Degradation Of Polyethylene Terephthalate Research Articles

Selective Degradation of Polyethylene Terephthalate Plastic Waste Using Iron Salt Photocatalysts.

Plastic pollution poses a significant challenge to environmental conservation. Efficient recycling of plastic is a key strategy to address this issue. Polyethylene terephthalate (PET), commonly found in plastic bottles, represents a substantial portion of plastic waste. Consequently, the efficient degradation and recycling of PET is crucial for the sustainable development of society. However, the implementation of methods for PET depolymerization and recycling typically necessitates alkaline/acidic pre-treatment and significant energy input for heating. Here, we propose a gentle, and highly efficient photocatalysis approach for selectively degrading PET plastic waste into terephthalic acid (TPA) in high yield (up to 99%) using cost-effective iron salts. Notably, this method achieved excellent selectivity with high TON and TOF values, applying oxygen or air as environmentally friendly oxidants. In addition, the solvent can be recycled without compromising the TPA yield, and large-scale reactions can be performed smoothly.

Structural dynamics of the Ca2+-regulated cutinase towards structure-based improvement of PET degradation activity

We previously revealed the structural basis of Ca2+ dependent regulation of a polyethylene terephthalate (PET)-degrading enzyme, Cut190, and proposed a unique reaction cycle in which the enzyme repeatedly binds and releases Ca2+. Here, we report crystal structures of Cut190 mutants with high thermal stability complexed with PET-like ligands that contain aromatic rings. The structural information has allowed us to perform further computational analyses using a PET-trimer bound model. Our multicanonical molecular dynamics simulations and subsequent analyses of the free energy landscapes revealed a novel intermediate form that occurs during the enzymatic reaction cycle. Furthermore, the computational analyses were used to investigate the effect of the point mutations F77L and F81L in the Ca2+-binding site, which showed that the former stabilizes the engaged and open forms to improve transition between the open and active forms, while the latter extremely increases the open form. Subsequent experiments showed that the F77L mutation increased the activity, while the F81L mutation decreased the activity. Our computational analysis has enabled us to explore the dynamics of Cut190 on a completely new level, providing key insights into how the balance between the various conformations influences the reaction cycle and ultimately how to improve the reaction cycle.

Enhanced degradation of polyethylene terephthalate (PET) microplastics by an engineered Stenotrophomonas pavanii in the presence of biofilm

Polyethylene terephthalate (PET) microplastics pose significant environmental and human health risks due to their resistance to degradation and accumulation in ecosystems. In this study, we engineered Stenotrophomonas pavanii JWG-G1, a robust biofilm-forming bacterium, to overexpress the PET hydrolase (DuraPETase) for PET microplastics degradation at ambient temperature. Nine endogenous PET hydrolases were identified through genome sequencing of S. pavanii, and were successfully expressed in Escherichia coli BL21(DE3). Among them, hydrolase Est_B achieved 100% degradation of bis(2-hydroxyethyl) terephthalate (BHET) at an initial concentration of 0.23 mg/mL at 30 °C within 4 h, identifying it as a novel BHETase. However, the PET degradation performance of all endogenous PET hydrolases was inferior to that of DuraPETase. The engineered strain overexpressing DuraPETase demonstrated a significant enhancement in PET degradation, achieving a 38.04 μM total product release of high-crystallinity PET microplastics after 30 days at 30 °C. The degradation extent was greater than that of low biofilm-forming engineered strains, attributing to the aggregation of DuraPETase on the PET surface in the presence of biofilm. Additionally, this engineered strain also maintained PET degradation activity across various water environments and demonstrated effectiveness in degrading other polyester plastics. This is the first report demonstrating that an engineered strain of Stenotrophomonas species is capable of simultaneously secreting exogenous hydrolase and degrading polyester microplastics, representing a novel approach in the development of engineered bacteria with potential applications in bioreactor systems and environmental remediation.

