Biodegradacion

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JECE 539 113
Journal of Environmental Chemical Engineering xxx (2015) xxxxxx
Contents lists available at ScienceDirect
Journal of Environmental Chemical Engineering

journal homepage: www.elsevier.com/locate/jece
1 Review
2 Microbial degradation of low density polyethylene (LDPE): A review

3 Q1 Sudip Kumar Sen, Sangeeta Raut *
4 Q2 Department of Biotechnology, Gandhi Institute of Engineering and Technology, Gunupur, Rayagada, Odisha 765 022, India
A R T I C L E I N F O A B S T R A C T
Article history: Biodegradation is considered to take place throughout three stages: biodeterioration, biofragmentation
Received 22 July 2014 and assimilation, without neglect the participation of abiotic factors. However, most of the techniques
Accepted 8 January 2015 used by researchers in this area are inadequate to provide evidence of the nal stage: assimilation. In this
review, we describe the different stages of biodegradation and we state several techniques used by some
Keywords: authors working in this domain. Validate assimilation (including mineralization) is an important aspect
Microorganisms to guarantee the real biodegradability of items of consumption (in particular friendly environmental new
Plastics
materials). Since LDPE is considered to be practically inert, efforts were made to isolate unique
Waste
Oxodegradable
microorganisms capable of utilizing LDPEs. Recent data showed that biodegradation of LDPE waste with
Biodegradable selected microbial strains became a viable solution. Among biological agents, microbial enzymes are one
of the most powerful tools for the biodegradation of LDPEs. Activity of biodegradation of most enzymes is
higher in fungi than in bacteria. It is important to consider fungal degradation of LDPE in order to
understand what is necessary for biodegradation and the mechanisms involved. This requires
understanding of the interactions between materials and microorganisms and the biochemical changes
involved. Widespread studies on the biodegradation of LDPEs have been carried out in order to overcome
the environmental problems associated with LDPE waste. This paper reviews the current research on the
biodegradation of LDPEs and also use of various techniques for the analysis of degradation in vitro.
2015 Published by Elsevier Ltd.
Contents
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Benets and challenges of bio- and oxo-degradable LDPEs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Biodegradable LDPEs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Oxo-degradable LDPEs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Microorganisms involved in LDPE degradation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Isolation and screening of LDPE degrading fungal strains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Effect of fungal activity on LDPE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Functional groups on the surface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Hydrophobicity/hydrophilicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Crystallinity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Molecular weight distribution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Surface topography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Mechanical properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Consumption of the polymer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Mechanisms of LDPE biodegradation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Conclusion and perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
Uncited references . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
* Corresponding author. Tel.: +91 9438450789.

