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UNLABELLED: Biodegradable plastics are urgently needed to replace petroleum-derived polymeric materials and prevent their accumulation in the environment. To this end, we isolated and characterized a halophilic and alkaliphilic bacterium from the Great Salt Lake in Utah. The isolate was identified as a Halomonas species and designated CUBES01. Full-genome sequencing and genomic reconstruction revealed the unique genetic traits and metabolic capabilities of the strain, including the common polyhydroxyalkanoate (PHA) biosynthesis pathway. Fluorescence staining identified intracellular polyester granules that accumulated predominantly during the strains exponential growth, a feature rarely found among natural PHA producers. CUBES01 was found to metabolize a range of renewable carbon feedstocks, including glucosamine and acetyl-glucosamine, as well as sucrose, glucose, fructose, and further glycerol, propionate, and acetate. Depending on the substrate, the strain accumulated up to ~60% of its biomass (dry wt/wt) in poly(3-hydroxybutyrate), while reaching a doubling time of 1.7 h at 30°C and an optimum osmolarity of 1 M sodium chloride and a pH of 8.8. The physiological preferences of the strain may not only enable long-term aseptic cultivation but also facilitate the release of intracellular products through osmolysis. The development of a minimal medium also allowed the estimation of maximum polyhydroxybutyrate production rates, which were projected to exceed 5 g/h. Finally, also, the genetic tractability of the strain was assessed in conjugation experiments: two orthogonal plasmid vectors were stable in the heterologous host, thereby opening the possibility of genetic engineering through the introduction of foreign genes. IMPORTANCE: The urgent need for renewable replacements for synthetic materials may be addressed through microbial biotechnology. To simplify the large-scale implementation of such bio-processes, robust cell factories that can utilize sustainable and widely available feedstocks are pivotal. To this end, non-axenic growth-associated production could reduce operational costs and enhance biomass productivity, thereby improving commercial competitiveness. Another major cost factor is downstream processing, especially in the case of intracellular products, such as bio-polyesters. Simplified cell-lysis strategies could also further improve economic viability.

Polyhydroxyalkanoates (PHAs) are a well-known group of biodegradable and biocompatible bioplastics that are synthesised and stored by microorganisms as carbon and energy reservoirs. Extracellular PHA depolymerases (ePhaZs), secreted by a limited range of microorganisms, are the main hydrolytic enzymes responsible for their environmental degradation. Pseudomonas sp. GK13, initially identified as P. fluorescens GK13, produces PHA and a prototypic ePhaZ that specifically degrades mcl-PHA. In this study, a comprehensive characterization of strain GK13 was performed. The whole genomic sequence of GK13 was consolidated into one complete chromosome, leading to its reclassification as P. solani GK13. We conducted a detailed in silico examination of the bacteria genomic sequence, specifically targeting PHA metabolic functions. From the different growth conditions explored, PHA accumulation occurred only under carbon/nitrogen (C/N) imbalance, whereas ePhaZ production was induced even at balanced C/N ratios in mineral media. We extend our study to other bacteria belonging to the Pseudomonas genus revealing that the ePhaZ production capacity is closely associated with mcl-PHA synthesis capacity, as also suggested by metagenomic samples. This finding suggests that these types of microorganisms could contribute to the carbon economy of the microbial community, by storing PHA in carbon-rich times, and sharing it with the rest of the population during times of carbon scarcity through PHA hydrolysis. The conclusion pointed that carbon cycle metabolism performed by P. solani GK13 may contribute to the environmental circular economy at a microscopic scale. [Display omitted] • The model ePhaZ-producing Pseudomonas sp. GK13 was reclassified as P. solani GK13. • Whole genome analysis revealed canonical clusters for PHA and alginate synthesis. • ePhaZ, unlike PHA, is produced even in balanced C/N ratios. • In Pseudomonas genus, ePhaZ and mcl-PHA production capacities are associated. • P. solani GK13 contributes to the microbial carbon circular economy. [ABSTRACT FROM AUTHOR]

Since polyhydroxyalkanoates (PHAs) are biodegradable, bioresourced and biocompatible, they are versatile biopolymers having huge applicability in food packaging, biomedicine, drug delivery systems, production of biofuels and many other fields. With the use of waste carbon sources or renewable feedstocks, their process of production can be made cost effective. Moreover, just by exploiting PHA synthesis enzymes, they can also be produced in vivo i.e., without employing microbial cell. Thanks to this adaptable PHA nanogranular technology, it also has become possible to immobilize a huge variety of enzymes, creating a novel and promising set of immobilization techniques. In comparison to soluble enzymes, immobilized enzymes have a number of significant benefits, like increased stability, higher catalytic activity, extended shelf life, recycling for repeated use and easier purification of the product, however, showing some limitations (like improper orientation, substrate diffusion and enzyme deactivate etc) as well. Due to directed display and a high surface area to volume ratio, the PHA based enzyme immobilization platform offers flexibility, stability and excellent functionality. The nano or micro beads of PHA produced in vitro , act as matrix for enzyme immobilization being both economically appealing and potentially applicable in a variety of process circumstances. The developments in synthetic biology and material science to produce more effective, adaptable PHA-enzyme systems for industrial and environmental applications, are future prospects for enzyme immobilization on PHA. It is important to optimize enzyme-PHA interactions for enhanced efficiency and cost effective industry level production processes. This review highlights the importance and great utility of PHA for enzyme immobilization. It mainly focuses on the in vivo and in vitro enzyme immobilization studies done so far, using latest technologies to enhance the properties of immobilized enzymes. [Display omitted] • Polyhydroxyalkanoates (PHAs) being biodegradable, biocompatible and highly versatile, are promising candidates for enzyme immobilization. • There exist a lot of scope for development of various applications of PHA-immobilized enzymes in different industries. • This review addresses the two approaches for enzyme immobilization i.e. in vivo and in vitro , detailing advantages and challenges of each method. • It evaluates efficiency, scalability, enzyme activity, cost-effectiveness and environmental impact of both approaches in various sectors. [ABSTRACT FROM AUTHOR]