Enhancing PET degradation efficiency through fusing carbohydrate-binding modules in LCC-ICCG

Polyethylene terephthalate (PET) is a largely-produced polymer worldwide. However, its extensive waste generation and resistance to degradation pose significant environmental concerns. Consequently, there is considerable interest in researching enzymatic degradation of PET. Relevant studies have shown that the addition of a carbohydrate binding module (CBM) can increase the affinity between the enzyme and the substrate, enhancing the enzyme's degradation ability. In order to develop more efficient PET hydrolytic enzymes, this study introduced carbohydrate binding domains (CBMs) from different families with different substrate affinities into the PET-degrading enzyme LCC-ICCG. High crystallinity PET powder and amorphous PET film were used as substrates to characterize the degradation efficiency of the modified enzymes, aiming to explore the enzyme with the optimal degradation ability. The results showed that the fusion of type B CBM reduced the degradation rate and Tm value of the enzyme towards PET, while the introduction of type A and type C CBMs significantly improved the degradation rate of the enzyme towards the film-like substrate. The degradation rate and Tm value of PET were also enhanced, especially with the fusion enzyme LCC-ICCG-CBM9-2, which showed an over 10-fold increase in the degradation rate compared to the original enzyme LCC-ICCG. Therefore, this study demonstrates that by introducing type A and type C CBMs, the degradation rate and thermal stability of LCC-ICCG towards film-like PET can be improved, addressing the issue of its low activity and enabling more effective PET degradation. This research provides support for plastic degradation technology and contributes to environmental conservation efforts.

Unravelling biochemical and molecular mechanism of a carboxylesterase from Dietzia kunjamensis IITR165 reveal novel activities against polyethylene terephthalate

Plastics and plasticizers accumulate in the ecological niches affecting biodiversity, and human and environmental health. Bacteria degrading polyethylene terephthalate (PET) were screened and PETases involved in PET degradation were characterized. Here, we identified a carboxylesterase Dkca1 of 48.44 kDa molecular mass from Dietzia kunjamensis IITR165 shown to degrade amorphous PET film into bis(2-hydroxyethyl) terephthalate (BHET) and terephthalic acid (TPA) formed 64.35 μM and 35.26 μM, respectively within 96 h at 37 °C as revealed by LC-MS analysis showed significant PET hydrolase activity similar to reported PETases. SEM analysis confirms the surface erosion as cavities and holes. Dkca1 also hydrolysed BHET and dibutyl phthalate (DBP) at a concentration of 1 mM within 3 h indicating its versatility. Fluorescence quenching shows Dkca1 protein has a maximum affinity (Kd) towards BHET (86.55 μM) than DBP (134.2 μM). The protein demonstrated high stability under temperatures above 40 °C and at the pH range of 6.0–9.0. Moreover, Amino acid composition showed that the Dkca1 enzyme belongs to family VII carboxylesterase containing conserved catalytic triad of Ser183-Glu289-His378 with pentapeptide motif GXSAG and an oxyanion hole H103GGG106, sharing 37.47 % and 32.44 % similarity with a PET hydrolase TfCa from Thermobifida fusca and PAE hydrolase CarEW from Bacillus sp. K91, respectively. A docking study revealed that ligand PET, BHET, and DBP showed favourable binding in the catalytic pocket of the Dkca1 protein.

An integrated approach for plastic polymer degradation by the gut bacterial resident of superworm, Zophobas morio (Coleoptera:Tenebrionidae).

The potential of superworm to remove certain plastic polymers has recently been noted. In this study, aerobic bacterial strains were isolated from the gut of Zophobas morio larvae which were fed with polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP), and polystyrene (PS) polymers. Strains P2 (Leminorella), P6 (Bacillus), P9 (Bacillus), and P5 (Citrobacter) were associated with the highest PS (2.7%), PP (1.3%), PET (1.1%), and PE (0.42%) weight loss after 28days, respectively. Pretreatments including thermal treatment (80°C for 10days), weathering (4months in the free environment), and nitric and sulfuric acids (1 N, 10days) improved the degradation of PE (1.3%), PET (1.9%), PP (5.2%), and PS (8.3%) by the same strains, respectively. Further analyses on the PS degradation by Leminorella sp. P2 revealed acid pretreatment promoted the formation of the C = C, C = O, and O-H functional groups. Surface irregularities, as well as a 3.6-fold increase in surface roughness, were observed in the PS film subjected to biodegradation. The contact angle dropped from 98.4° to 42.2° following the biodegradation. Bacterial depolymerization was confirmed by the 8.7% and 3.4% reduction of Mn and Mw and the change in polydispersity from 1.65 to 1.75. The results suggest that Zophobas morio microbiota in combination with abiotic pretreatment can be considered for plastic waste management.

