Enzymatic Recycling Research Articles

Toward Microbial Recycling and Upcycling of Plastics: Prospects and Challenges.

Annually, 400 Mt of plastics are produced of which roughly 40% is discarded within a year. Current plastic waste management approaches focus on applying physical, thermal, and chemical treatments of plastic polymers. However, these methods have severe limitations leading to the loss of valuable materials and resources. Another major drawback is the rapid accumulation of plastics into the environment causing one of the biggest environmental threats of the twenty-first century. Therefore, to complement current plastic management approaches novel routes toward plastic degradation and upcycling need to be developed. Enzymatic degradation and conversion of plastics present a promising approach toward sustainable recycling of plastics and plastics building blocks. However, the quest for novel enzymes that efficiently operate in cost-effective, large-scale plastics degradation poses many challenges. To date, a wide range of experimental set-ups has been reported, in many cases lacking a detailed investigation of microbial species exhibiting plastics degrading properties as well as of their corresponding plastics degrading enzymes. The apparent lack of consistent approaches compromises the necessary discovery of a wide range of novel enzymes. In this review, we discuss prospects and possibilities for efficient enzymatic degradation, recycling, and upcycling of plastics, in correlation with their wide diversity and broad utilization. Current methods for the identification and optimization of plastics degrading enzymes are compared and discussed. We present a framework for a standardized workflow, allowing transparent discovery and optimization of novel enzymes for efficient and sustainable plastics degradation in the future.

Mechanism-Based Design of Efficient PET Hydrolases.

Polyethylene terephthalate (PET) is the most widespread synthetic polyester, having been utilized in textile fibers and packaging materials for beverages and food, contributing considerably to the global solid waste stream and environmental plastic pollution. While enzymatic PET recycling and upcycling have recently emerged as viable disposal methods for a circular plastic economy, only a handful of benchmark enzymes have been thoroughly described and subjected to protein engineering for improved properties over the last 16 years. By analyzing the specific material properties of PET and the reaction mechanisms in the context of interfacial biocatalysis, this Perspective identifies several limitations in current enzymatic PET degradation approaches. Unbalanced enzyme-substrate interactions, limited thermostability, and low catalytic efficiency at elevated reaction temperatures, and inhibition caused by oligomeric degradation intermediates still hamper industrial applications that require high catalytic efficiency. To overcome these limitations, successful protein engineering studies using innovative experimental and computational approaches have been published extensively in recent years in this thriving research field and are summarized and discussed in detail here. The acquired knowledge and experience will be applied in the near future to address plastic waste contributed by other mass-produced polymer types (e.g., polyamides and polyurethanes) that should also be properly disposed by biotechnological approaches.

Carbios will build plant for enzymatic recycling of PET

Carbios will build plant for enzymatic recycling of PET

Opportunities and challenges for integrating the development of sustainable polymer materials within an international circular (bio)economy concept

HighlightsThe production and consumption of commodity polymers have been an indispensable part of the development of our modern society. Owing to their adjustable properties and variety of functions, polymer-based materials will continue playing important roles in achieving the Sustainable Development Goals (SDG)s, defined by the United Nations, in key areas such as healthcare, transport, food preservation, construction, electronics, and water management. Considering the serious environmental crisis, generated by increasing consumption of plastics, leading-edge polymers need to incorporate two types of functions: Those that directly arise from the demands of the application (e.g. selective gas and liquid permeation, actuation or charge transport) and those that enable minimization of environmental harm, e.g., through prolongation of the functional lifetime, minimization of material usage, or through predictable disintegration into non-toxic fragments. Here, we give examples of how the incorporation of a thoughtful combination of properties/functions can enhance the sustainability of plastics ranging from material design to waste management. We focus on tools to measure and reduce the negative impacts of plastics on the environment throughout their life cycle, the use of renewable sources for their synthesis, the design of biodegradable and/or recyclable materials, and the use of biotechnological strategies for enzymatic recycling of plastics that fits into a circular bioeconomy. Finally, we discuss future applications for sustainable plastics with the aim to achieve the SDGs through international cooperation.Leading-edge polymer-based materials for consumer and advanced applications are necessary to achieve sustainable development at a global scale. It is essential to understand how sustainability can be incorporated in these materials via green chemistry, the integration of bio-based building blocks from biorefineries, circular bioeconomy strategies, and combined smart and functional capabilities.Graphic abstract