Plastic production was almost 400.3 million tons worldwide in 2022, while bioplastics only accounted for 0.56% of the global plastic production. Polyhydroxyalkanoates (PHAs) are biodegradable and renewable polymers derived from bio-based sources that have the potential to replace traditional plastics. PHAs are naturally synthesized by bacteria and serve as carbon and energy reserves. However, the high production costs associated with PHAs have limited their commercialization and industrialization. Molecular dynamics (MD) simulations provide a comprehensive view of the interactions and binding between proteins and ligands, enabling a deeper understanding of the pharmacophoric interactions occurring within the binding site. By capturing the dynamic behavior of the ligand within the binding site, MD simulations can provide information about the conformational space occupied by the ligand over time, which can help determine the stability of the protein–ligand complex. MD simulation of PHA synthases from Pseudomonas aeruginosa and P. putida was performed to understand the kinematics of the binding interaction of the enzymes with ligands. The binding pockets for the enzymes were identified through conservation and superimposition. Comparison between PHA variants from P. putida was done to understand the metabolic activation of the enzymes and the mode of action followed by them. MD simulation was used to achieve the trajectories for each variant of PHA synthase, and different peaks were observed with different substrate molecules used. The highest stability was observed for PHA C1 from P. putida with 3-hydroxybutyral CoA. The second ligand which was studied, 3-hydroxyacyl CoA, showed the best interactions with PHA C2 from P. putida. PHA synthase from P. aeruginosa showed intermediate interaction with both ligands. Implementation of these enzymes has high-value applications in the development of bioplastics. [ABSTRACT FROM AUTHOR]

The aim of this study is to investigate the anaerobic treatability and generation of potential valuable by-products of agro-food industry wastewaters where bulgur production is carried out. Among the agro-food products, bulgur which is a wheat product (Triticum spp.) and rich in fibers, is one of the ancient foods of Anatolia. The most critical issue in bulgur production is high energy and water consumption for cooking where wheat starch gelatinizes and results in a considerable amount of high strength wastewater. In the raw bulgur wastewater sample, 81% of total chemical oxygen demand (tCOD) was in soluble form which was readily fermentable and acidic by nature. Batch anaerobic study results indicated almost complete tCOD removal (99%) which decreased from 7820 to 100 mg/L. The cumulative methane yield was determined as 275 mL/g tCODfed. Hence, for such industrial wastewaters with high level of organic pollution, anaerobic biotechnology stands out as one of the most effective treatment options. Fermented bulgur wastewater was also tested for the production of polyhydroxyalkanoates (PHAs). It was determined that 37% of the tCOD could be recovered as PHAs without hindering the biogas generation. In this context, it was shown that PHAs and biogas production could be simultaneously achievable with a smart organic carbon utilization strategy which will also contribute to reduce energy requirement and carbon footprint of the industrial bulgur production process as well as create environmentally friendly solutions through resource recovery. [ABSTRACT FROM AUTHOR]

To promote the formation of granular sludge with high polyhydroxyalkanoates (PHAs) synthesis ability, an anaerobic dynamic feeding process (AnDF) was proposed. This process combines the feast-famine mode with an anaerobic plug flow feeding process and involving variations in cycle length and settling time. The effects of lactic acid (LA) content (0 %, 20 %, and 40 % COD) on sludge granulation and PHAs production were investigated using three AnDF reactors (R1, R2, and R3). The results showed that the AnDF process feeding with LA not only effectively promoted sludge granulation but also improved its PHAs synthesis ability. The granules were quickly observed in R3 after 50 days of cultivation, with an average diameter of 0.69 mm. The maximum PHAs content reached 47.0 wt% in R3, representing a 30.09 % increase compared to R1. Additionally, extracellular polymeric substances (EPS)-producing bacteria observed in granular sludge may be the prime drivers of the formation of PHAs-producing granular sludge (PHAGS), which was defined as granular sludge with an average particle size larger than 0.30 mm and PHAs content above 40 % cell dry weight (CDW) of sludge samples. [Display omitted] • The concept of PHAs-producing granular sludge (PHAGS) was proposed. • Lactic acid feeding is benefit for sludge granulation and PHAs synthesis. • The rapid PHAGS formation was obtained by anaerobic dynamic feeding process. • The formation of PHAGS may be related to the enriched EPS-producing bacteria. [ABSTRACT FROM AUTHOR]

