More Like this

1. Environmental Consortium Containing Pseudomonas and Bacillus Species Synergistically Degrades Polyethylene Terephthalate Plastic

   https://doi.org/https://doi.org/10.1128/mSphere.01151-20.  

   Cameron Roberts; Sabrina Edwards; Morgan Vague; Rosa León-Zayas; Henry Scheffer; Gayle Chan; Natasja A. Swartz; Jay L. Mellies (September 2022, mSphere)

   Katherine McMahon, University of (Ed.)

   Plastics, such as polyethylene terephthalate (PET) from water bottles, are polluting our oceans, cities, and soils. While a number of Pseudomonas species have been described that degrade aliphatic polyesters, such as polyethylene (PE) and polyurethane (PUR), few from this genus that degrade the semiaromatic poly- mer PET have been reported. In this study, plastic-degrading bacteria were isolated from petroleum-polluted soils and screened for lipase activity that has been associ- ated with PET degradation. Strains and consortia of bacteria were grown in a liquid carbon-free basal medium (LCFBM) with PET as the sole carbon source. We moni- tored several key physical and chemical properties, including bacterial growth and modi!cation of the plastic surface, using scanning electron microscopy (SEM) and attenuated total re"ectance-Fourier transform infrared spectroscopy (ATR-FTIR) spec- troscopy. We detected by-products of hydrolysis of PET using 1H-nuclear magnetic resonance (1H NMR) analysis, consistent with the ATR-FTIR data. The full consortium of !ve strains containing Pseudomonas and Bacillus species grew synergistically in the presence of PET and the cleavage product bis(2-hydroxyethyl) terephthalic acid (BHET) as sole sources of carbon. Secreted enzymes extracted from the full consor- tium were capable of fully converting BHET to the metabolically usable monomers terephthalic acid (TPA) and ethylene glycol. Draft genomes provided evidence for mixed enzymatic capabilities between the strains for metabolic degradation of TPA and ethylene glycol, the building blocks of PET polymers, indicating cooperation and ability to cross-feed in a limited nutrient environment with PET as the sole carbon source. The use of bacterial consortia for the biodegradation of PET may provide a partial solution to widespread planetary plastic accumulation. 

   more » « less
2. Breakdown of polyethylene therepthalate microplastics under saltwater conditions using engineered Vibrio natriegens

   https://doi.org/10.1002/aic.18228  

   Li, Tianyu; Menegatti, Stefano; Crook, Nathan (September 2023, AIChE Journal)

   Abstract Poly(ethylene terephthalate) (PET) is a highly recyclable plastic that has been extensively used and manufactured. Like other plastics, PET resists natural degradation, thus accumulating in the environment. Several recycling strategies have been applied to PET, but these tend to result in downcycled products that eventually end up in landfills. This accumulation of landfilled PET waste contributes to the formation of microplastics, which pose a serious threat to marine life and ecosystems, and potentially to human health. To address this issue, our project leveraged synthetic biology to develop a whole‐cell biocatalyst capable of depolymerizing PET in seawater environments by using the fast‐growing, nonpathogenic, moderate halophileVibrio natriegens. By leveraging a two‐enzyme system—comprising a chimera ofIsPETase andIsMHETase fromIdeonella sakaiensis—displayed onV. natriegens, we constructed whole‐cell catalysts that depolymerize PET and convert it into its monomers in salt‐containing media and at a temperature of 30°C. 

   more » « less
3. Two-Step Chemo-Microbial Degradation of Post-Consumer Polyethylene Terephthalate (PET) Plastic Enabled by a Biomass-Waste Catalyst

   https://doi.org/10.3390/bioengineering10111253  

   Shingwekar, Deepika; Laster, Helen; Kemp, Hannah; Mellies, Jay L (November 2023, Bioengineering)