Synthesis, antibacterial activity, and enzymatic decomposition of bio-polyurethane foams containing propolis

Polyurethane foam (PUF) is formed by reacting sugar, propolis, and polyisocyanate, and then blowing with CO2, formed by reacting polyisocyanate with water. An optimum foaming rate of 1440% was found for 15 wt.% of sugar and 20 wt.% of propolis, and the synthesis and cell structure of polyurethane were analyzed by scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR) spectroscopy. Under the optimized conditions, the apparent density and thermal conductivity of the PUF-containing propolis were 0.04 – 0.147 g/mL and 0.028 – 0.029 W/(mK), respectively. The bacterial reduction rate was 92.7% in the PUF containing 20 wt.% propolis. PUF can be broken down by enzymatic reactions in water using invertase. The degradation reaction of invertase was kinetically analyzed. This study showed that eco-friendly polymers can be synthesized using sugar and propolis and decomposed through antibacterial activity and enzymatic green recycling.

Oxidized Forms of Ergothioneine Are Substrates for Mammalian Thioredoxin Reductase.

Ergothioneine (EGT) is a sulfur-containing amino acid analog that is biosynthesized in fungi and bacteria, accumulated in plants, and ingested by humans where it is concentrated in tissues under oxidative stress. While the physiological function of EGT is not yet fully understood, EGT is a potent antioxidant in vitro. Here we report that oxidized forms of EGT, EGT-disulfide (ESSE) and 5-oxo-EGT, can be reduced by the selenoenzyme mammalian thioredoxin reductase (Sec-TrxR). ESSE and 5-oxo-EGT are formed upon reaction with biologically relevant reactive oxygen species. We found that glutathione reductase (GR) can reduce ESSE, but only with the aid of glutathione (GSH). The reduction of ESSE by TrxR was found to be selenium dependent, with non-selenium-containing TrxR enzymes having little or no ability to reduce ESSE. In comparing the reduction of ESSE by Sec-TrxR in the presence of thioredoxin to that of GR/GSH, we find that the glutathione system is 10-fold more efficient, but Sec-TrxR has the advantage of being able to reduce both ESSE and 5-oxo-EGT directly. This represents the first discovered direct enzymatic recycling system for oxidized forms of EGT. Based on our in vitro results, the thioredoxin system may be important for EGT redox biology and requires further in vivo investigation.

Carbios to industrially scale up its enzymatic PET depolymerization-based recycling

On January 19, Carbios announced it will accelerate the industrial development of its enzymatic recycling process (C-ZYME™) with the construction of its reference unit in Europe. It has shortlisted two PET producers in Europe to host this unit.

Enzymatic recycling of polymacrolactones

The enzymatic recycling of polymacrolactones using the N435 biocatalyst provides macrocyclic oligoesters that could be repolymerized.

Front Cover: Comparative Performance of PETase as a Function of Reaction Conditions, Substrate Properties, and Product Accumulation (ChemSusChem 1/2022)

The Front Cover shows the application of poly(ethylene terephthalate) (PET) hydrolase enzymes on post-consumer waste plastic, towards the development of an enzymatic PET recycling strategy. More information can be found in the Full Paper by E. Erickson et al.

Comparative Performance of PETase as a Function of Reaction Conditions, Substrate Properties, and Product Accumulation.