The high accumulation of plastic waste in the environment has led to great interest in biodegradable polymers, such as polylactic acid (PLA) or polyhydroxyalkanoates (PHAs). Their benefits, combined with the application of electrospinning technology, represent an innovative proposal for the food packaging industry. This article provides a comprehensive review of the latest developments of PLA- and PHA-biopolyester-based electrospun materials for food packaging applications, summarizing the reported technologies, material properties, applications, and invention patents. In addition, the legislation used to assess their biodegradability is also detailed. Electrospun packaging materials are largely developed through uniaxial, coaxial, emulsion, multiaxial, and needleless techniques. PLA- and PHA-biopolyester-based electrospun materials can be obtained as single and multilayer packaging structures, and the incorporation of natural extracts, organic compounds, and nanoparticles has become a great strategy for designing active food packaging systems. The biodegradability of electrospun materials has mainly been evaluated in soil, compost, and aquatic systems through ASTM and ISO normatives. In this review, the dependence of the biodegradation process on the polymer type, conditions, and test methods is clearly reviewed. Moreover, these biodegradable electrospun materials have shown excellent antioxidant and antimicrobial properties, resulting in a great method for extending the shelf life of fruits, bread, fish, and meat products. [ABSTRACT FROM AUTHOR]

In recent years, the prevalence of bone diseases is showing an increasing trend, which is mainly attributed to the aging of the global population. However, bone repair and regeneration are still an unsolved problem in the treatment of bone diseases, which include a series of biological events. Photocrosslinked natural hydrogel‐based microspheres (PNHMs) are spherical particles composed of photocrosslinked natural hydrogel components. Due to their morphology, injectability, and biocompatibility, PNHMs are widely used in tissue regeneration, particularly for bone defects. In this article, we review the available photocrosslinked natural materials for PNHMs, and then introduce their preparation methods. After that, we summarize the important advanced functionalities of PNHMs. For example, PNHMs can be loaded with different active ingredients to exert anti‐inflammatory, antibacterial, and antioxidant effects, while also realizing lubrication due to their specific shape and size distribution. In addition, the function of capturing ions can be realized via coordination. When applied in bone tissue engineering, PNHMs can promote angiogenesis and osteogenesis, with great potential in the treatment of bone diseases. Finally, the challenges and prospects of PNHMs for bone repair are discussed. [ABSTRACT FROM AUTHOR]

A homo-aromatic polyester was produced from glucose for the first time. Aromatic polyhydroxyalkanoate (PHA) was produced to the highest titer yet achieved. 3D model-based rational engineering of PHA synthase was used to enhance aromatic PHA production. Polymer production was enhanced by utilizing heterologous phasins. Pilot-scale fermentative production of aromatic PHAs was successful. We report the development of a metabolically engineered bacterium for the fermentative production of polyesters containing aromatic side chains, serving as sustainable alternatives to petroleum-based plastics. A metabolic pathway was constructed in an Escherichia coli strain to produce poly[ d -phenyllactate(PhLA)], followed by three strategies to enhance polymer production. First, polyhydroxyalkanoate (PHA) granule-associated proteins (phasins) were introduced to increase the polymer accumulation. Next, metabolic engineering was performed to redirect the metabolic flux toward PhLA. Furthermore, PHA synthase was engineered based on in silico simulation results to enhance the polymerization of PhLA. The final strain was capable of producing 12.3 g/l of poly(PhLA), marking it the first bio-based process for producing an aromatic homopolyester. Additional heterologous gene introductions led to the high level production of poly(3-hydroxybutyrate- co -11.7 mol% PhLA) copolymer (61.4 g/l). The strategies described here will be useful for the bio-based production of aromatic polyesters from renewable resources. Graphical abstract [Display omitted] The production of polyhydroxyalkanoates (PHAs) containing aromatic repeating units has been studied for decades, but has yet to be demonstrated in large-scale fermentation. This study reports the successful production of aromatic polyesters from glucose in high titers that was performed both in lab-scale (5 l) and pilot-scale (30 l) fermenters. Therefore, the current Technology Readiness Level (TRL) of this technology lies between 4 and 5. To advance this technology for industrialization, demonstration in industry-scale fermenters and the development of large-scale downstream processes will be necessary. Nevertheless, given the ongoing development of various fermentation technologies and purification processes for other PHAs, we anticipate that the industrialization of aromatic PHAs is feasible, potentially replacing petroleum-based aromatic polymers for use as commodity, engineering, and biomedical plastics. Lee et al. report the fermentative production of aromatic polyesters, which can serve as sustainable alternatives to petroleum-based plastics. This represents the first instance of homo-aromatic polyester production using glucose. Further pathway engineering facilitated the production of poly(3-hydroxybutyrate- co - d -phenyllactate), achieving the highest titer of aromatic polyester reported using microorganisms thus far. [ABSTRACT FROM AUTHOR]

Poly (3-hydroxybutyrate) (PHB) is a valuable biopolymer that is produced in industrial quantity but is not widely used in applications due to some drawbacks. The addition of cellulose nanofibers (CNF) as a biofiller in PHB/CNF nanocomposites may improve PHB properties and enlarge its application field. In this work, n-octyltriethoxy silane (OTES), a medium-chain-length alkyl silane, was used to surface chemically modify the CNF (CNF_OTES) to enhance their hydrophobicity and improve their compatibility with PHB. The surface functionalization of CNF and nanodimension were emphasized by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, thermogravimetric analysis, atomic force microscopy, dynamic light scattering, and water contact angle (CA). Surface modification of CNF with OTES led to an increase in thermal stability by 25 °C and more than the doubling of CA. As a result of the higher surface hydrophobicity, the CNF_OTES were more homogeneously dispersed in PHB than unmodified CNF, leading to a PHB nanocomposite with better thermal and mechanical properties. Thus, an increase by 122% of the storage modulus at 25 °C, a slight increase in crystallinity, a better melting processability, and good thermal stability were obtained after reinforcing PHB with CNF_OTES, paving the way for increasing PHB applicability. [ABSTRACT FROM AUTHOR]

