Production of PETase by engineered Yarrowia lipolytica for efficient poly(ethylene terephthalate) biodegradation.

There has been a growing interest in poly(ethylene terephthalate) PET degradation studies in the last few years due to its widespread use and large-scale plastic waste accumulation in the environment. One of the most promising enzymatic methods in the context of PET degradation is the use of PETase from Ideonella sakaiensis, which has been reported to be an efficient enzyme for hydrolysing ester bonds in PET. In our study, we expressed a codon-optimized PETase gene in the yeast Yarrowia lipolytica. The obtained strain was tested for its ability to degrade PET directly in culture, and a screening of different supplements that might raise the level of PET hydrolysis was performed. We also carried out long-term cultures with PET film, the surface of which was examined by scanning electron microscopy. The efficiency of PET degradation was tested based on the concentration of degradation products released, and the results showed that supplementation of the culture with olive oil resulted in 66 % higher release of terephthalic acid into the medium compared to the mutant culture without supplementation. The results indicate the possibility of ethylene glycol uptake by both strains, and, additionally, the PETase produced by the newly engineered strain hydrolyses MHET. The structure of the PET film after culture with the modified strain, meanwhile, had numerous surface defects, cracks, and deformations.

Metabolic engineering of Yarrowia lipolytica for poly(ethylene terephthalate) degradation.

Polyethylene terephthalate (PET) is the most widely used plastic, whose global production scale causes serious problems due to it being highly non-biodegradable. The present work provides a novel approach to plastic degradation studies, which involves direct degradation of PET in the culture of a modified Y. lipolytica yeast strain extracellularly producing cutinase from Fusarium solani. In this study, we successfully accomplished a scale-up of the degradation process in culture, which is promising from the perspective of wider application of the developed method in the future. Additionally, we tested the effect of various supplements, which may increase the PET degradation efficiency in the culture of the Y. lipolytica pAD CUT_FS strain. The ability of PET decomposition was verified by the amount of the released degradation products, such as terephthalic acid (TPA) and mono-(2-hydroxyethyl)-terephthalic acid (MHET), during cultivation. We observed that the quantities of TPA and MHET released during the PET degradation process were increasing daily, and were 1.51 gL-1 and 0.45 gL-1, respectively after 240 h of the bioreactor fermentation. Analysis of the PET film by electron microscopy indicated that there was abundant damage on the surface of the material. This study also demonstrated that the engineered Y. lipolytica strain is able to degrade PET at 28 °C during fermentation. The results obtained in this study using amorphous PET powder provide a wide range of possibilities for application of the cutinase-secreting strain of Y. lipolytica on the more difficult to degrade highly crystalline PET films, PET bottles and PET melts.

Study of Thermal, Mechanical and Barrier Properties of Biodegradable PLA/PBAT Films with Highly Oriented MMT.

In order to improve the properties of biodegradable polylactide (PLA), mixtures with polybutylene adipate-co-terephthalate (PBAT) were prepared. PLA is a bio-based and renewable biodegradable material, made from starch. PBAT is a biodegradable polyester for compostable film. In order to improve the composite properties, two types of additives were implemented via melt mixing, a chain extender (CE) and montmorillonite (MMT). CE was used as an interfacial modifier to enhance the adhesion between components. Montmorillonite is a widely studied clay added to polymer nanocomposites. Due to the lamellar structure, it improves the barrier properties of materials. PLA/PBAT films were oriented in the extrusion process and the amounts of filler introduced into the PLA/PBAT nanocomposites were 1.0, 3.0, and 5.0%. The improvement in the PLA barrier properties by the addition of PBAT and 5% of MMT was confirmed as the oxygen permeability decreased almost by a factor of 3. The addition of the biodegradable polymer, chain extender, montmorillonite, and the implemented orientation process resulted in a decrease in composite viscosity and an increase in the PLA crystallinity percentage (up to 25%), and the wettability tests confirmed the synergic behavior of the selected polymer blend.

A comprehensive assessment of microbiome diversity in Tenebrio molitor fed with polystyrene waste.