Fast co-pyrolysis of corncob with plastics: Evaluation of thermal behavior using deconvolution procedure, kinetic analysis and product characterization

In this study, the co-pyrolysis of corncob (CC) with polyethylene terephthalate (PET) and polyvinyl chloride (PVC) was first carried out at different blending ratios in a concentrating photothermal TGA (CP-TGA) to gain insights into their thermal behavior, pyrolysis characteristics and kinetic analysis under high heating rate conditions (≥500°C/min). The change in comprehensive pyrolysis index (CPI) for CC/PET and CC/PVC blends was 514 % and 125 %, respectively, when their blending ratio was increased from 25 wt% to 75 wt%. The decomposition rate (Rp %/min) for CC/PET and CC/PVC blends with an increased heating rate showed a favorable co-pyrolysis process. Rapid heating leads to complexity and simultaneous overlapping of the co-pyrolysis components; therefore, the devolatilization phase (active pyrolysis zone) was divided into pseudo-reactions using the Fraser-Suzuki deconvolution function. The deconvolution results of the blend samples fitted well with experimental results (R2 ≥ 0.997). The three pseudo components of CC in the CC25PET75% blend relatively reduced the needed activation energy (E) for PET degradation to about 130 %. With the blending ratio increase, the synergistic activity of pseudo hemicellulose presented trends of an increasing E in CC/PET blends, whereas CC/PVC blends showed decreasing trends. The relative ΔEa deviation revealed that most blend samples would promote an active synergistic reaction. The resultant co-pyrolysis oil from photothermal fixed bed pyrolyzer contained promising bio-renewable platform chemicals such as levoglucosenone, furfural, D-allose, etc. The synergistic interactions between pseudo-lignin and PET contributed to a significant reduction in activation energy and higher selectivity of acetic acid due to the degradation of the ester bonds in PET and the carboxyl groups in lignin. The results of this work could serve as a guide for selecting appropriate reaction parameters for optimal co-pyrolysis synergism.

Advances in the structure and function of MHETase

Polyethylene terephthalate (PET) is one of the widely used plastics, but its waste pollution has become a global environmental issue. The discovery of polyethylene terephthalate hydrolase (PETase) has provided a green and environmentally friendly approach for PET degradation. However, PETase produces intermediate products that inhibit the enzyme's further activity, leading to a decrease in enzyme efficiency. Mono(2-hydroxyethyl) terephthalate hydrolase (MHETase) works synergistically with PETase to further degrade the intermediate product MHET into ethylene glycol (EG) and terephthalic acid (TPA). MHETase exhibits extremely high specificity for MHET and is crucial for the complete degradation of PET. This article comprehensively reviews MHETase from various perspectives, including its three-dimensional structure, substrate binding, and catalytic mechanism. It demonstrates the structural features and key residues associated with the enzyme's degrading activity and discusses the progress in enzyme engineering modifications. Additionally, the study envisions the development of a two-enzyme PET degradation system by combining MHETase with PETase, aiming to provide valuable references for designing and developing more efficient PET hydrolytic enzyme systems.

Targeted aggregation of PETase towards surface of Stenotrophomonas pavanii for degradation of PET microplastics

