastic polymers have become a fundamental
23000, Algeria element in our daily life worldwide, raising global plastic
2
Laboratory of Microbiology and Molecular Biology, Badji production to 400 million tons annually [2].
Mokhtar-Annaba University, [Link] 12, Annaba. 23000, Previous studies have highlighted the likelihood of plas-
Algeria tics exposition and subsequent impacts on diverse aquatic
3
Laboratory of Characterization and Valorization of Natural organisms [3–5]. However, recent investigations suggest
Resources, University Mohamed El Bachir El Ibrahimi of that polyethylene contamination in the terrestrial environ-
Bordj Bou Arreridj, 34000, Algeria ments may pose an even greater threat than in marine sys-
4
Univ Brest, INRAE, Laboratoire Universitaire de tems [6, 7], prompted increased attention toward terrestrial
Biodiversité et Écologie Microbienne, F-29280 Plouzané, ecosystems [8]. The accumulation of persistent plastics in
France
the environment, particularly polyethylene as the most pro-
5
Institut Universitaire de France, Paris, France duced plastic polymer (~ 26% of the global production),
6
Laboratory of Molecular Toxicology, University of Jijel, poses a significant global to terrestrial ecosystems [9, 10].
Jijel, Ouled Aïssa 18000,, Algeria

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3450 Brazilian Journal of Microbiology (2024) 55:3449–3463

Universal plastic waste has reached about 6,300 million continuous culture conditions. The aim of this study was to
tonnes in recent years, with 79% ending up in landfills or the assess the ability of a fungal strain isolated from a landfill
natural environment. In addition, recent estimates indicate located in northeastern Algeria to degrade plastic bag films.
that 12,000 million tons of plastic waste will be accumu- This was accomplished through batch and continuous cul-
lated on Earth by 2050 [2]. tures, with two primary aims: (i) to address LDPE disposal
Of the various types of polyethylene, low-density poly- challenges and enhance its biodegradation rates, and (ii) to
ethylene (LDPE) stands out as the most commonly pro- develop a cost-effective treatment approach for soil contam-
duced, accounting for 14% of global production. As a result, inated by plastic materials.
LDPE is also the most prevalent in nature as waste [1].
LDPE was the pioneering plastic adopted for widespread
commercial packaging use in the late 1940s [11], renowned Methodology
for its lightness, strength and durability, leading to a wide
range of applications in crafting various containers, dispens- Fungal strain
ing bottles, plastic bags, and various laboratory items [12].
Multiple processes contribute to plastic degradation, The fungal strain Rhizopus arrhizus SLNEA1 was isolated
including abiotic degradation (photodegradation and from a soil sample collected from a landfill in northeast-
thermo-oxidation), biotic degradation (microorganism ern Algeria after decimal dilution and inoculation into Petri
action), or a synergistic combination thereof. Enzymatic dishes containing potato dextrose agar (PDA). The plates
degradation provides a promising green and innovative were incubated at 27 °C for one week [21]. The isolated
route for recycling polymer waste [13]. strain was further purified on solid PDA.
Over 400 microbes have been identified as capable of
degrading plastic [2]. Besides, several studies have high- Morphological and molecular identification
lighted the remarkable ability of certain fungal strains to
degrade LDPE including the genera of Alternaria, Aspergil- The selected fungal strain was initially identified based on
lus, Fusarium and Penicillium [14–17]. its macroscopic and microscopic characteristics [22]. Sub-
Saxena and Pareek (2017) reported the degradation sequently, DNA extraction from a 7-day-old PDA medium
of LDPE films by Penicillium chrysogenum, Aspergillus culture was performed using a commercial NucleoSpin
fumigatus, A. niger and A. flavus with respectively 8%, 7%, Plant II fungal genomic DNA extraction kit (Macherey-
9% and 7% of weight loss after 30 days of incubation [18]. Nagel, Germany), following the manufacturer’s instruc-
Khruengsai et al. (2021) demonstrated the degradation capa- tions. To amplify the Internal Transcribed Spacer (ITS) and
bility of five fungal strains on LDPE films: Diaporthe itali- Large Subunit (LSU) regions, primer pairs ITS1 (5’ CTT
ana, Thyrostroma jaczewskii, Collectotrichum fructicola, GGT CAT TTA GAG GAA GTA A 3’) / ITS4 (5’ ​T​C​C​T​
Stagonosporopsis citrulli, and Aspergillus niger, achiev- C​C​G​C​T​T​A​T​T​G​A​T​A​T​G​C 3’) and LR0R (5’ ​A​C​C​C​G​C​T​G​A​
ing weight losses of 43.90%, 46.34%, 48.78%, 45.12%, C​T​T​A​A​G​C 3’) / LR5 (5’ ​T​C​C​T​G​A​G​G​G​A​A​A​C​T​T​C​G 3’)
and 28.78%, respectively. Additionally, scanning electron were used, respectively [23]. The cycling protocol included
microscopy (SEM) analyses of the LDPE films confirmed an initial denaturation step at 95 °C for 5 min, followed by
biodegradation through morphological changes such as 35 cycles of denaturation at 95 °C for 30 s, an hybridization
cracks, veins, and holes on the film surface. The same step at 52–55 °C for 30 s, an extension at 72 °C for 45 s, and
authors highlighted the fungus C. fructicola as a promising a final extension step at 72 °C for 7 min. PCR results were
candidate for LDPE biodegradation, suggesting its poten- separated through electrophoresis on a 1.5% (w/v) agarose
tial application in fungal-based plastic degradation systems gel and then stained with GelRed (Biotium, United States).
[19]. Moreover, the study conducted by Khruengsai et al. Imaging was performed using a Gel doc system from Bio-
(2023) demonstrates the ability of the fungus Neopestalo- rad (USA) under UV light. Following the purification of the
tiopsis phangngaensis to utilize LDPE as a carbon source, PCR products using the NucleoSpin Gel and PCR Clean-
resulting in a significant biodegradation rate (54.34% weight up kit from Macherey-Nagel (Germany), the sequencing
loss) without any pre-treatment. Hence, N. phangngaensis has been carried out according to the Sanger technique with
holds promise as an alternative agent for LDPE degradation the BigDye v3.1 kit from Applied Biosystems, employing
in environment settings [20]. the same primers used in the PCR. Subsequently, acquired
To date, all fungal studies have been conducted in vitro sequences were analyzed using the CHROMAS PRO soft-
using batch cultures. Additionally, research on fungi in ware [23, 24].
Algeria has been limited. This study marks the direst inves- The edited sequences were compared with the DNA data-
tigation into LDPE biodegradation by fungal species under base reference in GenBank, including Rhizopus type species