E-mail address: research.sangeeta@gmail.com (S. Raut).
http://dx.doi.org/10.1016/j.jece.2015.01.003
2213-3437/ 2015 Published by Elsevier Ltd.
Please cite this article in press as: S. Kumar Sen, S. Raut, Microbial degradation of low density polyethylene (LDPE): A review, J. Environ. Chem.
Eng. (2015), http://dx.doi.org/10.1016/j.jece.2015.01.003
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2 S. Kumar Sen, S. Raut / Journal of Environmental Chemical Engineering xxx (2015) xxxxxx
5 Introduction Table 1
Properties of low density polyethylene.
6 The discovery of the chemical process for developed synthetic Property Value Range/comments
7 polymers (plastics) from crude oil was a breakthrough, in Density, g/cc 0.91 0.9100.925 glee
8 chemistry and in material sciences, and paved the way to the Hardness, shore D 44 4146 Shore D
9 production of one of the most resourceful group of materials ever Tensile strength, yield MPa 10 416 MPa: ASTM D638
10 Tensile strength, ultiMate MPa 25 740 MPa
produced. Low density polyethylene (LDPE) was originally
11 Modulus of elasticity, GPa 0.2 0.070.3 GPa; in tension;
prepared by the high pressure polymerization of ethylene. Its ASTM D638
12 comparatively low density arises from the presence of a small Flexural modulus, GPa 0.4 00.7 GPa: ASTM D790
13 amount of branching in the chain (on about 2% of the carbon Coefcient of thermal expansion, linear 30 2040 mm/m 1C; ASTM
14 atoms) (Fig. 1). Chemically LDPE is un-reactive at room tempera- 20 C, pm/m- C D696
15 Melting point, C 115
ture although it is slowly attacked by strong oxidizing agents and
16 some solvents will cause softening or swelling. It may be used at
17 temperatures up to 95 C for short periods and at 80 C,
18 paper, textiles and other plastics. However, LDPE is hardly 54
continuously. Low-density polyethylene is an incompletely crys-
19 degraded after disposal, which pollutes the environment and 55
talline solid with a degree of crystallinity in the 5060% range that
20 disturbs the ecosystem [12]. Steady with its inert nature, a 56
leads to several properties such as opacity, tensile strength, tear
21 57
strength, rigidity and chemical resistance, exibility even at a low polyethylene sheet that was kept back in moist soil for a time
22 period of 12 years showed no conrmation of weight loss [13]. In 58
temperature [1,2]. Typical properties of LDPE are given in Table 1.
23 other study, only partial degradation of polyethylene lm has been 59
The most common LDPE types are: linear low density polyethylene
24 observed after a long incubation of 32 years inside the soil [14]. It 60
(LLDPE) and branched low density polyethylene (BLDPE). They
25 also poses an ever increasing ecological threat to terrestrial and 61
differ in their density, degree of branching and availability of
26 marine wild life [15,16]. There are reports that suggest that 62
functional groups on the surface. Low density polyethylene (LDPE)
27 polyethylene causes blockages in the intestines of sh, birds and 63
has been commonly used due to its versatile characteristics and
28 marine mammals. In addition, entanglement in or ingestion of this 64
usefulness. These traits facilitated the application of plastics to
29 waste has endangered hundreds of different species [17,18]. The 65
almost all industrial, agricultural or domestic market. Each year, an
30 increasing levels of LDPE waste, decreasing landll capacity and 66
estimated 500 billion to 1 trillion plastic bags are consumed
31 very slow rate of LDPE degradation in the environment have 67
worldwide [3]. In Japan, the percentage of municipal plastic
32 caused the research tendency to decrease the amount of waste. 68
wastes, as a fraction of municipal solid waste (MSW), that was
33 Degradation of polyethylene can be classied as abiotic or biotic, 69
landlled in the early 1980s was estimated to be 45%, incineration
34 the previous being dened as deterioration caused by natural 70
was 50%, and the other 5% was subjected to separation and
35 factors such as temperature, UV irradiation, while the latter is 71
recycling (Plastic Waste Management Institute, 1985). In USA, more
36 dened as biodegradation caused by the participation of micro- 72
than 15% of the total MSW was incinerated in 1990; only about 1%
37 organisms that modify and consume the polymer leading to 73
of post-consumer plastics were recycled [46]. Since the use of
38 changes in its properties. It is evident from the recent studies that 74
plastics is gradually increasing, the problem of post-consumer
39 the speed of biotic degradation of low-density polyethylene (LDPE) 75
recycling of these materials has become a pivotal issue for
40 can be enhanced by its prior oxidation [19,20,3,21]. It is probable 76
economic and environmental reasons [7]. With the excessive use of
41 that the oxidation of polyethylene generates carbonyl groups that 77
plastics, an increasing pressure is being placed on capacities
42 can be used by microorganisms for its degradation [2224]. All 78
available for plastic waste disposal, the need for biodegradable
43 these reports signify that polyethylene is very much recalcitrant in 79
plastics and biodegradation of plastic wastes has gained consider-
44 comparison to natural degradation. Biodegradation of LDPE/ 80
able importance in the last few years [8]. As per a recent estimate of
45 cellulose blends, starchPE, plasticPE have also been established 81
the Central Pollution Control Board, New Delhi, India, 8 million
46 by several workers [2528]. During degradation process, LDPE 82
tons of plastic products are consumed every year in India alone. A
47 provided carbon which is the sole energy source for micro- 83
study on plastic waste generation in 60 major Indian cities revealed
48 organisms specically, showed that small fragments were 84
that approximately 15,340 tons/day of plastic waste is generated in
49 85
the country [9]. Reasonable transparency of thin lms, free from consumed faster than larger ones [29].
50 Recently, efforts have focused on the biodegradation of LDPE 86
odor and toxicity, better ductility, low water vapor permeability
51 wastes due to the disadvantages of other methods such as cost and 87
and heat seal ability are also the peculiarities of LDPE [10,11]. It is
52 pollution. Biodegradation is the ability of microorganism to 88
used for packaging applications, making trays and plastic bags for
53 inuence abiotic degradation through physical, chemical or 89
food and non-food items. It is also used as a protective coating on
enzymatic action [3032]. This problem can be overcome by 90
proper introduction of natural polymers such as starch, chitosan or 91
cellulose in the matrix of polyethylene [33]. It is important to 92
highlight that although the damage to LDPE is classied by only 93
one of these two damage modes, in nature it is typical that both act 94
cooperatively [34]. In practice, biodegradation is seldom due to a 95
single cause, but a combined effect of heat, UV light, micro- 96
organisms, stress and water [30]. More recently it has been 97
demonstrated in soil burial tests that the use of suitable additives 98
in polyethylene lms induced substantial oxidation with conse- 99
quent fragmentation, drop in molecular weight, increase in wet 100
ability, ultimately followed by high mineralization (6070%) and 101
xation of about 810% of carbon into cell biomass [32,35]. The low 102
rate of biodegradation of plastics is usually due to lack of water 103
solubility and due to the size of the polymer molecules which 104
prevents it to get transported directly into the cells [36,37]. The 105
Fig. 1. (a) Low density polyethylene (b) structure of polyethylene.
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S. Kumar Sen, S. Raut / Journal of Environmental Chemical Engineering xxx (2015) xxxxxx 3
106 two major problems with polyethylene are its high hydrophobicity as initiators of thermal and photo oxidation of polyethylene lms 167
107 (due to the presence of only CH2 groups) and its high molecular promote the fragmentation of the tested samples eventually 168
108 weight (more than 30 kDa). The biotic mechanism reported for the followed by microbial attack. Oxo-degradable LDPEs are claimed to 169
109 degradation of high molecular weight polymers are due to the provide a potential solution to littering issues. The very few 170
110 extra cellular enzymes produced by microorganisms which positive biodegradation results obtained with oxo-degradable 171
111 degrade the main polymeric chain and result in intermediates of plastics were achieved in unrealistic testing environments and 172
112 lower molecular weight with modied mechanical properties, could not be repeated. Oxo-degradable plastics do not meet the 173
113 making it more accessible for the microbial assimilation [38]. requirements of industrial and/or home compostability set out in 174
114 Thermal or radiation treatments on polyethylene reduce the various established standards. Oxo-degradable plastic bags are also 175
115 polymeric chain size and form oxidized groups such as carboxyl, available today to substitute traditional LDPE plastic bags. The oxo- 176
116 carbonyl and hydroxyl. These treatments modify the properties degradable plastic bags are not biodegradable but are designed to 177
117 (crystallinity level, morphological changes) of the original polymer break down into small pieces after exposure to oxygen. The smaller 178
118 and facilitate the polymer biodegradation [25]. pieces may lead to environmental problems if they are consumed 179
119 The biodegradation of polyethylene has been reported in a by animals or if the small pieces are scattered over the ground. The 180
120 number of research studies published over the last 30 years; oxo-biodegradable additives are usually incorporated in conven- 181
121 however, there is general agreement that the process under normal tional plastics such as polyethylene (PE), polypropylene (PP), 182
122 conditions is extremely slow [3943]. But, LDPE biodegradation is polystyrene (PS), polyethylene terephtalate (PET) and sometimes 183
123 less focused in previous reviews. Till date, most of the published also polyvinyl chloride (PVC) at the moment of conversion into 184
124 reviews on LDPE degradation having focuses on abiotic mecha- nal products. Narayan [53] has pointed out that oxo-degradable 185
125 nisms of deterioration of polyethylene have been described fragments may act to concentrate pesticide residues in the soil, as 186
126 extensively [40], thus this review will be focusing on the has been shown for PE and polypropylene (PP) remains found in 187
127 biodegradation of LDPE and mechanisms associated with this the marine environment [54,55]. There are also concern that 188
128 process. Although there is enough evidence that proves biodegra- degraded fragments may become cross-linked and hence persist in 189
129 dation of LDPE, there is still a lack of knowledge on the complete the environment [56]. Research into the toxicological impact of 190
130 metabolic pathways involved in the process and in the structure oxo-degradable additives [57] found no evidence of toxicity to 191
131 and identity of all the enzymes involved. Only some advances have tomato, cucumber or cress seeds. There is evidence that plastic 192
132 been made in this regard and even then the conclusions outlined debris in the marine environment can degrade to give ne particles 193
133 require verication [4447]. Taking into account the above that then become ingested and accumulate in marine organisms 194
134 mentioned facts, present communication is an attempt to focus [58,59]. No evidence was found that oxo-degradable fragments 195
135 current and last 30 years of achievement in the eld of LDPE have a harmful bio-accumulative effect but neither was there 196
136 biodegradation. The present review describes three different evidence that they do not. There is very little evidence for the fate 197
137 topics, the rst being a comprehensive summary of the micro- of oxo-degradable fragments and this is an area identied as 198
138 organisms reportedly involved with LDPE biodegradation; second, requiring further research. 199
139 Q3 the effects of these microorganisms on LDPE properties will be
140 presented; and third, an outline of the degradation process of LDPE Microorganisms involved in LDPE degradation 200
141 based on published literature will be discussed.
Biodegradation of LDPE is complex and not fully understood. In 201
142 Benets and challenges of bio- and oxo-degradable LDPEs order to elucidate the potential mechanisms, two different 202
strategies have been followed in the literature. In the rst 203

143 Biodegradable LDPEs approach, degradation studies have been performed using pure 204
strains able to degrade LDPE [20,36,37,4547,6067]. This 205