Polyethylene terephthalate (PET) is one of the most widely used plastics, but its fragmentation into microplastics poses significant environmental challenges. The recycling of PET microplastics is hindered by their low solubility and widespread dispersion in the environment, making microbial in-situ degradation a promising solution. However, existing PET-degrading strains exhibited the limited effectiveness, primarily due to the diffusion of secreted hydrolases away from the PET surface. In this study, Stenotrophomonas pavanii JWG-G1 was engineered to achieve the targeted aggregation of PET hydrolase PETase on the cell surface by fusing it with an endogenous anchor protein. This approach aims to maximise the local concentration of PETase around PET, thereby increasing the overall rate of PET degradation. The PETase surface-aggregated system, S. pavanii/PaL-PETase, demonstrated the highest degradation efficiency, achieving 63.3 % degradation of low-crystallinity PET (lcPET) and 27.3 % degradation of high-crystallinity PET bottles (hcPET) at 30 °C. This represents the highest degradation rate reported for a displayed whole-cell system at ambient temperature. Furthermore, this system exhibited broad-spectrum degradation activity against various polyesters. These findings suggest that this system offers a promising, eco-friendly solution to PET and other polyester pollution, with potential implications for environmental bioremediation strategies.

Harnessing Nature-Inspired Catechol Amino Acid to Engineer Sticky Proteins and Bacteria.

3,4-Dihydroxy-L-phenylalanine (DOPA) serves as a post-translational modification amino acid present in mussel foot proteins. Mussels exploit the exceptional adhesive properties of DOPA to adhere to a wide range of surfaces. This study presents the development of sticky proteins and bacteria through the site-specific incorporation of DOPA using Genetic Code Expansion Technology. Through the optimization of the DOPA incorporation system, proteins containing DOPA demonstrate significantly improved binding abilities to various organic and metallic materials. The material-binding capabilities of DOPA to combat different types of biofoulings are harnessed by integrating it into intrinsically disordered proteins. Beyond the creation of adhesive proteins for anti-biofouling purposes, this highly efficient DOPA incorporation system is also applied to engineer adhesive bacteria, resulting in a remarkable increase in their binding capability to diverse materials including 400 folds of improvement to polyethylene terephthalate (PET). This substantial enhancement in PET binding of these bacteria has allowed to develop a unique approach for PET degradation, showcasing the innovative application of Genetic Code Expansion in cell engineering.

High terephthalic acid purity: Effective polyethylene terephthalate degradation process based on pH regulation with dual-function hydrolase

Eco-friendly enzymatic-recycling has been widely utilized in tackling plastic pollution. However, the limited activity on the polyethylene terephthalate (PET) degradation product mono-hydroxyethyl terephthalate (MHET) leads to the formation of heterogeneous hydrolysis products, resulting in PET downcycling. Herein, by applying a dual-function PET hydrolase IsPETasePA with balanced PET and MHET degradation efficiency, an effective PET hydrolysis process was developed to enhance the terephthalic acid (TPA) product purity. Firstly, the impact of pH on the catalytic activity of IsPETasePA revealed that the pH reduction caused by TPA generation hindered the complete conversion of MHET to TPA. Further investigation of the catalytic mechanism showed that the pH-induced protonation of His208 in the catalytic triad destabilized the interaction between IsPETasePA and MHET. Thus, by introducing pH regulation strategy on the bifunctional IsPETasePA, the single-enzyme process could achieve high-purity TPA recovery (>99 %). Overall, this work ensured the high-quality PET enzymatic-recycling for effectively addressing plastic pollution.

Degradation and Migration in Olive Oil Packaged in Polyethylene Terephthalate under Thermal Treatment and Storage Conditions

The research addresses challenges in food safety related to the migration of contaminants from plastics to food. It focused on the physicochemical and sensory degradation of olive oils packaged in polyethylene terephthalate (PET) and subjected to thermal exposure at 40 °C and 60 °C for several weeks and a subsequent 12 months of storage, as well as the stability and migration of compounds from the PET packaging itself. Olive oils (OO) from Spanish supermarkets (a mixture of refined and virgin, with commercial identifications of mild and intense) were selected and subjected to thermal treatments at 40 °C and 60 °C for 1, 2, and 3 weeks, followed by 12 months of storage. The treatments were conducted through the following two independent experiments: Experiment A focused on immediate analysis post-thermal treatment, while Experiment B included a 12-month storage period post-thermal treatment. The presence of antimony (Sb) was analyzed using acid digestion with nitric acid (HNO3) and high-resolution inductively coupled plasma mass spectrometry (HR-ICP-MS), while the metals cadmium (Cd), copper (Cu), lead (Pb), and iron (Fe) were analyzed using inductively coupled plasma mass spectrometry (ICP-MS). The PET characterization was assessed using Fourier transform infrared spectroscopy in the mid-infrared range (FT-IR/MIR), carbonyl index, and differential scanning calorimetry (DSC). The results showed increases in the acidity index by 0.29%, the peroxide value by 25.92%, and the K268 coefficient by 51.22% between the control sample and the most severe treatments, with more pronounced effects observed after 12 months. Sensory quality declined, with reduced intensity of the “fruity” attribute and increased presence of the “rancid” defect. PET degradation was reflected in an increase in the carbonyl index and greater structural amorphization. Fe was the predominant metal, and Sb concentration increased after thermal treatments. The lack of studies on the raw consumption of oils packaged in PET and the concerns about the migration of compounds from the packaging to the food highlight the relevance of this research. This study provides new insights into the effects of thermal exposure and storage on the migration of PET contaminants into oils, contributing to the development of strategies to ensure food safety and product quality.