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Brazilian Journal of Microbiology (2024) 55:3449–3463 3451

([Link]/Genbank), using MEGA7 and the out- MgSO4, 0.2 g NaCl, 0.01 g CuSO4, 1 g CaCO3 and 1 g
group Phycomyces blakesleeanus. A maximum likelihood ZnSO4 in 1000 ml of distilled water [27]. The pH of the
(ML) approach, employing 1000 bootstrap replicates and medium was then adjusted to 6.0.
the Tamura-Nei model in MEGA7, was utilized to construct
the phylogenetic tree of the combined ITS-LSU sequences Biodegradation in batch culture
derived from the alignment results. The sequences gener-
ated in this study have been submitted to GenBank under In batch cultures, 250 ml sterile Erlenmeyer flasks were
the accession numbers OR584306 and OR584305 for ITS employed, each filled with 150 ml of sterile liquid MM and
(588 bp) and LSU (649 bp), respectively [25]. a 0.2 g LDPE film. Five agar plugs (45 × 106 cells/ml) of the
tested fungal strain grown on PDA during seven days were
Biodegradation tests added to each flask to assess fungal degradation capability.
The fungal cultures were then incubated at 25 ± 2 °C for 90
The fungal strain Rhizopus arrhizus SLNEA1 was evalu- days with continuous rotation at 150 rpm in a shaker incuba-
ated for its capacity to degrade LDPE films. Biodegrada- tor (Bühler Incubator Hood) [28].
tion experiments were conducted in liquid minimal medium
containing LDPE films as the sole carbon source [19]. Biodegradation in continuous culture
Untreated LDPE films were used as controls. All trials were
performed in triplicates. Data from each experiment were Under sterile continuous culture, a 45 cm tall glass column
presented as the mean ± standard deviation. (inner diameter 8.5 cm, inner surface 56.75 cm2) was used,
filled with 1000 ml of MM as the culture medium. Thirty-
Preparation of LDPE films three agar plugs (5 mm diameter, 7 days old) from the tested
fungal strain and six LDPE films (F1, F2, F3, F4, F5 and
In this study, white plain plastic shopping bags were used as F6) each weighing 0.2 g were introduced into the system
LDPE material for biodegradation studies. The LDPE films (Fig. 1). A peristaltic pump (MasterFlex; Cole-Parmer) con-
were cut into small pieces of known weight, then disinfected trolled the recirculation of the aqueous phase. Compressed
with 70% ethanol for 30 min and subsequently transferred air, regulated by a Flowmeter (AALBORG) at 20 L/min
to sterile distilled water for 20 min [26]. was supplied by a compressor. The reactor temperature was
maintained at 25 °C, and the culture was incubated for three
Preparation of liquid minimal medium months. Every 30 days, the films were aseptically collected
and weighted [29].
A liquid minimal medium (MM) was formulated by dis-
solving 16 g K2HPO4, 2 g KH2PO4, 1 g (NH4)2SO, 0.475 g

Fig. 1 Experimental setup for

continuous culture of Rhizopus
arrhizus SLNEA1 in LDPE-
enriched condition [24]

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Evaluation of the biodegradation ability Results

After the incubation period, LDPE films were extracted Isolation and identification of indigenous soil
from the fungal cultures, disinfected with 70% ethanol for mycoflora
30 min, and then gently cleaned to remove any fungal bio-
film adhering to the LDPE surface [19, 20]. Subsequently, The polluted soil sample used in this study yielded mul-
the films were rinsed with sterile distilled water and air- tiple fungal isolates, among which SLNEA1 emerged as
dried at room temperature for analysis. the dominant strain. After incubation of this fungal strain
on PDA, macroscopic examination revealed a white thal-
Percentage of weight loss lus with a central brown to black region, exhibiting a fluffy
appearance. Additionally, a loose structured and extensive
The weight of each recovered LDPE film was recorded mycelium invaded the entire Petri dish (Fig. 2). While the
using an analytical balance (Radwag AS 60/220/C/2). microscopic examination revealed the presence of brown
The weight loss of an LDPE film (W) was tested on the to black sporocysts on the surface of the mycelium. These
basis of its weight before (W0: initial weight) and after deg- sporocysts, characterized by their globular shape and promi-
radation (W1: final weight) [30]: nent columella, released conidia, also known as sporocys-
tospores (Fig. 3). Based on these observations, the fungal
W (%) = [(W0 −W1 ) /W0 ] × 100 isolate SLNEA1was identified at the genus level as Rhizo-
pus sp.
The values obtained were then compared to controls. The morphological characteristics, both macroscopic and
microscopic, enabled the identification of this isolate only
Scanning electron microscope (SEM) analysis at the genus level. Subsequent analysis using the ITS and
LSU sequence data as a query in the Basic Local Alignment
LDPE films colonized by Rhizopus arrhizus SLNEA1 were Search Tool (BLAST) revealed that the isolate SLNEA1
analyzed by Scanning Electron Microscopy (QUANTA 250) exhibited a very high homology; where the percent iden-
to check the changes on the surface of the polymer. LDPE tity was 99.48 % for LSU with the Rhizopus arrhizus CBS
films were fixed with a scotch coal support and placed in an 112.07T type strain, and 99.66 % for ITS with the Rhizopus
ionized chamber with water spray. The later were captured arrhizus isolate YRR3. The query coverage of the sequences
at 1000x, 1500x, and 2000x magnification [30]. was 98%, and the E-value was zero.
The elemental composition of LDPE films was also ana- Bootstrap Test of Phylogeny, based on Tamura-Nei
lyzed using Energy Dispersive X Ray Spectroscopy (EDS) model, was performed using a Maximum Likelihood analy-
[31]. sis of the combined ITS- LSU sequence data from Rhizopus
and Phycomyces species.
Fourier transform infrared spectroscopy (FTIR) As depicted in Fig. 4, the concatenated phylogenetic
tree of ITS-LSU regions validates the classification of the
The surface functionalization of LDPE films was assessed SLNEA1 within Rhizopus arrhizus sensu lato (Rhizopus
by Fourier transform infrared spectroscopy (FTIR) using an arrhizus species complex), positioned within the Rhizopus
INVENIO-R (329) FTIR spectrometer (Bruker, Germany), arrhizus group. This finding is supported by a significant
with 30 scans at a resolution of 4 cm-1 for each sample. maximum likelihood (ML) value of 90%.
Spectra were collected over a range of wavenumbers from
227 to 5000 cm-1, with different peaks corresponding to Evaluation of fungal potential for LDPE film
functional groups present in each sample. Both control and biodegradation
degraded samples underwent FTIR analysis [30].
Additionally, the carbonyl index (CI) value was deter- The selected fungal strain SLNEA1 identified as Rhizopus
mined. The CI is defined as the ratio between the integrated arrhizus was tested for its capability to degrade shopping
band absorbance of the carbonyl (C = O) peak from 1.850 to white plastic bags, in both batch and continuous cultures.
1.650 cm− 1 and that of the methylene (CH2) scissoring peak
from 1.500 to 1.420 cm− 1. The CI values were calculated Biodegradation in batch culture
using the following equation [32]:
LDPE films were subjected to a 90-day batch culture incu-
Areaunderband1850 − 1650cm − 1 bation with Rhizopus arrhizus SLNEA1 to evaluate its deg-
CI =
Areaunderband1500 − 1420cm − 1 radation potential. Figure 5 displays the weight percentage

