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org/ on May 4, 2018

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PLASTIC DEGRADATION BY MICROORGANISM

Weight loss (mg)

10
20
30
40
Presented by:- Khushbu S Soni and Pragyan Nayak 50

(Review Poster) PET

film
60
0 20 40 60 80
St. Xavier's College, Ahmedabad Cultivation time (days)
Current Biology

2
1 Magazine 1 2 3
0

INTRODUCTION METHODOLOGY 6
• In the current study many bacterias20were isolated from
RESULT 10

Weight loss (mg)

3
4
1. Plastics are synthetic polymers derived • Initially, 250 PET debris-contaminated environmental samples including sediment, 6 4
different sites and after primary and secondary screening 4 of
from fossil oil and have been widely used as Correspondence A
soil, wastewater and activated sludge from PET bottle recycling site was collected, B 5 C
5 them showed positive results and identified
30
40 as Bacillus sp. ,
building materials of various consumer Polyethylene
then screening for microorganisms thatB
could use low crystallinity PET film as a Pseudomonas sp. , Staphylococcus 50sp. and Ideonella
products. Polyethylene(PE), major carbon source was done. one sediment sample 8
contained a 7microbial sakaiensis.
bio-degradation by 7 1.2 60
8
10 9 10• Wax moth such as G. mellonella was also identified 0 20 40
which 60
was
polypropylene(PP) and poly ethylene consortium formed on PET film upon culturing and induced morphological 9

terephthalate(PET) are the major caterpillars

changes in of PETthe film. Using living dilutions consortium no 46. was cultured with 13% capable of degrading plastic. When 100 wax worms of G.
Cultivation time (days)

Fig. 1. Microbial growth on PET. The degradation of PET film (60 mg, 20 × vitamins) medium
mellonella werewas in changed
contactweekly.
with (D F) SEM images
PEtoshopping bag forof I. sakaiensis
12 hours,
constituents of plastic production. waxPET, moth and Galleria
there was isolation of bacterium capable
by microbialof degrading PET. It was

mg cm-2
15 × 0.2 mm) consortium no. 46 at 30°C
1.0 is shown in (A) to (C). cells grown on PET film for 60 hours. Scale bars, 1 mm. Arrow heads in the left
it resulted in mass loss of 92mg.
2. P l a s t i c p r o d u c t i o n h a s i n c r e a s e d mellonella identified as Ideonella sakaiensis 201-F6. Then corresponding I. sakaiensis
The MLE (modified lettuce and egg) medium (10 mL) was changed biweekly. panel of (D) indicate contact points of cell appendages and the PET film surface.
(A) Growth of no. 46 on PET film after 20 days. (B) SEM image of degraded PET • During 20 are
Magnifications daysshowninterval Staphylococcus
in the right panel. Arrows in (F) sp. showed
indicate appendages10%
exponentially in the past 50 years, on proteins were purified and incubated with film PET
after 70 film
days. at
The 30°C
inset shows for 18
intact hours.
PET film. ScaleProminent
bar, 0.5 mm. (C) Time between the cell and the PET film surface. (G) SEM image of a degraded PET
~100 wax worms 0.2
RE S EAR CH | R E P O R T S degradation of 40 micron PE and Bacillus sp. showed 13.3%
contrary plastics causes the environmental Paolo Bombelli1,
pitting developed on film surface.

Christopher J. Howe1,*,
92 mg mass loss course of PET film degradation by no. 46. PET film degradation by I. sakaiensis film surface after washing out adherent cells. The inset shows intact PET film.
degradation of 10 micron PE and Pseudomonas sp. showed
~12 hours exposure 201-F6 at 30°C is shown in (D) to (H).The0.0 YSV (yeast extract–sodium carbonate– Scale bar, 1 mm. (H) Time course of PET film degradation by I. sakaiensis.
pollution by getting accumulated in the • azoa,
and In another
mixture
Federica of bacteria,
Bertocchini method,
yeast-like theproto-
2,3, cells, and
* worm homogenate
Biotechnology Informationof mothdatabase
taxonomy G. mellonella was
There are currently smeared
few known
Untreated on of
examples
Treated minimum degradation by weight loss.
environment because of the long branched whereas the culture fluid was almost trans-
left(Fig.in1A).contact with PE the films.
under identifier 1547922). In addition to being
Galvenetric analysis of treated
esterases, lipases, or cutinases that are capable
samples confirmed a • Ideonella sakaiensis 201-F6 degraded the PET film surface at
parent This consortium degraded D found in the culture fluid,
1196cells 11
were observed
MARCH 2016 on
• VOL of E
351hydrolyzing
ISSUE 6278 PET (8, 9). To explore the genes sciencemag.org SCIENCE
chains and it’s stable nature. Plastics–1significant
PET film surface (fig. S1)
are synthetic polymers loss ofderived
at a rate of 13% PE1.0over 14 hours of treatment compared to untreated
0.13 mg cm –2
the film (Fig. 1D) and appeared to be connected involved in PET hydrolysis in I. sakaiensis 201- –2 –1
day at 30°C (Fig. 1C), and 75% of the degraded to each other by appendages (Fig. 1E). Shorter F6, we assembled a draft sequence of its genome a rate of 0.13 mg cm day at 30°C, and 75% of the
3. In 27 EU countries plus Norway and fromPET samples.