Efficient degradation and mineralization of polyethylene terephthalate microplastics by the synergy of sulfate and hydroxyl radicals in a heterogeneous electro-Fenton-activated persulfate oxidation system

The presence of polyethylene terephthalate (PET) microplastics (MPs) in waters has posed considerable threats to the environment and humans. In this work, a heterogeneous electro-Fenton-activated persulfate oxidation system with the FeS2-modified carbon felt as the cathode (abbreviated as EF-SR) was proposed for the efficient degradation of PET MPs. The results showed that i) the EF-SR system removed 91.3 ± 0.9 % of 100 mg/L PET after 12 h at the expense of trace loss (< 0.07 %) of [Fe] and that ii) dissolved organics and nanoplastics were first formed and accumulated and then quickly consumed in the EF-SR system. In addition to the destruction of the surface morphology, considerable changes in the surface structure of PET were noted after EF-SR treatment. On top of the emergence of the O-H bond, the ratio of C-O/C=O to C-C increased from 0.25 to 0.35, proving the rupture of the backbone of PET and the formation of oxygen-containing groups on the PET surface. With the verified involvement and contributions of SO4•- and •OH, three possible paths were proposed to describe the degradation of PET towards complete mineralization through chain cleavage and oxidation in the EF-SR system.

Unveiling the crystal structure of thermostable dienelactone hydrolase exhibiting activity on terephthalate esters

Dienelactone hydrolase (DLH) is one of numerous hydrolytic enzymes with an α/β-hydrolase fold, which catalyze the hydrolysis of dienelactone to maleylacetate. The DLHs share remarkably similar tertiary structures and a conserved arrangement of catalytic residues. This study presents the crystal structure and comprehensive functional characterization of a novel thermostable DLH from the bacterium Hydrogenobacter thermophilus (HtDLH). The crystal structure of the HtDLH, solved at a resolution of about 1.67 Å, exhibits a canonical α/β-hydrolase fold formed by eight β-sheet strands in the core, with one buried α-helix and six others exposed to the solvent. The structure also confirmed the conserved catalytic triad of DHLs formed by Cys121, Asp170, and His202 residues. The HtDLH forms stable homodimers in solution. Functional studies showed that HtDLH has the expected esterase activity over esters with short carbon chains, such as p-nitrophenyl acetate, reaching optimal activity at pH 7.5 and 70 °C. Furthermore, HtDLH maintains more than 50 % of its activity even after incubation at 90 °C for 16 h. Interestingly, HtDLH exhibits catalytic activity towards polyethylene terephthalate (PET) monomers, including bis-1,2-hydroxyethyl terephthalate (BHET) and 1-(2-hydroxyethyl) 4-methyl terephthalate, as well as other aliphatic and aromatic esters. These findings associated with the lack of activity on amorphous PET indicate that HtDLH has characteristic of a BHET-degrading enzyme. This work expands our understanding of enzyme families involved in PET degradation, providing novel insights for plastic biorecycling through protein engineering, which could lead to eco-friendly solutions to reduce the accumulation of plastic in landfills and natural environments.

Regulation of the expression of MHETase and TPA degradation genes involved in the degradation of PET in Ideonella sakaiensis.

