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Correspondence from polyethylene degradation. A homogenate, is missing from the

simple inspection of the attenuated Bombelli et al. study.
Polyethylene total reflectance infrared (ATR–IR)
spectra for pure ethylene glycol and
It appears highly doubtful that
amphiphilic/lipophilic proteins can
bio-degradation ethylene glycol on a polyethylene be removed using the reported
film (Figure 1) — a simple control washing procedure with deionized
by caterpillars? experiment missing in the water, whereas the completely water
publication — indeed shows this soluble ethylene glycol should remain
Carina Weber, Stefan Pusch, signal. However, the characteristic on the polymer surface. To test this
and Till Opatz* absorbances at 1,085 and 1,033 cm–1 hypothesis, we treated a polyethylene
are missing in the published infrared film with egg yolk as well as ground
In their recent paper on the spectrum. A single infrared absorption pork (Figure 1). Here, an absorption
degradation of polyethylene by at 3,300 cm–1 is not sufficient proof around 3,300 cm–1, usually attributed
caterpillars of the wax moth Galleria for the presence of ethylene glycol, to N–H stretching vibrations from
melonella, Bombelli et al. [1] report which must also display all other proteins, was found. Furthermore,
various experiments, including spectral features characteristic for the carbonyl region is in almost full
microscopic and spectroscopic this compound. We hypothesize that agreement with the published infrared
data which the authors believe the infrared signals in question may spectrum, exhibiting absorbances at
support the chemical digestion of the rather be caused from residual protein 1,744, 1,651, and 1,545 cm–1, which
polymers by these insects. While the contamination on the polyethylene are again typical for lipids and proteins
biodegradation of mostly inert artificial surface, which could also explain the [2]. Thus, the band at 1,744 cm–1
polymers is definitely a very interesting additional surface roughness after wax corresponds to the C=O stretching
research field, we must respectfully moth treatment. Another alternative mode of an ester as present in a
disagree with the methodology and explanation would be abrasion through triglyceride or other membrane lipid,
conclusions from this paper. the mechanical application and the band around 1,651 cm–1 is the
The authors state that the infrared removal of the worm homogenate. amide-I band (C=O stretching mode
absorbance at 3,300 cm–1 originates Indeed, a control experiment, for of a peptidic bond), and the slightly
from ethylene glycol, resulting example, with heat-inactivated worm weaker band at 1,545 cm–1 is the
100
90
80
Transmittance [%]
70
60
50
40
30

1646.22
1745.13

1647.97
1543.07
3311.19
3296.02

2848.49

1744.00
1651.23

1471.45
2874.82

1033.48
3367.61

2938.48
2916.04

1463.45

1205.05
1162.66
1083.51

861.16
881.93

719.12
729.50

3500 3000 2500 2000 1500 1000

Wavenumber [cm-1]

Figure 1. Infrared spectra of ethylene glycol and possible protein/lipid contaminants.

ATR–IR spectra of pure ethylene glycol (green), pure polyethylene (black), egg yolk-treated polyethylene (yellow), ground pork-treated polyethylene
(blue), and ethylene glycol-treated polyethylene (red).