fossil oil and largely resistant to

T(%)
Untreated Untreated
film carbon was catabolized into CO2 at 28°C
biodegradation. Polyethylene (PE) and
appendages
0.8 were observed between the cells and (table S1). One identified open reading frame degraded PET film carbon was catabolised into CO2 at 28°C .
Switzerland up to 38% of plastic is • (fig. Lastly,
S2). soil samples were collected
Treated
the film; thesefrom areas
might assist in thenear
deliverypetrol
of se- pump,
(ORF), hospital
ISF6_4831, encodes aand Treated
putative local
lipase that
polypropylene (PP)dilutions
represent ~92% of

Abs
Using limiting of consortium no. 46 0.6
creted enzymes into the film (Fig. 1, D and F). shares 51% amino acid-C-H sequence identity and
discarded in landfills carrying a heavy totalthat areas
plastic in Dehradun
were production.
cultured with PETPE film toand
enrich dilutions
is largely for The PET were made.extensively
film was damaged Further (Fig. dilutions
1G) catalyticwere
residuesspread andfrom
with a hydrolase PETher-
0.4
environmental impact. Also large quantities utilized in packaging,
strips
microorganisms that arerepresenting
of 3*3cma bacterium werecapa-
nutritionally dependent
cut 30°C
and placed on nutrient agarexhibits
and almost completely degraded after 6 weeks at
plate. after incubation
mobifida fusca (TfH) (fig. S4 and table S2) that
-C=O CONCLUSION
~40% on PET, we successfully
of total demandisolatedfor plastic 0.2 (Fig. 1H). In the course of subculturing no. PET-hydrolytic activity (10). We purified the
of PET have been introduced into the ble ofmicrobial
degrading growthPET.
and assimilating was seen 46,
The strain onwePE foundstrips. Thenthatmicrobes
a subconsortium lost its PET were isolated
corresponding recombinant using zone
I. sakaiensis proteins By observing the results it can be concluded that G.
products (www.plasticseurope.org) 0.0
environment through its production and represents a novel species of the genus Ideonella,
with over clearance method, variousI. sakaiensis
a trillion plastic bags used
degradation
4000
capability. This subconsortium lacked
morphological
3500 3000 and2500biochemical (fig. S5) and
1800 incubated them
tests and were
1500 1200 with PET900 film at
mellonella, Ideonella sakaiensis and Bacillus sp. possess
for which we propose the name Ideonella sakaiensis (fig. S3), indicating that I. sakaiensis 30°C for 18 hours. Prominent pitting developed on
disposal, resulting in accumulation of PET in every year [1]. Plastic production has cm cm
-1 -1
201-F6identified
(deposited inas theBacillus
National Center sp.,
for Pseudomonas
is functionally involved insp. and Staphylococcus
PET degradation. the film surface (Fig.sp.2A). Mono(2-hydroxyethyl) greater potential then other bacterias in degradation of
increased exponentially in the past F G
the ecosystem across the globe. 50 years (Figure S1A in Supplemental polyethylene, poly ethylene terephthalate. Microbes all
4. Therefore new solutions of plastic Information, published with this article Untreated Thermobifida group Treated around have great potential to degrade plastics and solving
degradation are urgently needed. one such online). In the 27 EU countries plus 4OYY Z[nm] ADV92528 TfH
Z[nm] (Humicola insolens) (T. fusca) the future environmental problems caused by plastics.
Norway and Switzerland up to 38% 500.0
ADV92526
solution is by microbial degradation.
0.0
ADV92527
(T. cellulosilytica)
of plastic is discarded in landfills, -6.0
0.0
(T. cellulosilytica)
5. Microbial degradation of plastic is carried 8.0 -10.0 AFA45122.1 ADV92525
with the rest utilized for recycling 982 997991

Downloaded from http://science.sciencemag.org/ on May 4, 2018

-5.0 7.0 -9.0 (T. alba)
(T. halotolerans)
out by enzymatic activities leading to (26%) and energy recovery (36%) via -4.0 6.0 -8.0 9.0

combustion (www.plasticseurope.
-3.0
4.0
5.0
Y[ m] X[ m]
-7.0 1000
8.0
BAO42836.1
7.0
BIBLIOGRAPHY
breakdown of polymer into monomers and X[ m] -2.0 -6.0
1) BIODEGRADATION OF POLYTHENE BY BACTERIA
668 (Saccharomonospora
6.0 Y[ m] viridis)
org), carrying a heavy environmental -1.0
3.0
1000
-5.0
1000 5.0
oligomers that the bacteria can absorb impact. Therefore, new solutions FsC0.0 2.0
1000
-4.0
986 4.0 ISOLATED FROM SOIL( Gauri Singh*, Ashok Kumar Singh
for plastic degradation are urgently
using the carbon in them as food and Kalpana Bha) International Journal of Research and
needed. We report the MHETfast bio- O O Figure 1. Polyethylene degradation by Galleria mellonella.