Ideonella sakaiensis is a bacterium that can degrade and consume polyethylene terephthalate (PET), a plastic material that was previously considered non-biodegradable. The degradation of PET requires two enzymes, namely poly (ethylene terephthalate) hydrolase (PETase) and mono (2-hydroxyethyl) terephthalate hydrolase (MHETase), which break down PET into terephthalate (TPA) and ethylene glycol (EG), which serve as carbon sources for the bacterium. Previous studies have focused on the enzymatic properties, structure, and mechanism of action of PETase and MHETase. However, the regulation of PETase and MHETase gene expression has not been investigated. This study identified a protein that binds to the MHETase promoter DNA, MHETase gene-regulating protein (MRP) in I. sakaiensis. PET or TPA induced the expression of PETase and MHETase genes. Furthermore, the induction of the MHETase gene was abolished by the deletion of the mrp gene, while the expression of the PETase gene was maintained. In addition, the genes involved in TPA metabolism were not induced in the mrp mutant. Furthermore, the growth of the PET and TPA deteriorated due to mrp mutation. Also, MRP binds to the promoter regions of the MHETase gene and TPA metabolizing genes, but not to the PETase gene promoter. These results suggest that MRP is a transcription factor that activates MHETase and TPA-metabolizing genes.

Enhanced activation of peroxymonosulfate by ZIF-67/g-C3N4 S-scheme photocatalyst under visible light assistance for degradation of polyethylene terephthalate

Photocatalyst-activated peroxymonosulfate (PMS) degradation of pollutants is already widely used for wastewater treatment under visible light. Polyethylene terephthalate (PET) is widely used in daily life, but waste plastics have an irreversible negative impact on the environment. In this paper, the ZIF-67/g-C3N4 S-scheme heterojunction catalyst was synthesized as a photocatalyst to achieve a good effect on PET degradation in coordination with PMS. The results indicated that PET could be degraded up to 60.63 ± 2.12 % under the combined effect of catalyst, PMS, and light. In this experiment, the influence of catalyst-to-plastic ratio, PMS concentration, aqueous pH, and inorganic anions on plastic degradation by the photocatalytic synergistic PMS system was discussed, and the excellent performance of this system for degrading PET was highlighted through a comparative test. Electron spin resonance (ESR) and free radical quenching experiments demonstrated that SO4•− contributes the largest amount to the PET degradation performance. Furthermore, results from gas chromatography and liquid chromatography-mass spectrometry (LC-MS) indicated that the plastic degradation products include CO, CH4, and organic small-molecule liquid fuels. Finally, a possible mechanism for the light/PMS system to degrade PET in water was suggested. This paper provides a feasible solution to treat waste microplastics in water.

Biodegradation of combined pollutants of polyethylene terephthalate and phthalate esters by esterase-integrated Pseudomonas sp. JY-Q with surface-co-displayed PETase and MHETase

The waste pollution problem caused by polyethylene terephthalate (PET) plastics poses a huge threat to the environment and human health. As plasticizers, Phthalate esters (PAEs) are widely used in PET production and become combined pollutants with PET. Synthetic biology make it possible to construct engineered cells for microbial degradation of combined pollutants of PET and PAEs. PET hydroxylase (PETase) and monohydroxyethyl terephthalate hydroxylase (MHETase) isolated from Ideonella sakaiensis 201-F6 exhibit the capability to depolymerize PET. However, PET cannot enter cells, thus enzymatic degradation or cell surface displaying technology of PET hydrolase are the potential strategies. In this study, Pseudomonas sp. JY-Q was selected as a chassis strain, which exhibits robust stress tolerance. First, a truncated endogenous outer membrane protein cOmpA and its variant Signal (OprF)-cOmpA were selected as anchor motifs for exogenous protein to display on the cell surface. These anchor motifs were fused at the N-terminal of PET hydrolase and MHETase and transformed into Pseudomonas sp. JY-Q, the mutant strains successfully display the enzymes on cell surface, after verification by green fluorescent protein labeling and indirect immunofluorescence assay. The resultant strains also showed the catalytic activity of co-displaying PETase and MHETase for PET biodegradation. Then, the cell surface displaying PET degradation module was introduced to a JY-Q strain which genome was integrated with PAEs degrading enzymes and exhibited PAEs degradation ability. The resultant strain JY-Q-R1-R4-SFM-TPH have the ability of degradation PET and PAEs simultaneously. This study provided a promising strain resource for PET and PAEs pollution control.

