Supporting Information

Biodegradation and Mineralization of Polystyrene by

Plastic-Eating Mealworms. 2. Role of Gut
Microorganisms

Yu Yang1, Jun Yang1, *, Wei-Min Wu2, Jiao Zhao3, Yiling Song4, Longcheng Gao1,

Ruifu Yang3, Lei Jiang1, *

1
Key Laboratory of Bio-Inspired Smart Interfacial Science and Technology of

Ministry of Education, School of Chemistry and Environment, Beihang University,

Beijing 100191, P. R. China.

2
Department of Civil and Environmental Engineering, William & Cloy Codiga

Resource Recovery Research Center, Center for Sustainable Development & Global

Competitiveness, Stanford University, Stanford, California 94305-4020, USA.

3
Shenzhen Key Laboratory of Bioenergy, BGI-Shenzhen, Shenzhen 518083, P. R.

China.

4
School of Biological Science and Medical Engineering, Beihang University, Beijing

100191, P. R. China.

7 pages, 4 figures, and 2 tables

1
Supplementary Figures

Figure S1. Inhibition of the growth of bacteria extracted from the guts of mealworms
in the presence of 6 different antibiotics. (a) Ampicillin with strong halo of bacterial
colony inhibition (1.6 ± 0.2 cm). (b) Chloramphenicol without distinct halo of
bacterial colony inhibition. (c) Erythromycin without distinct halo of bacterial colony
inhibition. (d) Gentamicin with strong halo of bacterial colony inhibition (2.4 ± 0.1
cm). (e) Tetracycline with strong halo of bacterial colony inhibition (2.3 ± 0.1 cm). (f)
Vancomycine without distinct halo of bacterial colony inhibition. The central disk in
each plate was the purified water control and the surrounding disks were added
respective antibiotic at dosage of 30 µg per disk. Based on above results, we selected
gentamicin as the antibiotic to feed the mealworm larvae in order to eliminate gut
bacteria.

2
Figure S2. Change in bacterial cell number colonized on the PS film samples
incubated with thirteen different isolated bacterial cultures after 28 days. The strain
YT2 showed the best performance among all isolates.

3
Figure S3. 16S RNA gene-based Neighbor-joining phylogenetic tree of strain YT2.
The numbers above the branches are support value obtained from 1,000 bootstrap
replicates. Percentages of 16S RNA gene nucleotide sequence identities between
strain YT2 and each of the comparison species are shown in parentheses. The results
indicated that the isolated strain YT2 belongs to Exiguobacterium genus.

4
Figure S4. Gas chromatograph of extract from the culture inoculated with strain YT2
and the uninoculated control after 60 days.

5
Supplementary Tables

Table S1 Thirteen bacterial strains isolated from the PS-enrichment from

Styrofoam-eating mealworm gut content with PS as sole carbon source.
Strain Possible genus Highest-homology organism Identity, %

YT2 Exiguobacterium sp. Exiguobacterium indicum HHS31(T) 99

YT3 Chryseobacterium sp. Chryseobacterium vietnamense GIMN1.005(T) 97

YHE1 Klebsiella pneumonia Klebsiella pneumoniae JCM 1662(T) 99

YHE2 Bacillus cereus Bacillus cereus ATCC 14579(T) 99

YHE3 Morganella morganii Morganella morganii DSM 14850(T) 97

YHE4 Enterobacter hormaechei Enterobacter hormaechei ATCC 49162(T) 99

YHE5 Proteus vulgaris Proteus vulgaris ATCC 29905(T) 99

YHE9 Enterobacter cancerogenus Enterobacter cancerogenus LMG 2693(T) 99

YHE12 Enterococcus viikkiensis Enterococcus viikkiensis IE3.2(T) 97

YHE13 Enterococcus faecalis Enterococcus faecalis JCM 5803(T) 99

YHE14 Bacillus stratosphericus Bacillus stratosphericus 41KF2a(T) 99

YHE27 Citrobacter freundii Citrobacter freundii DSM 30039(T) 99

YHE28 Enterococcus gallinarum Enterococcus gallinarum CECT970(T) 99

6
Table S2. GC/MS identification of water soluble products released from PS pieces in
the presence and absence of the activities of strain YT2 after 60 days. (Control=in the
absence of strain YT2)

Retention Proposed Similarity,

Mass Potentially proposed chemical Control YT2
time, min formula %
3.479 116 C6H12O2 2-Pentanone, 4-hydroxy-4-methyl- 80 − +
4.877 132 C6H12O3 (R)-(-)-2,2-Dimethyl-1,3-dioxolane-4-methanol 91 − +
10.392 286 C16H30O4 Oxalic acid, butyl 6-ethyloct-3-yl ester 72 − +
*
13.048 206 C14H22O Phenol, 2,4-bis(1,1-dimethylethyl)- 97 − +
1,2-Benzenedicarboxylic acid, butyl
18.023* 278 C16H22O4 96 − +
2-methylpropyl ester

* indicates the phenyl derivatives.