Mechanisms of Polyethylene Terephthalate Pellet Fragmentation into Nanoplastics and Assimilable Carbons by Wastewater Comamonas

Wilkes, Rebecca A.; Zhou, Nanqing; Carroll, Austin L.; Aryal, Ojaswi; Teitel, Kelly P.; Wilson, Rebecca S.; Zhang, Lichun; Kapoor, Arushi; Castaneda, Edgar; Guss, Adam M.; Waldbauer, Jacob R.; Aristilde, Ludmilla

doi:10.1021/acs.est.4c06645

Mechanisms of Polyethylene Terephthalate Pellet Fragmentation into Nanoplastics and Assimilable Carbons by Wastewater Comamonas

Journal Article · Thu Oct 03 00:00:00 EDT 2024 · Environmental Science and Technology

DOI:https://doi.org/10.1021/acs.est.4c06645· OSTI ID:2455044

Wilkes, Rebecca A. ^[1]; Zhou, Nanqing ^[1]; Carroll, Austin L. ^[2]; Aryal, Ojaswi ^[1]; ^[1]; Wilson, Rebecca S. ^[1]; Zhang, Lichun ^[3]; Kapoor, Arushi ^[1]; Castaneda, Edgar ^[1]; ^[2]; ^[3]; ^[1]

Northwestern University, Evanston, IL (United States)
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
University of Chicago, IL (United States)

Here, Comamonadaceae bacteria are enriched on poly- (ethylene terephthalate) (PET) microplastics in wastewaters and urban rivers, but the PET-degrading mechanisms remain unclear. Here, we investigated these mechanisms with Comamonas testosteroniKF-1, a wastewater isolate, by combining microscopy, spectroscopy, proteomics, protein modeling, and genetic engineering. Compared to minor dents on PET films, scanning electron microscopy revealed significant fragmentation of PET pellets, resulting in a 3.5-fold increase in the abundance of small nanoparticles (<100 nm) during 30-day cultivation. Infrared spectroscopy captured primarily hydrolytic cleavage in the fragmented pellet particles. Solution analysis further demonstrated double hydrolysis of a PET oligomer, bis(2-hydroxyethyl) terephthalate, to the bioavailable monomer terephthalate. Supplementation with acetate, a common wastewater co-substrate, promoted cell growth and PET fragmentation. Of the multiple hydrolases encoded in the genome, intracellular proteomics detected only one, which was found in both acetate-only and PET-only conditions. Homology modeling of this hydrolase structure illustrated substrate binding analogous to reported PET hydrolases, despite dissimilar sequences. Mutants lacking this hydrolase gene were incapable of PET oligomer hydrolysis and had a 21% decrease in PET fragmentation; re-insertion of the gene restored both functions. Thus, we have identified constitutive production of a key PET-degrading hydrolase in wastewater Comamonas, which could be exploited for plastic bioconversion.

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Research Organization:: Northwestern Univ., Evanston, IL (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Energy Efficiency Office. Advanced Materials & Manufacturing Technologies Office (AMMTO); U.S. National Science Foundation (NSF); USDOE

Grant/Contract Number:: AC05-00OR22725; CHE-2109097

OSTI ID:: 2455044

Alternate ID(s):: OSTI ID: 2475496; OSTI ID: 2498430

Journal Information:: Environmental Science and Technology, Vol. 58, Issue 43; ISSN 0013-936X

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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Mechanisms of Polyethylene Terephthalate Pellet Fragmentation into Nanoplastics and Assimilable Carbons by Wastewater Comamonas

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