Chemicals from packaging materials might be transferred into food resulting in consumer exposure. Identifying these migrated chemicals is highly challenging and crucial to perform their safety assessment, usually starting by the understanding of the chemical composition of the packaging material itself. This study explores the use of the Molecular Networking (MN) approach to support identification of the extracted chemicals. Two formulations of bioplastics were analyzed using Liquid Chromatography hyphenated to High-Resolution Mass Spectrometry. Data processing and interpretation using a conventional manual method was performed as a point of comparison to understand the power of MN. Interestingly, only the MN approach facilitated the identification of unknown chemicals belonging to a novel oligomer series containing the azelaic acid monomer. The MN approach provided a faster visualization of chemical families in addition to the highlight of unrelated chemicals enabling to prioritize chemicals for further investigation improving the safety assessment of packaging materials.

The industrial production of polyhydroxyalkanoates (PHAs) faces several limitations that hinder their competitiveness against traditional plastics, mainly due to high production costs and complex recovery processes. Innovations in microbial biotechnology offer promising solutions to overcome these challenges. The modification of the biosynthetic pathways is one of the main tactics; allowing for direct carbon flux toward PHA formation, increasing polymer accumulation and improving polymer properties. Additionally, techniques have been implemented to expand the range of renewable substrates used in PHA production. These feedstocks are inexpensive and plentiful but require costly and energy-intensive pretreatment. By removing the need for pretreatment and enabling the direct use of these raw materials, microbial biotechnology aims to reduce production costs. Furthermore, improving downstream processes to facilitate the separation of biomass from culture broth and the recovery of PHAs is critical. Genetic modifications that alter cell morphology and allow PHA secretion directly into the culture medium simplify the extraction and purification process, significantly reducing operating costs. These advances in microbial biotechnology not only enhance the efficient and sustainable production of PHAs, but also position these biopolymers as a viable and competitive alternative to petroleum-based plastics, contributing to a circular economy and reducing the dependence on fossil resources.

With the rising demand for sustainable renewable resources, microorganisms capable of producing bioproducts such as bioplastics are attractive. While many bioproduction systems are well-studied in model organisms, investigating non-model organisms is essential to expand the field and utilize metabolically versatile strains. This investigation centers on Rhodopseudomonas palustris TIE-1, a purple non-sulfur bacterium capable of producing bioplastics. To increase bioplastic production, genes encoding the putative regulatory protein PhaR and the depolymerase PhaZ of the polyhydroxyalkanoate (PHA) biosynthesis pathway were deleted. Genes associated with pathways that might compete with PHA production, specifically those linked to glycogen production and nitrogen fixation, were deleted. Additionally, RuBisCO form I and II genes were integrated into TIE-1\'s genome by a phage integration system, developed in this study. Our results show that deletion of phaR increases PHA production when TIE-1 is grown photoheterotrophically with butyrate and ammonium chloride (NH4Cl). Mutants unable to produce glycogen or fix nitrogen show increased PHA production under photoautotrophic growth with hydrogen and NH4Cl. The most significant increase in PHA production was observed when RuBisCO form I and form I & II genes were overexpressed, five times under photoheterotrophy with butyrate, two times with hydrogen and NH4Cl, and two times under photoelectrotrophic growth with N2 . In summary, inserting copies of RuBisCO genes into the TIE-1 genome is a more effective strategy than deleting competing pathways to increase PHA production in TIE-1. The successful use of the phage integration system opens numerous opportunities for synthetic biology in TIE-1.IMPORTANCEOur planet has been burdened by pollution resulting from the extensive use of petroleum-derived plastics for the last few decades. Since the discovery of biodegradable plastic alternatives, concerted efforts have been made to enhance their bioproduction. The versatile microorganism Rhodopseudomonas palustris TIE-1 (TIE-1) stands out as a promising candidate for bioplastic synthesis, owing to its ability to use multiple electron sources, fix the greenhouse gas CO2, and use light as an energy source. Two categories of strains were meticulously designed from the TIE-1 wild-type to augment the production of polyhydroxyalkanoate (PHA), one such bioplastic produced. The first group includes mutants carrying a deletion of the phaR or phaZ genes in the PHA pathway, and those lacking potential competitive carbon and energy sinks to the PHA pathway (namely, glycogen biosynthesis and nitrogen fixation). The second group comprises TIE-1 strains that overexpress RuBisCO form I or form I & II genes inserted via a phage integration system. By studying numerous metabolic mutants and overexpression strains, we conclude that genetic modifications in the environmental microbe TIE-1 can improve PHA production. When combined with other approaches (such as reactor design, use of microbial consortia, and different feedstocks), genetic and metabolic manipulations of purple nonsulfur bacteria like TIE-1 are essential for replacing petroleum-derived plastics with biodegradable plastics like PHA.