The escalating environmental concerns associated with conventional plastic packaging have accelerated the development of sustainable alternatives, making food packaging a focus area for innovation. Bioplastics, particularly polyhydroxyalkanoates (PHAs), have emerged as potential candidates due to their biobased origin, biodegradability, and biocompatibility. PHAs stand out for their good mechanical and medium gas permeability properties, making them promising materials for food packaging applications. In parallel, zinc oxide (ZnO) nanoparticles (NPs) have gained attention for their antimicrobial properties and ability to enhance the mechanical and barrier properties of (bio)polymers. This review aims to provide a comprehensive introduction to the research on PHA/ZnO nanocomposites. It starts with the importance and current challenges of food packaging, followed by a discussion on the opportunities of bioplastics and PHAs. Next, the synthesis, properties, and application areas of ZnO NPs are discussed to introduce their potential use in (bio)plastic food packaging. Early research on PHA/ZnO nanocomposites has focused on solvent-assisted production methods, whereas novel technologies can offer additional possibilities with regard to industrial upscaling, safer or cheaper processing, or more specific incorporation of ZnO NPs in the matrix or on the surface of PHA films or fibers. Here, the use of solvent casting, melt processing, electrospinning, centrifugal fiber spinning, miniemulsion encapsulation, and ultrasonic spray coating to produce PHA/ZnO nanocomposites is explained. Finally, an overview is given of the reported effects of ZnO NP incorporation on thermal, mechanical, gas barrier, UV barrier, and antimicrobial properties in ZnO nanocomposites based on poly(3-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyvalerate), and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). We conclude that the functionality of PHA materials can be improved by optimizing the ZnO incorporation process and the complex interplay between intrinsic ZnO NP properties, dispersion quality, matrix–filler interactions, and crystallinity. Further research regarding the antimicrobial efficiency and potential migration of ZnO NPs in food (simulants) and the End-of-Life will determine the market potential of PHA/ZnO nanocomposites as active packaging material. [ABSTRACT FROM AUTHOR]

Polyhydroxyalkanoates (PHAs) are thermoplastic biopolyester with high biodegradability and biocompatibility. This review explores the potential of PHAs as promising alternatives to conventional petrochemical plastics, offering a solution to reduce plastic pollution. The strategic selection of conditions, including type of microorganism, carbon substrates and extraction methods, enables the effective PHAs production for maximum yield and cost-effectiveness. Recent advancements in characterization techniques, each with its own advantages and limitations, are explored to offer insights into PHA concentration, composition, and purity. This review highlights the versatility of PHAs in various applications, focusing on medicine, environmental remediation, industrial processes, and agriculture, where they can offer significant environmental benefits by replacing non-biodegradable materials. Despite production cost challenges, PHAs demonstrate promise for sustainable polymer development. This review contributes to advancing PHAs as a viable solution for eco-friendly applications across diverse industries. • Diverse cultivation and extraction strategies were explored to optimize PHAs production. • Characterization methods for PHAs include microscopy, spectroscopy, and chromatography. • PHAs are versatile and sustainable alternatives used in medicine, environmental remeidation, industry and agriculture. [ABSTRACT FROM AUTHOR]

[Display omitted] Monodisperse nanoparticles of biodegradable polyhydroxyalkanoates (PHAs) polymers, copolymers of 3-hydroxybutyrate (3HB) and 4-hydroxybutyrate (4HB), are synthesized using a membrane-assisted emulsion encapsulation and evaporation process for biomedical resorbable adhesives. The precise control over the diameter of these PHA particles, ranging from 100 nm to 8 μm, is achieved by adjusting the diameter of emulsion or the PHA concentration. Mechanical properties of the particles can be tailored based on the 3HB to 4HB ratio and molecular weight, primarily influenced by the level of crystallinity. These monodisperse PHA particles in solution serve as adhesives for hydrogel systems, specifically those based on poly(N , N -dimethylacrylamide) (PDMA). Semi-crystalline PHA nanoparticles exhibit stronger adhesion energy than their amorphous counterparts. Due to their self-adhesiveness, adhesion energy increases even when those PHA nanoparticles form multilayers between hydrogels. Furthermore, as they degrade and are resorbed into the body, the PHA nanoparticles demonstrate efficacy in in vivo wound closure, underscoring their considerable impact on biomedical applications. [ABSTRACT FROM AUTHOR]