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The weight of the control LDPE film remained

unchanged. However, all LDPE films incubated with R.
arrhizus SLNEA1 exhibited a weight loss throughout the
entire incubation period.
After one month of incubation, the weight of F1, F2,
F3, F4, F5 and F6 decreased by 23.91%, 17.74%, 23.59%,
11.67%, 25.61%, and 25.56%, respectively. By the end
of the second month, the weight reduction percentages of
each film were 24.75%, 20.09%, 24.19%, 12.88%, 28.80%,
and 25.75%, respectively. After three months, F5 showed
the highest weight loss with 29.74%, while the weight loss
percentages of F1, F2, F3 and F6 were 24.80%, 27.76%,
25.41%, and 26.03%, respectively. Notably, F4 exhib-
ited the lowest percentage of weight loss with a value of
16.50%. Statistical analysis using ANOVA tests was per-
formed to assess significant differences among LDPE films
and across various incubation periods (time). The mean val-
ues obtained exhibited statistically significant differences
(p-value < 2e-16 ***). Further examination of the ANOVA
outcomes based on the least significant differences (LSD)
Fig. 2 Macroscopic appearance of Rhizopus arrhizus SLNEA1 on and revealed substantial disparities among the films, with
PDA after 7 days of incubation at 27 °C
only F1 and F3 yielding similar results (Fig. 1a, supple-
mentary material). Additionally, the LDS test indicated
evolution of LDPE films over time. After 30 days of incuba- significant differences among the 30, 60, and 90-day incu-
tion with the fungal isolate, the LDPE film weight decreased bation period (p-value < 2e-16***) (Fig. 1b, supplementary
by 23.77%. However, no further weight loss was observed material).
after 60 and 90 days of incubation (23.77%). In contrast, the
control LDPE film showed no weight loss. Highlighting LDPE biodegradation by the tested
fungal strain
Biodegradation in continuous culture
To confirm the degradation of LDPE films by Rhizopus
LDPE films, labelled F1 to F6, were subjected to continuous arrhizus SNLEA1 in both batch and continuous cultures,
culture incubation with the selected fungal strain Rhizopus SEM, EDS and FTIR analysis were performed. However,
arrhizus SLNEA1 for three months. Figure 6 displays the in the continuous culture, only the LDPE film (F5), which
evolution of the weight of LDPE films after 30, 60 and 90 exhibited the highest weight loss, underwent these analyses.
days of incubation.
Fig. 3 Microscopic appearance
of Rhizopus arrhizus SLNEA1
strain under optical microscopy
(10x and 40x magnifications)
after 7 days of incubation

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Fig. 4 Phylogenetic tree gener-

ated by maximum likelihood
analysis of combined ITS- LSU
sequence data of Rhizopus and
Phycomyces species. The tree
was rooted using Phycomyces
blakesleeanus CBS 284.35. ML
bootstrap support values greater
than 50% are shown near the
nodes. The isolate of interest
SLNEA1 is indicated in bold

Characterization of LDPE films by SEM and EDS Analysis

The LDPE films were examined under magnifications of

1000x, 1500x, and 2000x using a Scanning Electron Micro-
scope to observe surface morphology before and after deg-
radation. The control LDPE films revealed a smooth surface
with no significant changes at all magnification levels
(Fig. 7). Conversely, LDPE films used in both batch and
continuous cultures displayed various structural alterations,
including surface fragility, damaged layers, presence of
grooves, cracks and pits. These observations suggest fungal
activities led to the breakdown of the polyethylene carbon
chains (Figs. 8, 9 and 10).

EDS analysis of LDPE films

In this study, the selected LDPE films were exposed to an

Fig. 5 Evolution of the weight of LDPE film after 30, 60 and 90 days
electron beam inside a scanning electron microscope (SEM)
of exposure to Rhizopus arrhizus SLNEA1 in a batch culture com- for Energy Dispersive X-ray Spectroscopy (EDS) analysis.
pared to the control The results revealed the presence of four key elements -
carbon, oxygen, titanium and calcium - in all LDPE films,
albeit in varying percentages (Figs. 11, 12 and 13). LDPE
bags typically contain polyethylene [C2H4]n, titanium

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Fig. 6 Evolution of the weight of