In Silico Analysis and Biochemical Characterization of Streptomyces PET Hydrolase with Bis(2-Hydroxyethyl) Terephthalate Biodegradation Activity.

Polyethylene terephthalate (PET), one of the most widely used plastics in the world, causes serious environmental problems. Recently, scientists have been focused on the enzymatic degradation of PET, an environmentally friendly method that offers an attractive approach to the degradation and recycling of PET. In this work, PET hydrolase from Streptomyces sp. W2061 was biochemically characterized, and the biodegradation of PET was performed using the PET model substrate bis (2- hydroxyethyl terephthalate) (BHET). PET hydrolase has an isoelectric point of 5.84, and a molecular mass of about 50.31 kDa. The optimum pH and temperature were 7.0 and 40°C, respectively. LC-MS analysis of the enzymatic products showed that the PET hydrolase successfully degraded a single ester bond of BHET, leading to the formation of MHET. Furthermore, in silico characterization of the PET hydrolase protein sequence and its predicted three-dimensional structure was designed and compared with the well-characterized IsPETase from Ideonella sakaiensis. The structural analysis showed that the (Gly-x1-Ser-x2-Gly) serine hydrolase motif and the catalytic triad (Ser, Asp, and His) were conserved in all sequences. In addition, we integrated molecular dynamics (MD) simulations to analyze the variation in the structural stability of the PET hydrolase in the absence and presence of BHET. These simulations showed the formation of a stable complex between the PET hydrolase and BHET. To the best of our knowledge, this is the first study on Streptomyces sp. W2061 to investigate the BHET degradation activity of PET hydrolase, which has potential application in the biodegradation of plastics in the environment.

Biochemical Characterisation of Sis: A Distinct Thermophilic PETase with Enhanced NanoPET Substrate Hydrolysis and Thermal Stability.

Polyethylene terephthalate (PET) degradation by enzymatic hydrolysis is significant for addressing plastic pollution and fostering sustainable waste management practices. Identifying thermophilic and thermostable PET hydrolases is particularly crucial for industrial bioprocesses, where elevated temperatures may enhance enzymatic efficiency and process kinetics. In this study, we present the discovery of a novel thermophilic and thermostable PETase enzyme named Sis, obtained through metagenomic sequence-based analysis. Sis exhibits robust activity on nanoPET substrates, demonstrating effectiveness at temperatures up to 70 °C and displaying exceptional thermal stability with a melting temperature (Tm) of 82 °C. Phylogenetically distinct from previously characterised PET hydrolases, Sis represents a valuable addition to the repertoire of enzymes suitable for PET degradation.

Upgrading and Enhancement of Recycled Polyethylene Terephthalate with Chain Extenders: In-Depth Material Characterization.

Chemical chain extenders (CEs) can be used to restore the properties of recycled low-molecular-weight polyethylene terephthalate (PET). The aim of this work is to investigate the influence of the type and concentration of the CEs Joncryl and pyromellitic dianhydride (PMDA) on the viscosity and other rheological properties with a unique combination of different methods based on industrial samples originating from recycled PET bottles and trays. The resulting chain-extended thermoplastics were characterized by a combination of differential scanning calorimetry, viscometry, cone plate rheometry, pyrolysis-gas chromatography-mass spectroscopy, optical photothermal infrared spectroscopy, 13C solid-state- and 1H NMR liquid spectroscopy, and size exclusion chromatography. For a recycled PET mixture containing bottle and tray materials, our investigations have shown that a significantly better effect for chain elongation can be achieved with Joncryl compared to PMDA. This can presumably be attributed to water molecules formed during the use of PMDA, which accelerate the degradation of PET. The storage modulus values are therefore significantly higher for the samples with Joncryl compared to PMDA. The results of this study show that chain extension with Joncryl proceeds better compared to the reaction with PMDA.