The present study aimed to i) assess the disintegration of a novel bio-packaging during aerobic composting (2 and 6 % tested concentrations) and evaluate the resulting compost ii) analyse the ecotoxicity of bioplastics residues on earthworms; iii) study the microbial communities during composting and in \'earthworms\' gut after their exposure to bioplastic residues; iv) correlate gut microbiota with ecotoxicity analyses; v) evaluate the chemico-physical characterisation of bio-packaging after composting and earthworms\' exposure. Both tested concentrations showed disintegration of bio-packaging close to 90 % from the first sampling time, and compost chemical analyses identified its maturity and stability at the end of the process. Ecotoxicological assessments were then conducted on Eisenia fetida regarding fertility, growth, genotoxic damage, and impacts on the gut microbiome. The bioplastic residues did not influence the earthworms\' fertility, but DNA damages were measured at the highest bioplastic dose tested. Furthermore bioplastic residues did not significantly affect the bacterial community during composting, but compost treated with 2 % bio-packaging exhibited greater variability in the fungal communities, including Mortierella, Mucor, and Alternaria genera, which can use bioplastics as a carbon source. Moreover, bioplastic residues influenced gut bacterial communities, with Paenibacillus, Bacillus, Rhizobium, Legionella, and Saccharimonadales genera being particularly abundant at 2 % bioplastic concentration. Higher concentrations affected microbial composition by favouring different genera such as Pseudomonas, Ureibacillus, and Streptococcus. For fungal communities, Pestalotiopsis sp. was found predominantly in earthworms exposed to 2 % bioplastic residues and is potentially linked to its role as a microplastics degrader. After composting, Attenuated Total Reflection analysis on bioplastic residues displayed evidence of ageing with the formation of hydroxyl groups and amidic groups after earthworm exposure.

Waste cooking oil is a common byproduct in the culinary industry, often posing disposal challenges. This study explores its conversion into the valuable bioplastic material, medium-chain-length polyhydroxyalkanoate (mcl-PHA), through microbial biosynthesis in controlled bioreactor conditions. Twenty-four bacterial isolates were obtained from oil-contaminated soil and waste materials in Mahd Ad-Dahab, Saudi Arabia. The best PHA-producing isolates were identified via 16S rDNA analysis as Neobacillus niacini and Metabacillus niabensis, with the sequences deposited in GenBank (accession numbers: PP346270 and PP346271). This study evaluated the effects of various carbon and nitrogen sources, as well as environmental factors, such as pH, temperature, and shaking speed, on the PHA production titer. Neobacillus niacini favored waste cooking oil and yeast extract, achieving a PHA production titer of 1.13 g/L, while Metabacillus niabensis preferred waste olive oil and urea, with a PHA production titer of 0.85 g/L. Both strains exhibited optimal growth at a neutral pH of 7, under optimal shaking -flask conditions. The bioreactor performance showed improved PHA production under controlled pH conditions, with a final titer of 9.75 g/L for Neobacillus niacini and 4.78 g/L for Metabacillus niabensis. Fourier transform infrared (FT-IR) spectroscopy and gas chromatography-mass spectrometry (GC-MS) confirmed the biosynthesized polymer as mcl-PHA. This research not only offers a sustainable method for transforming waste into valuable materials, but also provides insights into the optimal conditions for microbial PHA production, advancing environmental science and materials engineering.

Titanium dioxide (TiO2) is a common pigment used in food packaging to provide a transparent appearance to plastic packaging materials. In the present study, poly(butylene adipate-co-terephthalate) (PBAT) incorporated with lignin-TiO2 nanoparticles (L-TiO2) eco-friendly composite films was prepared by employing an inexpensive melting and hot-pressing technique. The P-L-TiO2 composite films have been studied using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscope (SEM), Thermogravimetric analysis (TGA), and Differential scanning calorimetry (DSC) analysis. The FTIR results and homogeneous, dense SEM images confirm the interaction of L-TiO2 with the PBAT matrix. It has also been found that the addition of L-TiO2 nanoparticles can increase the crystallinity, tensile strength, and thermal stability of PBAT. The addition of L-TiO2 increased the tensile strength and decreased the elongation at break of films. The maximum tensile strength of the film, achieved with 5 wt% L-TiO2, was 47.0 MPa, compared with 24.3 MPa for pure PBAT film. The composite film with 5 wt% L-TiO2 has outstanding oxygen and water vapor barrier properties. As the content of lignin-TiO2 increases, the antimicrobial activity of the composite films also increases; the percentage of growth of all the tested bacteria Staphylococcus aureus (S. aureus), and Escherichia coli (E. coli) is significantly reduced. Strawberries were packed to evaluate the suitability of produced composite films as packaging materials, as they effectively preserved pigments from accumulation and extended the shelf-life as compared to commercial polyethylene packaging film.

