Journal of Applied Microbiology ISSN 1364-5072

REVIEW ARTICLE

Degradation and metabolism of synthetic plastics and

associated products by Pseudomonas sp.: capabilities and
challenges
R.A. Wilkes and L. Aristilde
Department of Biological and Environmental Engineering, College of Agriculture and Life Sciences, Cornell University, Ithaca, NY, USA

Keywords
biodegradation, biofilm, bioremediation,
environmental, Pseudomonads.
Correspondence
Ludmilla Aristilde, 214 Riley-Robb Hall, Cor-
nell University, Ithaca, NY 14850, USA.
E-mail: ludmilla@cornell.edu
2017/0147: received 20 January 2017, revised
17 March 2017 and accepted 10 April 2017
doi:10.1111/jam.13472
Summary
Synthetic plastics, which are widely present in materials of everyday use, are
ubiquitous and slowly-degrading polymers in environmental wastes. Of special
interest are the capabilities of microorganisms to accelerate their degradation.
Members of the metabolically diverse genus Pseudomonas are of particular interest
due to their capabilities to degrade and metabolize synthetic plastics. Pseudomonas
species isolated from environmental matrices have been identified to degrade
polyethylene, polypropylene, polyvinyl chloride, polystyrene, polyurethane,
polyethylene terephthalate, polyethylene succinate, polyethylene glycol and polyvinyl
alcohol at varying degrees of efficiency. Here, we present a review of the current
knowledge on the factors that control the ability of Pseudomonas sp. to process these
different plastic polymers and their by-products. These factors include cell surface
attachment within biofilms, catalytic enzymes involved in oxidation or hydrolysis of
the plastic polymer, metabolic pathways responsible for uptake and assimilation of
plastic fragments and chemical factors that are advantageous or inhibitory to the
biodegradation process. We also highlight future research directions required in
order to harness fully the capabilities of Pseudomonas sp. in bioremediation
strategies towards eliminating plastic wastes.

(<5 mm). Microplastics can be degraded further by micro-

Introduction
Plastic waste is a global issue, rapidly escalating, with
approximately 311 million tons of plastic produced world-
wide in 2014 (PlasticsEurope 2015; Neufeld et al. 2016).
The synthetic plastics that constitute about 80% of total
global plastic usage are polyethylene (PE), polypropylene
(PP), polyvinyl chloride (PVC), polystyrene (PS), polyur-
ethane (PU) and polyethylene terephthalate (PET) (Plas-
ticsEurope 2015) (Fig. 1a). Other less common plastics of
similar structures but with more hydrolysable functional
groups include polyethylene succinate (PES), polyethylene
glycol (PEG) and polyvinyl alcohol (PVA) (Fig. 1b). In
environmental matrices, the degradation of these synthetic
plastics is very slow (Devi et al. 2016). This resistance to
degradation can be circumvented by physicochemical envi-
ronmental factors and microbial capabilities (Devi et al.
2016). Long-term exposure to sunlight and physical abra-
sion contributes to the formation of microplastics
organisms. Consequently, biological degradation and sub-
sequent metabolism have the potential to eliminate plastics
from contaminated environments (Kolvenbach et al.
2014).
Species of the genus Pseudomonas, which are ubiqui-
tous in both aquatic and terrestrial environments, have
been touted for the bioremediation of crude oil, simple
hydrocarbons, naphthalene, toluene and other hydropho-
bic polymers (Timmis 2002; Tribedi et al. 2012; Dash
et al. 2013; Wierckx et al. 2015). In addition, Pseu-
domonas species are recognized as a valuable platform for
biotechnology (Poblete-Castro et al. 2012; Tiso et al.
2015; Wierckx et al. 2015; Nikel et al. 2016). Due to their
diverse metabolic capabilities and genetic plasticity, Pseu-
domonas species are attractive for synthetic biology (Dos
Santos et al. 2004; Nikel et al. 2016). Through genetic
manipulations, a Pseudomonas strain was engineered to
oxidize aromatic, aliphatic, terpenic and polyaromatic