144 Q4 Bio-degradable plastics were originally developed in order to approach has the advantage of using pure strains, which is a 206
145 solve specic waste issues related either to agricultural lms or convenient way to investigate metabolic pathways or to evaluate 207
146 collection and separation of food waste. The majority of the effect of different environmental conditions on LDPE degrada- 208
147 biodegradable LDPEs meet the requirements of well-recognized tion. A disadvantage of this approach is that it ignores the 209
148 standards of industrial composting. Solid proof of biodegradation possibility that LDPE biodegradation can be the result of a 210
149 is available through certication by accredited laboratories and cooperative process between different species. These limitations 211
150 institutes for biodegradable plastics. One of the major strategy to are avoided by the second approach, in which the use of complex 212
151 facilitate dissolution and subsequent degradation is by direct environments and microbial communities are applied 213
152 degradation of LDPE by microorganisms using only the polymer as [6870,23,71,32,7275]. Table 2 summarizes some of the different 214
153 sole carbon source [3]. Previous studies have reported on microenvironments that have been employed to study LDPE 215
154 biodegradation of polyethylene by bacterial [48,37] and fungal biodegradation using mixed and complex microbial communities. 216
155 species [4850]. There are few reports that deal with microbial Marine water, soil or compost are examples of the environments 217
156 degradation of polyethylene materials [31,51,52,8]. Fungi survive whereby LDPE has been investigated under the second approach. 218
157 environments with low nutrient availability, low pH and low The structure of a microbial community isolated on an LDPE 219
158 moisture as well. The use of biodegradable polymers is increasing surface during biodegradation experiments can also be inuenced 220
159 at a rate of 30% per year in some markets worldwide. by the type of polymer used as the substrate. In several studies it 221
has been proven that the physicochemical nature of a surface 222

160 Oxo-degradable LDPEs
Table 2
161 Oxo-degradable plastics are made of petroleum-based poly- Different experimental environments used in the study of LDPE biodegradation.
162 mers (usually polyethylene (PE)) and contain special additives that
163 Experimental environment References
cause them to degrade. The very low propensity to oxidation and
164 further degradation followed by biodegradation of conventional Marine exposure conditions [23,30,6971,73,74]
165 Composting conditions [32]
polyethylene is widely accepted. UV, heat exposure, oxidation with Soil burial conditions [68,72,75]
166 nitric acid as well as the addition of pro degrading systems acting
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223 determines the ability of microorganisms to form biolm Table 3

