Abstract
Plastics are a serious cause of environmental pollution, and microplastics pose a threat to human health. To solve this problem, the plastic-degrading mechanism of insect larvae is being investigated. The aim of this study was to examine the metabolic pathways involved in polyethylene metabolism, the interaction between the host and microorganisms, and the role of superworms in promoting plastic degradation in polyethylene-fed superworms. Through host transcriptomic analysis, we identified 429 up-regulated and 777 down-regulated genes and analyzed their functions using the Kyoto Encyclopedia of Genes and Genomes and Gene Ontology databases. We found that insects promote the degradation of polyethylene through two main mechanisms. First, polyethylene metabolites activate the lipid metabolism pathway in insects, promoting the synthesis of carboxylic ester hydrolases and accelerating polyethylene degradation. Second, insect larvae generate reactive oxygen species (ROS) which are critical for insect immune responses and for the initial oxidation of polyethylene. In metagenomic analysis, bacterial species, such as Citrobacter sp. and Raoultella sp., which are known to be involved in the degradation of polyethylene and its metabolites, were more abundant in the guts of insects that consumed polyethylene. In addition, increases in the concentration of peroxide in the gut and the activity of esterase (lipase) acting on lipophilic substrates were observed. Furthermore, we suggest that xenobiotic metabolism is critical for polyethylene metabolism in superworm guts. In particular, enzymes involved in xenobiotic metabolism phase 2, such as glutathione S-transferase and uridine diphosphate glycosyltransferase, convert lipophilic plastic degradation intermediates into water-soluble forms and promote polyethylene degradation.
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Data Availability
The RNA sequence data have been deposited in the NCBI database under ID codes PRJNA954984, SRR24150553–SRR24150558.
References
Tournier V, Duquesne S, Guillamot F, Cramail H, Taton D, Marty A, Andre I (2023) : Enzymes’ Power for Plastics Degradation. Chem Rev
Ali SS, Elsamahy T, Al-Tohamy R, Zhu D, Mahmoud YA, Koutra E, Metwally MA, Kornaros M, Sun J (2021) Plastic wastes biodegradation: mechanisms, challenges and future prospects. Sci Total Environ 780:146590
Yang J, Yang Y, Wu WM, Zhao J, Jiang L (2014) Evidence of polyethylene biodegradation by bacterial strains from the guts of plastic-eating waxworms. Environ Sci Technol 48(23):13776–13784
Yang Y, Yang J, Wu WM, Zhao J, Song Y, Gao L, Yang R, Jiang L (2015) Biodegradation and Mineralization of Polystyrene by Plastic-Eating Mealworms: part 1. Chemical and physical characterization and isotopic tests. Environ Sci Technol 49(20):12080–12086
Peng BY, Chen Z, Chen J, Yu H, Zhou X, Criddle CS, Wu WM, Zhang Y (2020) Biodegradation of polyvinyl chloride (PVC) in Tenebrio molitor (Coleoptera: Tenebrionidae) larvae. Environ Int 145:106106
Yang Y, Wang J, Xia M (2020) Biodegradation and mineralization of polystyrene by plastic-eating superworms Zophobas atratus. Sci Total Environ 708:135233
Wang Y, Luo L, Li X, Wang J, Wang H, Chen C, Guo H, Han T, Zhou A, Zhao X (2022) Different plastics ingestion preferences and efficiencies of superworm (Zophobas atratus Fab.) And yellow mealworm (Tenebrio molitor Linn.) Associated with distinct gut microbiome changes. Sci Total Environ 837:155719
Cassone BJ, Grove HC, Elebute O, Villanueva SMP, LeMoine CMR (2020) Role of the intestinal microbiome in low-density polyethylene degradation by caterpillar larvae of the greater wax moth, Galleria mellonella. Proc Biol Sci 287(1922):20200112
Yang Y, Yang J, Wu WM, Zhao J, Song Y, Gao L, Yang R, Jiang L (2015) Biodegradation and Mineralization of Polystyrene by Plastic-Eating Mealworms: part 2. Role of gut microorganisms. Environ Sci Technol 49(20):12087–12093