582 Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
R.A. Wilkes and L. Aristilde
(a)
PE
PP
PVC
Figure 1 Structures of (a) the common PS
synthetic plastics polyethylene (PE),
polypropylene (PP), polyvinyl chloride (PVC),
polystyrene (PS), polyethylene terephthalate
(PET) and polyurethane (PU), and (b) the
more hydrolysable plastics polyethylene
succinate (PES), polyethylene glycol (PEG) and PET
polyvinyl alcohol (PVA). The R-groups in PU
can represent a number of functional groups
that are used to subclassify PUs as aromatic,
R R
aliphatic, polyester, polyether or PU
polycaprolactone.
hydrocarbons (Friello et al. 2001). With respect to plastic
degradation, the species of the genus Pseudomonas are
amongst the most cited degraders of various extents for a
wide range of plastic polymers. Complete biologically
mediated elimination of plastic polymers first requires
breakdown of the polymer into smaller oligomers and,
eventually, monomers that can pass through the cell mem-
brane followed by assimilation and subsequent intracellular
metabolism (Lucas et al. 2008; Singh and Sharma 2008;
Kolvenbach et al. 2014). Here, we present a review of the
capabilities of species of the genus Pseudomonas to degrade
and metabolize synthetic plastics. Within the realm of their
established ability to degrade organic contaminants, under-
standing the constitutive capabilities of Pseudomonas sp. to
degrade plastic polymers can be instrumental in employing
new bioremediation strategies.
Relevance of Pseudomonas sp. to plastic
biodegradation
Polyethylene, which is the most widely produced syn-
thetic plastic, is used in plastic bags, water and milk bot-
tles, food packaging and toys (Shah et al. 2008; Sangale
et al. 2012). A long-chain polymer saturated with ethy-
lene bonds, PE is highly hydrophobic (Fig. 1). When PE
has branching chains that prevent tight packing into a
crystalline structure, it is characterized as low-density PE
(LDPE). On the other hand, when there is little to no
branching and the molecules can stack and form strong
Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
Plastic degradation by Pseudomonas
O
(b) O
PES O n
n
O
O
PEG H OH
n n
CI OH
PVA
n
n
n
O O
O O n
O O
O N N O n
H H
intermolecular forces, PE is considered a high-density PE
(HDPE). From the 15 HDPE-degrading bacterial species
isolated from a marine ecosystem, Pseudomonas sp. were
found to be the most efficient followed by Arthrobacter
sp. (Balasubramanian et al. 2010). The addition of pro-
oxidant additives was shown to increase the hydrophilic-
ity of the long-chain polymer of PE, resulting in chain
scission of the polymer and the generation of carbonyl
functional groups and low molecular weight (MW) com-
ponents (Chiellini et al. 2006; Singh and Sharma 2008).
After pretreatment with nitric acid, P. aeruginosa was able
to degrade 0
25 g of LDPE by 50
5% in 2 months
(Rajandas et al. 2012). However, no chemical pretreat-
ment was needed for Pseudomonas sp. AKS2 to degrade
LDPE films, albeit only 5% of the total mass of 300 mg
was degraded within 45 days (Tribedi and Sil 2013c).
Also without any pretreatment, an uncharacterized Pseu-
domonas sp. was found to degrade 28
6% of 5% dry
weight of low MW PE (MW = 1700 Da) in a sterilized
compost condition after 40 days (Yoon et al. 2012).
Therefore, the extent to which PE and related plastics are
biodegraded depends both on the structural arrangement
of the plastic polymer and the type of Pseudomonas
strains exposed to the polymer. Of particular interest to
bioremediation strategies of PE are the mechanisms
through which these strains can degrade PE-containing
plastics without pretreatment.
Plastics with similar structures to PE, but with more
hydrolysable functional groups, include PVA, PES and
583
Plastic degradation by Pseudomonas
PEG. Polyvinyl alcohol, which has similar carbon–carbon
linkages to PE, is more water soluble than PE because of
the presence of the hydroxyl functional group (Shimao
2001). As a result, PVA is more easily degraded than PE.
Since Pseudomonas sp. O-3 was reported to degrade PVA
in 1973 (Suzuki et al. 1973), the majority of reported
PVA-degrading bacteria belong to the Pseudomonas genus
(Shimao 2001). Similar to PE in structure but with added
hydrolysable ester bonds, PES is a plastic polymer that is
considered more amenable to biodegradation; PES is typ-
ically found in shopping bags and agricultural films (Tri-
bedi and Sil 2013a). Pseudomonas sp. AKS2 can
maximally degrade PES at a rate of 1
65 mg day 1 (Tri-
bedi et al. 2012). Furthermore, bioaugmentation of soil
microcosms with Pseudomonas sp. AKS2 resulted in
enhanced PES biodegradation (Tribedi and Sil 2013a). In
addition to PVA and PES, PEG is more biodegradable
than PE due to the presence of ether bonds and a hydro-
xyl end group. Complete biodegradation of PEG using P.
stutzeri JA1001 has been demonstrated with PEG MWs of
up to 14 000 Da in concentrations of 0
2% (w/v) after
30 h (Obradors and Aguilar 1991). While PEG is not as
commonly used as PE, it is still a pervasive plastic that is
found in products such as pharmaceuticals, lubricants,
cosmetics and inks (Gu 2003).
Polystyrene, which is both lightweight and stiff, serves
as an effective thermal insulation in disposable cups,
packaging materials and laboratory equipment (Shah
et al. 2008). The PS structure is characterized by phenyl
functional groups along the hydrocarbon chain. Although
reports of PS biodegradation are scarce, the PS polymer
can be eventually broken down to compounds such as
styrene, toluene and benzene that are metabolizable in
Pseudomonas sp. (Shah et al. 2008; Devi et al. 2016).
Additionally, it was shown that Pseudomonas sp. NCIM
2220 was able to degrade a heteropolymer made of PS
with maleic anhydride anchored with minute amounts of
lactose, sucrose or glucose (Galgali et al. 2002). More-
over, a high impact PS composed of a mixture of PS and
polybutadiene can be degraded by an unclassified Pseu-
domonas sp., but there was only a 10% weight loss to the
200-mg high-impact PS film (Mohan et al. 2016).
Polyurethanes, which are characterized by urethane
bonds formed from the condensation of polyisocyanate
and polyol, have varied structures (aromatic-, aliphatic-,
polycaprolactone-, polyether- or polyester-type PU)
depending on their usage (Shah et al. 2008; Cregut et al.
2013). They are found in tires, sponges, refrigerator insu-
lation, furniture cushions, gaskets, bumpers and paints
(Shah et al. 2008). The presence of carbamate bonds in
PU renders it insoluble in common solvents including
water, acetone and ethanol. Additionally, the durable
properties of PU, which mediate its role as a flame
584 Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
R.A. Wilkes and L. Aristilde
retardant and antimicrobial, reduce the degradation
effects of temperature, pH, chemical agents and micro-
organisms (Cregut et al. 2013; Biffinger et al. 2015).
Despite these PU characteristics that are adverse to
biodegradation, several Pseudomonas species including P.
fluorescens, P. aeruginosa, P. cepacia, P. protegens and P.
chlororaphis have been identified as PU degraders (Cregut
et al. 2013). It was reported that P. aeruginosa AKS9 can
utilize both PU diol and Impranil DLNTM, a commercial
variety of polyester PU, as a sole carbon source (Mukher-
jee et al. 2011). In addition, P. aeruginosa strain MZA-85
was found to degrade and metabolize polyester PU (Shah
et al. 2013). Extensive degradation of Impranil DLN was
also accomplished by P. protegens Pf-5 and other P. prote-
gens strains (Hung et al. 2016).
The polymers with the least amount of reported
biodegradation by Pseudomonas species are PP, PVC and
PET. Polypropylene is found in bottle caps, medicine
bottles, car seats and disposable syringes and is the sec-
ond most produced plastic after PE (Shah et al. 2008;
PlasticsEurope 2015). Polyvinyl chloride is the third
highest produced plastic and is commonly found in
shower curtains, raincoats, bottles, garden hoses and shoe
soles (Shah et al. 2008; PlasticsEurope 2015). Both PP
and PVC are highly hydrophobic and resistant to chemi-
cal abrasion (Shah et al. 2008). Polyethylene terephtha-
late, which is also both thermally and chemically stable,
has a structure amenable for use in water and soda bot-
tles, electronics, automotive parts and textile fibres
(Webb et al. 2013). The global production of PET is
increasing and is approximated to be 50% of synthetic
plastic products (Webb et al. 2013). Both with and with-
out chemical pretreatments, PP is difficult to be biode-
graded (Arkatkar et al. 2009). Following pretreatment
with ultraviolet radiation, P. azotoformans and P. stutzeri
were able to survive on PP as a sole carbon source, but
only minimal weight loss of the plastic polymer resulted
after 1 year (Arkatkar et al. 2010). The PVC monomer,
vinyl chloride, was able to serve as a sole carbon source
to support the growth of P. putida strain AJ (Danko
et al. 2004). Pseudomonas species have not been very
effective for PET biodegradation. A lipase isolated from
an unspecified Pseudomonas sp. was unable to catalyse
PET degradation (M€ uller et al. 2005). Likewise, an extra-
cellular lipase from an unspecified Pseudomonas sp. was
not effective at degrading a mixture with poly(e-caprolac-
tone) and >50% PET (Jun et al. 1994). However, a cuti-
nase from P. mendocina had high affinity to low
crystallinity PET and reduced the weight of the film by
5%—the initial weight of the film was not reported
(Ronkvist et al. 2009).
Taken collectively, the aforementioned studies highlight
the diverse capabilities of Pseudomonas species to degrade
R.A. Wilkes and L. Aristilde
and metabolize the different synthetic plastic polymers.
In the following sections, we review the different factors
that need to be considered in the biotechnological applica-
tion of these species towards the complete biodegradation
of synthetic plastics: (i) biofilm formation, (ii) enzymatic
degradation, (iii) intracellular metabolism, (iv) challenges
to biotechnological applications, (v) favourable chemical
and biological environments, and (vi) future research
needs.
Importance of biofilms to biodegradation
The ability of bacterial cells to attach to and degrade
plastic polymers is dependent on the structure of the
polymer surface (Donlan 2002). The addition of hydro-
philic functional groups to plastic polymers is often
required to promote cell surface attachment due to the
typical hydrophilic nature of cell surfaces, which impair