224 Bacterial strains associated with polyethylene biodegradation.
structures [7679]. LDPE has more branching (on about 2% of
225 the carbon atoms) than HDPE, so its intermolecular forces Genus Species References
226 (instantaneous-dipole induced-dipole attraction) are weaker, its Pseudomonas sp. [67,47,65]
227 tensile strength is lower, and its resilience is higher. Also, since its aeruginosa [66,143]
228 molecules are less tightly packed and less crystalline (because of uorescens [74]
229 Paenibacillus macerans [74]
the side branches), its density is lower. LDPE contains the chemical
230 Rahnella aquatilis [74]
elements carbon and hydrogen. Ralstonia sp. [143]
231 It is important to highlight that LDPE can be also found mixed Rhodococcus erythropolis [143]
232 with additives such as pro-oxidants or starch [80,41], both of these rhodochrous [64,63,116]
233 being applied to improve the biodegradability of the polymer. The ruber [46,37,36]
234 Staphylococcus cohnii [74]
presence of these additives can affect the types of microorganisms
epidermidis [21]
235 colonizing the surfaces of these polymers. xylosus [74]
236 Over the past 50 years, a number of strains have been identied Stenotrophomonas sp. [143]
237 for their ability to interact with LDPE causing some kind of Streptomyces badius [45]
238 setonii [45]
deterioration; this has been done based on the two approaches
239 viridosporus [45]
mentioned before, and using different kinds of LDPE. The richness Bacillus amyloliquefaciens [74]
240 of microorganisms able to degrade LDPE is so far limited to brevies [122]
241 19 genera of bacteria and 12 genera of fungi; however, these cereus [74,3,150,118]
242 numbers are likely to increase based on the more sensitive circulans [122]
243 halodenitricans [3]
isolation and characterization techniques based on sequencing of
mycoides [74,100]
244 rDNA. This technology allows a broader approach to assessing the pumilus [74,3,150]
245 composition of a community, including the non-culturable fraction sphericus [118,29]
246 of microorganisms that is invisible by traditional microbiology thuringiensis [74]
247 methods yet that constitutes up to the 90% of the real biodiversity Brevibacillus borstelensis [62]
248 Delftia acidovorans [143]
in an ecosystem [81]. Several studies have suggested partial Flavobacterium sp. [143]
249 biodegradation of polyethylene after UV irradiation [24], thermal Micrococcus luteous [74]
250 treatment [82,83] or oxidation with nitric acid [84]. It is also lylae [74]
251 reported that there is a synergistic effect between photo-oxidation Microbacterium paraoxydans [66]
252 Nocardia asteroides [63,116]
and biodegradation of polyethylene [30]. An increase in the
253 Acinetobacter baumannii [74]
biodegradation of polyethylene was observed with increase in the Arthrobacter sp. [65,150]
254 time of exposure to UV [62]. Biodegradation in the un-inoculated parafneus [19,60]
255 treatments were slow and were about 7.6% and 8.6% of minerali- viscosus [74]
256 zation for the non-UV-irradiated and UV-irradiated LDPE respec-
257 tively after 126 days. In contrast, in the presence of the selected
258 microorganisms, the biodegradation was much more efcient and
photodegraded LDPE-Mn with B. borstelensis, 15.7%, and similar 289
259 the percentages of biodegradation were 29.5% and 15.8% for the
to that obtained by Fontanella et al. [64], 1624% using Rhodococcus 290
260 UV-irradiated and non-UV-irradiated lms, respectively.
261 rhodochrous at 58 C on photo- and thermo-degraded LDPE 291
The percentage decrease in the carbonyl index was higher for 292
262 containing a mixture of Mn/Fe pro-oxidants [87]. The ability of
the UV-irradiated LDPE when the biodegradation was performed in 293
263 Bacillus species to utilize PE, with and without pro-oxidant
soil inoculated with the selected fungal and bacterial isolates [85]. 294
264 additives, was also evaluated [87]. Biodegradation resulting from
Tables 3 and 4 present an extensive list of the microorganisms that 295
265 the utilization of polyethylene as a nutrient (carbon source) may be
in some way have been related with LDPE colonization, biodegra- 296
266 more efcient if the degrading microorganism forms a biolm on
dation or both. This list has to be approached carefully because in 297
267 the polyethylene surface. However, the hydrophobicity of the
some studies not all the tests required to prove LDPE biodegrada- 298
268 polyethylene interferes with the formation of a microbial biolm.
tion were performed. As the microorganisms possess different 299
269 Attempts to facilitate colonization of polyethylene by adding non-
characteristics, so the degradation varies from one microorganism 300
270 ionic surfactants to the culture medium promoted the biodegra-
to another [86]. Recently, several microorganisms have been 301
271 dation of polyethylene [88,89]. Gilan et al. [36] isolated a strain
reported for degradation of plastics. The bacterial species
272 Q5 identied from the polyethylene bags tested were Bacillus sp.,
273 Staphylococcus sp., Streptococcus sp., Diplococcus sp., Micrococcus
274 sp., Pseudomonas sp. and Moraxella sp. Among the fungal species Table 4
275 identied, Aspergillus niger,Amauroclopius ornatus, Aspergillus Fungal strains associated with polyethylene biodegradation.
276 nidulans, Janibacter cremeus, Aspergillus avus, Aspergillus candidus Genus Species References
277 Q6 and Aspergillus glaucus were the predominant species [48].
278 Aspergillus niger [91,20,119]
Brevibacillus borstelensis strain isolated from soil, a thermophilic versicolor [149,69]
279 bacterium, recovered for the degradation of branched avus [121,63]
280 low-density polyethylene by utilizing it as the sole carbon source Cladosporium cladosporioides [63,116]
281 and energy source. The incubation of polyethylene lm with B. Chaetomium sp. [152]
282 Fusarium redolens [70,69,23]
borstelensis revealed the reduction in molecular weight of
283 Glioclodium virens [91]
polyethylene by 30% [62]. Mucor circinelloides [121]
284 A maximum biodegradation of 1724% reduction in gravimetric Penicillum simplicissimum [61]
285 weight after irradiation of polyethylene containing pro-oxidants pinophilum [91,20]
286 followed by 90 days of incubation with the bacterium frequentans [100]
Phanerochaete chrysosporium [91,71,144]
287 B. borstelensis at 50 C was observed. The lower value of Verticillium lecanii [69]
288 mineralization found in this study was in the case of
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302 Rhodococcus ruber that was found to colonize and degrade a shorter period of time since all the previous reports showed 368
303 polyethylene. The ability of this bacterium to form biolm on activity only after a minimum period of 34 months. 369
304 polyethylene was attributed to the hydrophobicity of its cell
305 surface. Addition of a small amount of mineral oil to the culture Isolation and screening of LDPE degrading fungal strains 370
306 medium increased biolm formation and the subsequent biodeg-
307 radation of the polyethylene, presumably by increasing the Although several microorganisms are involved in degradation 371
308 hydrophobic interactions between the bacterial biolm and the of LDPE, it remains a challenging task to obtain a strain for 372
309 polymer. Gilan et al. (2004) and Sivan et al. (2006) [36,37] isolated commercial and eco-friendly degradation of LDPE. Over the years, 373
310 a biolm producing strain of R. ruber (C208) that degraded PE at a culturable, LDPE degrading microbes have been isolated from a 374
311 rate of 0.86% per week. Kathiresan and Bingham [90], reported that wide variety of sources such as soil, waste water, compost, garbage, 375
312 bacteria caused the biodegradation ranging from 2.19 to 20.54% for etc. Moreover, efcient screening techniques are a prerequisite for 376
313 polyethylene and 0.568.16% for plastics. A. glaucus was more isolation of novel strains. Kumar et al. (2010) [102] reported the 377
314 active in degrading 28.8% of polyethylene and 7.26% of plastics techniques for isolation and screening of potential fungal strains to 378
315 within a month. This may be documented to the thickness of the degrade LDPE in-vitro. Identication of microorganisms is based 379
316 polyethylene that is 5-times thinner than the plastics. Some on their cellular fatty acid methyl ester (FAME) proles. 380
317 studies have investigated the PE biodegradation process using A very simple semi-quantitative method is the so-called clear- 381
318 fungal isolates, such as Phanerochaete chrysosporium [71], A. niger zone test. This is an agar plate test in which the polymer is 382
319 [20,91], and other strains of the Aspergillus genus including dispersed as very ne particles within the synthetic medium agar; 383
320 A. terreus, A. fumigatus [92] and A. avus [27]. The most convenient this results in the agar having an opaque appearance. After 384
321 method to determine the degradation is to measure the weight inoculation with microorganisms, the formation of a clear halo 385
322 loss. The microbial enzymes catalyzed the depolymerization and around the colony indicates that the organisms are at least able to 386
323 thus there was weight reduction of polyethylene. Among the fungi depolymerize the polymer, which is the rst step of biodegrada- 387
324 strains, identied as Aspergillus sp. (FSM-3, 5, 6, 8) and the rest one tion. This method is usually applied to screen organisms that can 388
325 was Fusarium sp. (FSM-10), FSM-10 (9%) and FSM-3 showed degrade a certain polymer [103,104] but it can also be used to 389
326 maximum (8%), both FSM-6, 8 exhibited moderate (7%) and FSM- obtain semi-quantitative results by analyzing the growth of clear 390
327 5 showed less (5%) LDPE weight reduction and thus degradation zones [105]. Usha et al. (2011) [106] isolated Aspergillus nidulans 391
328 after 60 days of incubation [93]. A. niger showed degradation of and A. avus by enrichment culture using sabouraud dextrose agar 392