Invited for this month's cover is the BOTTLE Consortium, featuring Gregg Beckham's laboratory from NREL and John McGeehan's laboratory from the University of Portsmouth. The cover image shows the application of poly(ethylene terephthalate) (PET) hydrolase enzymes on post-consumer waste plastic, towards the development of an enzymatic PET recycling strategy. The Full Paper itself is available at 10.1002/cssc.202101932.

Assessment of IsPETase-Assisted Depolymerization of Terephthalate Aromatic Polyesters and the Effect of the Thioredoxin Fusion Domain

Terephthalate polyesters such as poly(ethylene terephthalate) (PET) have been massively produced over the last few decades due to their attractive properties in multiple applications. However, due to their limited biodegradability, they have accumulated in landfills and oceans, posing an environmental threat. Enzymatic recycling technologies are predicted to generate long-term socioeconomic benefits. In the present work, we compared the IsPETase (from Ideonella sakaiensis 201-F6) activity on a series of polyesters, including poly(butylene) terephthalate (PBT), poly(hexamethylene) terephthalate (PHT) and Akestra™, with PET. The IsPETase showed remarkable activity toward PET (39% degradation of the original polyester) that was higher than that toward Akestra™ (0.13%), PBT (0.25%) and PHT (0.13%) after 72 h. Thus, based on experimental data and computational analysis, we report insights into IsPETase activity on a series of terephthalate-based polyesters. Aside from that, the fusion domain (Trx) effect in the production and activity of a recombinant Trx-IsPETase is reported.

Techno-economic, life-cycle, and socioeconomic impact analysis of enzymatic recycling of poly(ethylene terephthalate)

<h2>Summary</h2> Esterases have emerged as important biocatalysts for enzyme-based polyester recycling of poly(ethylene terephthalate) (PET) to terephthalic acid (TPA) and ethylene glycol (EG). Here, we present process modeling, techno-economic, life-cycle, and socioeconomic impact analyses for an enzymatic PET depolymerization-based recycling process, which we compare with virgin TPA manufacturing. We predict that enzymatically recycled TPA (rTPA) can be cost-competitive and highlight key areas to achieve this. In addition to favorable long-term socioeconomic benefits, rTPA can reduce total supply chain energy use by 69%–83% and greenhouse gas emissions by 17%–43% per kg of TPA. An economy-wide assessment for the US estimates that the TPA recycling process can reduce environmental impacts by up to 95% while generating up to 45% more socioeconomic benefits, also relative to virgin TPA production. Sensitivity analyses highlight impactful research opportunities to pursue toward realizing biological PET recycling and upcycling.

Enzymatic textile recycling - best practices and outlook.

Recently, textiles and their end-of-life management have become the focus of public and political attention. In the European Union the revised waste framework directive defines textiles as municipal waste and stipulates their separate collection by 2025. In the context of these developments, this paper summarises briefly the current state-of-the-art in textile recycling. It is evident that recycling methods are not yet fully developed. This is especially the case with multi-material textiles, which are composed of two or more polymers that are incompatible for recycling. In the practical part of the communication, results are presented which show that enzymatic hydrolysis is a suitable process for recycling textiles made of cotton and polyester. After a complete removal of cotton, the remaining pure polyester fibres undergo a re-granulation and post-condensation step. The so obtained recycled polyester is fed back into the textile processing chain and finally towels are obtained. The main steering parameters of the enzymatic hydrolysis process are described. The study proves that solutions in accordance with the Circular Economy in the textile sector are available but an industrial implementation has not yet been realised.