attraction to the hydrophobic polymers. Thus, greater
surface roughness and hydrophilicity of the polymer
were shown to facilitate both enhanced attachment of
bacterial colonies and accessibility of secreted extracellu-
lar enzymes to polymer surface (Sanin et al. 2003; Tri-
bedi and Sil 2013c; Nauendorf et al. 2016). In cases of
plastic polymers including PE, which have high
hydrophobicity and MW, the formation of a biofilm
either requires the polymer to be altered by oxidation
reactions or supplemented with chemicals in order to
increase the surface interactions with bacterial cells
(Shah et al. 2008; Sivan 2011). Biofilm-forming bacterial
species with relatively high hydrophobic cell surfaces
have improved cell surface attachment to unmodified
plastic polymers (Gilan et al. 2004; Tribedi and Sil
2013b; Devi et al. 2016). Accordingly, biofilm-adapted
Pseudomonas sp. AKS2 cells were found to have greater
cell surface hydrophobicity and LDPE-degrading ability
than planktonic cells (Tribedi et al. 2015). Furthermore,
it was determined that the cells in biofilms secreted
exopolysaccharides that aid in attachment to the plastic
polymer (Tribedi and Sil 2013c). Although it was evi-
dent that the ability of Pseudomonas sp. AKS2 to form
biofilms promoted the degradation of LDPE (Tribedi
and Sil 2013c). Tribedi et al. (2015) proposed that this
ability to degrade LDPE may not be transferrable to
other plastic polymers due to structural differences as
detailed in the previous section.
Furthermore, the nutritional environment of the
growth media may influence the extent of the biofilm
formation (Sivan 2011). The composition of extracellu-
lar polymer matrices can change depending on the
growth conditions and can play an important role in
the attachment properties of bacteria (McEldowney and
Fletcher 1986; Durmaz and Sanin 2001; Sanin et al.
Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
Plastic degradation by Pseudomonas
2003). Cell surface hydrophobicity was found to be cor-
related positively with cell attachment, biofilm forma-
tion and PES weight loss (Tribedi and Sil 2013b). In
addition, low glucose content and high ammonium sul-
phate concentrations resulted in the greatest cell surface
hydrophobicity for Pseudomonas sp. AKS2 grown on
PES (Tribedi and Sil 2013b). Conversely, in the pres-
ence of organic carbon-rich marine sediments, biofilm
formation was diminished and there was minimal to no
degradation of PE (Nauendorf et al. 2016). Therefore,
environmental and nutritional conditions that favour
the genesis of biofilms on plastic polymers are impor-
tant stimuli for the degradation of synthetic plastics by
Pseudomonas sp.
Plastic degradation by extracellular and
intracellular enzymes
In general, enzymatic degradation involves two impor-
tant processes that can be measured by weight loss and
additions of functional groups. The reduction in MW
of the polymer enables the catalytic effects of enzymes
that can only operate on smaller molecules and facili-
tates the transport of smaller molecules through the cell
membrane (Shah et al. 2008). Chemical or biological
oxidation reactions are often necessary to increase the
hydrophilicity of the polymer by providing a functional
group such as alcohol or carbonyl groups that can
enhance bacterial attachment and degradation (Alberts-
son et al. 1995; Lucas et al. 2008; Arkatkar et al. 2010).
Degradative products with carbonyl functional groups
can be metabolized inside the cell through b-oxidation
and the tricarboxylic acid (TCA) cycle (Shah et al.
2008; Restrepo-Florez et al. 2014).
Extracellular enzymes such as depolymerases and
hydrolases act on large plastic polymers to break them
down into smaller molecules (Shah et al. 2008). Hydroly-
tic cleavage can occur either at the polymer chain termi-
nus (exo-attack) or somewhere along the polymer chain
(endo-attack) (Lenz 1993). The two different modes of
attack create different products. Exo-attack results in
small oligomers or monomers that the bacteria can
assimilate into the cell. On the other hand, endo-attack
primarily reduces the MW of the polymer, whereby the
resulting products are not likely to be assimilable without
further degradation (Lenz 1993). An extracellular depoly-
merase from a Pseudomonas sp. was effective in breaking
down a brominated high-impact PS (Mohan et al. 2016).
Degradation of PEG by P. stutzeri JA1001 involved a sin-
gle intracellular PEG dehydrogenase that produced gly-
oxylic acid (Obradors and Aguilar 1991). Alkane
hydroxylases from the AlkB family in Pseudomonas sp. E4
were involved in the degradation of PE with MW up to
585
Plastic degradation by Pseudomonas
27 000 Da (Yoon et al. 2012). Furthermore, the extracel-
lular PVA oxidase found in a number of Pseudomonas
sp., including Pseudomonas sp. O-3, P. vesicularis PD and
Pseudomonas sp. VM15C, can oxidize PVA into a dike-
tone structure (Kawai and Hu 2009).
Esterases, lipases and cutinases are hydrolases that are
instrumental in plastic degradation (Table 1) (Ruiz et al.
1999; Sangale et al. 2012; Novotny et al. 2015; Mohan
et al. 2016). Hydrolases are important for enzymatic poly-
mer cleavage wherein ester bonds are broken through a
nucleophilic attack on carbonyl carbon atoms created
by previous oxidation reactions (Devi et al. 2016). The
degradation of PES by Pseudomonas sp. AKS2 in a bioaug-
mented soil was facilitated by hydrolase and dehydrogenase
activity as determined by enzyme assays (Tribedi and Sil
2013a). Esterases can hydrolyse esters, either already pre-
sent in the polymer or produced through oxidation reac-
tions, into alcohols, phenols and acids. For instance, an
esterase from Pseudomonas sp. AKS2 was able to break the
ester bonds in PES to generate succinic acid, a TCA cycle
metabolite (Tribedi et al. 2012). Following the activity of a
PVA oxidase, which introduced acetyl groups in the PVA,
P. vesicularis PD was able to assimilate the altered PVA
into the cell and hydrolyse it further with an intracellular
esterase (Kawai and Hu 2009). Polyurethane-degrading
enzymes are thought to be primarily extracellular esterases
or proteases that are either membrane-bound or secreted
extracellularly (Mukherjee et al. 2011; Cregut et al. 2013;
Shah et al. 2013). The term polyurethanase is often used to
describe enzymes responsible for the degradation of PU
(Ruiz et al. 1999; Stern and Howard 2000). However, this
term is used without definitive confirmation of the hydrol-
ysis of the carbamate bond. Therefore, it is recommended
that polyurethanase should be reported as hydrolases or
esterases (Biffinger et al. 2014). To this point, an extracel-
lular enzyme from P. chlororaphis with esterase and pro-
tease activities was shown to degrade successfully polyester
PU (Ruiz et al. 1999). This enzyme was also classified as a
serine hydrolase because it could be inhibited by phenyl-
methane sulfonyl fluoride (Ruiz et al. 1999). Polyurethane
was degraded significantly by Pseudomonas sp. lipase but
only partially degraded by a recombinant esterase from P.
fluorescens (Biffinger et al. 2015). The production of high
amounts of extracellular esterases and lipases in P. aerugi-
nosa was reported to facilitate the degradation of aro-
matic–aliphatic polyesters and polyesteramides (Novotny
et al. 2015). An extracellular cutinase from P. mendocina
acting on a PET film with 7% crystallinity caused a 5%
weight loss of the film after 96 h and produced tereph-
thalic acid (TPA) and ethylene glycol (EG) as the sole
products (Ronkvist et al. 2009). These products can subse-
quently be incorporated into intracellular metabolism
(Fig. 2). Previous research has focused on extracellular
586 Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
R.A. Wilkes and L. Aristilde
enzymes primarily involved in the first steps of plastic
degradation. However, much remains unknown regarding
what happens intracellularly to assimilable plastic oligo-
mers or monomers after they are transported across the
bacterial cellular membrane.
From extracellular degradation to potential
intracellular metabolism
The intracellular metabolism of plastic-associated prod-
ucts has not yet been evaluated explicitly. However, mea-
surements of oxygen depletion and carbon dioxide
generation are consistent with growth and metabolism of
plastic degradation products (Lucas et al. 2008). Follow-
ing the breakdown of plastic polymers into smaller com-
pounds, some of these compounds can be potentially
processed through the bacterial metabolism to be ulti-
mately mineralized as CO2 or be exploited for the biosyn-
thesis of valuable products through metabolic pathways
(Shah et al. 2008; Sangale et al. 2012; Devi et al. 2016).
For instance, PE degradation is proposed to form acetic
acid, which can be processed through the TCA cycle
(Fig. 3) (Lenz 1993; Restrepo-Fl orez et al. 2014). The
degradation of high-impact PS by an unclassified Pseu-
domonas sp. led to the formation of intermediates such
as phenyl ethanol, which can enter the aromatic catabo-
lism pathway (Mohan et al. 2016). In P. aeruginosa
MZA-85, cell-bound esterases hydrolyse ester linkages in
PU to produce adipic acid and 1,4-butanediol, which can
be utilized as sole carbon sources by feeding into the
TCA cycle (Shah et al. 2013). Following the initial PET
hydrolysis with a cutinase from P. mendocina, the TPA
and EG produced can be transported into the cell (Fig. 2)
(Ronkvist et al. 2009). In bacterial species, a TPA trans-
porter is implicated in the transport of TPA into the cell
(Hosaka et al. 2013). Once inside the cell, the TPA can
then be broken down intracellularly into protocatechuate,
which undergoes ring cleavage via the aromatic catabo-
lism pathway, before generating TCA cycle intermediates
(Fig. 2) (Karegoudar and Pujar 1985; Wang et al. 1995;
Novotny et al. 2015; Yoshida et al. 2016).
PVA degradation has been well studied and many
PVA degradation pathways for different bacteria (e.g.
Sphingopyxis species, Alcaligenes species and Pseudomonas
species) have been proposed (Kawai and Hu 2009).
These pathways commonly include a cleavage of the
main polymer chain by an extracellular dehydrogenase
or oxidase with subsequent aldolase and hydrolase reac-
tions in order to modify the compounds into products
such as acetic acid and hydroxyl fatty acids that can be
incorporated eventually into the TCA cycle and b-oxida-
tion respectively (Fig. 4) (Shimao 2001; Kawai and Hu
2009).
R.A. Wilkes and L. Aristilde Plastic degradation by Pseudomonas