329 LDPE up to 5.8% in 1 month while A. japonicas showed more medium having polyethylene powder at a nal concentration of 393
330 capability to degrade LDPE up to 11.11% in 1 month under 0.1% (w/v) and the mixture was sonicated for 1 h at 120 rpm in 394
331 laboratory conditions [94]. These ndings corroborate the fact that shaker. After sonication the medium was sterilized at 121 C and 395
332 fungi have better biodegradation efciency than bacteria. Research pressure for 15 lbs/inch2 for 20 min. The organisms, producing 396
333 works have shown that most of the constituents of plastics can be zone of clearance around their colonies were selected for further 397
334 degraded by microbes and the plastic lms can be treated by analysis. The fungus was identied after staining them with cotton 398
335 microbial systems. Acrylonitrile bers are attacked by species of blue by following the keys Raper and Fennell (1987) [107]. Webb 399
336 Aspergillus, Penicillium, Stachybotrys, and Nigrespora. Pullularia et al. (2000) [108] studied the fungal colonization and biodeterio- 400
337 pullulans can degrade polycaprolactone and other aliphatic ration of plasticized polyvinyl chloride in in situ and ex situ 401
338 polyesters. N-alkenes, alkenes and other aliphatic hydrocarbons conditions. These results suggested that microbial succession may 402
339 are readily utilized by yeasts and fungi. Since a wide variety of fungi occur during the long periods of exposure in in situ. They have 403
340 grow and degrade plastics and their polymers, only they have to be identied Aureobasidium pullulans as the principal colonizing 404
341 upgraded [95]. It has been shown that the members of order fungus and a group of yeasts and yeast-like fungi, including 405
342 Xylariales belonging to class Ascomycetae such as Xylaria also grow Rhodotorula aurantiaca and Kluyveromyces spp. To test the ability of 406
343 on the plastic strips (as a source of carbon) [96]. Microorganism for fungal strains to attack polyethylene lms, strains were cultured on 407
344 biological decomposition of polyethylene and plastics are isolated mineral base agar (MBA) medium described by ASTM G21-90 408
345 and tested for their ability in in-vivo and in-vitro condition by (1996). This carbon free synthetic medium was supplemented with 409
346 Nayak et al. [97]. 0.02% yeast extract. pH was adjusted to 6.5. Degradative ability of 410
347 The faster growth of fungal biomass in soil compared to bacteria the fungal isolates was examined by the quantitative assessment of 411
348 [98], and the growth extension and penetration into other growth on polyethylene sample as per ASTM G21-90 (1996) 412
349 locations through the distribution of hyphae makes them more specications. A standard rating system by ASTM for the evaluation 413
350 preferable than bacteria for degradation of LDPE. In most studies, of fungal growth on polymeric materials, for biodegradation is 414
351 fungi were considered for the degradation of LDPE due to their 0 = no visible growth, l = less than 10% growth, 2 = 1030% growth, 415
352 ability to form hydrophobic proteins that can attach to the polymer 3 = 3060% growth, 4 = 60100% growth covering surface of 416
353 surface [99,100], their generation of degrading enzymes that are polymer lm. However, these methods are mainly limited to 417
354 well-matched to the insoluble LDPE [8] and survive in environ- culturable microbes and the full LDPE degrading potential of the 418
355 ments with low nutrient, pH and moisture availability. Since a wide microbes (culturable and nonculturable microorganisms) is not 419
356 variety of fungi grow and degrade plastics and their polymers, only being fully examined. Researchers are now taking the efforts on 420
357 they have to be upgraded [101]. It has been recently shown that the identication and exploitation of LDPE degrading genes from 421
358 members of order Xylariales belonging to class Ascomycetae such unculturable microbes using metagenomic approach. Kathiresan 422
359 as Xylaria also grow on the plastic strips (as a source of carbon) (2003) [48] has reported isolating fungi from the mangrove soil 423
360 [96]. Microorganisms for biological decomposition of polyethylene which has the potential to degrade polyethylene materials. 424
361 and plastics are isolated and tested for their ability in in-vivo and Yamada-Onodera et al. (2001) [61] isolated a strain of fungus 425
362 in-vitro condition by Nayak et al. [97]. Thus, characterization of Penicillium simplicissimum YK to bio-degrade polyethylene without 426
363 LDPE degradation by waste-source fungi is major objective of this additives. El-Shafei et al. (1998) [27] investigated the ability of 427
364 review because of their compatibility with a waste-rich environ- fungi to attack degradable polyethylene consisting of disposed 428
365 ment (such as landll and composting) that contains a variety of polyethylene bags containing 6% starch. 429
366 discarded polymers. Still a lot has to be done to isolate the right Paul Das and Kumar (2014) [109] isolated four Aspergillus sp. 430
367 kind of microbial strain that could promote degradation of LDPE in (FSM-3, 5, 6, 8) and one Fusarium sp. (FSM-10) from soil samples by 431
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432 spread plate technique using mineral salt medium (g/l: K2HPO4 1.0, potential for environmental and health consequences [113]. In 478
433 KH2PO4 0.2, NaCl 1.0, CaCl22H2O 0.002, H3BO3 0.005, NH4 (SO4)2 most studies, fungi have been investigated for the biodegradation 479
434 1.0, MgSO47H2O 0.5, CuSO45H2O 0.001, ZnSO4H2O 0.001, of LDPE because these organisms produce degrading enzymes [8] 480
435 MnSO4H2O 0.001, Fe2(SO4)3.6H2O 0.01, Agar 15) supplemented and, extracellular polymers, such as polysaccharides, which can 481
436 with 3% LDPE powder as carbon source. The developed fungal mats help to colonize the polymer surface [20], and the distribution and 482
437 were subcultured on Saborauds dextrose agar to get pure culture penetration ability of the fungal hyphae is an advantage. Some 483
438 and preserved in slant at 4 C. The identication of the fungal studies have investigated the polyethylene biodegradation process 484
439 isolate was performed by recognizing the diagnostic morphologi- using fungal isolates, such as Phanerochaete chrysosporium [71], 485
440 cal features of genera using macroscopic and microscopic A. niger [20,91], and other strains of the Aspergillus genus including 486
441 examinations [85]. In addition, the molecular identication A. terreus, A. fumigatus [92] and A. avus [27]. 487
442 methods using PCR to amplify a segment of the rRNA operon In order to establish the extent of biodegradation of the 488
443 encompassing the 5.8S rRNA gene and anking internal tran- polymer, seven different characteristics are usually monitored: 489
444 scribed spacers (ITS) is now in progress at the Iranian Biological functional groups on the surface, hydrophobicity/hydrophilicity, 490
445 Resource Center (IBRC). Esmaeili et al. (2013) [85] isolated A. niger crystallinity, surface topography, mechanical properties, molecular 491
446 from soils of landlls (in which PE wastes had been buried for weight distribution and consumption of the polymer. So far there 492
447 different periods) on mineral medium containing PE powder have been no studies in the literature that prove incorporation of 493
448 (5% ethylene oligomer) as the sole source of carbon. Mishra et al. LDPEs carbon into a microorganisms macromolecular structure 494
449 (2013) [110] isolated, Chrysonilia, Aspergillus and Penicillium using such as its DNA or polysaccharides. 495
450 synthetic medium on the basis of microscopic examination and
451 morphological characteristics. The fungal strain was identied Functional groups on the surface 496
452 with the help of Manual of soil Fungi [111]. Further this
453 taxonomic identication was conrmed to Agharkar Research An increase in the growth of fungus and some structural 497
454 Institute, Pune. During second set of experiment more fungal forms changes as observed by FTIR, were observed in case of treated PE 498
455 were isolated by using same synthetic medium where plastic was which according to Jacinto indicated the breakdown of polymer 499
456 sole carbon source instead of glucose. About 4 different forms were chain and presence of oxidation products of PE. Non-degraded 500
457 found growing on powder of PVC and granules of LDPE and HDPE. polyethylene exhibits almost zero absorbancy at those wave 501
458 These forms were species of genus, Aspergillus, Penicillium. numbers (http://www.dasma.dlsu.edu.ph/ofces/ufro/sinag/ 502
459 Fusarium and Chaetomium. Jacinto.htm). Absorbance at 17101715 cm1 (corresponding to 503
460 LDPE pieces buried in soil mixed with sewage sludge were carbonyl compound), 1640 cm1 and 830880 cm1 (correspond- 504
461 examined microscopically after 10 months, fungal attachment was ing to CC), which appeared after UV and nitric acid treatment, 505
462 found on the surface of the plastic, indicating possible utilization of decreased during cultivation with microbial consortia [115] 506
463 plastic as a source of nutrient [112] (Shah, 2007). The isolated (Hasan et al., 2007). Typical degradation of modied LDPE-starch 507
464 fungal strains were identied as Fusarium sp. AF4, Aspergillus mixed and formation of bands at 16201640 cm1 and 508
465 terreus AF5 and Penicillum sp. AF6. The ability of the fungal strains 840880 cm1 was also reported by [61] Yamada-Onodera et al. 509
466 to form a biolm on polyethylene was attributed to the gradual (2001), attributed to oxidation of polyethylene (Fig. 2b). Overall, 510
467 decrease in hydrophobicity of its surface [36]. polyethylene degradation is a combined photo- and bio-degrada- 511
tion process. First, either by abiotic oxidation (UV light exposure) 512
468 Effect of fungal activity on LDPE or heat treatment, essential abiotic precursors are obtained. 513
Secondly, selected thermophilic microorganisms degrade the 514