source(metabolism). Aerobic metabolism degradation of PE by larvae of the wax C (A)

HO CH CH O C 2 2 OH
Plastic bag after exposure to ~100 wax worms for 12 hours. (B) Magnification LCC of the area
Development in Pharmacy and Life Sciences February -
leads to production of carbon dioxide and moth Galleria mellonella, producing indicated in A. (C) Gravimetric analysis of homogenate-treated versus untreated polyethylene March, 2016, Vol. 5, No.2, pp 2056-2062 ISSN (P):
water whereas in aerobic metabolism
O
ethyleneHOglycol.
C
O
C OH MECHANISM
(PE), showing a reduction (13%) of mass per unit of area in the former. (D,E) FTIR analysis of
the homogenate-treated and control PE films. (F,G) Atomic Force Microscopy on homogenate- 2393-932X, ISSN (E): 2278-0238
PE Talking
comprises
TPA aabout linear BHET
backbone
what drives the(G)microorganisms to feed on plastics lies inmaps theeach).
10 mV

treated and untreated (F) PE film (representative examples of 3 topographic 2) A bacterium that degrades and assimilates poly(ethylene
O O

production of carbon dioxide, water and of carbon atoms (Figure S1B), which HO CH CH O C 2 2 C O CH CH OH 2 2 PETase
mechanism. terephthalate).(Shosuke Yoshida, Kazumi Hiraga, Toshihiko
methane takes place as end products. is resistant to degradation. Although18h (FTIR) analysis of treated samples of another plastic, poly(ethylene
PE is believed not to be susceptible 0h revealed formation of an absorbance terephthalate) (PET) by a microbial Takehana, Ikuo Taniguchi, Hironao Yamaji, Yasuhito Maeda,
to•bio-degradation,
Wax worms of a few moth G. mellonella
attempts peak around feed3,300 on PE cm-1as it has the similar
, a signature consortium CH2 –CH2a newly
including
0.1
frequentisolated Kiyotsuna Toyohara, Kenji Miyamoto, Yoshiharu Kimur and
have hydrocarbon
been
18 made,
19
bond
20 as21PE is
that
22 the23most
Retention time (min)
is
24
also forfound
ethylene in the
glycol, beeswax
confi rming on
PE
ADH43200.1
(Bacillus which
subtilis) they
bacterium, feed. The
Ideonella wax
sakaiensis, worms was
Kohei Oda) Science 351 (6278), 1196-1199. DOI: 10.1126/
common packaging plastic. Slow degradation. More recently, Yang described recently [5]. Although PET is
grow to their pupa stage eating
(weeks/months) PE biodegradation
beeswax.

et al. reported bacterial degradation a resistant material, one might expect science.aad6359
• been observed, given appropriate of secreting
has Ideonella sakaiensis
pNP-aliphatic esters (a) works
PET-film (b) by PE over b/a
several an weeksenzyme
BHET
[4]. known as its
hcPET
PETase
biodegradation which
10 tosplits
be easier certainthan 3) Polyethylene bio-degradation by caterpillars of the wax
chemical
conditions. bonds(esters)
ForpNP-butyrate
example,
pNP-acetate (C2) modest in PET, leaving
However, no smaller
production molecules
of ethylene that PE, the
as PETbacteria
has a can
polyester absorb,
backbone
otal released compounds (mM)

PETase (C4) moth Galleria mellonella. (Paolo Bombelli1, Christopher J.

degradation ofpNP-caproate
using carbon PE wasin(C6) observed
them as afterfoodglycolsource. from the Latestbiodegradation
study reveals was thatandmodified
can be hydrolysed. 100 PETase Weworkedreport LCC

nitric acid treatment and(C8)incubation

pNP-caprylate
described. The authors reported that
PETase
here the fast biodegradation PETase of PE Howe1,*,and Federica Bertocchini 2,3,*) Current Biology 27,
for 3well
TfH as in
months the molecule
a liquid culture involved
of the in biodegradation
PE the reaction depended is very accessible,
onTfH
the bymaking
the wax worm, it10-1easy the for enzyme
caterpillar larva R283–R293, April 24, 2017
PETase to attack
fungus Penicillium simplicissimum even buried PET molecules

activity of microorganisms present of the wax moth Galleria mellonella TfH

of 4) phys.org/news/2018-04-plastic-eating-bacteria-worka-
10-2
[2]. Slow
LCC PE degradation was in the gut of the larvae of the Indian the snout moth (Pyralidae) family of
TPA b/a (C2)
LCC
chemist.html
also recorded after 4 to 7 months MHETmealmoth b/a Plodia interpunctella (two
(C4) Lepidoptera.
TPA 10-3 FsC
FsC to the bacterium Nocardia
exposure bacterial strains, Bacillus sp. YP1 FsC and WhenMHET a PE film was left in direct
BHET b/a (C6)
b/a (C8)