Kim HR, Lee HM, Yu HC, Jeon E, Lee S, Li J, Kim DH (2020) Biodegradation of Polystyrene by Pseudomonas sp. Isolated from the gut of Superworms (Larvae of Zophobas atratus). Environ Sci Technol 54(11):6987–6996
Luo L, Wang Y, Guo H, Yang Y, Qi N, Zhao X, Gao S, Zhou A (2021) Biodegradation of foam plastics by Zophobas atratus larvae (Coleoptera: Tenebrionidae) associated with changes of gut digestive enzymes activities and microbiome. Chemosphere 282:131006
Lou Y, Ekaterina P, Yang SS, Lu B, Liu B, Ren N, Corvini PF, Xing D (2020) Biodegradation of Polyethylene and Polystyrene by Greater Wax Moth Larvae (Galleria mellonella L.) and the Effect of Co-diet Supplementation on the Core Gut Microbiome. Environ Sci Technol 54(5):2821–2831
Brandon AM, Gao SH, Tian R, Ning D, Yang SS, Zhou J, Wu WM, Criddle CS (2018) Biodegradation of Polyethylene and Plastic Mixtures in Mealworms (Larvae of Tenebrio molitor) and Effects on the gut Microbiome. Environ Sci Technol 52(11):6526–6533
Cassone BJ, Grove HC, Kurchaba N, Geronimo P, LeMoine CMR (2022) Fat on plastic: metabolic consequences of an LDPE diet in the fat body of the greater wax moth larvae (Galleria mellonella). J Hazard Mater 425:127862
Sanluis-Verdes A, Colomer-Vidal P, Rodriguez-Ventura F, Bello-Villarino M, Spinola-Amilibia M, Ruiz-Lopez E, Illanes-Vicioso R, Castroviejo P, Aiese Cigliano R, Montoya M et al (2022) Wax worm saliva and the enzymes therein are the key to polyethylene degradation by Galleria mellonella. Nat Commun 13(1):5568
Kundungal H, Gangarapu M, Sarangapani S, Patchaiyappan A, Devipriya SP (2019) Efficient biodegradation of polyethylene (HDPE) waste by the plastic-eating lesser waxworm (Achroia grisella). Environ Sci Pollut Res Int 26(18):18509–18519
Brandon AM, Garcia AM, Khlystov NA, Wu WM, Criddle CS (2021) Enhanced bioavailability and Microbial Biodegradation of Polystyrene in an Enrichment Derived from the gut microbiome of Tenebrio molitor (Mealworm Larvae). Environ Sci Technol 55(3):2027–2036
Zhong Z, Nong W, Xie Y, Hui JHL, Chu LM (2022) Long-term effect of plastic feeding on growth and transcriptomic response of mealworms (Tenebrio molitor L). Chemosphere 287(Pt 1):132063
Kong HG, Kim HH, Chung JH, Jun J, Lee S, Kim HM, Jeon S, Park SG, Bhak J, Ryu CM (2019) The Galleria mellonella Hologenome supports microbiota-independent metabolism of Long-Chain Hydrocarbon Beeswax. Cell Rep 26(9):2451–2464e2455
Peng BY, Li Y, Fan R, Chen Z, Chen J, Brandon AM, Criddle CS, Zhang Y, Wu WM (2020) Biodegradation of low-density polyethylene and polystyrene in superworms, larvae of Zophobas atratus (Coleoptera: Tenebrionidae): broad and limited extent depolymerization. Environ Pollut 266(Pt 1):115206
Langmead B, Trapnell C, Pop M, Salzberg SL (2009) Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol 10(3):R25
Li B, Dewey CN (2011) RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinformatics 12:323
Li G, Shi M, Zhao S, Li D, Long Y, Yang C, Zhu Y (2020) RNA-Seq comparative analysis reveals the response of Enterococcus faecalis TV4 under fluoride exposure. Gene 726:144197
Li G, Shi M, Zhao S, Long Y, Zhu Y (2019) Toxicity response of silkworm intestine to Bacillus cereus SW7-1 pathogen. Sci Total Environ 692:1282–1290
Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Pena AG, Goodrich JK, Gordon JI et al (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7(5):335–336
Navas-Molina JA, Peralta-Sanchez JM, Gonzalez A, McMurdie PJ, Vazquez-Baeza Y, Xu Z, Ursell LK, Lauber C, Zhou H, Song SJ et al (2013) Advancing our understanding of the human microbiome using QIIME. Methods Enzymol 531:371–444
Kuczynski J, Stombaugh J, Walters WA, Gonzalez A, Caporaso JG, Knight R (2012) : Using QIIME to analyze 16S rRNA gene sequences from microbial communities. Curr Protoc Microbiol Chap. 1:Unit 1E 5.
