Enzymatic polyethylene biorecycling: Confronting challenges and shaping the future.

Polyethylene (PE) is a widely used plastic known for its resistance to biodegradation, posing a significant environmental challenge. Recent advances have shed light on microorganisms and insects capable of breaking down PE and identified potential PE-degrading enzymes (PEases), hinting at the possibility of PE biorecycling. Research on enzymatic PE degradation is still in its early stages, especially compared to the progress made with polyethylene terephthalate (PET). While PET hydrolases have been extensively studied and engineered for improved performance, even the products of PEases remain mostly undefined. This Perspective analyzes the current state of enzymatic PE degradation research, highlighting obstacles in the search for bona fide PEases and suggesting areas for future exploration. A critical challenge impeding progress in this field stems from the inert nature of the C-C and C-H bonds of PE. Furthermore, breaking down PE into small molecules using only one monofunctional enzyme is theoretically impossible. Overcoming these obstacles requires identifying enzymatic pathways, which can be facilitated using emerging technologies like omics, structure-based design, and computer-assisted engineering of enzymes. Understanding the mechanisms underlying PE enzymatic biodegradation is crucial for research progress and for identifying potential solutions to the global plastic pollution crisis.

Solid-State Enzymatic Hydrolysis of Mixed PET/Cotton Textiles.

Waste polyester textiles are not recycled due to separation challenges and partial structural degradation during use and recycling. Chemical recycling of polyethylene terephthalate (PET) textiles through depolymerization can provide a feedstock of recycled monomers to make "as-new" polymers. While enzymatic PET recycling is a more selective and more sustainable approach, methods in development, however, have thus far been limited to clean, high-quality PET feedstocks, and require an energy-intensive melt-amorphization step ahead of enzymatic treatment. Here, high-crystallinity PET in mixed PET/cotton textiles could be directly and selectively depolymerized to terephthalic acid (TPA) by using a commercial cutinase from Humicola insolens under moist-solid reaction conditions, affording up to 30±2 % yield of TPA. The process was readily combined with cotton depolymerization through simultaneous or sequential application of the cellulase enzymes CTec2®, providing up to 83±4 % yield of glucose without any negative influence on the TPA yield.

Novel irreversible peptidic inhibitors of transglutaminase 2.

Transglutaminase 2 (TG2), also referred to as tissue transglutaminase, plays crucial roles in both protein crosslinking and cell signalling. It is capable of both catalysing transamidation and acting as a G-protein, these activities being conformation-dependent, mutually exclusive, and tightly regulated. The dysregulation of both activities has been implicated in numerous pathologies. TG2 is expressed ubiquitously in humans and is localized both intracellularly and extracellularly. Targeted TG2 therapies have been developed but have faced numerous hurdles including decreased efficacy in vivo. Our latest efforts in inhibitor optimization involve the modification of a previous lead compound's scaffold by insertion of various amino acid residues into the peptidomimetic backbone, and derivatization of the N-terminus with substituted phenylacetic acids, resulting in 28 novel irreversible inhibitors. These inhibitors were evaluated for their ability to inhibit TG2 in vitro and their pharmacokinetic properties, and the most promising candidate 35 (k inact/K I = 760 × 103 M-1 min-1) was tested in a cancer stem cell model. Although these inhibitors display exceptional potency versus TG2, with k inact/K I ratios nearly ten-fold higher than their parent compound, their pharmacokinetic properties and cellular activity limit their therapeutic potential. However, they do serve as a scaffold for the development of potent research tools.

Mechanoenzymatic Reactions Involving Polymeric Substrates or Products.

Mechanoenzymology is an emerging field in which mechanical mixing is used to sustain enzymatic reactions in low-solvent or solvent-free mixtures. Many enzymes have been reported that thrive under such conditions. Considering the central role of biopolymers and synthetic polymers in our society, this minireview highlights the use of mechanoenzymology for the synthesis or depolymerization of oligomeric or polymeric materials. In contrast to traditional in-solution reactions, solvent-free mechanoenzymology has the advantages of avoiding solubility issues, proceeding in a minimal volume, and reducing solvent waste while potentially improving the reaction efficiency and accessing new reactivity. It is expected that this strategy will continue to gain popularity and find more applications.