LDPE films (F1 to F6) after 30,
60 and 90 days of exposure to
Rhizopus arrhizus SLNEA1 in a
continuous culture compared to
the control

dioxide (TiO2) and calcium carbonate (CaCO3), which serve higher intensity (Fig. 15b, c and d). However, new peaks
as white pigments to ensure opacity. emerged. These included peaks of relatively low intensity
The EDS analysis indicated that control LDPE films at 3739.815, 3850.144, 2358.704, 2329.775, 1836.047,
primarily consist of carbon, with 94.83% of the compo- 1739.886, 1697.187 cm− 1, along with a medium intensity
sition. Additionally, small amounts of oxygen, calcium, band at 1425.251 cm− 1 observed in LDPE film exposed to
and titanium were detected at 4.04%, 0.93%, and 0.21%, R. arrhizus SLNEA1 in batch culture. These spectra corre-
respectively (Fig. 11). LDPE films cultured with R. arrhi- sponded, respectively, to O-H, C ≡ C, C = O bands stretch-
zus SLNEA1 in both batch and continuous cultures exhib- ing and the asymmetric bending vibration of C-H bonds in
ited a similar composition, albeit with varying percentages, methylene (-CH2) group.
particularly in carbon and oxygen mass. Specifically, the Additionally, new bands emerged in the LDPE film (F5)
carbon and oxygen content for batch culture were 69.26% exposed to R. arrhizus SLNEA1 in continuous culture after
and 19.87%, respectively, while for continuous culture, they one month, observed at 3648.309, 2359.765, 2159.014,
were 56.84% and 26.30%, respectively (Figs. 12 and 13). 1836.047, 1741.545, 1648.971, 1424.068, 1263.247, and
1107.858 cm− 1. These bands were assigned respectively to
FTIR and CI analysis of LDPE films stretching vibration of O-H, C-C (triple bonds), C = O (anhy-
dride), C = C, CH2 bending vibration and C-O stretching
The selected LDPE films underwent FTIR analysis to con- vibration. After 3 months of incubation, analysis of the same
firm LDPE sample degradation by revealing structural film revealed the presence of the same peaks with higher
changes in both control and degraded LDPE films (Figs. 14 intensities. However, the spectra obtained at 3648, 2359 and
and 15). 1107 shifted to 3650.105, 2357.926, and 1105.495, respec-
The FTIR spectrum of the control LDPE film exhib- tively. In addition, the band obtained at 2159 cm− 1 disap-
ited characteristic peaks at 2912.478, 2845.155, 1463.002, peared. However, new peaks of medium intensity appeared
1432.965, and 718.746 cm− 1 (Fig. 15a). The strong bands in the area 3347–4912 cm− 1 and at 1262.660 cm− 1. These
at 2912.478 and 2845.155 correspond respectively to asym- peaks correspond respectively to the elongation of the O-H
metric and symmetric vibrations of aliphatic C-H stretches in bonds and the elongation vibration of the C-O bonds (car-
alkanes molecules. The intense peaks obtained at 1463.002, boxylic acids and derivatives). Low-intensity peaks also
1432.965 and 718.746 cm− 1 were respectively assigned occurred at 2208.931, 2161.667; 1841.833, 1648.742, and
to asymmetric scissoring vibration of C-H bonds (-CH2) 800.906 cm− 1, attributed respectively to the elongation of
and the rocking vibration of a methylene group (CH₂). C ≡ C bonds (terminal alkynes), C = O, C = C, and the defor-
Additionally, medium bands were observed in the range mation of C-H aromatic bonds (characteristic bands of the
of 227–874 cm− 1 corresponding to the backbone structure substitution type).
vibrations of the polymer. These vibrations are primarily The carbonyl index (CI) of the selected LDPE films
related to the stretching, bending and twisting motions of was calculated and depicted in Table 1. The control LDPE
the C-C and C-H bonds within the polymer chains. (µ-LDPE) film exhibited the lowest CI value of 0.50. LDPE
Similarly, LDPE films exposed to R. arrhizus SLNEA1 films exposed to R. arrhizus SLNEA1 in batch culture
in both batch (α- LDPE) and continuous (γ- LDPE) cul- (α-LDPE) and continuous culture (γ-LDPE) showed higher
tures showed the same peaks as the control but with CI values after 30 and 90 days, raising 0.70, 0.80, and 0.76,

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Fig. 7 Control LDPE film under Scanning Electron Microscopy at 1000x (a), 1500x (b) and 2000x (c) magnification

Fig. 8 LDPE film exposed to Rhizopus arrhizus SLNEA1 in batch culture after 30 days of incubation under Scanning Electron Microscopy at
1000x (a), 1500x (b) and 2000x (c) magnification

Fig. 9 LDPE film (F5) exposed to Rhizopus arrhizus SLNEA1 in continuous culture after 30 days of incubation under Scanning Electron Micros-
copy at 1000x (a), 1500x (b) and 2000x (c) magnification

Fig. 10 LDPE film (F5) exposed to Rhizopus arrhizus SLNEA1 in continuous culture after 90 days of incubation under Scanning Electron Micros-
copy at 1000x (a), 1500x (b) and 2000x (c) magnification

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Fig. 11 Energy dispersive X-ray spectroscopy analysis of control LDPE film

Fig. 12 Energy dispersive X-ray spectroscopy analysis of LDPE film exposed to Rhizopus arrhizus SLNEA1 in batch culture

Fig. 13 Energy dispersive X-ray spectroscopy analysis of LDPE film (F5) exposed to Rhizopus arrhizus SLNEA1 in continuous culture

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Fig. 14 FTIR spectra of LDPE films exposed to Rhizopus arrhizus SLNEA1 in batch and continuous cultures compared to the control

respectively. These increases in CI indicate the degradation Thus, this study aims to evaluate the capacity of a fungal
of the LDPE films, confirming the breakdown of the long strain isolated from a landfill located in northeastern Alge-
carbon chains and the oxidation of the chemical structure by ria to degrade LDPE films in both batch and continuous
the tested fungal strain, R. arrhizus SLNEA1. cultures. The ultimate goal is to develop a cost-effective
method for mitigating plastic pollution.
In this study, Rhizopus arrhizus SLNEA1 strain was iso-
Discussion lated and identified based to its morphological appearance,
microscopic features, and phylogenetic analysis. This fun-
The accumulation of plastic waste, including LDPE bags, gus demonstrated the capability to utilize LDPE films as the
poses a significant threat to ecosystems, impacting human sole carbon and energy sources. While reports on the ability
and animal health, as well as the natural environment, and of R. arrhizus to degrade LDPE are limited, some strains
even influencing the global carbon cycling [33]. Therefore, affiliated to the genus Rhizopus are believed to possess the
the degradation of LDPE by microorganisms, including capacity to degrade complex polymers. For instance, Harrat
fungi, is highly encouraged for proper disposal, aiming et al. (2022) demonstrated a 20% weight loss of LDPE in
convert carbon atoms in recalcitrant plastics into environ- a batch culture with a Rhizopus sp. strain [36]. Similarly,
mentally harmless biological components [34]. Currently, Tokiwa et al. (1986) highlighted the degradation potential of
landfills store up to 42% of worldwide plastic waste [35]. R. arrhizus on polycaprolactone (PCL) [37]. Furthermore,