Clostridium ragsdalei was found to produce (S)-2-hydroxyisovalerate (2-HIV) as a novel product in addition to acetate, ethanol, and d-2,3-butanediol in heterotrophic (d-xylose), autotrophic (CO), and mixotrophic (d-xylose + CO) conditions. Mixotrophic batch processes in stirred-tank bioreactors with continuous gassing resulted in improved production of this alpha-hydroxy acid compared to batch processes solely with either d-xylose or CO. The maximal CO uptake rate was considerably reduced in mixotrophic compared to autotrophic processes, resulting in a concomitant decreased total CO2 production. Simultaneous conversion of 9.5 g L−1 d-xylose and 320 mmol CO enabled the production of 1.8 g L−1 2-HIV in addition to 1.1 g L−1 d-2,3-butanediol, 2.0 g L−1 ethanol, and 1.8 g L−1 acetate. With reduced initial d-xylose (3.1 g L−1), l-valine production started when d-xylose was depleted, reaching a maximum of 0.4 g L−1 l-valine. Using l-arabinose or d-glucose instead of d-xylose in mixotrophic batch processes reduced the 2-HIV production by C. ragsdalei. Considerable amounts of meso-2,3-butanediol (0.9–1.3 g L−1) were produced instead, which was not observed with d-xylose. The monomer 2-HIV can form polyesters that make the molecule attractive for application as bioplastic (polyhydroxyalkanoates) or new composite material. [ABSTRACT FROM AUTHOR]

Polyhydroxyalkanoates (PHA) compose a family of biodegradable and biocompatible polyesters, and are a potential alternative to the use of petrochemical plastics. Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] is an extensively studied copolymer member of the PHA family due to its medical applications. Usually, 4HB monomer incorporation depends on the supply of structurally related carbon sources, such as 4-hydroxybutyric acid, ɣ-butyrolactone (GBL), and 1,4-butanediol, although it can occur in some bacteria when supplying unrelated carbon sources, such as glucose. This review is focused on the bacterial cell factories and on the economic aspects of P(3HB-co-4HB) production, discussing strategies to broadly enable its biotechnological production on an industrial scale. Articles discussing the genetic engineering and cultivation conditions of major promising bacterial platforms to enhance P(3HB-co-4HB) production (Cupriavidus necator, Escherichia coli, Burkholderia sacchari, Halomonas bluephagenesis, and others) are reviewed, describing and comparing the advantages and disadvantages of each one. Economic aspects of large-scale P(3HB-co-4HB) manufacturing in the context of a biorefinery are analyzed, highlighting the main bottlenecks of the bioprocess. Improvement of bacterial conversion rate of GBL to 4HB and the use of cheaper substrates for 4HB incorporation to the copolymer, through the development of novel pathways and newly isolated strains, should be the main targets for future studies. [ABSTRACT FROM AUTHOR]

Medium chain‐length polyhydroxyalkanoates (mcl‐PHAs) are efficiently synthesized from fatty acids. It was hypothesized that under non‐axenic conditions, the anti‐microbial properties of fatty acids would reduce growth of microbial competitors and allow Pseudomonas putida LS46 to dominate the population resulting in non‐sterile mcl‐PHA production. Non‐sterile batch and fed‐batch cultures produced similar final mcl‐PHA content, monomer composition, and productivity during 24–27 h cultivations when compared to sterile control cultures for initial octanoic acid concentrations ranging from 5 to 40 mM at an initial pH of 6.5. In the absence of a P. putida inoculum, growth was eventually observed in non‐sterile medium after a lag period of up to 120 h, the length of which was dependent on the octanoic acid concentration. The efficacy of prolonged non‐sterile cultivation was tested using a sequential batch reactor (SBR). After 28 days (672 h), 1.62 g L−1 of total biomass was observed, of which the mcl‐PHA content was 47%. This resulted in a PHA titer of 0.77 g L−1, and a volumetric PHA productivity of 32 mg L−1 h−1. The polymer composition remained stable throughout at 87.0 ± 2.7% C8 monomers, and short‐chain length (scl‐) monomers were not detected. This study is the first of its kind to report that purely mcl‐PHAs were produced after prolonged periods in a non‐sterile environment and demonstrate that medium chain‐length fatty acids exert a strong selective pressure toward organisms that synthesize mcl‐PHA. This suggests an opportunity for mcl‐PHA production in open continuous cultivation, which could reduce both fixed and operating costs. [ABSTRACT FROM AUTHOR]

AbstractThe development and commercialization of bio-based and biodegradable polyhydroxyalkanoates (PHAs) biopolymers could be crucial for the transition toward a sustainable circular economy. However, despite potential traditional and novel applications in the packaging, textiles, agriculture, automotive, electronics, and biomedical industries, the commercialization of PHAs is limited by their current market competitiveness. This review provides the first critical assessment of the current pure culture pilot-scale PHA literature, which could be crucial in translating promising laboratory-scale developments into industrial-scale commercial PHA production. It will also complement reviews of mixed microbial cultures currently dominating pilot-scale PHA literature. Pure culture fermentations could provide advantages, such as ease of characterizing microbial producers’ behavior, higher PHA productivities, and better alignment with existing PHA commercialization and industrial biotechnology approaches. Key aspects, including producer organisms, fermentation volumes and schemes, control schemes, optimization, and properties of the polymers produced, are discussed in-depth, to elucidate important trends, achievements, and knowledge gaps. Furthermore, specific ways for future pilot-scale studies to help address current PHA commercialization challenges are also identified. The insights, and recommendations provided will be extremely beneficial for the future development of PHA production, at both pilot and commercial scales, whilst also being beneficial to the production of other microbial polymers and industrial biotechnology as a whole. [ABSTRACT FROM AUTHOR]