469 Once the organisms get attached to the surface, it starts low molar mass oxidation products to complete the biodegrada- 515
470 growing by using the polymer as the carbon source. In the primary tion [116,91]. Manzur et al. (2004) [91] reported that the segments Q7 516
471 degradation, the main chain cleaves leading to the formation of formed by the rupturing of the chains because of the biological 517
472 low-molecular weight fragments (oligomers), dimers or mono- treatment could cause the formation of the vinyl group and the 518
473 mers [113]. The degradation is due to the extra cellular enzyme increase in the double bond index (DBI). In addition, the increase in 519
474 secreted by the organism (Table 5). These low molecular weight the DBI can be attributed to biotic dehydrogenation [32]. Kumar 520
475 compounds are further utilized by the microbes as carbon and and Jha (2013) [117] reported that polyethylene biodegradation Q8 521
476 energy sources. The resultant breakdown fragments must be was further conrmed by an increase in the keto carbonyl bond 522
477 completely used by the microorganisms, otherwise there is the index, the ester carbonyl bond index and the vinyl bond index, 523
Table 5
Some of the useful fungal and bacterial enzymes for biodegradation of plastics. Q14
Source Enzyme Microorganisms Plastic act as substrate References

Fungal Glucosidases Aspergillus avus Polycaprolactone (PCL) [158]
Unknown Penicillium funiculosum Polyhydroxybutyrate (PHB) [158]
Amycolaptosis sp. Polylactic acid (PLA) [159]
Streptomyces sp. PHB, PCL [158]
Cutinase Aspergillus oryzae Polybutylene succinate (PBS) [160]
Fusarium sp. PCL [159]
Catalase, Protease Aspergillus niger PCL [158]
Urease Trichoderma sp. Polyurethane [161]
Manganese peroxidase Phanerochaete chrysosporium Polyethylene [59]
Serine hydrolase Pestalotiopsis microspora Polyurethane [162]
Bacterial Lipase Rhizopus delemar PCL [158]
Rhizopus arrizus Polyethylene adipate (PEA), PBS, PCL [158]
Unknown Firmicutes sp. PHB, PCL, and PBS [158]
Protobacteria sp.
Serine hydrolase Pseudomonas stutzeri Polyhydroxyalkanoate (PHA) [159]
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JECE 539 113
Fig. 2. (a) Tensile strength depending on the time of exposure to UV radiation from LDPE-modied starch mixed (b) FTIR spectrum of LDPE-modied starch mixed.
524 which were calculated from FT-IR spectra. Although the FTIR the thought they are more accessible to microorganisms. Due to 568
525 ndings discussed might seem contradictory at rst glance they consumption of amorphous portions there is an initial increase in 569
526 reveal the degradation of LDPE to be a complex process that can percentage crystallinity [119,60,82,20,118,46]. Once the accessible 570
527 differ for different microorganisms and different communities. amorphous regions have been depleted, microorganisms will start 571
528 What is certainly true is that incubation with microorganisms consuming the smaller crystals present [82], resulting in an 572
529 generates changes in the concentrations of functional groups at the increase in the proportion of larger crystals [60,82,118]. Yet there is 573
530 surface of an LDPE substrate either because of their consumption insufcient research to date to state denitively what happens 574
531 or production. In a complex microbial community in which abiotic after the amorphous regions are consumed. 575
532 factors are also affecting the chemistry of the polymer the net
533 effect observed (accumulation or consumption of functional Molecular weight distribution 576
534 groups) will depend on the balance of rates of oxidation and
535 degradation, which in turn will depend on the nature of the As a result of consumption of the lower molecular weight 577
536 microorganisms present. chains, an increase in the average molecular weight is observed 578
537 The study of the chemistry of LDPE surface turns out to be very after microbial attack [45,61,62,46] (Table 6). However, this result 579
538 important, because oxidized groups are more easily degraded by is not universal, with some authors observing only a slight change 580
539 microorganisms [60] and because oxidized groups modulate in the molecular weight distribution [116,64]. Some others have 581
540 microbial attachment by increasing the hydrophilicity of the concluded that the main factor affecting the molecular weight is 582
541 surface [67], which implies that LDPE degradation will be boosted the effect of abiotic factors such as UV irradiation rather than direct 583
542 if a more oxidized surface is used as substrate. microbial attack [64]. Ohtaki et al. (1998) [120] tested LDPE bottles 584
exposed in aerobic soil for over 30 years, and observed some 585
543 Hydrophobicity/hydrophilicity evidences of biodegradation as reduction in molecular weight by 586
time of ight mass spectrometry (TOF-MS). For the determination 587

