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bers of the filamentous fungi Fusarium oxysporum and F. solani, which have been shown
to grow on a mineral medium containing PET
yarns [although no growth levels were specified (5, 6)]. Once identified, microorganisms with
the enzymatic machinery needed to degrade PET
could serve as an environmental remediation
strategy as well as a degradation and/or fermentation platform for biological recycling of PET
waste products.
We collected 250 PET debriscontaminated environmental samples including sediment, soil,
wastewater, and activated sludge from a PET
bottle recycling site (7). Using these samples,
we screened for microorganisms that could use
low-crystallinity (1.9%) PET film as the major
carbon source for growth. One sediment sample
contained a distinct microbial consortium that
formed on the PET film upon culturing (Fig. 1A)
and induced morphological change in the PET
film (Fig. 1B). Microscopy revealed that the consortium on the film, termed no. 46, contained
1
PET
10
20
30
40
50
60
0
film
20
40
60
80
4
5
6
3
6
5
8
9 10
10
20
30
40
50
60
8
9 10
20
40
60
Fig. 1. Microbial growth on PET. The degradation of PET film (60 mg, 20
15 0.2 mm) by microbial consortium no. 46 at 30C is shown in (A) to (C).
The MLE (modified lettuce and egg) medium (10 mL) was changed biweekly.
(A) Growth of no. 46 on PET film after 20 days. (B) SEM image of degraded PET
film after 70 days. The inset shows intact PET film. Scale bar, 0.5 mm. (C) Time
course of PET film degradation by no. 46. PET film degradation by I. sakaiensis
201-F6 at 30C is shown in (D) to (H).The YSV (yeast extractsodium carbonate
1196
BIODEGRADATION
RE S EAR CH | R E P O R T S
a mixture of bacteria, yeast-like cells, and protozoa, whereas the culture fluid was almost transparent (Fig. 1A). This consortium degraded the
PET film surface (fig. S1) at a rate of 0.13 mg cm2
day1 at 30C (Fig. 1C), and 75% of the degraded
PET film carbon was catabolized into CO2 at 28C
(fig. S2).
Using limiting dilutions of consortium no. 46
that were cultured with PET film to enrich for
microorganisms that are nutritionally dependent
on PET, we successfully isolated a bacterium capable of degrading and assimilating PET. The strain
represents a novel species of the genus Ideonella,
for which we propose the name Ideonella sakaiensis
201-F6 (deposited in the National Center for
Thermobifida group
4OYY
(Humicola insolens)
TfH
ADV92528
(T. fusca)
ADV92526
ADV92527
(T. cellulosilytica)
(T. cellulosilytica)
ADV92525
AFA45122.1
982 997991
(T. alba)
(T. halotolerans)
1000
BAO42836.1
(Saccharomonospora viridis)
668
1000
1000
FsC
986
1000
MHET
HO CH2 CH2
LCC
O
HO C
10 mV
O
C OH
O C
C OH
BHET
TPA
HO CH2 CH2
PETase
C O CH2 CH2 OH
O C
18h
0h
19
20
21
22
23
0.1
ADH43200.1
(Bacillus subtilis)
24
PET-film (b)
b/a
BHET
PETase
10
hcPET
pNP-acetate (C2)
pNP-butyrate (C4)
pNP-caproate (C6)
pNP-caprylate (C8)
PETase
TfH
TfH
LCC
TPA
MHET
BHET
FsC
40
80 120
Apparent kcat (sec-1)
0.1
0.2
0.3 -5 -4 -3 -2 -1 0
LCC
b/a(C2)
b/a(C4)
b/a(C6)
b/a(C8)
log10Ratio
0.4
0.8
SCIENCE sciencemag.org
TPA
MHET
FsC
0
0.010
0.005
0.015
18
LCC
100
PETase
10-1
TfH
10-2
10-3
FsC
10-4
20
30
40
50
60
70
80
Temperature (C)
1197
R ES E A RC H | R E PO R TS
Fold change
(relative to maltose)
-1
10
10
10
10
10
( CH
CH2
O C
)n
*
**
* **
0077
**
* **
HO CH2 CH2
0228
0230
0229
O
C OH
O C
MHETase
O
HO C
****
****
**
**
**
****
**
****
0227
**
**
****
****
PCA degradation
0627
**
**
0076
0626
PETase
(4831)
**
0224
**
**
4831
TPA degradation
(0224)
C OH
TPATP (0076/0077)
O2
NADPH + H+
TPADO (0227/0228/0230)
NADP+
Pca34
DCDDH
(0626/0627)
(0229)
TPA-Na
PET film
BHET
COOH
COOH
COOH
H
OH
HO OC
OH
NADP+
OH
CO2
NADPH + H+
O2
HOO C
HOO C
OH
ORF#
(ISF6_XXXX)
Table 1. Kinetic parameters of MHETase. The kinetic parameters were determined in pH 7.0 buffer at
30C. Because the enzymatic PET hydrolysis involves a heterogeneous reaction, MichaelisMenten
kinetics were not applied to PETase. ND, not detected (activity was below the detection limit of the assay).
Substrate
kcat (s1)
Km (mM)
MHET
PET film
31 0.8*
ND
7.3 0.6*
.....................................................................................................................................................................................................................
.....................................................................................................................................................................................................................
BHET
0.10 0.004
.....................................................................................................................................................................................................................
pNP-aliphatic
esters
(pNP-acetate,
pNP-butyrate)
ND
.....................................................................................................................................................................................................................
Aromatic esters (ethyl gallate, ethyl ferulate, chlorogenic acid hydrate)
ND
.....................................................................................................................................................................................................................
RE S EAR CH | R E P O R T S
SUPPLEMENTARY MATERIALS
www.sciencemag.org/content/351/6278/1196/suppl/DC1
Materials and Methods
Supplementary Text
Figs. S1 to S14
Tables S1 to S5
References (1839)
15 October 2015; accepted 29 January 2016
10.1126/science.aad6359
NEURODEVELOPMENT
hromosomal aberrations at 22q13 that delete or inactivate one SH3 and multiple
ankyrin repeat domains 3 (SHANK3) allele
are genetic hallmarks of Phelan-McDermid
syndrome (PMDS). De novo mutations in
1199
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