The combination of paclitaxel (PTX) with other chemotherapy drugs (e.g., gemcitabine, GEM) or genetic drugs (e.g., siRNA) has been shown to enhance therapeutic efficacy against tumors, reduce individual drug dosages, and prevent drug resistance associated with single-drug treatments. However, the varying solubility of chemotherapy drugs and genetic drugs presents a challenge in co-delivering these agents. In this study, nanoparticles loaded with PTX were prepared using the biodegradable polymer material poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx). These nanoparticles were surface-modified with target proteins (Affibody molecules) and RALA cationic peptides to create core-shell structured microspheres with targeted and cationic functionalization. A three-drug co-delivery system (PTX@PHBHHx-ARP/siRNAGEM) were developed by electrostatically adsorbing siRNA chains containing GEM onto the microsphere surface. The encapsulation efficiency of PTX in the nanodrug was found to be 81.02%, with a drug loading of 5.09%. The chemogene adsorption capacity of siRNAGEM was determined to be 97.3%. Morphological and size characterization of the nanodrug revealed that PTX@PHBHHx-ARP/siRNAGEM is a rough-surfaced microsphere with a particle size of approximately 150 nm. This nanodrug exhibited targeting capabilities toward BT474 cells with HER2 overexpression while showing limited targeting ability toward MCF-7 cells with low HER2 expression. Results from the MTT assay demonstrated that PTX@PHBHHx-ARP/siRNAGEM exhibits high cytotoxicity and excellent combination therapy efficacy compared to physically mixed PTX/GEM/siRNA. Additionally, Western blot analysis confirmed that siRNA-mediated reduction of Bcl-2 expression significantly enhanced cell apoptosis mediated by PTX or GEM in tumor cells, thereby increasing cell sensitivity to PTX and GEM. This study presents a novel targeted nanosystem for the co-delivery of chemotherapy drugs and genetic drugs. [ABSTRACT FROM AUTHOR]

Polyhydroxyalkanoates (PHA) is a carbon neutral material that contributes to reducing greenhouse gas emissions due to manufactured from biomass and easily degraded by the enzymatic effects of microorganisms in the natural environment. However, PHA exhibits poorer mechanical properties and processability compared with petroleum‐based plastics. This study used cellulose nanofiber (CNF) to improve limit of PHA. Moreover, CNF was silylated to reduce polarity difference with PHA and enhance the dispersibility of nanocellulose at PHA. At a result, the silylation process was successfully performed by Si‐CH3 stretching peaks in Fourier transform infrared spectroscopy spectra and hydrophobicity of TEOS‐MTES‐CNF (TECNF) was confirmed by observed water contact angle (147°). In addition, nanostructure of TECNF was maintained during silylation and drying process through field emission scanning electron microscopy. Moreover, the PHA/TECNF composite showed enhanced processability, and tensile strength was increase almost 37% (0.52 MPa) compared with PHA. Oxygen transmission rates (300 cc/m2۰day) and single lap shear strength (225 kPa) were determined to be at least equivalent or superior to those of commercial packaging materials. Therefore, TECNF could be considered as a reinforcing agent, nucleating agent, and plasticizer in PHA. Also, this composite has possibilities to using as environmentally friendly packaging materials. [ABSTRACT FROM AUTHOR]

Background/Objectives: Owing to the growing resistance of methicillin-resistant Staphylococcus aureus (MRSA) to conventional antibiotics, the development of innovative therapeutic strategies for the treatment of MRSA-infected cutaneous wounds poses a significant challenge. Methods: Here, by using polyhydroxyalkanoates (PHA), emerging biodegradable and biocompatible polymers naturally produced by various microorganisms, we developed clindamycin-loaded PHA nanoparticles (Cly-PHA NPs) as a novel approach for the treatment of MRSA-infected cutaneous wounds. Results: Cly-PHA NPs were characterized in terms of mean particle size (216.2 ± 38.9 nm), polydispersity index (0.093 ± 0.03), zeta potential (11.3 ± 0.5 mV), and drug loading (6.76 ± 0.19%). Owing to the sustained release of clindamycin over 2 days provided by the PHA, Cly-PHA NPs exhibited potent antibacterial effects against MRSA. Furthermore, Cly-PHA NPs significantly facilitated wound healing in a mouse model of MRSA-infected full-thickness wounds by effectively eradicating MRSA from the wound bed. Conclusions: Therefore, our results suggest that Cly-PHA NPs offer a promising approach for combating MRSA infections and accelerating cutaneous wound healing. [ABSTRACT FROM AUTHOR]

The current study exploited cheese whey supernatant (CWS) as a renewable resource for polyhydroxyalkanoates (PHAs) formation. Structure identification and investigating the influence of lipid membranes' chemistry on PHA structuration were detailed. Approximately 66 bacterial isolates from dairy products companies in Egypt were screened for PHA production, and the bacterial isolate AZU‐A5, identified as Bacillus licheniformis strain AZU‐A5, showed the highest production PHA 0.93 g L−1) using whey lactose. The highest PHA rate in the individual design was 1.59 g L−1 with a recovery yield of 33.08% (w w−1), while the production rate in the statistical design reached 1.75 g L−1 PHA and 51.60% PHA content. Structurally, the PHA was identified as polyhydroxy‐3R‐butyrate (R‐PH3B). The fibrous texture by SEM highlighted the self‐assembly of PHB. The CD analysis of the PHA films suggested favorable hydrophobic interactions between lipids and PHB. Higher lipid contents not only caused a decrement in the CD signal of PHB but also bathochromic effects occurred. The chain length of lipids exerted high impacts on interactions (CD) and structuration of PHB (Δλ). The unsaturation showed little influence on CD and negligible effect on Δλ, while the head group exerted no effect on CD with a considerable impact on Δλ. [ABSTRACT FROM AUTHOR]