   Polyethylene terephthalate (PET) pollution has significant environmental consequences; thus, new degradation methods must be explored to mitigate this problem. We previously demonstrated that a consortium of three Pseudomonas and two Bacillus species can synergistically degrade PET in culture. The consortium more readily consumes bis(2-hydroxyethyl) terephthalate (BHET), a byproduct created in PET depolymerization, compared to PET, and can fully convert BHET into metabolically usable monomers, namely terephthalic acid (TPA) and ethylene glycol (EG). Because of its crystalline structure, the main limitation of the biodegradation of post-consumer PET is the initial transesterification from PET to BHET, depicting the need for a transesterification step in the degradation process. Additionally, there have been numerous studies done on the depolymerization reaction of PET to BHET, yet few have tested the biocompatibility of this product with a bacterial consortium. In this work, a two-step process is implemented for sustainable PET biodegradation, where PET is first depolymerized to form BHET using an orange peel ash (OPA)-catalyzed glycolysis reaction, followed by the complete degradation of the BHET glycolysis product by the bacterial consortium. Results show that OPA-catalyzed glycolysis reactions can fully depolymerize PET, with an average BHET yield of 92% (w/w), and that the reaction product is biocompatible with the bacterial consortium. After inoculation with the consortium, 19% degradation of the glycolysis product was observed in 2 weeks, for a total degradation percentage of 17% when taking both steps into account. Furthermore, the 10-week total BHET degradation rate was 35%, demonstrating that the glycolysis products are biocompatible with the consortium for longer periods of time, for a total two-step degradation rate of 33% over 10 weeks. While we predict that complete degradation is achievable using this method, further experimentation with the consortium can allow for a circular recycling process, where TPA can be recovered from culture media and reused to create new materials. 

   more » « less
4. Dihydroxyterephthalate—A Trojan Horse PET Counit for Facile Chemical Recycling

   https://doi.org/10.1002/adma.202210154  

   Lee, Ting‐Han; Forrester, Michael; Wang, Tung‐ping; Shen, Liyang; Liu, Hengzhou; Dileep, Dhananjay; Kuehl, Baker; Li, Wenzhen; Kraus, George; Cochran, Eric (March 2023, Advanced Materials)

   Here, low-energy poly(ethylene terephthalate) (PET) chemical recycling in water: PET copolymers with diethyl 2,5-dihydroxyterephthalate (DHTE) undergo selective hydrolysis at DHTE sites, autocatalyzed by neighboring group participation, is demonstrated. Liberated oligomeric subchains further hydrolyze until only small molecules remain. Poly(ethylene terephthalate-stat-2,5-dihydroxyterephthalate) copolymers were synthesized via melt polycondensation and then hydrolyzed in 150–200 °C water with 0–1 wt% ZnCl2, or alternatively in simulated sea water. Degradation progress follows pseudo-first order kinetics. With increasing DHTE loading, the rate constant increases monotonically while the thermal activation barrier decreases. The depolymerization products are ethylene glycol, terephthalic acid, 2,5-dihydroxyterephthalic acid, and bis(2-hydroxyethyl) terephthalate dimer, which could be used to regenerate virgin polymer. Composition-optimized copolymers show a decrease of nearly 50% in the Arrhenius activation energy, suggesting a 6-order reduction in depolymerization time under ambient conditions compared to that of PET homopolymer. This study provides new insight to the design of polymers for end-of-life while maintaining key properties like service temperature and mechanical properties. Moreover, this chemical recycling procedure is more environmentally friendly compared to traditional approaches since water is the only needed material, which is green, sustainable, and cheap. 

   more » « less
5. Hydrophobic Residues Promote Interfacial Activation of Candida rugosa Lipase: A Study of Rotational Dynamics

   https://doi.org/10.1021/acs.langmuir.4c02174  

   Waggett, Ava; Pfaendtner, Jim (August 2024, Langmuir)

   Microbial lipases constitute a class of biocatalysts with the ability to cleave ester linkages of long-chain triglycerides. This property makes them particularly attractive for industrial applications ranging from food processing to pharmaceutical preparation. Among such enzymes, Candida rugosa lipase (CRL) is one of the most frequently used in biotransformation. A notable feature of CRL, among many lipases, is its propensity for interfacial activation: these enzymes exhibit elevated catalytic rates when acting at the interface between aqueous and hydrophobic phases. Notably, this phenomenon can be attributed to the presence of a mobile lid domain, which in its closed state occludes the enzyme active site. To advance our understanding of interfacial activation, we explore the dynamics of CRL rotation at the octane–water interface in this work. To do so, we employ molecular dynamics and umbrella sampling to evaluate the free energy of rotation of the enzyme at the interface. We identify a global minimum in the rotational landscape that coincides with lid opening at the interface. Additionally, we investigate the role of surface residues outside the lid domain as they serve to instigate rotation of the lid toward the aqueous phase. In doing so, we identify a patch of leucine residues which when mutated to glycine impose a barrier to rotation that maintains the enzyme in the inactive (closed lid) state on the order of 1 μs. Importantly, this study presents a novel quantification of the rotational free energy corresponding to CRL lid opening at the octane–water interface. The accompanying mutagenesis study likewise clarifies the role of hydrophobic surface residues in the transition. As such, this work provides valuable insight into the phenomenon of interfacial activation that might open up new avenues for manipulating the microenvironment of industrially relevant lipases, affording enhanced control over the enzyme state. 

   more » « less