This study evaluates the anaerobic mesophilic mono- and co-digestion of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH) plastic bottles as a proxy for rigid packaging materials. Initial tests showed a 97.3 ± 0.2 % reduction in weight and an observable alteration in the surface (thinning, color fading and pitting) of the PHBH bottles after eight weeks. Subsequent tests showed that PHBH squares (3 × 3 cm) produced 400 NmL-CH4/g-VSfed, at a slower rate compared to powdered PHBH but with similar methane yield. Co-digestion experiments with food waste, swine manure, or sewage sludge showed successful digestion of PHBH alongside organic waste (even at a high bioplastic loading of 20 % volatile solids basis), with methane production comparable to or slightly higher than that observed in mono-digestion. Molecular analyses suggested that the type of co-substrate influenced microbial activity and that methane production was mainly driven by hydrogenotrophic methanogenesis. These results suggest the potential for integrating rigid PHBH packaging into anaerobic digesters.

The development and application of an electrochemical sensor is reported for detection of poly(3-hydroxybutyrate) (P3HB) - a bioplastic derived from agro-industrial residues. To overcome the challenges of molecular imprinting of macromolecules such as P3HB, this study employed methanolysis reaction to break down the P3HB biopolymer chains into methyl 3-hydroxybutyrate (M3HB) monomers. Thereafter, M3HB were employed as the target molecules in the construction of molecularly imprinted sensors. The electrochemical device was then prepared by electropolymerizing a molecularly imprinted poly (indole-3-acetic acid) thin film on a glassy carbon electrode surface modified with reduced graphene oxide (GCE/rGO-MIP) in the presence of M3HB. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), scanning electron microscopy with field emission gun (SEM-FEG), Raman spectroscopy, attenuated total reflection Fourier-transform infrared (ATR-FTIR) and X-ray Photoelectron Spectroscopy (XPS) were employed to characterize the electrode surface. Under ideal conditions, the MIP sensor exhibited a wide linear working range of 0.1 - 10 nM and a detection limit of 0.3 pM (n = 3). The sensor showed good repeatability, selectivity, and stability over time. For the sensor application, the bioproduction of P3HB was carried out in a bioreactor containing the Burkholderia glumae MA13 strain and sugarcane byproducts as a supplementary carbon source. The analyses were validated through recovery assays, yielding recovery values between 102 and 104%. These results indicate that this MIP sensor can present advantages in the monitoring of P3HB during the bioconversion process.

The bioplastics sector promotes environmentally friendly means of cutting down on the usage of fossil fuels, plastic waste, and environmental pollution. Plastic contamination has detrimental effects on both ecological systems and the global food supply. The approach we present here to resolve this issue involves the integration of sericin and gelatin, obtained from cocoon and fish waste, respectively, with nano-reinforced cellulose crystals, to develop a biodegradable and compostable plastic material. The use of cocoon and fish wastes for the extraction of sericin and gelatin presents an environmentally beneficial approach since it contributes to waste reduction. The sericin level found in silk cocoon waste was determined to be 28.08%, and the gelatin amount in fish waste was measured to be 58.25%. The inclusion of sericin and gelatin in bioplastics was accompanied by the incorporation of glycerol, vinegar, starch, sodium hydroxide, and other coloring agents. Fourier transform infrared (FTIR) examination of bioplastics revealed the presence of functional groups that corresponded to the sericin and gelatin components. The tensile strength of the bioplastic material was measured to be 27.64 MPa/psi, while its thickness varied between 0.072 and 0.316 mm. The results of burial experiments indicated that the bioplastic material had a degradation rate of 85% after 14 days. The invention exhibits potential as a viable alternative for packaging, containment, and disposable plastic materials. The use of this sustainable approach is recommended for the extraction of sericin and gelatin from silk cocoons and fish waste, with the intention of using them as raw materials for bioplastic production.

At the International Symposium on Biodegradable Polymers (ISBP2022) in Sion, Switzerland, experts from academia and industry underscored the remarkable progress in biobased and biodegradable polymers (BBPs) since their initial commercialization around 50 years ago. Despite significant advancements, the technology readiness level (TRL), market adoption, and industrialization of BBPs is not yet competitive to conventional plastics. In this perspective, we summarize the challenges and requirements for advancing the development and industrialization of BBPs, drawing insights from international experts coming from academia and industry, who had participated in the survey and podium discussion during the ISBP2022. In fact, BBPs grapple with persistent and emerging challenges throughout the value chain. These challenges can be grouped into four areas and involve i) the pursuit of sustainable feedstocks together with efficient production and downstream processes as well as recycling technologies and infrastructure; ii) meeting or revisiting product requirements by industry, markets, and consumers; iii) navigating a non-level playing field in their sustainability assessment (LCA) compared to conventional plastics; and iv) struggling with underdeveloped and partially biased policy and financial frameworks as well as lacking clear definitions, terminologies and communication.