Table 1 List of Pseudomonas sp. and associated enzymes for plastic degradation

Plastic Microorganism Enzyme/s Reference

Polyethylene (PE)
LDPE Pseudomonas sp. AKS2 Hydrolase Tribedi and Sil (2013c)
LMWPE Pseudomonas sp. E4 Alkane hydroxylase Yoon et al. (2012)
Polystyrene (PS)
High impact Pseudomonas sp. Esterase Mohan et al. (2016)
Vinyl chloride P. putida AJ Alkene monooxygenase Danko et al. (2004)
Polyurethane (PUR)
Polyester P. chlororaphis Polyurethanase Ruiz et al. (1999)
Polyester P. aeruginosa Esterase Mukherjee et al. (2011)
Polyester P. aeruginosa MZA-85 Esterase Shah et al. (2013)
Polyester Pseudomonas sp. Lipase Biffinger et al. (2015)
Polyester P. fluorescens Esterase Biffinger et al. (2015)
Polyester P. fluorescens Protease Howard and Blake (1998)
Polyester P. protegens BC2-12 Lipase Hung et al. (2016)
Polyester P. protegens CHA0 Lipase Hung et al. (2016)
Polyester P. protegens Pf-5 Lipase Hung et al. (2016)
Polyester P. fluorescens A506 and Pf0-1 Lipase Hung et al. (2016)
Polyester P. chlororaphis Lipase Stern and Howard (2000)
Polyester P. fluorescens Esterase/protease Vega et al. (1999)
Poly(ethylene terephthalate) (PET) Pseudomonas sp. Lipase Mu€ller et al. (2005)
Polyethylene succinate (PES) Pseudomonas sp. AKS2 Esterase Tribedi et al. (2012)
Polyethylene glycol (PEG) P. stutzeri PEG dehydrogenase Obradors and Aguilar (1991)
Polyvinyl alcohol (PVA) P. vesicularis PD Esterase Kawai and Hu (2009)

Unlike the previous plastics that undergo an initial
extracellular degradation, PEG is thought to enter directly
the periplasmic space through porins followed by the
intracellular oxidation by PEG dehydrogenase in P. stut-
zeri JAlOOl (Obradors and Aguilar 1991). This catalytic
mechanism produces glyoxylic acid, which can be
invested into metabolism via the glyoxylate shunt in the
TCA cycle (Obradors and Aguilar 1991). An important
next step in elucidating the metabolism of plastic
degradative products in Pseudomonas sp. is to study the
metabolic network required to process the different
degradative products intracellularly.
Challenges in biotechnological applications
The main constraints to plastic biodegradation include
polymer characteristics such as high MW, lack of favour-
able functional groups and crystallinity (Devi et al. 2016).
High MW is one of the strongest deterrents to biodegra-
dation because high MW plastic polymers are less suscep-
tible to microbial attack, resulting in less yield of
oligomers or monomers required for subsequent degrada-
tion or metabolism (Shah et al. 2008; Singh and Sharma
2008; Yoon et al. 2012). Long-chain plastic polymers also
cannot typically pass through the cell membrane and
have to be first acted upon by extracellular enzymes. For
instance, PE with a MW of 500 Da is considered the
maximum weight that can pass through the cell mem-
brane (Yoon et al. 2012). Additionally, synthetic plastics
are typically highly hydrophobic with stable functional
groups such as alkane and phenyl (Shah et al. 2008).
Therefore, oxidation and hydrolysis of these polymers are
necessary to increase hydrophilicity prior to microbial
attack (Shah et al. 2008). Furthermore, amorphous
regions of plastics with greater branching structure are
more prone to microbial attack than crystalline plastic
polymers with more rigid shape (Singh and Sharma
2008).
The extent of biodegradation can also be affected by
abiotic factors such as pH, temperature and humidity
(Gu 2003). It was reported that high humidity and tem-
perature conditions can promote microbially mediated
hydrolysis (Devi et al. 2016). Furthermore, reactions by
plastic-degrading enzymes require favourable conditions
(Lucas et al. 2008). For instance, a dual functioning PU-
degrading enzyme from P. chlororaphis was found to have
an optimal pH of 8
5 for esterase activity, but an optimal
pH of 7 for protease activity (Ruiz et al. 1999).
Additives to plastic polymers can act as inhibitors
(Kolvenbach et al. 2014). For instance, PU-containing
materials can be accompanied by highly toxic additives
such as dibutyl tin dilaurate that act as antimicrobials
(Cregut et al. 2013). In addition, other carbon sources
can interfere with the metabolism of plastics. It was

Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology 587
Plastic degradation by Pseudomonas R.A. Wilkes and L. Aristilde