Recently it was demonstrated that mealworm (Tenebrio molitor) larvae consume and biodegrade polystyrene. Thus, in this study a breeding investigation with various types of polystyrene was performed to follow the changes in the gut microbiome diversity. Polystyrene used for packaging purposes (PSp) and expanded polystyrene (EPS) were perceived as more favorable and attacked more frequently by mealworms compared to raw polystyrene (PS) and material commercially available for parcels (PSp). Although our studies showed that larvae could bite and chew selected materials, they are not able to degrade and use them for consumption purposes. In a next-generation sequencing experiment, among all samples, seven classes, Gammaproteobacteria, Bacilli, Clostridia, Acidobacteria, Actinobacteria, Alphaproteobacteria and Flavobacteria, were indicated as the most abundant, whereas the predominant genera were Enterobacter, Lactococcus and Enterococcus. Additionally, we isolated three bacteria strains able to use diverse types of bioplastic as a sole carbon source. The strains with biodegradable activity against bioplastic were identified as species of the genera Klebsiella, Pseudomonas and Serratia. The presence of a bacterial strain able to degrade bioplastic may suggest a potential niche for further investigations.

The Impact of Filler Geometry on Polylactic Acid-Based Sustainable Polymer Composites.

Recently, biocomposites have emerged as materials of great interest to the scientists and industry around the globe. Among various polymers, polylactic acid (PLA) is a popular matrix material with high potential for advanced applications. Various particulate materials and nanoparticles have been used as the filler in PLA based matrix. One of the extensively studied filler is cellulose. However, cellulose fibres, due to their hydrophilic nature, are difficult to blend with a hydrophobic polymer matrix. This leads to agglomeration and creates voids, reducing the mechanical strength of the resulting composite. Moreover, the role of the various forms of pure cellulose and its particle shape factors has not been analyzed in most of the current literature. Therefore, in this work, materials of various shapes and shape factors were selected as fillers for the production of polymer composites using Polylactic acid as a matrix to fill this knowledge gap. In particular, pure cellulose fibres (three types with different elongation coefficient) and two mineral nanocomponents: precipitated calcium carbonate and montmorillonite were used. The composites were prepared by a melt blending process using two different levels of fillers: 5% and 30%. Then, the analysis of their thermomechanical and physico-chemical properties was carried out. The obtained results were presented graphically and discussed in terms of their shape and degree of filling.

Evaluation of the Rosemary Extract Effect on the Properties of Polylactic Acid-Based Materials.

New multifunctional materials containing additives derived from natural resources as powdered rosemary ethanolic extract were obtained by melt mixing and processed in good conditions without degradation and loss of additives. Incorporation of powdered rosemary ethanolic extract (R) into poly(lactic acid) (PLA) improved elongation at break, rheological properties, antibacterial and antioxidant activities, in addition to the biocompatibility. The good accordance between results of the chemiluminescence method and radical scavenging activity determination by chemical method evidenced the increased thermoxidative stability of the PLA biocomposites with respect to neat PLA, with R acting as an antioxidant. PLA/R biocomposites also showed low permeability to gases and migration rates of the bioactive compounds and could be considered as high-performance materials for food packaging. In vitro biocompatibility based on the determination of surface properties demonstrated a good hydrophilicity, better spreading and division of fibroblasts, and increased platelet cohesion. The implantation of PLA/R pellets, was proven to possess a good in vivo biocompatibility, and resulted in similar changes in blood parameters and biochemical responses with the control group, suggesting that these PLA-based materials demonstrate very desirable properties as potential biomaterials, useful in human medicine for tissue engineering, wound management, orthopedic devices, scaffolds, drug delivery systems, etc. Therefore, PLA/R-based materials show promising properties for applications both in food packaging and as bioactive biomaterials.

Improvement of Interaction in a Composite Structure by Using a Sol-Gel Functional Coating on Carbon Fibers.

The modification of carbon fibers for improving adhesion between fibers and an epoxy resin in composite materials has become the focus of attention. In this work the carbon fiber coating process has been devised in a way preventing the stiffening and clumping of fibers. To improve interactions between coated fibers and a resin in composites, four types of silica coatings with different organic functional groups (3-aminopropyl-coating 1, 3-mercaptopropyl-coating 2, 2-(3,4-epoxycyclohexyl) ethyl-coating 3, methyl-coating 4) were obtained. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) were used to distinguish the changes of a carbon fibers surface after coating deposition. The thickness of the obtained coatings, including the diversity of thickness, was determined by transmission electron microscopy (TEM). The increase in surface free energy (SFE) of modified fibers, including the distinction between the polar and dispersive parts, was examined by wettability measurements using a tensometric test. The developed coating preparation process allowed to cover fibers separately with nanoscale silica layers, which changed their morphology. The introduction of organic functional groups resulted in surface free energy changes, especially an increase in specific polar surface energy components.