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Fig. 15 FTIR insights into

chemical changes in LDPE films
exposed to Rhizopus arrhi-
zus SLNEA1 in batch (b) and
continuous culture after 30 and
90 days of incubation (c and d
respectively) compared to the
control (a). (i) represents the
intensity of the peaks

Table 1 The carbonyl indexes of µ- LDPE, α- LDPE and γ- LDPE after

biodegradation fungal growth and enzyme secretion for biodegradation
Sample CI average throughout the incubation period. This contrasts with batch
µ- LDPE film 0.504906 culture, where nutrient depletion and degradation product
α- LDPE film after 30 days of incubation 0.709265 accumulation lead to a short exponential phase, followed by
γ- LDPE film after 30 days of incubation 0.809938 cell death and cessation of the biodegradation process after
γ- LDPE film after 90 days of incubation 0.763616 one month. To date, the degradation of LDPE by fungi in
continuous culture has not been studied. However, Kumar
Awasthi et al. (2017) observed an 8.4 ± 3% decrease in et al. (2016) reported the degradation of commercial plastics
LDPE weight using a fungal lab isolate, Rhizopus oryzae in soil using a bioreactor and found that the bacterial consor-
NS5 [38]. Additionally, Saxena et al. (2022) reported a 6% tium consisting of species such as Anabaena sp., Burkhold-
weight loss of LDPE after incubation with Rhizopus nigri- eria sp., Flavobacterium sp., Pseudomonas aeruginosa, P.
cans in a controlled environment over 30 days [39]. fluorescens, P. putida, P. stutzeri and Vibrio alginolyticus
In our experiments, we observed a higher percentage of could degrade LDPE films, resulting in a 19.8% weight loss
weight loss in γ-LDPE films compared to α-LDPE films. after 100 days of incubation [40].
Specifically, LDPE films exposed to R. arrhizus SLNEA1 The weight loss observed in LDPE samples can be attrib-
in batch culture exhibited a weight loss of 23.77% after 30 uted to the breakdown of the carbon backbone by fungal
days of incubation, with no further weight loss observed enzymes. Fungi are known to produce a wide array of
thereafter. Conversely, LDPE films exposed to R. arrhi- enzymes capable of cleaving the chemical bonds of plas-
zus SLNEA1 in continuous culture exhibited a weight loss tic polymers, thereby utilizing them as a carbon source.
higher than 23% after 90 days of incubation. Among the dif- Upon adhesion and colonization onto the LDPE film sur-
ferent films tested, only F4 displayed a lower rate of biodeg- face (a process known as biodeterioration), the fungal strain
radation (16.50%), possibly due to limited contact between secretes extracellular enzymes such as laccases, esterases,
the LDPE film and the fungal strain. Notably, F5 exhibited peroxidases, cutinases, lipases, proteases, and lignocellulo-
the highest weight reduction, raising 29.74%. This signifi- lytic enzymes. These enzymes facilitate the depolymeriza-
cant loss can be attributed to the continuous nutrient supply tion of the long carbon chains of the polymer into simpler
and biomass removal in the culture conditions, facilitating and more degradable oligomers, dimers and monomers,

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3460 Brazilian Journal of Microbiology (2024) 55:3449–3463

further transported across the cellular membrane, intro- formation signify polymer oxidation by fungal enzymes, a
duced into the fungal cell, and assimilated into appropriate clear indication of LDPE degradation [52]. Also, the bands
metabolic pathway(s) [41–43]. Several studies have identi- observed at 1424, 1425 and 1463 cm-1 corresponding to
fied fungal enzymes involved plastic polymer degradation. C = C bending and the presence of alkene groups may result
For instance, the degradation of polyethylene by the laccase from hydrolysis reactions facilitated by fungal enzymes
enzyme produced by Pleurotus ostreatus and Aspergillus [53]. FTIR analysis underscores significant degradation of
flavus has been reported by Gomez-Mendez et al. (2018) α-LDPE and γ-LDPE films by R. arrhizus SLNEA1, likely
[44] and Zhang et al. (2020) [45], respectively. Enzymes through enzymatic hydrolysis reactions promoting micro-
produced by representatives of the genus Rhizopus have bial action [19]. The increased CI value in biodegraded
also been studied. De Souza et al. (2015) reported that Rhi- samples compared to control LDPE film confirms oxida-
zopus is an important fungal species capable of producing tive changes resulting from LDPE breakdown by the fungal
proteases for plastic polymer degradation [46]. Addition- strain [54]. Moreover, prolonged exposure of γ-LDPE film
ally, Walter et al. (1995) demonstrated the involvement of for 90 days resulted in a slight decrease in carbonyl index
lipases excreted by a Rhizopus delemer strain in plastic deg- due to biotic biodegradation through the formation of ester
radation [47]. groups [55]. When LDPE is oxidized, carbonyl groups are
Structural alterations on the surface of LDPE films due introduced into the polymer chain, leading to the formation
to biodegradation were assessed using conventional tech- of carboxylic acids and other low molecular weight prod-
niques such as SEM, EDS, and ATR-FTIR. The degradation ucts. These alterations render the material more hydrophilic,
patterns observed via SEM were consistent with findings enhancing its potential for biodegradation and accelerating
reported by Khruengsai et al. (2021) [19, 39, 48], demon- the rate of biodegradation [56].
strating a transition from a smooth to a rougher surface on Finally, LDPE films exposed to R. arrhizus SLNEA1 in
LDPE films exposed to R. arrhizus SLNEA1. Additionally, continuous culture for 90 days exhibited better results com-
EDS analysis provided clear evidence of oxidation, con- pared to α-LDPE exposed to R. arrhizus SLNEA1 in batch
firming the modification and breakdown of polymer carbon culture over the same incubation period, either through
chains by the tested fungal strain. weight loss, SEM and EDS analyses, ATR-FTIR or car-
Prior to degradation, the carbon content in µ-LDPE bonyl index. This difference can be attributed to the high
was notably high (94.83%). However, following degrada- production of primary metabolites in continuous culture
tion by R. arrhizus SLNEA1, the percentage of carbon in facilitated by nutrient supplementation. Primary metabo-
LDPE films decreased, estimated at 69.26% and 56.84% in lites are crucial products synthesized during the exponential
α-LDPE and γ-LDPE, respectively. Conversely, the oxygen phase of microbial growth and play an integral role in meta-
content increased along the degradation process, highlight- bolic processes [57]. Filamentous fungi such as Rhizopus
ing oxidation processes. This reduction in carbon content species possess the capability to degrade polymers and con-
can be attributed to its utilization by the fungal isolate, vert simple sugars into primary metabolites [58, 59]. This
resulting in the introduction of oxygen into the polymer enhanced metabolic activity in continuous culture likely
matrix, by the formation of oxygen-containing compounds leads to more efficient degradation of LDPE films by R.
such as hydroxyl (-OH) and carbonyl (C = O) groups, thus arrhizus SLNEA1, contributing to the observed differences
explaining the observed increase in elemental oxygen. in degradation performance between batch and continuous
FTIR analysis was employed to detect chemical changes cultures.
in the composition of LDPE films (α-LDPE and γ-LDPE)
exposed to R. arrhizus SLNEA1, confirming their degra-
dation by fungal enzymes. Indeed, chemical transforma- Conclusion
tions and the emergence of new peaks were evident. The
increased intensity of absorption bands at 2912, 2845, 1463, Biodegradation stands out as a promising strategy for
and 718 cm-1 aligns with findings reported by El-sayed et addressing plastic pollution. This study demonstrates
al. (2021) [49]. The new peaks corresponding to stretching the effectiveness of the fungal strain Rhizopus arrhizus
vibrations of the C = O bond and the O-H bond could be SLNEA1 in degrading LDPE films in both batch and contin-
attributed to organic carboxylic acid groups generated via uous cultures. Different methods, including weight loss (%),
hydrolysis reactions by fungal enzymes [50]. Moreover, scanning electron microscopy (SEM), Energy Dispersive
peaks at 1424, 1425, 1107, and 1105 cm-1 obtained from X-ray Spectroscopy (EDS) and attenuated total reflectance-
CH2 bending vibration and C-O stretching are indicative of Fourier transform infrared (ATR-FTIR), were employed to
polymer crystallization [51]. Additionally, spectra obtained confirm biodegradation. Interestingly, the degradation rates
at 1836 and 1841 cm-1, attributed to carbonyl groups differed between the two culture modes, with continuous