In this study, specific additives were incorporated in polyhydroxyalcanoate (PHB) and polylactic acid (PLA) blend to improve its compatibility, and so enhance the cell metabolic activity of scaffolds for tissue engineering. The formulations were manufactured through material extrusion (MEX) additive manufacturing (AM) technology. As additives, petroleum-based poly(ethylene) with glicidyl metacrylate (EGM) and methyl acrylate-co-glycidyl methacrylate (EMAG); poly(styrene-co-maleic anhydride) copolymer (Xibond); and bio-based epoxidized linseed oil (ELO) were used. On one hand, standard geometries manufactured were assessed to evaluate the compatibilizing effect. The additives improved the compatibility of PHB/PLA blend, highlighting the effect of EMAG and ELO in ductile properties. The processability was also enhanced for the decrease in melt temperature as well as the improvement of thermal stability. On the other hand, manufactured scaffolds were evaluated for the purpose of bone regeneration. The mean pore size and porosity exhibited values between 675 and 718 μm and 50 and 53%, respectively. According to the results, the compression stress was higher (11–13 MPa) than the required for trabecular bones (5–10 MPa). The best results in cell metabolic activity were obtained by incorporating ELO and Xibond due to the decrease in water contact angle, showing a stable cell attachment after 7 days of culture as observed in SEM. [ABSTRACT FROM AUTHOR]

Although efforts have been undertaken to produce polyhydroxyalkanoates (PHA) with various monomers, the low yield of PHAs because of complex metabolic pathways and inhibitory substrates remains a major hurdle in their analyses and applications. Therefore, we investigated the feasibility of mass production of PHAs containing 5-hydroxyvalerate (5HV) using δ-valerolactone (DVL) without any pretreatment along with the addition of plant oil to achieve enough biomass. We identified that PhaCBP-M-CPF4, a PHA synthase, was capable of incorporating 5HV monomers and that C. necator PHB−4 harboring phaCBP-M-CPF4 synthesized poly(3HB-co-3HHx-co-5HV) in the presence of bean oil and DVL. In fed-batch fermentation, the supply of bean oil resulted in the synthesis of 49 g/L of poly(3HB-co-3.7 mol% 3HHx-co-5.3 mol%5HV) from 66 g/L of biomass. Thermophysical studies showed that 3HHx was effective in increasing the elongation, whereas 5HV was effective in decreasing the melting point. The contact angles of poly(3HB-co-3HHx-co-5HV) and poly(3HB-co-3HHx) were 109 and 98°, respectively. In addition, the analysis of microbial degradation confirmed that poly(3HB-co-3HHx-co-5HV) degraded more slowly (82% over 7 days) compared to poly(3HB-co-3HHx) (100% over 5 days). Overall, the oil-based fermentation strategy helped produce more PHA, and the mass production of novel PHAs could provide more opportunities to study polymer properties. [ABSTRACT FROM AUTHOR]

Upcycling food waste into PHAs offers a promising solution to reduce environmental impact and generate value-added products for drug delivery, scaffold materials, and tissue engineering. Lignocellulosic food waste requires pretreatment and detoxification, whereas waste cooking oil can be fermented directly to produce PHAs. Cupriavidus necator excels in PHA production from lignocellulosic feedstock, showing tolerance to inhibitors. Pseudomonas spp. stand out for waste cooking oil utilization owing to their capacity for esterase and lipase production, enabling PHA accumulation. Butyl acetate is the optimal solvent for PHA recovery owing to its high yield, purity, and molecular weight, with minimal toxicity. PHAs can be recycled through methods such as extrusion, pyrolysis, gasification, and anaerobic digestion within a closed-loop cycle. This review emphasizes the urgent need for food waste upcycling as a response to the mounting global food waste crisis. Focusing on polyhydroxyalkanoates (PHAs) as an alternative to traditional plastics, it examines the potential of various food wastes as feedstock for microbial fermentation and PHA production. The upcycling of food waste including cheese whey, waste cooking oil, coffee waste, and animal fat is an innovative practice for food waste management. This approach not only mitigates environmental impacts but also contributes to sustainable development and economic growth. Downstream processing techniques for PHAs are discussed, highlighting their role in obtaining high-quality materials. The study also addresses sustainability considerations, emphasizing biodegradability and recycling, while acknowledging the challenges associated with this path. [ABSTRACT FROM AUTHOR]

