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Department of Biology, Sangmyung University, 7 Hongji-dong Chongro-gu, Seoul 110-743, South Korea
b
Department of Polymer Science and Engineering, Inha University, Incheon 402-751, South Korea
Received 20 October 2003; received in revised form 15 December 2003; accepted 2 January 2004
Abstract
Biodegradation of plastics was tested in the compost stored at 20 (C, 4 (C and 20 (C for dierent periods. Viable cells in the
compost stored at 20 (C were expected to be fewer than those in the compost stored at 4 (C and 20 (C, because microbes may be
under stress or even be killed due to the formation of ice crystals at the subzero temperature. Mesophilic bacteria and mesophilic
actinomycetes were fewer in number in the compost stored at 20 (C than in the compost stored at the other two lower temperatures
contrary to expectation. In contrast, both thermophilic bacteria and thermophilic actinomycetes were fewest in the compost stored
at 20 (C as was expected, indicating that thermophilic microbes were more susceptible to stress in the freezing conditions than the
mesophilic ones. Activity of the exo-enzymes plausibly excreted by the microbes in the compost decreased as a result of the storage.
Nevertheless, biodegradation of cellulose in the compost was almost independent of the storage time and temperature. In contrast,
biodegradability of both polycaprolactone (PCL) and poly(butylene succinate) (PBS) depended strongly on the storage conditions.
From the point of view that the existing standards for biodegradation tests of plastics in compost accept reproducibility of cellulose
biodegradability as a criterion for the validity of the biodegradation tests, a new standard of the compost preparation should be
provided to guarantee more reliable results on the biodegradability of plastics.
2004 Elsevier Ltd. All rights reserved.
Keywords: Compost; Storage; Biodegradation; Bacteria; Actinomycetes
1. Introduction
Synthetic plastics give rise to water clogging phenomenon in soil burial sites and thus cause rain water overow, provoking serious contamination of nearby rivers
and sea [1]. Moreover, accumulation of synthetic plastics
in cultivation soil reduces crop production. Many animals
and sh are killed due to swallowing of debris of synthetic
plastics, which in turn disrupts the ecosystem.
Recycling of plastics is not economically feasible in
many cases due to deterioration of mechanical properties and excessive cost. Development of biodegradable
plastics is under investigation because it may provide
a solution to the above-mentioned environmental problems. Food garbage can be composted together with
biodegradable packaging materials, which may contribute to a signicant reduction of municipal solid waste.
Evaluation of biodegradability of plastics is usually
carried out in mature compost, because many kinds of
microbes are present in the compost and the evaluation
may represent eventual fate of the plastics in nature.
Evaluation of biodegradability of plastics appeared
to be satisfactorily reproducible in a compost made from
cow fodder [2]. The compost for the biodegradability
tests was harvested 1e2 weeks after the onset of the
maturation stage. The harvest time was kept constant
because the content of organic matters and the microbial community should be changed with time.
As more than 45 days are required to produce fresh
compost, the biodegradation tests can only be started
with a signicant delay in many cases.
412
2. Methods
2.1. Composting
Compost enzyme activity was measured using APIZYM kit (API Analytab products, Plainview NY, USA).
APIZYM is a commercial semi-quantitative micromethod designed for systematic and rapid study of 19
enzymatic reactions. It consists of a series of microcupules containing dehydrated chromogenic substrates
for the 19 dierent enzymes and a control (a microcupule without any substrates for the enzymes).
Enzyme extracts were prepared by mixing 10 g of the
compost sample with 90 ml of sterilized distilled water
for 30 min. The suspension solution was allowed to
413
70
compost temperature
ambient temperature
60
50
40
30
20
10
0
0
10
20
30
40
Time (Day)
Fig. 1. Temperature prole during the composting process.
Table 1
Physicochemical characteristics of the compost as a function of the storage conditions
Fresh
compost
65.0
8.5
44.0
3.1
14.1
Storage time
30 days
60 days
90 days
Storage temperature
Storage temperature
Storage temperature
20 (C
4 (C
20 (C
20 (C
4 (C
20 (C
20 (C
4 (C
20 (C
64.2
8.3
43.2
3.1
14.1
65
8.4
43.9
3.12
14.1
62.7
8.3
41.6
3.3
12.6
64.9
8.2
42.7
3.0
14.4
64.3
8.2
44.0
3.1
14.1
62.6
7.8
41.6
4.0
10.4
59.8
8.2
42.1
2.8
14.9
58.5
7.8
43.7
3.1
14.1
59.3
7.7
39.4
3.6
10.9
414
109
108
107
fresh compost
-20/30/2
4/30/2
20/30/2
-20/60/2
4/60/2
20/60/2
-20/90/2
4/90/2
20/90/2
106
1010
1010
105
109
108
107
fresh compost
-20/30/2
4/30/2
20/30/2
-20/60/2
4/60/2
20/60/2
-20/90/2
4/90/2
20/90/2
106
105
109
108
107
fresh compost
-20/30/2
4/30/2
20/30/2
-20/60/2
4/60/2
20/60/2
-20/90/2
4/90/2
20/90/2
106
105
Fig. 3. Eect of storage conditions on mesophilic actinomycete
population in an animal fodder compost. The symbol N1/N2/N3
means that the compost has been stored at N1 (C for N2 days, and
then conditioned at 17 (C for N3 days.