The Current State of Research on PET Hydrolyzing Enzymes Available for Biorecycling

This short paper reviews two groups of enzymes designated as polyethylene terephthalate (PET) hydrolases: one consists of thermophilic cutinases from thermophilic microorganisms (actinomycetes and a fungus) and the other consists of mesophilic cutinases, the representative of which is IsPETase from a mesophilic bacterium. From the viewpoint that PET hydrolysis requires a high temperature close to the glass transition temperature (65–70 °C in water) of PET, mesophilic cutinases are not suitable for use in the enzymatic recycling of PET since their degradation level is one to three orders of magnitude lower than that of thermophilic cutinases. Many studies have attempted to increase the thermostability of IsPETase by introducing mutations, but even with these modifications, the mesophilic cutinase does not reach the same level of degradation as thermophilic cutinases. In addition, this kind of trial contradicts the claim that IsPETase works at ambient temperature. As plastic pollution is an urgent environmental issue, scientists must focus on feasible thermophilic enzymes for the enzymatic processing of disposed PET, rather than on mesophilic cutinases. Thermophilic and mesophilic cutinases must be evaluated precisely and comparatively, based on their features that enable them to hydrolyze PET, with the aim of enzymatic PET disposal. The level of thermophilic cutinases has already reached their optimal level in PET biorecycling. The optimal level may be reached through the processing of PET waste, by amorphization and micronization into readily hydrolysable forms and the improvement of PET hydrolases by engineering higher degradation ability and low-cost production. Here I summarize the critical points in the evaluation of PET hydrolases and discuss the biorecycling of PET.

Characterization of the enzymatic degradation of polyurethanes.

For decades, polyurethanes (PUR) have mainly been synthesized for long-term applications and are therefore highly persistent in the environment. Proper waste disposal approaches, including recycling techniques, must be developed to limit the accumulation of PUR in the environment. Evaluation of enzymatic polyurethane degradation is needed for the development of enzymatic recycling. A series of techniques has been carefully implemented to monitor the biotic and abiotic degradation of PUR. Both the degraded polymer and the degradation products are analyzed to obtain a complete overview of the degradation.

Stimuli-responsive DNA microcapsules for homogeneous electrochemiluminescence sensing of tumor exosomes

A homogeneous electrochemiluminescence (ECL) sensing system was designed for highly sensitive detection of tumor exosomes by combining stimuli-responsive DNA microcapsules with target recycling amplification. In this work, CdS quantum dots (QDs) as ECL emitters were sealed in the inner pores of DNA microcapsules with crosslinking DNA shells. In the presence of target exosomes, S4-combined exosomes were formed and then hybridized with the crosslinking base sequences of DNA shells. Thus, DNA microcapsules were effectively unlocked, leading to the release of CdS QDs. The target exosomes-dependent amount of release CdS QDs could be conveniently monitored by ECL technique. With enzymatic recycling binding of target amplification, the proposed homogeneous ECL sensor possessed a wide detection range, high sensitivity and selectivity in complex biological samples.

Recycling of Chrysoporthe cubensis enzymes to overcome the adsorption in lignin and to improve sugarcane bagasse saccharification

Biomass enzymatic hydrolysis efficiency shows some limitations, such as the non-specific adsorption of enzymes in lignin. We evaluated the adsorption of proteins and enzymes from Chrysoporthe cubensis into alkaline and acid pretreated sugarcane bagasse, containing 8.1 and 33.8% of lignin, respectively. The data from Langmuir isotherm showed that at 4 °C, proteins were more adsorbed by acid-treated bagasse, 10.5 mg g−1, while higher adsorption affinity, 2.0 mL mg−1, was determined from alkali-treated bagasse. Similarly, under hydrolytic conditions (50 °C), higher amounts of proteins were adsorbed into acid-treated bagasse, confirming the predominant effect of lignin on protein adsorption. Enzymatic recycling rounds during alkali-treated bagasse saccharification showed that the addition of fresh substrate and enzymes promoted the higher release of glucose and xylose, 40.3 and 38.3 g L−1, respectively. However, the addition of only fresh substrates showed to be efficient for C. cubensis enzymes recovery and substrate hydrolysis, which reduces enzyme consumption and costs.

Process strategies to improve biocatalytic depolymerization of post-consumer PET packages in bioreactors, and investigation on consumables cost reduction.