O O

n
PE
O O
n
PET * *
OH O

O O
*
OH
n
O OH
HO *
MHET O O

O O

HO + OH
* n
OH *
HO OH
EG TPA O O

* +
HO OH OH
n Acetic acid

HO COOH

PCA
Biosynthetic Biosynthetic
pathways Glyoxylate pathways

TCA TCA
cycle CO2 CO 2
cycle

Figure 2 Proposed pathway for the degradation of polyethylene
terephthalate (PET). Through complete enzymatic hydrolysis, PET can
be degraded directly to terephthalic acid (TPA) and ethylene glycol
(EG) as indicated by the solid arrow on the far right. Intermediary
products from incomplete hydrolysis of PET include mono-(2-hydro-
xyethyl) terephthalate (MHET), shown between the dashed parenthe-
sies, and bis-(2-hydroxyethyl) terephthalate (BHET) that can later be
hydrolysed into TPA and EG. A TPA transporter can bring TPA into
the cell where a series of reactions converts it to protocatechuic acid
(PCA) followed by integration into metabolism through the tricar-
boxylic acid (TCA) cycle. Similarly, EG can be processed through the
glyoxylate shunt in the TCA cycle. The dashed arrows represent multi-
ple steps. An asterisk indicates where new functional groups were
added to the polymer during the different enzymatic steps. (Adapted
from Wang et al. (1995); Ronkvist et al. (2009); and Yoshida et al.
(2016)).
shown that the presence of glucose exerted repression on
PES-degrading enzymes from Pseudomonas sp. AKS2
(Tribedi et al. 2012). In the absence of glucose, the PES
Figure 3 Proposed biodegradation pathway of polyethylene (PE). The
pathway involves several steps of oxidation, dehydrogenation and car-
bon–carbon bond breaking to produce acetic acid that can be inte-
grated into the TCA cycle. Small aliphatic hydrocarbons (approximately
up to 20 carbon atoms) could also be transported directly into the bac-
terial cell before subsequent breakdown. The dashed arrows represent
when more than one reaction occurred in the scheme. An asterisk indi-
cates where new functional groups were added to the polymer during
the different enzymatic steps. (Adapted from Lenz (1993)).
film underwent a 45% weight loss, whereas with the addi-
tion 0
25% glucose, the weight loss was decreased to 25%
(Tribedi et al. 2012). Similarly, P. protegens Pf-5 no
longer exhibited Impranil hydrolysis when grown in a
medium containing glucose (Hung et al. 2016). Catabo-
lite repression was also seen, but at a lesser extent, when
P. protegens Pf-5 was grown in the presence of pyruvate,
fructose, gluconate, mannitol and glycerol (Hung et al.

588 Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
R.A. Wilkes and L. Aristilde Plastic degradation by Pseudomonas

OH OH OH OH
Favourable chemical and biological environments
Plastic polymers can take decades to degrade without pre-
n treatment to increase the hydrophilicity of the polymer
PVA surface. Degradation of plastics increases after pretreat-
*
OH OH O OH ment with thermal oxidation, UV photooxidation, oxida-
tive chemicals or surfactants (Gilan et al. 2004; Arkatkar
et al. 2010; Devi et al. 2016). Photodegradation can cause
chain scission of plastic molecules via free radical-
* mediated mechanisms that create smaller molecules that
OH O O OH can be taken up by microorganisms (Devi et al. 2016).
Furthermore, analysis of PP via infrared spectroscopy indi-
cated that pretreatment with short UV radiation intro-
duced keto carbonyl peaks while thermal pretreatment
introduced ester carbonyl peaks (Arkatkar et al. 2010). It
OH O O OH was proposed that the ester bonds might be preferentially
attacked by P. stutzeri because PP pretreated with thermal
+
* * oxidation had enhanced weight loss over PP pretreated
CH3 HO with short UV (Arkatkar et al. 2010). By contrast, neither
O UV nor thermal pretreatment increased the ability of
P. azotoformans to degrade PP (Arkatkar et al. 2010).
Oxidative chemicals such as sulphuric acid, nitric acid,
OH hydrochloric acid and hydrogen peroxide were also suc-
Acetic acid cessful at creating hydroxyl group radicals that oxidize
polymer surfaces (Arkatkar et al. 2010; Sen and Raut
2015). Nitric acid is a common pretreatment method that
oxidizes the polymer and establishes carbonyl groups and
Biosynthetic double bonds into the chain backbone (Rajandas et al.
pathways 2012; Sen and Raut 2015). Similarly, the addition of pro-
oxidant additives was shown to increase the hydrophilicity
of the long-chain polymer of PE by introducing carbonyl
TCA functional groups and generating lower MW components
cycle CO2
(Chiellini et al. 2006; Singh and Sharma 2008). In addition
to chemical oxidants, surfactants are used to promote
microbial attacks by increasing the hydrophilicity of the
polymer surface. Nonionic surfactants such as Tween 80
Figure 4 Predicted pathway for polyvinyl alcohol (PVA) degradation. have been utilized to increase P. aeruginosa adhesion to
Extracellular depolymerization reactions via a dehydrogenase or oxi- LDPE (Albertsson et al. 1993). However, in the case of
dase first cleave PVA. Hydrolase or aldolase reactions further break Pseudomonas sp. AKS2 that has naturally high cell surface
down the products into acetic acid that is metabolized in the TCA hydrophobicity, Tween 80 reduced cell attachment to and
cycle. The dashed arrow on the far right indicates an alternative aldo-
degradation of LDPE while mineral oil that promotes
lase reaction mechanism. An asterisk indicates where new functional
groups were added to the polymer during the different enzymatic
hydrophobic interactions increased cell attachment and
steps. The dashed arrows represent multiple steps. (Adapted from degradation ability (Tribedi and Sil 2013c). Therefore, pre-
Kawai and Hu (2009) and Shimao (2001)). treatments have species-specific effects and have to be
adjusted to each degradation scenario.
The addition of alternative substrates can stimulate the
2016). In contrast, citrate and succinate both were found degradation ability of Pseudomonas sp. (Singh and
to stimulate Impranil-clearing activity (Hung et al. 2016). Sharma 2008). Biodegradation can be enhanced by the
The inhibitory or stimulatory effects of alternative sub- addition of a biodegradable additive such as starch, which
strates on different types of plastic degradation are not provides a nutrient source that can be easily metabolized
clearly defined and should be better understood to by the bacterial cells (Singh and Sharma 2008). Accord-
inform the construction of optimal conditions for ingly, a PE film blended with hydroxypropylated starch
biodegradation. exhibited higher degradation by P. aeruginosa ATCC

Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology 589
Plastic degradation by Pseudomonas
13388 compared to nonblended PE films (Kim 2003).
However, this type of sugar additive to the polymer
structure can reduce the mechanical strength and usabil-
ity of the plastic (Kim 2003). Therefore, it is preferable to
have a nutrient stimulant present externally of the plastic
polymer rather than blended into the polymer structure.
External enhancement of PU degradation by P. protegens
Pf-5 occurred with the addition of citrate or succinate
into the growth media (Hung et al. 2016). When Pseu-
domonas sp. A was exposed to nitrogen-starved and car-
bon-rich conditions, there was a significant reduction (by
about 60%) in both cell surface hydrophobicity and cell
attachment after 100 h (Sanin et al. 2003). This was
attributed to the increased production of extracellular
polysaccharides due to metabolic overflow of excess car-
bon (Sanin et al. 2003). This extracellular increase in
polysaccharide concentration thus increases the
hydrophilicity of the cell surface (Sanin et al. 2003). On
the other hand, an increase in the protein content of the
extracellular matrix during growth on carbon-depleted
and nitrogen-rich media was accompanied instead by a
small increase in cell surface hydrophobicity and attach-
ment (Sanin et al. 2003). A similar effect was observed
with Pseudomonas sp. AKS2, whereby the addition of
ammonium sulphate to the media resulted in an increase
in cell surface hydrophobicity (Tribedi and Sil 2013b).
Trace elements can also aid in the breakdown and assimi-
lation of plastics by influencing the efficiency and func-
tion of metabolic enzymes. Furthermore, manganese and
iron can act as a pro-oxidants of polymers (Singh and
Sharma 2008). However, some trace metals such as cobalt
and nickel acted as inhibitors of the enzymes involved in
PVA degradation in Pseudomonas sp. O-3 (Suzuki 1976).
These reports highlight the importance of manipulating
nutrient availability and the composition of metabolic
overflow products in order to enhance the attachment
between bacterial cells and plastic surfaces.
Symbiotic bacterial consortium can be more effective
in degrading plastics. A consortium of two known PU
degraders, Bacillus subtilis MZA-75 and P. aeruginosa
MZA-85, resulted in the greatest weight loss of the
250 mg film of polyester PU and the highest esterase
activity compared to the individual strains (Shah et al.
2016). Microbial interactions in a consortium can also
promote degradation capabilities. Although incapable of
using PVA individually as a sole carbon source, P. putida
VM15A and Pseudomonas sp. VM15C together were able
to grow symbiotically on PVA (Shimao 2001). This was
achieved due to the production of the growth factor
pyrroloquinoline quinone by P. putida VM15A, which in
turn facilitated the metabolism of PVA by Pseudomonas
sp. VM15C (Shimao 2001). In another symbiotic exam-
ple, a Flavobacterium sp. was able to degrade PEG when
590 Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
R.A. Wilkes and L. Aristilde
co-cultured with a Pseudomonas sp, which removes toxic
by-products of PEG degradation produced by Flavobac-
terium sp. (Gu 2003).
Future research needs
Pseudomonas species, which have been touted historically
for contaminant remediation due to their ability to
degrade oil contaminants, also have the potential to
degrade and metabolize plastic wastes. Biodegradation of
structurally different plastics and their associated by-
products is species-dependent due to the required set of
enzymes. A comprehensive understanding of the enzymes
involved in the degradation of different plastics as well as
the identification of their extracellular versus intracellular
localization will inform bioengineering approaches for
optimizing plastic biodegradation. For instance, the
hydrocarbon degradation genotype of P. putida was used
to transform a native marine bacterium in order to con-
fer capabilities to degrade hydrocarbons (Latha and
Lalithakumari 2001). A similar approach, which would
remove the potential for issues associated with the intro-
duction of novel species to the environment, can be
applied for promoting plastic biodegradation by native
microbial species. This genetic engineering approach pro-
vides the opportunity to combine strategically the effects
of mutually beneficial enzymes as well as enzymes that
can act on different plastics.
Plastic polymers can be broken down to varying
degrees both physically and biologically with minimal
generation of compounds amenable to metabolism inside
the cells. Intermediate products produced from the first
steps of biodegradation can interfere with future steps
needed for uptake and subsequent intracellular metabo-
lism (Kolvenbach et al. 2014; Barth et al. 2016). For
instance, the MHET by-product of PET hydrolysis is
inhibitory of enzymes found in Thermobifida fusca KW3,
and glyoxylic acid inhibits Flavobacterium sp. during PEG
degradation (Gu 2003; Barth et al. 2016). Potential inhi-
bitory effects of plastic degradation by-products on Pseu-
domonas species remain to be fully understood. A
complete elucidation of the degradation pathways of plas-
tics from the initial oxidation and hydrolysis reactions to
the ultimate generation of CO2 and H2O via intracellular
metabolism is important for determining the rate-limiting
steps. Moreover, the identification of the intermediate
compounds that block further metabolism remains largely
unknown. These different pieces of information could
also aid in the design of plastics that are more susceptible
to degradation when released in the environment. Fur-
thermore, an attractive option as a sustainable approach
in bioremediation strategies is the investment of funnel-
ing plastic metabolism into the biosynthetic pathways of
R.A. Wilkes and L. Aristilde
value-added products. An understanding of the metabolic
network in plastic metabolism is a necessary prerequisite
for implementing such strategy.
Many bacterial candidates from the environment that
have been screened for plastic degradation ability were
genetically classified at the species level. However, several
strains of Pseudomonas sp. capable of degrading and
metabolizing plastics still remain to be characterized. As a
result of this limitation, it is challenging to build on previ-
ous research with uncharacterized bacteria, thus leading to
isolated studies with each research group working with
their own strains. For plastic bioremediation to be indus-
trially applicable, better genetic characterization and avail-
ability of efficient biodegraders are warranted.
Of particular interest are future research studies that
concentrate on elucidating novel mechanisms to enhance
bacterial attack on the most commonly used plastics that
are highly resilient and unlikely to degrade naturally in
the environment. To this end, we have identified the fol-
lowing four research focus areas for taking advantage of
the capabilities of Pseudomonas species to degrade and
metabolize synthetic plastics: (i) molecular to nanoscale
interactions important for cell surface attachment to plas-
tic surfaces within biofilms, (ii) the catalytic mechanisms
of the enzymes responsible for oxidation or hydrolysis of