Edgar RC (2010) Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26(19):2460–2461
Rognes T, Flouri T, Nichols B, Quince C, Mahe F (2016) VSEARCH: a versatile open source tool for metagenomics. PeerJ 4:e2584
Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glockner FO (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41(Database issue):D590–596
Price MN, Dehal PS, Arkin AP (2009) FastTree: computing large minimum evolution trees with profiles instead of a distance matrix. Mol Biol Evol 26(7):1641–1650
Caporaso JG, Bittinger K, Bushman FD, DeSantis TZ, Andersen GL, Knight R (2010) PyNAST: a flexible tool for aligning sequences to a template alignment. Bioinformatics 26(2):266–267
Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73(16):5261–5267
McDonald D, Clemente JC, Kuczynski J, Rideout JR, Stombaugh J, Wendel D, Wilke A, Huse S, Hufnagle J, Meyer F et al (2012) The Biological Observation Matrix (BIOM) format or: how I learned to stop worrying and love the ome-ome. GigaScience 1:7–7
Soergel DA, Dey N, Knight R, Brenner SE (2012) Selection of primers for optimal taxonomic classification of environmental 16S rRNA gene sequences. ISME J 6(7):1440–1444
Saito M, Seki M, Iida K, Nakayama H, Yoshida S (2007) A novel agar medium to detect hydrogen peroxide-producing bacteria based on the prussian blue-forming reaction. Microbiol Immunol 51(9):889–892
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Adlan NA, Sabri S, Masomian M, Ali MSM, Rahman R (2020) Microbial Biodegradation of Paraffin Wax in malaysian crude oil mediated by degradative enzymes. Front Microbiol 11:565608
Zadjelovic V, Erni-Cassola G, Obrador-Viel T, Lester D, Eley Y, Gibson MI, Dorador C, Golyshin PN, Black S, Wellington EMH et al (2022) A mechanistic understanding of polyethylene biodegradation by the marine bacterium Alcanivorax. J Hazard Mater 436:129278
Lamb CA, Dooley HC, Tooze SA (2013) Endocytosis and autophagy: Shared machinery for degradation. BioEssays 35(1):34–45
Esteves F, Rueff J, Kranendonk M (2021) The Central Role of Cytochrome P450 in Xenobiotic Metabolism-A brief review on a fascinating enzyme family. J Xenobiot 11(3):94–114
Zampolli J, Orro A, Manconi A, Ami D, Natalello A, Di Gennaro P (2021) Transcriptomic analysis of Rhodococcus opacus R7 grown on polyethylene by RNA-seq. Sci Rep 11(1):21311
Koirala BKS, Moural T, Zhu F (2022) Functional and structural diversity of insect glutathione S-transferases in Xenobiotic Adaptation. Int J Biol Sci 18(15):5713–5723
Mohanan N, Montazer Z, Sharma PK, Levin DB (2020) Microbial and enzymatic degradation of Synthetic Plastics. Front Microbiol 11:580709
Weers PM, Ryan RO (2006) Apolipophorin III: role model apolipoprotein. Insect Biochem Mol Biol 36(4):231–240
Borst P, Zelcer N, van Helvoort A (2000) ABC transporters in lipid transport. Biochim Biophys Acta 1486(1):128–144
Pols MS, Klumperman J (2009) Trafficking and function of the tetraspanin CD63. Exp Cell Res 315(9):1584–1592
Patnaik BB, Kang SM, Seo GW, Lee HJ, Patnaik HH, Jo YH, Tindwa H, Lee YS, Lee BL, Kim NJ et al (2013) Molecular cloning, sequence characterization and expression analysis of a CD63 homologue from the coleopteran beetle, Tenebrio molitor. Int J Mol Sci 14(10):20744–20767