1010
109
108
107
fresh compost
-20/30/2
4/30/2
20/30/2
-20/60/2
4/60/2
20/60/2
-20/90/2
4/90/2
20/90/2
106
105
Fig. 5. Eect of storage conditions on thermophilic actinomycete
population in an animal fodder compost. The symbol N1/N2/N3 means
that the compost has been stored at N1 (C for N2 days, and then
conditioned at 17 (C for N3 days.
415
Fresh
compost
60 days
90 days
4 (C
20 (C
4 (C
20 (C
4 (C
20 (C
Phosphatases
Alkaline phosphatase
Acid phosphatase
Naphthol-AS-BI-phosphohydrolase
5
4
3
5
3
3
5
4
2
3
3
2
5
3
2
3
2
2
5
3
2
Esterases
Esterase
Esteraseelipase
Lipase
2
3
3
2
2
2
2
2
3
1
2
2
1
0
2
1
2
1
1
2
1
Arylamidase
Leucine arylamidase
Valine arylamidase
Cystine arylamidase
4
2
1
3
2
1
3
2
0
2
1
0
3
0
0
2
1
0
3
0
0
Proteases
Trypsin
a-Chymotrypsin
1
1
0
0
0
0
0
0
0
0
0
0
0
0
Glycosyl hydrolases
a-Galactosidase
b-Galactosidase
b-Glucuronidase
a-Glucosidase
b-Glucosidase
N-acetyl-b-glucosaminidase
a-Mannosidase
a-Fucosidase
1
1
2
3
3
4
0
0
0
0
2
3
3
4
0
0
0
0
2
1
2
2
0
0
0
0
1
2
2
4
0
0
0
0
1
1
1
2
0
0
0
0
0
1
1
3
0
0
0
0
0
1
0
1
0
0
Low intensity (value of 1); moderate intensity (values of 2e4); high intensity (value of 5).
80
CO2 evolution
60
40
fresh compost
-20/30/2
4/30/2
20/30/2
20
0
0
10
15
20
25
30
Time (Day)
Fig. 6. Cellulose biodegradation in a compost stored for 30 days at
dierent temperatures. The symbol N1/N2/N3 means that the compost
has been stored at N1 (C for N2 days, and then conditioned at 17 (C
for N3 days.
416
100
100
80
80
CO2 evolution
CO2 evolution
60
40
fresh compost
-20/90/2
4/90/2
20/90/2
20
60
40
fresh compost
-20/90/2
4/90/2
20/90/2
20
0
0
10
15
20
25
30
Time (Day)
10
15
20
25
30
Time (Day)
100
100
80
CO2 evolution
80
CO2 evolution
fresh compost
-20/30/2
4/30/2
20/30/2
60
40
fresh compost
-20/30/2
4/30/2
20/30/2
20
60
40
20
0
0
10
15
20
25
30
Time (Day)
Fig. 8. PCL biodegradation in a compost stored for 30 days at dierent
temperatures. The symbol N1/N2/N3 means that the compost has been
stored at N1 (C for N2 days, and then conditioned at 17 (C for N3
days.
10
15
20
25
30
Time (Day)
Fig. 10. PBS biodegradation in a compost stored for 30 days at
dierent temperatures. The symbol N1/N2/N3 means that the compost
has been stored at N1 (C for N2 days, and then conditioned at 17 (C
for N3 days.
100
References
fresh compost
-20/90/2
4/90/2
20/90/2
80
CO2 evolution
417
60
40
20
0
0
10
15
20
25
30
Time (Day)
Fig. 11. PBS biodegradation in a compost stored for 90 days at
dierent temperatures. The symbol N1/N2/N3 means that the compost
has been stored at N1 (C for N2 days, and then conditioned at 17 (C
for N3 days.
Acknowledgements
This work was supported by an Interdisciplinary
Research Program grant R01-2002-000-00146-0 from
the Korea Science and Engineering Foundation.