plastic polymers extracellularly, (iii) the metabolic path-
ways that mediate uptake and initial catabolism of plastic
fragments intracellularly, and (iv) development of the
implementation of enhancing factors such as pretreat-
ments, microbial consortia and nutrient availability while
minimizing the effects of constraining factors such as
alternative carbon sources and inhibitory by-products.
Acknowledgements
Graduate student support for R.W. was provided by a
graduate fellowship from the College of Agriculture and
Life Sciences at Cornell University.
Conflict of Interest
No conflict of interest declared.
References
Albertsson, A.C., Sares, C. and Karlsson, S. (1993) Increased
biodegradation of LDPE with nonionic surfactant. Acta
Polym 44, 243–246.
Albertsson, A., Barenstedt, C., Karlsson, S. and Lindberg, T.
(1995) Degradation morphology differentiate degradable
product pattern and changes as means to abiotically and
biotically polyethylene. Polymer 36, 3075–3083.
Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
Plastic degradation by Pseudomonas
Arkatkar, A., Arutchelvi, J., Bhaduri, S., Uppara, P.V. and
Doble, M. (2009) Degradation of unpretreated and
thermally pretreated polypropylene by soil consortia. Int.
Biodeterior. Biodegrad 63, 106–111.
Arkatkar, A., Juwarkar, A.A., Bhaduri, S., Uppara, P.V. and
Doble, M. (2010) Growth of Pseudomonas and Bacillus
biofilms on pretreated polypropylene surface. Int.
Biodeterior Biodegrad 64, 530–536.
Balasubramanian, V., Natarajan, K., Hemambika, B., Ramesh,
N., Sumathi, C.S., Kottaimuthu, R. and Rajesh Kannan, V.
(2010) High-density polyethylene (HDPE)-degrading
potential bacteria from marine ecosystem of Gulf of
Mannar, India. Lett Appl Microbiol 51, 205–211.
Barth, M., Honak, A., Oeser, T., Wei, R., Belisario-Ferrari,
M.R., Then, J. Schmidt, J. et al. (2016) A dual enzyme
system composed of a polyester hydrolase and a
carboxylesterase enhances the biocatalytic degradation of
polyethylene terephthalate films. Biotechnol J 11, 1082–
1087.
Biffinger, J.C., Barlow, D.E., Pirlo, R.K., Babson, D.M.,
Fitzgerald, L.A., Zingarelli, S., Nadeau, L.J., Crookes-
Goodson, W.J. et al. (2014) A direct quantitative agar-
plate based assay for analysis of Pseudomonas protegens Pf-
5 degradation of polyurethane films. Int Biodeterior
Biodegrad 95, 311–319.
Biffinger, J.C., Barlow, D.E., Cockrell, A.L., Cusick, K.D.,
Hervey, W.J., Fitzgerald, L.A., Nadeau, L.J., Hung, C.S.
et al. (2015) The applicability of ImpranilâDLN for
gauging the biodegradation of polyurethanes. Polym
Degrad Stab 120, 178–185.
Chiellini, E., Corti, A., D’Antone, S. and Baciu, R. (2006)
Oxo-biodegradable carbon backbone polymers – oxidative
degradation of polyethylene under accelerated test
conditions. Polym Degrad Stab 91, 2739–2747.
Cregut, M., Bedas, M., Durand, M.J. and Thouand, G. (2013)
New insights into polyurethane biodegradation and
realistic prospects for the development of a sustainable
waste recycling process. Biotechnol Adv 31, 1634–3647.
Danko, A.S., Luo, M., Bagwell, C.E., Brigmon, R.L. and
Freedman, D.L. (2004) Involvement of linear plasmids in
aerobic biodegradation of vinyl chloride. Appl Environ
Microbiol 70, 6092–6097.
Dash, H.R., Mangwani, N., Chakraborty, J., Kumari, S. and
Das, S. (2013) Marine bacteria: potential candidates for
enhanced bioremediation. Appl Microbiol Biotechnol 97,
561–571.
Devi, R.S., Kannan, V.R., Natarajan, K., Nivas, D., Kannan, K.,
Chandru, S. and Antony, A.R. (2016) The role of
microbes in plastic degradation. In Environ Waste Manage
ed. Chandra, R. pp. 341–370 United States: CRC Press.
Donlan, R.M. (2002) Biofilms: microbial life on surfaces.
Emerg Infect Dis 8, 881–890.
Dos Santos, V.A.P., Heim, S., Moore, E.R.B., Str€atz, M. and
Timmis, K.N. (2004) Insights into the genomic basis of
591
Plastic degradation by Pseudomonas
niche specificity of Pseudomonas putida KT2440. Env
Microbiol 6, 1264–1286.
Durmaz, B. and Sanin, F.D. (2001) Effect of carbon to
nitrogen ratio on the composition of extracellular
polymers in activated sludge. Wat Sci Technol 44, 221–229.
Friello, D.A., Mylroie, J.R. and Chakrabarty, A.M. (2001) Use
of genetically engineered multi-plasmid microorganisms
for rapid degradation of fuel hydrocarbons. Int Biodeterior
Biodegrad 48, 233–242.
Galgali, P., Varma, A.J., Puntambekar, U.S., Gokhale, D.V.,
Tokiwa, Y., Fan, H., Hiraguri, Y., Kurane, R. et al. (2002)
Towards biodegradable polyolefins: strategy of anchoring
minute quantities of monosaccharides and disaccharides
onto functionalized polystyrene, and their effect on
facilitating polymer biodegradation. Chem Commun 33,
2884–2885.
Gilan, I., Hadar, Y. and Sivan, A. (2004) Colonization, biofilm
formation and biodegradation of polyethylene by a strain
of Rhodococcus ruber. Appl Microbiol Biotechnol 65, 97–
104.
Gu, J.D. (2003) Microbiological deterioration and degradation
of synthetic polymeric materials: recent research advances.
Int Biodeterior Biodegrad 52, 69–91.
Hosaka, M., Kamimura, N., Toribami, S., Mori, K., Kasai, D.,
Fukuda, M. and Masai, E. (2013) Novel tripartite aromatic
acid transporter essential for terephthalate uptake in
Comamonas sp. sstrain E6. Appl Environ Microbiol 7,
6148–6155.
Howard, G.T. and Blake, R.C. (1998) Growth of Pseudomonas
fluorescens on a polyester-polyurethane and the
purification and characterization of a polyurethane-
protease enzyme. Int. Biodeterior. Biodegrad. 42, 213–220.
Hung, C.-S., Zingarelli, S., Nadeau, L.J., Biffinger, J.C., Drake,
C.A., Crouch, A.L., Barlow, D.E., Russell, J.N. et al. (2016)
Carbon catabolite repression and impranil polyurethane
degradation in Pseudomonas protegens strain Pf-5. Appl
Environ Microbiol 82, 6080–6090.
Jun, H.S., Kim, B.O., Kim, Y.C., Chang, H.N. and Woo, S.I.
(1994) Synthesis of copolyesters containing poly (ethylene
terephthalate) and poly (e-caprolactone) units and their
susceptibility to Pseudomonas sp. lipase. J Env Polym
Degrad 2, 9–18.
Karegoudar, T.B. and Pujar, B.G. (1985) Degradation of
terephthalic acid by a Bacillus species. FEMS Microbiol Lett
30, 217–220.
Kawai, F. and Hu, X. (2009) Biochemistry of microbial
polyvinyl alcohol degradation. Appl Microbiol Biotechnol
84, 227–237.
Kim, M. (2003) Evaluation of degradability of
hydroxypropylated potato starch/polyethylene blend films.
Carbohydr Polym 54, 173–181.
Kolvenbach, B.A., Helbling, D.E., Kohler, H.E. and Corvini,
P.F.-X. (2014) Emerging chemicals and the evolution of
biodegradation capacities and pathways in bacteria. Curr
Opin Biotechnol 27, 8–14.
592 Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
R.A. Wilkes and L. Aristilde
Latha, K. and Lalithakumari, D. (2001) Transfer and
expression of a hydrocarbon-degrading plasmid pHCL
from Pseudomonas putida to marine bacteria. World J
Microbiol Biotechnol 17, 523–528.
Lenz, R.W. (1993) Biodegradable polymers. In Langer, R.S.,
Peppas, N.A. (eds)Biopolymers I. vol 107 pp. 1–40. Berlin
Heidelberg: Springer.
Lucas, N., Bienaime, C., Belloy, C., Queneudec, M., Silvestre,
F. and Nava-Saucedo, J.E. (2008) Polymer biodegradation:
mechanisms and estimation techniques – a review.
Chemosphere 73, 429–442.
McEldowney, S. and Fletcher, M. (1986) Effect of growth
conditions and surface characteristics of aquatic bacteria
on their attachment to solid surface. J Gen Microbiol 132,