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Brazilian Journal of Microbiology (2024) 55:3449–3463 3461

11. Selke SE, Hernandez RJ (2001) Packaging: polymers in flexible

culture exhibiting superior efficiency. This study marks the packaging. Encyclopedia Mater: Sci Technol 6652–6656. https://
first to underscore the efficacy of continuous culture, show- [Link]/10.1016/B0-08-043152-6/01176-1
casing higher LDPE fungal degradation rates. 12. Pramila R, Ramesh KV (2011) Biodegradation of low density
polyethylene (LDPE) by fungi isolated from marine water- a
Supplementary Information The online version contains SEM analysis. Afr J Microbiol Res 5(28):5013–5018. [Link]
supplementary material available at [Link] org/10.5897/AJMR11.670
024-01487-8. 13. Lu H, Diaz DJ, Czarnecki NJ, Zhu C, Kim W, Shroff R, Acosta
DJ, Alexander BR, Cole HO, Zhang Y, Lynd NA, Ellington AD,
Alper HS (2022) Machine learning-aided engineering of hydro-
Acknowledgements The study was supported by the National Higher
lases for PET depolymerization. Nature 604(7907):662–667.
School of Mines and Metallurgy, Annaba (Algeria), the General Direc-
[Link]
torate for Scientific Research and Technological Development (Alge-
14. Ameen F, Moslem M, Hadi S, Al-Sabri AE (2015) Biodegradation
ria) and the National Polytechnic School of Constantine -Malek Ben-
of low density polyethylene (LDPE) by Mangrove Fungi from
nabi (Algeria).
the Red Sea Coast. Progress Rubber Plast Recycling Technol
31(2):125–143. [Link]
Declarations 15. Munir E, Harefa RSM, Priyani N, Suryanto D (2018) Plastic
degrading fungi Trichoderma viride and Aspergillus nomius iso-
Conflict of interest On behalf of all authors, the corresponding author lated from local landfill soil in Medan. IOP Conference Series:
states that there is no conflict of interest. Earth and Environmental Science 126: 012145. [Link]
org/10.1088/1755-1315/126/1/012145
16. Das MP, Kumar S (2014) Microbial Deterioration of Low Density
Polyethylene by Aspergillus and Fusarium Sp. Int J ChemTech
References Res 6(1):299–305
17. Ojha N, Pradhan N, Singh S, Barla A, Shrivastava A, Khatua P,
1. Abraham J, Ghosh E, Mukherjee P, Gajendiran A (2016) Micro- Rai V, Bose S (2017) Evaluation of HDPE and LDPE degrada-
bial degradation of low density polyethylene. Environ Prog Sus- tion by fungus, implemented by statistical optimization. Sci Rep
tain Energy 36:147–154. [Link] 7:39515. [Link]
2. Ekanayaka AH, Tibpromma S, Dai D, Xu R, Suwannarach N, 18. Saxena A, Pareek A (2017) Possible degradation of LDPE and
Stephenson SL, Dao C, Karunarathna SC (2022) A Review of Biodegradable Polythene by Natural Fungal Flora. J Phytological
the Fungi That Degrade Plastic. Journal of Fungi (Basel) 25;8(8): Res 30(2):77–81
772. [Link] 19. Khruengsai S, Sripahco T, Pripdeevech P (2021) Low-density
3. Plummer C, Bourban P, Månson J (2001) Polymer Matrix polyethylene Film Biodegradation potential by Fungal species
composites: matrices and Processing. Ref Module Mater from Thailand. J Fungi 7:594. [Link]
Sci Mater Eng 7388–7396. [Link] 20. Khruengsai S, Sripahco T, Pripdeevech P (2023) Microbial
B978-0-12-803581-8.02386-9 degradation of low-density polyethylene by Neopestalotiopsis
4. Lu S, Qu R, Forcada J (2009) Preparation of magnetic poly- phangngaensis. J Gen Appl Microbiol 68(6):287–294. [Link]
meric composite nanoparticles by seeded emulsion polym- org/10.2323/jgam.2022.07.001
erization. Mater Lett 63:770–772. [Link] 21. Davet P, Rouxel F (1997) Détection et isolement des champi-
matlet.2008.12.045 gnons du sol. INRA, Paris
5. Rao JP, Geckeler KE (2011) Polymer nanoparticles: preparation 22. Pardo-Rodríguez ML, Zorro-Mateus PJP (2021) Biodegradation
techniques and size-control parameters. Prog Polym Sci 36:887– of polyvinyl chloride by Mucor sp. and Penicillium sp. isolated
913. [Link] from soil. Revista de Investigación Desarrollo e Innovación 11(2):
6. Horton AA, Walton A, Spurgeon DJ, Lahive E, Svendsen C 387–400. [Link]
(2017) Microplastics in freshwater and terrestrial environments: 23. Saidi A, Mebdoua S, Mecelem D, Al-Hoshani N, Sadrati N,
evaluating the current understanding to identify the knowledge Boufahja F, Bendif H (2023) Dual biocontrol potential of the
gaps and future research priorities. Sci Total Environ 586:127– entomopathogenic fungus Akanthomyces muscarius against Thau-
141. [Link] metopoea pityocampa and plant pathogenic fungi. Saudi J Biol
7. Mao R, Song J, Yan P, Ouyang Z, Wu R, Liu S, Guo X (2021) Sci 30(8):103719. [Link]
Horizontal and vertical distribution of microplastics in the Wul- 24. Hurdeal VG, Gentekaki E, Hyde KD, Nguyen TT, Lee HB
iangsuhai Lake sediment, northern China. Sci Total Environ (2021) Novel Mucor species (mucoromycetes, Mucoraceae) from
754:142426. [Link] northern Thailand. MycoKeys 84:57. [Link]
8. Chae Y, An YJ (2018) Current research trends on plastic pol- mycokeys.84.71530
lution and ecological impacts on the soil ecosystem: a review. 25. Sadrati N, Zerroug A, Demirel R, Harzallah D (2023) Anti-mul-
Environ Pollut 240:387–395. [Link] tidrug-resistant Staphylococcus aureus and anti-dermatophyte
envpol.2018.05.008 activities of secondary metabolites of the endophytic fungus
9. Rochman CM, Hoh E, Hentschel BT, Kaye S (2013) Long-Term Penicillium Brevicompactum ANT13 associated with the Alge-
Field Measurement of Sorption of Organic Contaminants to five rian endemic plant Abies Numidica. Arch Microbiol 205(4):110.
types of Plastic pellets: implications for Plastic Marine debris. [Link]
Environ Sci Technol 47(3):1646–1654. [Link] 26. Alshehrei F (2017) Biodegradation of low density polyethylene by
es303700s Fungi isolated from Red Sea Water. Int J Curr Microbiol Appl Sci
10. Huang D, Li X, Ouyang Z, Zhao X, Wu R, Zhang C, Lin C, Li 6(8):1703–1709. [Link]
Y, Guo X (2021) The occurrence and abundance of microplastics 27. Nakajima-Kambe T, Shigeno-Akutsu Y, Nomura N, Onuma F,
in surface water and sediment of the West River downstream, in Nakahara T (1999) Microbial degradation of polyurethane, poly-
the south of China. Sci Total Environ 756:143857. [Link] ester polyurthanes and polyether polyurthanes. Appl Microbiol
org/10.1016/[Link].2020.143857 Biotechnol 51:134–140. [Link]

