Biodegradation of plastics has been observed at rapid turnover rate by some insect larvae, especially those of Coleoptera, in particular Tenebrionidae. Tenebrio molitor larva is well studied and capable of biodegrading polystyrene (PS), polyethylene (PE), polypropylene (PP), and polyvinyl chloride (PVC) in their digestive intestine in synergy with their gut microflora. This chapter includes the methods, protocols, and procedures used to characterize biodegradation of plastics in T. molitor larvae and their gut microbiomes with polystyrene as the model feedstock. The methods used can be expanded to enable investigation of other plastics and/or insects.Environmental pollution with synthetic polymers (commonly named plastics) nowadays poses serious threats to the environment and human health. Unfortunately, most conventional plastics are highly recalcitrant even under conditions known to be favorable for microbial degradation. Expanding the knowledge regarding opportunities and limitations of the microbial degradability of plastics would largely contribute to the development of adequate decontamination and management strategies for plastic pollution. This chapter provides cultivation approaches to be applied for the characterization of eco-physiologically diverse asco- and basidiomycete fungi with respect to their ability to attack solid and water-soluble synthetic polymers with the help of quinone redox cycling-based Fenton-type reactions, which result in the production of highly reactive hydroxyl radicals. These reactive oxygen species are the strongest oxidants known from biological systems. However, their potential employment by fungi dwelling in diverse habitats as a biodegradation tool to attack synthetic polymers is still insufficiently explored.Many complex natural and synthetic compounds are degraded by microbial assemblages rather than single strains, due to usually limited metabolic capacities of single organisms. It can therefore be assumed that plastics can be more efficiently degraded by microbial consortia, although this field has not been as widely explored as plastic degradation by individual strains. In this chapter, we present some of the current studies on this topic and methods to enrich and cultivate plastic-degrading microbial consortia from aquatic and terrestrial ecosystems, including substrate preparation and biodegradation assessment. We focus on both conventional and biodegradable plastics as potential growth substrates. Cultivation methods for both aerobic and anaerobic microorganisms are presented.Enzymatic hydrolysis of polyethylene terephthalate (PET) is considered to be an environmentally friendly method for the recycling of plastic waste. Recently, a bacterial enzyme named IsPETase was found in Ideonella sakaiensis with the ability to degrade amorphous PET at ambient temperature suggesting its possible use in recycling of PET. However, applying the purified IsPETase in large-scale PET recycling has limitations, i.e., a complicated production process, high cost of single-use, and instability of the enzyme. Yeast cell surface display has proven to be an effectual alternative for improving enzyme degradation efficiency and realizing industrial applications. This chapter deals with the construction and application of a whole-cell biocatalyst by displaying IsPETase on the surface of yeast (Pichia pastoris) cells.Plastic pollution has become a serious issue on Earth. Although efficient industrial recycling processes exist, a significant fraction of plastic waste still ends up in nature, where it can endure for centuries. Slow mechanical and chemical decay lead to the formation of micro- and nanoplastics, which are washed from land into rivers and finally end up in the oceans. As such particles cannot be efficiently removed from the environment, biological degradation mechanisms are highly desirable. Several enzymes have been described that are capable of degrading certain plastic materials such as polyethylene terephthalate (PET). Such enzymes have a huge potential for future biotechnology applications. However, they require model systems that can be efficiently adapted to very specific conditions. Here, we present detailed instructions, how to convert the model diatom Phaeodactylum into a solar-fueled microbial cell factory for PETase expression, resulting in a whole cell catalyst for PET degradation at moderate temperatures under saltwater conditions.The diverse benefits of synthetic polymers is overshadowed by the amount of plastic waste and its whereabouts. The problem can only be tackled by reducing and recycling of plastics. In this respect, investigating the (microbial) degradation of each type of polymer currently used may provide further understanding that fosters the development of new feasible recycling technologies. Here, we present a strategy to isolate bacteria from environmental samples that are able to degrade hydrolysis products and building blocks of polyurethane (PUR). Protocols are presented to enrich bacteria on the primary diamines 2,4-diaminotoluene (TDA) and 4,4'-diaminodiphenylmethane (MDA) as well as an oligomeric PUR (Sigma Aldrich, proprietary composition). For TDA and the oligomeric PUR, methods are suggested to monitor their concentration in bacterial enrichment cultures.The enzymatic degradation of polyethylene terephthalate (PET) results in a hydrolysate consisting almost exclusively of its two monomers, ethylene glycol and terephthalate. To biologically valorize the PET hydrolysate, microbial upcycling into high-value products is proposed. https://www.selleckchem.com/products/thiamet-g.html Fatty acid derivatives hydroxyalkanoyloxy alkanoates (HAAs) represent such valuable target molecules. HAAs exhibit surface-active properties and can be exploited in the catalytical conversion to drop-in biofuels as well as in the polymerization to bio-based poly(amide urethane). This chapter presents the genetic engineering methods of pseudomonads for the metabolization of PET monomers and the biosynthesis of HAAs with detailed protocols concerning product purification.Biodegradation of synthetic polymers is recognized as a useful way to reduce their environmental load and pollution, loss of natural resources, extensive energy consumption, and generation of greenhouse gases. The potential use of enzymes responsible for the degradation of the targeted polymers is an effective approach which enables the conversion of the used polymers to original monomers and/or other useful compounds. In addition, the enzymes are expected to be applicable in industrial processes such as improving the surface structures of the polymers. Especially, conversion of the solid polymers to soluble oligomers/monomers is a key step for the biodegradation of the polymers. Regarding the hydrolysis of polyamides, three enzymes, 6-aminohexanoate-cyclic-dimer hydrolase (NylA), 6-aminohexanoate-dimer hydrolase (NylB), and 6-aminohexanoate-oligomer endo-hydrolase (nylon hydrolase, NylC), are found in several bacterial strains. In this chapter, we describe our approach for the screening of microorganisms which degrade nylons and related compounds; preparation of substrates; assay of hydrolytic activity for soluble and insoluble substrates; and X-ray crystallographic and computational approaches for analysis of structure and catalytic mechanisms of the nylon-degrading enzymes.