513–523.
Mohan, A.J., Sekhar, V.C., Bhaskar, T. and Nampoothiri, K.M.
(2016) Microbial assisted shigh impact polystyrene (HIPS)
degradation. Bioresour Technol 213, 204–207.
Mukherjee, K., Tribedi, P., Chowdhury, A., Ray, T., Joardar,
A., Giri, S. and Sil, A.K. (2011) Isolation of a
Pseudomonas aeruginosa strain from soil that can degrade
polyurethane diol. Biodegradation 22, 377–388.
M€uller, R.J., Schrader, H., Profe, J., Dresler, K. and
Deckwer, W.D. (2005) Enzymatic degradation of poly
(ethylene terephthalate): rapid hydrolyse using a
hydrolase from T. fusca. Macromol Rapid Commun 26,
1400–1405.
Nauendorf, A., Krause, S., Bigalke, N.K., Gorb, E.V., Gorb,
S.N., Haeckel, M., Wahl, M. and Treude, T. (2016)
Microbial colonization and degradation of polyethylene
and biodegradable plastic bags in temperate fine-grained
organic-rich marine sediments. Mar Pollut Bull 103, 168–
178.
Neufeld, L., Stassen, F., Sheppard, R. and Gilman, T. (2016)
The new plastics economy: rethinking the future of
plastics. In World Economic Forum. http://www3.weforum.
org/docs/WEF_The_New_Plastics_Economy.pdf
Nikel, P.I., Chavarrıa, M., Danchin, A. and de Lorenzo, V.
(2016) From dirt to industrial applications: Pseudomonas
putida as a synthetic biology chassis for hosting
harsh biochemical reactions. Curr Opin Chem Biol 34,
20–29.
Novotny, E., Erbanov, P., Sezimov, H., Malachov, K.R.,
Rybkov, Z., Malinov, L., Prokopov, I. and Brozek, J.
(2015) Biodegradation of aromatic-aliphatic copolyesters
and polyesteramides by esterase activity-producing
microorganisms. Int Biodeterior Biodegrad 97, 25–30.
Obradors, N. and Aguilar, J. (1991) Efficient biodegradation of
high-molecular- weight polyethylene glycols by pure
cultures of Pseudomonas stutzeri. Appl Environ Microbiol
57, 2383–2388.
PlasticsEurope (2015) Plastics—The Facts 2015. An analysis of
European plastics production, demand and waste data.
Available at: http://www.plasticseurope.org/Document/pla
stics—the-facts-2015.aspx.
R.A. Wilkes and L. Aristilde
Poblete-Castro, I., Becker, J., Dohnt, K., dos Santos, V.M. and
Wittmann, C. (2012) Industrial biotechnology of
Pseudomonas putida and related species. Appl Microbiol
Biotechnol 93, 2279–2290.
Rajandas, H., Parimannan, S., Sathasivam, K., Ravichandran,
M. and Su Yin, L. (2012) A novel FTIR-ATR
spectroscopy based technique for the estimation of low-
density polyethylene biodegradation. Polym Test 31,
1094–1099.
Restrepo-Florez, J.M., Bassi, A. and Thompson, M.R. (2014)
Microbial degradation and deterioration of polyethylene –
a review. Int Biodeterior Biodegrad 88, 83–90.
Ronkvist, A.M., Xie, W., Lu, W. and Gross, R.A. (2009)
Cutinase-catalyzed hydrolysis of poly(ethylene
terephthalate). Macromolecules 42, 5128–5138.
Ruiz, C., Main, T., Hilliard, N.P. and Howard, G.T. (1999)
Purification and characterization of two polyurethanase
enzymes from Pseudomonas chlororaphis. Int Biodeterior
Biodegrad 43, 43–47.
Sangale, M.K., Shahnawaz, M. and Ade, A.B. (2012) A review
on biodegradation of polythene: the microbial approach. J
Bioremediation Biodegrad 3, 1–9. doi:10.4172/2155-6199.
1000164
Sanin, S.L., Sanin, F.D. and Bryers, J.D. (2003) Effect of
starvation on the adhesive properties of xenobiotic
degrading bacteria. Process Biochem 38, 909–914.
Sen, S.K. and Raut, S. (2015) Microbial degradation of low
density polyethylene (LDPE): a review. J Environ Chem
Eng 3, 462–473.
Shah, A.A., Hasan, F., Hameed, A. and Ahmed, S. (2008)
Biological degradation of plastics: a comprehensive review.
Biotechnol Adv 26, 246–265.
Shah, Z., Hasan, F., Krumholz, L., Atkas, D. and Shah, A.A.
(2013) Degradation of polyester polyurethane by newly
isolated Pseudomonas aeruginosa strain MZA-85 and
analysis of degradation products by GC-MS. Int
Biodeterior Biodegrad 77, 114–122.
Shah, Z., Gulzar, M., Hasan, F. and Shah, A.A. (2016)
Degradation of polyester polyurethane by an indigenously
developed consortium of Pseudomonas and Bacillus species
isolated from soil. Polym Degrad Stab 134, 349–356.
Shimao, M. (2001) Biodegradation of plastics. Curr Opin
Biotechnol 12, 242–247.
Singh, B. and Sharma, N. (2008) Mechanistic implications of
plastic degradation. Polym Degrad Stab 93, 561–584.
Sivan, A. (2011) New perspectives in plastic biodegradation.
Curr Opin Biotechnol 22, 422–426.
Stern, R.V. and Howard, G.T. (2000) The polyester
polyurethanase gene (pueA) from Pseudomonas
chlororaphis encodes a lipase. FEMS Microbiol Lett 185,
163–168.
Suzuki, T. (1976) Purification and some properties of
polyvinyl alcohol-degrading enzyme produced by
Pseudomonas O-3. Agric Biol Chem 40, 497–504.
Journal of Applied Microbiology 123, 582--593 © 2017 The Society for Applied Microbiology
Plastic degradation by Pseudomonas
Suzuki, T., Ichihara, Y., Yamada, M. and Tonomura, K. (1973)
Some characteristics of Pseudomonas O-3 which utilizes
polyvinyl alcohol. Agric Biol Chem 37, 747–756.
Timmis, K.N. (2002) Pseudomonas putida: a cosmopolitan
opportunist par excellence. Env Microbiol 4, 779–781.
Tiso, T., Wierckx, N. and Blank, L.M. (2015) Non-pathogenic
Pseudomonas as platform for industrial biocatalysis. In
Industrial Biocatalysis ed. Grunwald, P. pp. 323–372.
Singapore: Pan Stanford Publishing.
Tribedi, P. and Sil, A.K. (2013a) Bioaugmentation of
polyethylene succinate-contaminated soil with
Pseudomonas sp. AKS2 results in increased microbial
activity and better polymer degradation. Env Sci Pollut Res
20, 1318–1326.
Tribedi, P. and Sil, A.K. (2013b) Cell surface hydrophobicity: a
key component in the degradation of polyethylene succinate
by Pseudomonas sp. AKS2. J Appl Microbiol 116, 295–303.
Tribedi, P. and Sil, A.K. (2013c) Low-density polyethylene
degradation by Pseudomonas sp. AKS2 biofilm. Env Sci
Pollut Res 20, 4146–4153.
Tribedi, P., Sarkar, S., Mukherjee, K. and Sil, A.K. (2012)
Isolation of a novel Pseudomonas sp. from soil that can
efficiently degrade polyethylene succinate. Env Sci Pollut
Res 19, 2115–2124.
Tribedi, P., Das Gupta, A. and Sil, A.K. (2015) Adaptation of
Pseudomonas sp. AKS2 in biofilm on low-density polyethylene
surface: an effective strategy for efficient survival and polymer
degradation. Bioresour Bioprocess 2, 1–10.
Vega, R.E., Main, T. and Howard, G. (1999) Cloning and
expression in Escherichia coli of a polyurethane-degrading
enzyme from Pseudomonas fluorescens. Int Biodeterior
Biodegrad 43, 49–55.
Wang, Y.Z., Zhou, Y. and Zylstra, G.J. (1995) Molecular
analysis of isophthalate and terephthalate degradation by
Comamonas testosteroni YZW-D. Env Heal Perspect 103, 9–
12.
Webb, H.K., Arnott, J., Crawford, R.J. and Ivanova, E.P.
(2013) Plastic degradations and its environmental
implications with special references to poly(ethylene
terepththalate). Polymers 5, 1–18.
Wierckx, N., Prieto, M.A., Pomposiello, P., de Lorenzo, V.,
O’Connor, K. and Blank, L.M. (2015) Plastic waste as a
novel substrate for industrial biotechnology. Microb
Biotechnol 8, 900–903.
Yoon, M., Jeong Jeon, H. and Nam Kim, M. (2012)
Biodegradation of polyethylene by a soil bacterium and
AlkB cloned recombinant cell. J Bioremediat Biodegrad 3,
1–8.
Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji,
H., Maeda, Y., Toyohara, K., Miyamoto, K. et al. (2016) A
bacterium that degrades and assimilates poly(ethylene
terephthalate). Science 351, 1196–1199.
593