Massive plastics production has raised concerns about low recycling rates and disposal of these materials in nature, causing environmental and economic impacts. Poly(ethylene terephthalate) (PET) is one of main polymers used for manufacture of plastic packaging (e.g. bottles, trays). Enzymatic recycling of PET has been a route of increasing study aiming at to recover its monomers (terephthalic acid and ethylene glycol), resulting in a circular production chain. In this study, investigation of pH control and fractionation of enzyme feeding were explored in post-consumed PET (PC-PET) hydrolysis reactions catalyzed by Humicola insolens cutinase (HiC) in stirred reactors. It was found that the unbuffered reaction provided of pH control by 0.5M NaOH addition showed 2.39-fold improvement in the released monomers (to a total of 26.3mM), comparatively to the Tris-HCl-buffered reaction. In addition, it was observed a possibility of reducing the enzyme loading used in the process by half, leading to an increase of 2.41-fold in the specific terephthalic acid concentration released per protein amount, whilst maintaining a high products concentration (97mM). A simplified cost analysis of reaction consumables was performed, and the data reported here demonstrates that these alternative process strategies contribute to costs reduction on the enzymatic depolymerization reactions of PET.

Effect of Occupational Exposure to Herbicides on Oxidative Stress in Sprayers.

BackgroundHerbicides such as glyphosate, paraquat, and 2,4-dichlorophenoxyacetic acid have been reported to cause adverse side effects through production of reactive oxygen species. However, there were no data representing the adverse effects of a mixture herbicide usage in farmers, especially the changes in oxidative marker and antioxidant defense. This study aimed to determine the urinary malondialdehyde (MDA) and glutathione (GSH) level in farmers using mixed herbicides.MethodsNinety-three farmers were recruited, and two spot urine samples (before and after work) were collected. The urinary MDA level was evaluated by thiobarbituric acid reactive substance assay, and the urinary GSH level was determined using the enzymatic recycling method.ResultsSixty-two percent of the participants were men, and 59% of the participants worked in a farm for 20–40 years. The common combinations of herbicide usage were glyphosate with 2,4-dichlorophenoxyacetic acid (36.5%). There was no significant difference between pre- and post-work urinary MDA and GSH levels among the 3 groups of herbicides. However, the urinary MDA levels in farmers using the combination of glyphosate and paraquat were significantly higher than those found in farmers using glyphosate alone. The associated factors with changes in MDA levels found that the exposure intensity index (B = 0.154), the cumulative exposure intensity index (B = 0.023), and wearing gloves while working (B = −2.347) were found to be significantly associated with MDA level.ConclusionThe results suggest that the combined use of glyphosate and paraquat caused a significant increase in urinary MDA levels. Moreover, intensity of exposure to herbicide and wearing gloves were associated with the level of MDA.

Catalytic synthesis of renewable p-xylene from biomass-derived 2,5-dimethylfuran: a mini review

In this work, the renewable synthesis of p-xylene (PX) from biomass-derived carbohydrates has been reviewed. PX is a crucial chemical feedstock and an essential starting material of polyethylene terephthalate (PET). PX can be produced selectively by the Diels-Alder reaction between ethylene and 2,5-dimethylfuran (DMF) followed by catalytic dehydration of the oxanorbornene adduct. DMF is primarily produced by the catalytic hydrogenation of 5-(hydroxymethyl)furfural (HMF), a furanic intermediate produced by the acid-catalyzed hydrolysis/dehydration of biomass-derived hexoses. With ethylene being sourced by dehydrating bioethanol, PET can be made biorenewable in its entirety. The atom economy and carbon efficiency of converting glucose into PX have been calculated. The existing literature (both theoretical and experimental) on the catalytic production of PX from DMF and ethylene are summarized, and future directions on this research have been proposed. The effect of Bronsted and Lewis acidity, porosity, and surface area of the heterogeneous catalysts on the selectivity and yield of PX have been highlighted. In addition, the techno-economic analysis of renewable PET, its future prospects based on the petroleum market, and the possibility of a circular economy of PET using chemical and enzymatic recycling strategies have been discussed.