13
3462 Brazilian Journal of Microbiology (2024) 55:3449–3463

28. Bourzama G, Iratni N, Ennaghra N, Ouled-Haddar H, Soumati degradation. J Appl Biology Biotechnol 11(3):61–69. [Link]
B, Ameur Soualem K, Bechiri K (2021) In vitro removal of elec- org/10.7324/JABB.2023.108929
tronic and electrical wastes by fungi isolated from soil at Annaba 44. Gomez-Mendez LD, Moreno-Bayona DA, Poutou-Piñales RA,
Area in Northeast of Algeria. Environ Nat Resour J 19(4):302– Salcedo-Reyes JC, Pedroza-Rodriguez AM, Vargas A, Bogoya
309. [Link] JM (2018) Biodeterioration of plasma pretreated LDPE sheets
29. Ouartsi N, Djeribi R, Boukachabia A, Menaa F, Gasmi K, Akacem by Pleurotus Ostreatus. PLoS ONE 13(9):e0203786. [Link]
D, Rouabhia M (2019) In vitro Biodegradation of Gliclazide by org/10.1371/[Link].0203786
Aeromonas hydrophila and Serratia odorifera bacteria. Environ 45. Zhang J, Gao D, Li Q, Zhao Y, Li L, Lin H, Bi Q, Zhao Y (2020)
Eng Sci 36(6):643–649. [Link] Biodegradation of polyethylene microplastic particles by the fun-
30. Dsouza GC, Sheriff RS, Ullanat V, Shrikrishna A, Joshi AV, Hire- gus aspergillus flavus from the guts of wax moth Galleria mel-
math L, Entoori K (2021) Fungal biodegradation of low-density lonella. Sci Total Environ 704:135931. [Link]
polyethylene using consortium of aspergillus species under con- scitotenv.2019.135931
trolled conditions. Heliyon 7:e07008. [Link] 46. De Souza PM, Bittencourt MLDA, Caprara CC, de Freitas M,
heliyon.2021.e07008 de Almeida RPC, Silveira D, Fonseca YM, Filho EXF, Junior
31. Ali SS, Quazi IA, Arshad M, Khan Z, Voice TC, Mehmood CT AP, Magalhães PO (2015) A biotechnology perspective of fun-
(2016) Photocatalytic degradation of low density polyethylene gal proteases. Brazilian J Microbiol 46(2):337–346. [Link]
(LDPE) films using titania nanotubes. Environ Nanatechnol Monit org/10.1590/s1517-838246220140359
Manage 5:44–53. [Link] 47. Walter T, Augusta J, Muller RJ, Widdecke H, Klein J (1995)
32. Almond J, Sugumaar P, Wenzel MN, Hill G, Wallis C (2020) Enzymatic degradation of a model polyester by lipase from Rhi-
Determination of the carbonyl index of polyethylene and poly- zopus delemar. Enzym Microb Technol 17:218–224. [Link]
propylene using specified area under band methodology with org/10.1016/0141-0229(94)00007-E
ATR-FTIR spectroscopy. E-polymers 20:369–381. [Link] 48. Gong Z, Jin L, Yu X, Wang B, Hu S, Ruan H, Sung YJ, Lee HG,
org/10.1515/epoly-2020-0041 Jin F (2023) Biodegradation of low density polyethylene by the
33. Gao R, Liu R, Sun C (2022) A marine fungus Alternaria alter- Fungus Cladosporium sp. Recovered from a Landfill Site. J Fungi
nata FB1 efficiently degrades polyethylene. J Hazard Mater 9:605. [Link]
431:128617. [Link] 49. El-Sayed MT, Rabie GH, Hamed EA (2021) Biodegradation
34. Anderson TA, Tsao R, Coats JR (1995) Consumption and deg- of low-density polyethylene (LDPE) using the mixed culture
radation of 3H-Polyethylene/Starch disks by Terrestrial iso- of Aspergillus carbonarius and A. fumigatus Environment,
pods. Bull Environ Contam Toxicol 54:214–221. [Link] Development and Sustainability 23: 14556–14584. [Link]
org/10.1007/BF00197433 org/10.1007/s10668-021-01258-7
35. Lin X, Zhang S, Yang S, Zhang R, Shi X, Song L (2023) A landfill 50. Bhatia M, Girdhar A, Tiwari A, Nayarisseri A (2014) Implica-
serves as a critical source of microplastic pollution and harbors tions of a novel Pseudomonas species on low density polyethyl-
diverse plastic biodegradation microbial species and enzymes: ene biodegradation: an in vitro to in silico approach. SpringerPlus
study in large-scale landfills, China. J Hazard Mater 457:131676. 3(1):497. [Link]
[Link] 51. Rivard C, Moens L, Roberts K, Brigham J, Kelley S (1995)