Arockiaraj J, Gnanam AJ, Muthukrishnan D, Thirumalai MK, Pasupuleti M, Milton J, Kasi M (2013) Macrobrachium rosenbergii cathepsin L: molecular characterization and gene expression in response to viral and bacterial infections. Microbiol Res 168(9):569–579
Mettlen M, Chen PH, Srinivasan S, Danuser G, Schmid SL (2018) Regulation of clathrin-mediated endocytosis. Annu Rev Biochem 87:871–896
Kuraishi T, Binggeli O, Opota O, Buchon N, Lemaitre B (2011) Genetic evidence for a protective role of the peritrophic matrix against intestinal bacterial infection in Drosophila melanogaster. Proc Natl Acad Sci U S A 108(38):15966–15971
Li J, Kim HR, Lee HM, Yu HC, Jeon E, Lee S, Kim DH (2020) Rapid biodegradation of polyphenylene sulfide plastic beads by Pseudomonas sp. Sci Total Environ 720:137616
Collins JW, Keeney KM, Crepin VF, Rathinam VA, Fitzgerald KA, Finlay BB, Frankel G (2014) Citrobacter rodentium: infection, inflammation and the microbiota. Nat Rev Microbiol 12(9):612–623
Wirth F, Goldani LZ (2012) Epidemiology of Rhodotorula: an emerging pathogen. Interdiscip Perspect Infect Dis 2012:465717
Chen WJ, Hsieh FC, Hsu FC, Tasy YF, Liu JR, Shih MC (2014) Characterization of an insecticidal toxin and pathogenicity of Pseudomonas taiwanensis against insects. PLoS Pathog 10(8):e1004288
Mikonranta L, Mappes J, Kaukoniitty M, Freitak D (2014) Insect immunity: oral exposure to a bacterial pathogen elicits free radical response and protects from a recurring infection. Front Zool 11(1):23
Yeom SJ, Le TK, Yun CH (2022) P450-driven plastic-degrading synthetic bacteria. Trends Biotechnol 40(2):166–179
Mittapalli O, Neal JJ, Shukle RH (2007) Antioxidant defense response in a galling insect. Proc Natl Acad Sci U S A 104(6):1889–1894
Wei R, Zimmermann W (2017) Microbial enzymes for the recycling of recalcitrant petroleum-based plastics: how far are we? Microb Biotechnol 10(6):1308–1322
Amezian D, Nauen R, Le Goff G (2021) Transcriptional regulation of xenobiotic detoxification genes in insects - an overview. Pestic Biochem Physiol 174:104822
Kim HR, Lee C, Shin H, Kim J, Jeong M, Choi D (2023) Isolation of a polyethylene-degrading bacterium, Acinetobacter guillouiae, using a novel screening method based on a redox indicator. Heliyon 9(5):e15731
Funding
This work was supported by the Tech Incubator Program for Startup (TIPS) funded by the Ministry of SMEs (Small and Medium Enterprises) and Startups, Republic of Korea (No. S3136229).
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Hong Rae Kim: Conceptualization, Investigation, Data Curation, Visualization, Writing – Original Draft, Chaerin Lee: Investigation, Hyeyoung Shin: Investigation, Hye Yeon Koh: Investigation, Sukkyoo Lee: Supervision, Writing – Review & Editing, Donggeon Choi: Validation, Investigation, Project administration, Writing – Review & Editing.
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Kim, H.R., Lee, C., Shin, H. et al. Transcriptomic Response of Superworm in Facilitating Polyethylene Biodegradation. J Polym Environ 32, 1658–1671 (2024). https://doi.org/10.1007/s10924-023-03029-z
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DOI: https://doi.org/10.1007/s10924-023-03029-z