544 Hydrophobicity is an important property of the surface in of molecular weight distribution, two different approaches have 588
545 biodegradation studies, because the relation between surface been used: the most common one is the use of size exclusion 589
546 hydrophobicity and microorganisms will determine the extent of chromatography techniques at high temperature 590
547 colonization on the polymer substrate. In general, it is accepted [45,60,19,61,32,116,63,64]. The other possibility is the use of 591
548 that more hydrophilic surfaces are more easily colonized by rheological measurements that correlate indirectly with the 592
549 microorganisms [7679]. If the extent of the oxidation process due molecular weight distribution [62]. Some results showing the 593
550 to the action of abiotic factors such as UV light or activity of extent of reduction based on the number-average molecular 594
551 enzymes is higher than the extent of consumption of functional weight (Mn) of polyethylene samples are presented in Table 6. 595
552 groups then an increase in the hydrophilicity will be observed. The
553 viability of the isolates growing on the LDPE surface is conrmed Surface topography 596
554 using a triphenyl tetrazolium chloride reduction test. The viability
555 is also correlated with a concomitant increase in the protein Development of microcolonies of different microorganisms on 597
556 density of the biomass. Hydrophobicity is usually determined by the surface of the polymer [116,36,63,37,64,121,67] as well as 598
557 the contact angle of the surface with a probe liquid such as water, penetration of hyphal structures [119,82,20] have been reported as 599
558 the more hydrophilic the surface the smaller the contact angle common features after microbial attack. There is enough evidence 600
559 Q9 with water [118,3]. A more advance approach to study hydrophi- which proves that some supercial damage will be observed after 601
560 licity of surfaces is the use of YoungeDupr equation (Eq. (1)), LDPE surfaces have been exposed to biodegradation [122,73,74]. 602
561 which allows the estimation of the energy of adhesion to the solid The structural changes in the form of pits and erosions observed 603
562 Q10 as well as its acid (gS+), basic (gS) and Vander Waals (gSLW)
563 components [72]. Table 6
Molecular number changes in different biodegradation studies.
564 Crystallinity
Substrate %D Molecular number (Mn) References
565 LDPE molecules are less tightly packed and less crystalline LDPE UV irradiated 34 [62]
566 LDPE 15 [46]
because of the side branches. It has been corroborated experimen- LDPE + starch 17 [45]
567 tally that, rst the amorphous regions are consumed because it is
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JECE 539 113
604 through scanning electron microscopy indicated surface damage of Since CO2 evolution out of the system can be monitored 655
605 PE incubated with Fusarium sp. AF4. That suggested that the fungal continuously it allow determining not only the total consumption 656
606 strains, especially Fusarium sp. AF4, was able to adhere to the of the polymer but also the rate of degradation [23,69,100,121]. 657
607 surface of LDPE and can cause surface damage [52]. In a study by Some authors have used this technique successfully to verify the 658
608 Bonhomme et al. (2003) [116], SEM evidence conrmed that ability of some strains to degrade polyethylene of very low 659
609 microorganisms (fungi) build up on the surface of the polymer molecular weight [47]. Results in weight reduction have to be read 660
610 (polyethylene) and after removal of the microorganisms; the with special care when LDPE mixed with starch is used; in this case 661
611 surface became physically pitted and eroded. The surface of the initial reduction in weight can be due to starch consumption rather 662
612 polymer after biological attack was physically weak and readily than LDPE usage. Table 8 presents the main results obtained for the 663
613 disintegrates under mild pressure. Otake et al. (1995) [14] reported extent of biodegradation found in different LDPE types prepared 664
614 the changes like whitening of the degraded area and small holes on without any oxidative treatment. 665
615 the surface of PE lm after soil burial for 32 years.
Mechanisms of LDPE biodegradation 666
616 Mechanical properties
Biodegradation is governed by different factors that include 667
617 Results of most of the studies on LDPE biodegradation, shows LDPE characteristics, type of organism, and nature of pretreatment. 668
618 that in this form of the substrate deterioration of the mechanical The polymer characteristics such as its mobility, tacticity, 669
619 properties such as breaking load, is common (Fig. 2a). Oxidation- crystallinity, molecular weight, the type of functional groups 670
620 induced changes in crystallinity and in the average molecular and substituents present in its structure, and plasticizers or 671
621 weight cause modication of the mechanical properties. Table 7 additives added to the polymer all play an important role in its 672
622 presents results showing changes to different mechanical proper- degradation [123,124]. 673
623 ties for polyethylene after biodegradation. Biodegradation of During degradation, LDPE is rst converted to its monomers, 674
624 modied polyethylene lms in soils led to signicant changes then these monomers are mineralized. LDPEs are too large to pass 675
625 (i.e., elongation at brake of 98%) in their mechanical properties that through cellular membranes, so they must rst be depolymerized 676
626 are caused by biochemical modications of the polyethylene. to smaller monomers before they can be absorbed and biode- 677
627 Compared to waste coal soil, lms underwent rapid biodegrada- graded within microbial cells. The initial breakdown of an LDPE can 678
628 tion in soils that were rich in organic matter. Fungi belonging to the result from a variety of physical and biological forces [125]. 679
629 genus, Gliocladium viride, Aspergillus awamori and Mortierella Physical forces, such as heating/cooling, freezing/thawing, or 680
630 subtilissima, are easily able to colonize polyethylene and polyeth- wetting/drying, can cause mechanical damage such as the cracking 681
631 ylene modied with Bionolle [74]. Although rheological analysis of polymeric materials [126]. The growth of many fungi can also 682
632 can be performed to determine the storage and loss modulus of the cause small-scale swelling and bursting, as the fungi penetrate the 683
633 polymer, in biodegradation studies authors have been preferred polymer solids [127]. LDPEs are also depolymerized by microbial 684
634 the use of a universal mechanical testing system (UMTS) for enzymes, after which the monomers are absorbed into microbial 685
635 determination of mechanical properties of a polymer specimen cells and biodegraded [128]. The enzymatic degradation of LDPEs 686
636 [68,118,74]. However, it is likely that a microorganisms effect will by hydrolysis is a two step process: rst, the enzyme binds to the 687
637 only be supercial in that case since the resolution of bulk testing LDPE substrate then subsequently catalyzes a hydrolytic cleavage. 688
638 methods commonly employed for such measurements diminish as LDPE can be degraded either by the action of intracellular and 689
639 the local surface related damage inuences less of the overall extracellular depolymerases in LDPE-degrading fungi. Intracellular 690
640 sample. degradation is the hydrolysis of an endogenous carbon reservoir by 691
the accumulating microbes themselves while extracellular degra- 692

641 Consumption of the polymer dation is the utilization of an exogenous carbon source not 693
necessarily by the accumulating microorganisms [129]. During 694

642 Biodegradation of LDPE lm was also reported as 0.2% weight degradation, extracellular enzymes from microorganisms break 695
643 loss in 10 years [23]. It is important to note that the rate and extent down complex polymers yielding short chains or smaller 696
644 of polymer consumption can be extensively inuenced by abiotic molecules, e.g., oligomers, dimers, and monomers that are smaller 697
645 factors that promote oxidation. Albertsson and Karlsson (1990) enough to pass the semi-permeable membranes. The process is 698
646 [70] proved that biodegradation rate can increase from 0.2% to 8.4% called depolymerization. These short chain molecules are then 699
647 by irradiating the samples with UV light before a biotic treatment. mineralized into end products, e.g., CO2, H2O, or CH4, the 700
648 Nevertheless, some studies have reported a reduction in the weight degradation is called mineralization, which are utilized as carbon 701
649 of samples determined either by gravimetric measurements and energy source [39]. There are several papers reporting both the 702
650 [62,37,118,72,74,67] or by CO2 evolution from the samples formation of carbonyl groups (oxidation) and reduction of 703
651 [70,23,69,100,121]. It is assumed that LDPE used by microorgan- molecular weight after treatment with UV light [30,69,70,63,64]. 704
652 isms as a carbon source will be nally converted to CO2 during Various methods are available to estimate the biodegradability of 705
653 respiration and can therefore be used as an indirect measurement LDPE. It is desirable to estimate the biodegradability of plastic 706
654 of the amount of LDPE that has been used by microorganisms. wastes under natural condition such as soil [71]. A standard test to 707
Table 7
Changes in mechanical properties due to microbial activity in different polyethylene samples.
Substrate Environment Time %D Elongation %D Tensile strength References