36. Harrat R, Bourzama G, Ouled-Haddar H, Boudjema S (2022) Starch esters as biodegradable plastics: effects of Ester group
In vitro and Ex situ Biodegradation of low-density polyethyl- chain length and degree of substitution on anaerobic biodeg-
ene by a Rhizopus sp strain isolated from a local Dumpsite in radation. Enzym Microb Technol 17:848–852. [Link]
North-East Algeria. Environ Nat Resour J 20(5):1–10. [Link] org/10.1016/0141-0229(94)00120-G
org/10.32526/ennrj/20/202200026 52. Cai L, Wang J, Peng J, Wu Z, Tan X (2018) Observation of the
37. Tokiwa Y, Suzuki T, Takeda K (1986) Hydrolysis of polyesters degradation of three types of plastic pellets exposed to UV irra-
by Rhizopus arrhizus lipase. Agric Biol Chem 50(5):1323–1325. diation in three different environments. Sci Total Environ 628–
[Link] 629:740–747. [Link]
38. Awasthi S, Srivastava N, Singh T, Tiwary D, Mishra PK (2017) 53. Esmaeili A, Pourbabaee AA, Alikhani HA, Shabani F, Esmaeili
Biodegradation of thermally treated low density polyethylene E (2013) Biodegradation of low-density polyethylene (LDPE)
by fungus Rhizopus oryzae NS 5. 3 Biotech 7(1):73. [Link] by mixed culture of Lysinibacillus xylanilyticus and aspergillus
org/10.1007/s13205-017-0699-4 Niger in soil. PLoS ONE 8:e71720. [Link]
39. Saxena A, Jain S, Pareek A (2022) Estimation of possible Bio- [Link].0071720
degradation of Polythene by Fungal isolates growing on poly- 54. Ratanakamnuan U, Aht-Ong D (2006) Photobiodegradation of
thene debris. Pollution 8(2):567–577. [Link] low-density polyethylene/banana starch films. J Appl Polym Sci
poll.2021.331370.1195 100:2725–2736. [Link]
40. Kumar BM, Noobia S, Mythri S (2016) Studies on Biodegrada- 55. Sudhakar M, Doble M, Murthy PS, Venkatesan R (2008) Marine
tion of Plastic Packaging materials in Soil Bioreactor. Indian J microbe-mediated biodegradation of low- and high-density poly-
Adv Chem Sci S1:297–299. [Link] ethylenes. Int Biodeterior Biodegrad 61(3):203–213. [Link]
CorpusID:220544519 org/10.1016/[Link].2007.07.011
41. Srikanth M, Sandeep TSRS, Sucharitha K, Godi S (2022) Biodeg- 56. Albertsson AC, Erlandsson B, Hakkarainen M, Karlsson S (1998)
radation of plastic polymers by fungi: a brief review. Bioresources Molecular Weight Changes and Polymeric Matrix Changes corre-
Bioprocess 9:42. [Link] lated with the formation of Degradation products in Biodegraded
42. Puliga F, Zuffi V, Baldo D, Cavatorta D, Zambonelli A, Fran- Polyethylene. J Environ Polym Degrad 6(4):187–195. [Link]
cioso O, Sanchez-Cortes S (2023) Cladosporium cladosporioides org/10.1023/A:1021873631162
(strain Clc/1): a candidate for low-density polyethylene degra- 57. Sanchez S, Demain AL (2008) Metabolic regulation and over-
dation. Chem Biol Technol Agric 10:50. [Link] production of primary metabolites. Microb Biotechnol 1(4):283–
s40538-023-00419-2 319. [Link]
43. Varshney S, Gupta V, Yadav AN, Rahi RK, Devki, Neelam 58. Tamano K, Yoshimi A (2021) Metabolic Engineering techniques
DK (2023) An overview on role of fungi in systematic plastic to increase the Productivity of primary and secondary metabolites

13
Brazilian Journal of Microbiology (2024) 55:3449–3463 3463

within filamentous Fungi. Front Fungal Biology 2:743070. Springer Nature or its licensor (e.g. a society or other partner) holds
[Link] exclusive rights to this article under a publishing agreement with the
59. Zaveri A, Edwards J, Rochfort S (2022) Production of primary author(s) or other rightsholder(s); author self-archiving of the accepted
metabolites by Rhizopus Stolonifer, Causal Agent of Almond manuscript version of this article is solely governed by the terms of
Hull Rot Disease. Molecules 27:7199. [Link] such publishing agreement and applicable law.
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