R744 Current Biology 27, R731–R745, August 7, 2017 © 2017 Elsevier Ltd.
Current Biology
Magazine
amide-II band (symmetrical stretch
of N–C=O). These four bands can
clearly be seen in the spectra depicted
in Figure 1D,E of Bombelli et al. [1].
The intensity of the peaks around
1,740 cm–1 and the associated band
around 1,170 cm–1 (corresponding to
the asymmetric stretching mode of the
C–O-bond in esters), relative to the
amide bands around 1650 and 1,545
cm–1, are determined by the lipid/
protein ratio in the respective sample
(the ground pork used obviously being
richer in lipids than egg yolk). The
adhesion of lipid/protein matrices to
surfaces is further demonstrated by
the fact that our infrared spectrometer
was difficult to clean after the
measurements, so we expect the
Galleria mellonella homogenate
to exhibit similar properties. The
HPLC-ESI mass spectra obtained by
rinsing the worm homogenate-treated
polyethylene with acetonitrile under
sonication do not show significant
peaks compared to the usual noise
level for this method, which can also
be seen from the respective spectrum
of the untreated control sample
(Figure S1G of [1]).
We cannot rule out the possibility
that the wax moth Galleria
mellonella might indeed be capable
of the chemical (rather than
physico-mechanical) destruction
of polyethylene but the present
publication does not provide
sufficient proof for this claim. In
our opinion, future studies need to
include 13C labeling experiments to
probe the formation of 13C-labeled
metabolites in the worms resulting
from the digestion of 13C–polyethylene,
ideally as a function of the time of
treatment/exposure. The weight loss
alone is clearly insufficient to prove
the proposed biodegradation of
polyethylene.
REFERENCES
1. Bombelli, P., Howe, C.J., and Bertocchini,
F. (2017). Polyethylene bio-degradation by
caterpillars of the wax moth Galleria mellonella.
Curr. Biol. 27, R292–R293.
2. Barth, A. (2007). Infrared spectroscopy of
proteins. Biochim. Biophys. Acta Bioenerg.
1767, 1073–1101.
Institute of Organic Chemistry, Johannes
Gutenberg University Mainz, Duesbergweg
10–14, 55128 Mainz, Germany.
*E-mail: opatz@uni-mainz.de
Correspondence
Response to
Weber et al.
Paolo Bombelli1,
Christopher J. Howe1,*,
and Federica Bertocchini2,3,*
A number of previous studies have
reported microbial degradation of
polyethylene [1,2]. Fourier transform
infrared spectroscopy analyses of
the products of degradation are, in
many cases, contradictory, especially
with regard to the relative intensities
of different signals, suggesting that
pathways are complex and may
differ among organisms [1,2]. A
detailed consideration of possible
degradation products and pathways
would have been beyond the
scope of our initial brief report [3].
Nevertheless, the peaks to which we
drew attention are consistent with
those generally described in other
studies.
Weber et al. [4] point out that
ethylene glycol has characteristic
absorbances at 1,085 and 1,038 cm–1
in addition to that at 3,300 cm–1. For
pure ethylene glycol those peaks are
quite sharp and defined, although
for ethylene glycol on polyethylene
they are rather less sharp (Figure 1
in [4]). In our Figure 1E (red trace)
[3], the spectrum for polyethylene
treated with worm homogenate
shows increased absorbance in
this region, although the increase
is modest in comparison with other
peaks in the spectrum, and broad
(as is also the case with the peak at
3,300 cm–1). A similar absorbance
increase around 1,050 cm–1 is seen
for the measurement close to the
site of breakdown by an intact worm,
compared to a measurement further
away (Figure S1E, red trace) [3]. We
acknowledge that the differences
are modest and merit further
investigation.
Weber et al. [4] suggest that the
peaks at 1,740 cm–1 to 1,545 cm–1 in
their experiments are due to residues
of organic material after treatment of
polyethylene with egg yolk or ground
pork. We note that the argument that
the different intensities of signals
ascribed to lipid and protein in egg
Current Biology 27, R731–R745, August 7, 2017 © 2017 Published by Elsevier Ltd. R745
yolk and pork can be accounted
for by a higher lipid content in pork
does not appear consistent with
USDA data. The mass ratio of lipid to
protein in egg yolk is 1.7:1, whereas
for fresh ground raw pork the ratio is
1.3:1 (https://ndb.nal.usda.gov/ndb/
search/list, accessed 12 June 2017).
Although we cannot completely
exclude the possibility that material
was left after treatment with worm
homogenate in our experiments, we
tried to remove residual material by
careful washing. We also observed
similar peaks in material at the site
of holes made by intact wax worms,
in the absence of homogenate (our
Figure S1E) [3].
It is not clear on what basis
Weber et al. [4] assert that weight
loss is “clearly insufficient” to prove
breakdown of the polyethylene.
However, the suggestion that the
weight loss is due to substantial
abrasion would seem to be
inconsistent with the argument that
the treatment actually leaves lipid
and protein behind. Nevertheless,
we agree that labeling studies
would be very desirable to confirm
polyethylene breakdown and identify
more accurately the nature of the
breakdown products, although in
our view the fact that polyethylene is
broken down at all is more significant
for our initial study than the precise
nature of the breakdown products.
REFERENCES
1. Restrepo-Florez, J-M., Bassi, A., and
Thompson, M.R. (2014). Microbial
degradation and deterioration of
polyethylene — a review. Int. Biodeterior.
Biodegradation 88, 83–90.
2. Yang, J., Yang, Y., Wu, W-M., Zhao, J. and
Jiang, L. (2014). Evidence of polyethylene
biodegradation by bacterial strains from the
guts of plastic-eating waxworms. Environ.
Sci. Tech. 48, 13776–13784.
3. Bombelli, P., Howe, C.J., and Bertocchini,
F. (2017). Polyethylene bio-degradation
by caterpillars of the wax moth Galleria
mellonella. Curr. Biol. 27, R292–R293.
4. Weber, C., Pusch, S., and Opatz, T. (2017).
Polyethylene bio-degradation by caterpillars?
Curr. Biol. 27, R744–R745.
1
Department of Biochemistry, University
of Cambridge, Downing Site, Tennis
Court Road, Cambridge, UK. 2Instituto
de Biomedicina y Biotecnologia de
Cantabria-CSIC-Universidad de Cantabria-
SODERCAN, Av.da A. Einstein, Santander,
Spain. 3Lead contact.
*E-mail: ch26@cam.ac.uk (C.J.H.);
bertocchinif@unican.es (F.B.)