The Utilization of Ideonella sakaiensis in Greater

Wax Moth (Galleria mellonella) larvae for

biodegradation of polyethylene plastic

Garcia, Justin Ferdinand; Sentasas, Davidbryan; Nepomuceno, Aimee

10-Roentgen

Manila Science High School, Taft Ave. cor Padre Faura St., Ermita, Manila, Philippines
ABSTRACT

Polyethylene terephthalate (pet) plastic is one of the most used types of plastic, used in plastic
bottles and other products. They are also the main cause of pollution and its accumulation in the
environment has become a global concern. Ideonella sakaiensis is one of the many
microorganisms that can digest pet plastic and is said that it can be found on the gut of the larvae
of the greater wax moth (galleria mellonella) which are a major pest of honeybees and a model
organism for research.this study aims to extract the bacterium ideonella sakaiensis from the
greater wax moth larvae and use it to help in the biodegration of pet plastic and reduce plastic
pollution.

INTRODUCTION

Poly(ethylene terephthalate) (PET) is used extensively worldwide in plastic products, and

its accumulation in the environment has become a global concern. Because the ability to
enzymatically degrade PET has been thought to be limited to a few fungal species,
biodegradation is not yet a viable remediation or recycling strategy.
Speaking globally, the plastic's rate of decomposition is an estimated 450 years (Harmon,
2012). Meaning by the year 2090, a whole continent can be covered with plastic, since plastic is
made with such a fast rate of speed.
In 2016, scientists from Japan tested different bacteria from a bottle recycling plant and
found that Ideonella sakaiensis 201-F6 could digest the plastic used to make single-use drinks
bottles, polyethylene terephthalate (PET). It works by secreting an enzyme (a type of protein that
can speed up chemical reactions) known as PETase. This splits certain chemical bonds (esters) in
PET, leaving smaller molecules that the bacteria can absorb, using the carbon in them as a food
source.
And according to a research, (Bombelli, 2017) Ideonella sakaiensis can be found in the
larvae of an insect called the Greater Wax Moth (Galleria mellonella). The Greater wax moth
can be found throughout the world and is one of the well known parasites of honeybees. The
larvae are used as a model organism for research because of their ability to digest plastic.
The researchers’ plan is to find larvae, extract Ideonella sakaiensis from the larvae’s gut
and get the results from the experimentation. The research aims to provide a healthier nature for
bacterium and find an alternative agent in biodegrading plastic. Getting Ideonella sakaiensis in
Greater Wax Moth larvae is better since they are considered to be parsites and extracting their
pupae would be a lot less expensive for the reporters.
METHODOLOGY
A) Gathering of Materials

The larvae of the moth were collected in a pet shop in Manila that sells the larvae for reptile
food. The larvae had been fed PET plastic. The plastic used is sterilized by washing it with
hot sterilized water.

B) Isolation of Ideonella sakaiensis

The isolation of the bacterium is done in the laboratories of Philippine Women’s
University at September 1, 2018 by acquiring the digestive tract of the larvae and extracting a
microbial consortium on a partly digested piece of PET plastic. The bacterium is identified
through gram testing,
C) Culturing of the Bacteria

The bacteria are cultured using NBRC no. 802 medium with pH 7.0. The researchers used
two petri dishes as the setup for the culture. It will be also used as the setup for the
experimentation. The colony will be left to cultivate for 5 days.

D) Experimentation
To find out how much plastic the bacteria can biodegrade, the researchers put 1.0 mg of
two types of PET samples, one is oriented PET film and the other is non-oriented. The
results will be based on the weight of each sample on both layouts in 24 hours with the
interval of 4 hours.

RESULTS AND DISCUSSION

1.05

1
PET film (Non-oriented)
0.95 PET sample (oriented)
Weight (mg)

0.9

0.85

0.8

0.75
4 8 12 16 20 24
Hours
Fig 1.1 Table of the biodegradation of the two PET films within 24 hours
As shown in Fig. 1.1, the rate of biodegradation in the non oriented PET film in 8 hours is 0.98
mg, in 12 hours 0.971 mg, in 16 hours 0.957 mg, 20 hours 0.92 mg and in 24 hours 0.89 mg. In
24 hours, the bacteria had digested 0.11 mg of non oriented film.
The rate of bio degradation in the oriented PET film in 8 hours is 0.97 mg, in 12 hours 0.969 mg,
in 16 hours 0.951 mg, 20 hours 0.91 mg and in 24 hours 0.864 mg. In 24 hours, the bacteria had
digested 0.136 mg of non oriented film.
The rates of biodegradation between the two samples are almost the same rate. However the
progress is much faster on oriented PET film.
Due to limited time and resources, the researchers cannot show a significant difference in the rate
of biodegradation of plastic on the bacterial samples.

CONCLUSION
The utilization of Ideonella sakaiensis as a means of biodegradation of polyethylene plastic is
feasible and the larvae of Galleria mellonella is one of the sources of this bacteria and can help
get rid of the pests of honeybees and pollution of plastic as well.
The researchers recommend making the duration of the experiment longer because more accurate
and significant results can be seen and more detailed discussions can be made. Another
recommendation is to research a way to either engineer or modify the organism so that it can be
used in other locations such as landfills. Another recommendation is to find other
microorganisms that can also digest plastic that is easy to find and cultivate.
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