LDPE Waste coal 225 +4% 16.4 [74]
Sea water 365 12% 15 [68]
Sterile seawater + B. sphericus 365 +2.7% 3.8 [118]
Mineral media + Pseudomonas sp. 45 NR 30 [118]
Forest soil 225 4% 16.4 [74]
Cater soil 225 1.5% 19.5 [74]
HDPE Sterile seawater + B. sphericus 365 +8.9 9.7 [118]
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Table 8
A brief summary of biodegradation of polymers.
Material Reported result Conditions Rate or duration of Reported degradation References

degradation
LDPE Partially Bioactive soil (rather shallow) 3237 years in soil 1. About 2/3 decrease of thickness [147]
biodegradable 2. Slow rate of oxidative degradation
LDPE Biodegradable Physicochemical treatments thermal Subjected to biodegradation Morphological, structural, surface [91]
treatment at 105 and 150 C or by a consortium of four fungi changes and mineralization.
accelerated aging treatment during 9 months
LDPE Biodegradable Polyethylene-degrading microorganism Incubation of polyethylene Degradation of polyethylene in the [62]
Brevibaccillus borstelensis strain 707 with B. borstelensis (30 days, presence of mannitol. Maximal
(isolated from soil) 50 C) biodegradation was obtained in
combination with photo-oxidation.
LDPE containing Biodegradable 1. Pre-thermally-oxidized at 55 C 800 weeks (600 days) in 1. Cumulative CO2 emissions [32]
totally degradable inoculum 1700 mg CO2 (70 mg/g soil)
plastic additives 2. Fragmented 2. 44% Mineralization
(TDPA) and pro-
oxidants
LDPE/starch Partially Usage of activated sludge 1 month in inoculum Not reported [146]
biodegradable

LDPE with pro- Environmentally Thermally-oxidized at 100 C 14 days 1. Drop in molecular weight [141]
oxidants LLDPE degradable 2. Carbonyl formation
LDPE/LLDPE Biodegradable Pre-thermally oxidized in an oven 40 140 days 1727% O2 consumption [156]
70 and then in compost
LDPE, LLDPE, HDPE, Thermally Addition of metals Accelerated aging. Thermal 1. Decrease in chemiluminensence [138]
UHMWPE degradable degradation caused by intensity
contact with metals 2. Increase in oxidationrates
LDPE, HDPE, LLDPE Degradable Articial accelerated weathering (UV- 1600 h using a xenon lamp 1. Reported density 0.96 g/cm3 [139]
and xenon arc radiation) of 65,000 W and 800 h using 2. 35% weight loss for HDPE, 5% for
UVB lamp 0.60 W/m2 LDPE, >5% for NP
irradiance at 313 nm
PE Biodegradable Pre-heated at 60 C in an air oven to 1. Sterilized by UV/ 1. Microbial growth [116]
simulate the effect of the compost inoculated 30 min]
environment. Incubated in the presence 2. Incubated for 6 months at 2. Erosion of the lm surface
of selected microorganisms 27 C in soil containing 85%
of water
PAH Biodegradable Buried in extremely acidic environment 28 days 1. 60% mineralization [153]
(coal runoff basin) 2. CO2 production from 010%
depending on the hydrocarbon
LDPE/starch (12%) Biodegradable Controlled biologically active soil 7 months Produced biomass 300 lg/l 7 g CO2/ [156]
50 ml 1727% O2 consumption
708 determine the biodegradation of plastic materials when exposed to that alkane hydroxylase performs the rst oxidation that leads to 736
709 soil was developed by the ASTM (2003) [133]. The microbial the subsequent degradation of a hydrocarbon [131]. Kumar and Jha Q11 737
710 degradation process of polymers is initiated by the secretion of (2013) [117] reported that the viability of the isolates growing on 738
711 enzymes which cause a chain cleavage of the polymer into the polyethylene surface can be conrmed using a triphenylte- 739
712 monomers. Metabolism of the split portions leads to progressive trazolium chloride reduction test. The viability is also correlated 740
713 enzymatic dissimilation of the macromolecules from the chain- with a concomitant increase in the protein density of the biomass. 741
714 ends; eventually, the chain fragments become short enough to be Name of the isolate Total CO2 evolved (g/L) by Modied Sturm 742
715 consumed by microorganisms [130]. By rapid and sensitive test 743
716 analytical technique using Fourier transform infrared coupled Aspergillus avus 4.4507 744
717 attenuated total reectance (FTIR-ATR) spectroscopy we can Mucor circinelloides 5.9328 745
718 quantify LDPE biodegradation. Biodegradation quantication can
719 be performed with FTIR-ATR spectroscopy using various concen- Conclusion and perspectives 746
720 trations of LDPE standards for comparison. It was observed that the
721 percentage of transmittance at 2920 cm1 was directly propor- This review has covered the major concerns about LDPE, their 747
722 tional to the concentration of LDPE. Results indicated that types, uses and degradability. It has looked at the disposal methods 748
723 M. paraoxydans degraded 61.0% of LDPE while P. aeruginosa and the standards used in assessing polymer degradation. Another 749
724 degraded 50.5% in 2 months [66]. Breaking down large polyethyl- area examined has been the developments in the biodegradation of 750
725 ene molecules can be accomplished by enzymatic action, as proven some of the new polymers, either alone or in blended lms. 751
726 by Santo et al. (2012) [46], who found that by incubation with the The above discussion illustrates conceptually different 752
727 enzyme laccase the molecular weight of a polyethylene was approaches to LDPE biodegradation where many groups have 753
728 reduced and its keto carbonyl index increased. These two factors drawn straight forward portrait on bacterial degradation of LDPE 754
729 were felt to indicate that both scission and oxidation reactions rather than fungal. LDPE biodegradation in solid waste environ- 755
730 were taking place by the same enzyme. In regards to the oxidation ments, show that isolated fungi have great potential for LDPE 756
731 process, Yoon et al. (2012) [47] isolated an alkane hydroxylase from biodegradation in the composting process. There is great potential 757
732 the AlkB family that was active to polyethylene samples with for the development of a process for degrading LDPE in a 758
733 molecular weights up to 27,000 Da. It is interesting to note that composting environment using fungi in the near future. Charac- 759
734 enzymes of this family have been described as microorganisms terization of genes responsible for the production of degrading 760
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Journal of Environmental Chemical Engineering xxx (2015) xxxxxx

Contents lists available at ScienceDirect

Journal of Environmental Chemical Engineering

2 Microbial degradation of low density polyethylene (LDPE): A review

* Corresponding author. Tel.: +91 9438450789.

proper introduction of natural polymers such as starch, chitosan or 91

cellulose in the matrix of polyethylene [33]. It is important to 92

highlight that although the damage to LDPE is classied by only 93

cooperatively [34]. In practice, biodegradation is seldom due to a 95

single cause, but a combined effect of heat, UV light, micro- 96

organisms, stress and water [30]. More recently it has been 97

demonstrated in soil burial tests that the use of suitable additives 98

in polyethylene lms induced substantial oxidation with conse- 99

quent fragmentation, drop in molecular weight, increase in wet 100

ability, ultimately followed by high mineralization (6070%) and 101

rate of biodegradation of plastics is usually due to lack of water 103

strategies have been followed in the literature. In the rst 203

strains able to degrade LDPE [20,36,37,4547,6067]. This 205

has been proven that the physicochemical nature of a surface 222

223 determines the ability of microorganisms to form biolm Table 3

Secondly, selected thermophilic microorganisms degrade the 514

Source Enzyme Microorganisms Plastic act as substrate References

time of ight mass spectrometry (TOF-MS). For the determination 587

the accumulating microbes themselves while extracellular degra- 692

necessarily by the accumulating microorganisms [129]. During 694

Substrate Environment Time %D Elongation %D Tensile strength References

Material Reported result Conditions Rate or duration of Reported degradation References

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