Synthetic plastics are in great demand in society due to their diversified properties, but they cause environmental pollution due to their non-biodegradable nature. Therefore, synthetic plastics are in need to be replaced with biodegradable plastics. Polyhydroxyalkanoates (PHAs), bacterial biopolymers are natural alternative to synthetic plastics. These are present inside the bacterial cytoplasm in granular form. Presently, the production cost of PHA is high due to expensive carbon substrates used in its biosynthesis. Therefore, this study focuses on the cost-effective production of PHA using waste carbon sources. Rice bran and sugarcane molasses were used as the carbon source for PHA production from Bacillus subtilis, Bacillus cereus, Alcaligenes sp. and Pseudomonas aeruginosa. PHA production from these bacterial strains was confirmed through Sudan Black-B screening. With rice bran, as carbon source, the highest PHA yield obtained was for P. aeruginosa, which yielded 93.7% and lowest was 35.5% for B. cereus. Surprisingly, B. cereus produced the highest cell dry mass (0.045 g/L) but its extracted PHA contents were lowest being only 0.02 g/L. Alcaligenes sp. with 0.031 g/L CDM yielded 87.1% PHA. B. subtilis had a CDM 0.029 g/L, 0.02 g/L PHA content and a yield of 69.10%. In the case of sugarcane molasses, P. aeruginosa produced 95% PHA yield, 0.02 g/L CDM, and 0.019 g/L PHA content. Alcaligenes sp. yielded 90.9% PHA, 0.011 g/L CDM, and 0.01 g/L PHA content. B. subtilis produced 91.6% PHA yield, 0.012 g/L CDM, 0.011 g/L PHA content; B. cereus produced 80% PHA yield, 0.015 g/L CDM, 0.012 g/L PHA content at 37 °C, pH 7. Higher concentrations of carbon sources increased the CDM and decreased the PHA yield. The maximum yield of PHA was obtained from sugarcane molasses. 24–48 h of incubation was optimal for B. subtilis and B. cereus, while for Alcaligenes and P. aeruginosa incubation time of 48–96 h was desirable for higher PHA yield. The extracted biopolymers were analyzed by Fourier transform infrared spectroscopy (FTIR), which identified the extracted biopolymers as poly-3-hydroxybutyrate P(3HB). The thermal properties of the extracted biopolymers, such as melting temperatures, were analyzed by differential scanning calorimetry (DSC), which confirmed the thermal stability. [ABSTRACT FROM AUTHOR]

Conventional petroleum-derived polymers are valued for their versatility and are widely used, owing to their characteristics such as cost-effectiveness, diverse physical and chemical qualities, lower molecular weight, and easy processability for large-scale production. However, the extensive accumulation of such plastics leads to serious environmental issues. To combat this existing situation, an alternative lies in the production of bioplastics from natural and renewable sources such as plants, animals, microbes, etc. Bioplastics obtained from renewable sources are compostable and susceptible to degradation caused by microbes hydrolyzing to CO2, CH4, and biomass. Also, certain additives are reinforced into the bioplastic films to improve their physicochemical properties and degradation rate. However, on degradation, the bio-microplastic (BM) produced could have positive as well as negative impact on the soil health. This article thus focuses on the degradation of various fossil based as well as bio based biodegradable plastics such as polyhydroxyalkanoates (PHA), polyhydroxy butyrate (PHB), polylactic acid (PLA), polybutylene succinate (PBS), polycaprolactone (PCL), and polysaccharide derived bioplastics by mechanical, thermal, photodegradation and microbial approaches. The degradation mechanism of each approach has been discussed in detailed for different bioplastics. How the incorporation or reinforcement of various additives in the biodegradable plastics effects their degradation rates has also been discussed. In addition to that, the impact of generated bio-microplastic on physicochemical properties of soil such as pH, bulk density, carbon, nitrogen content etc. and biological properties such as on genome of native soil microbes and on plant nutritional health have been discussed in detailed. [ABSTRACT FROM AUTHOR]

This review highlights recent innovations in food packaging, emphasizing the shift from conventional petroleum-based materials to bio-based alternatives and smart packaging systems. Bio-based materials, such as starch, cellulose, and polyhydroxyalkanoates (PHA), offer sustainable solutions due to their biodegradability and reduced environmental impact. These materials are positioned as eco-friendly alternatives to traditional plastics but face challenges related to production costs and scalability. Additionally, advancements in smart packaging technologies, including sensor and indicator systems, provide real-time food quality monitoring, enhancing food safety and reducing waste. Active packaging technologies, incorporating natural antioxidants and moisture control, extend product shelf life and improve food preservation. Furthermore, these biopolymers typically present a lower CO2 footprint, energy costs, and water consumption during production, compared to traditionally used synthetic plastics. The review identifies challenges, such as regulatory barriers and technological limitations, but also outlines significant opportunities for future research and innovation in the food packaging sector, aiming for more efficient, safer, and environmentally sustainable packaging solutions. [ABSTRACT FROM AUTHOR]

Background: Polyhydroxyalkanoates (PHAs) are naturally produced biopolymers with significant scientific and biotechnological potential. This study aimed to investigate the diversity of the PHA-producing bacterial community and PhaC genes in soil samples collected from a municipal solid waste disposal site known as "Qoshe" in Addis Ababa, Ethiopia, using a shotgun metagenomics approach. The SqueezeMeta pipeline was used to analyze the microbial community in the waste samples. A CD search against the TIGRFAM protein family database was performed to identify the complete-length multidomain sequences of PhaC genes and classify them into their respective classes. Statistical analysis and data visualization were performed using RStudio with R version 4.2.3. Results: The findings of this study suggest that known and unknown taxa likely contribute to the phaC genes of municipal solid waste. Taxonomic profiling of the metagenomic data revealed that the majority of the PHA-producing taxa belonged to the phylum Proteobacteria (80%), followed by Actinomycetota (16.5%). Furthermore, this study identified Thiomonas and unclassified Mycobacterium as the main contributors to class I PhaC genes. Class II PhaC genes are predominantly associated with the Pseudomonadaceae family, followed by unclassified Hyphomicrobials and Acidimicrobiales. Class III PhaC genes are abundantly related to the Methylococcaceae family, specifically the Methylocaldum genus. The analysis of PhaC gene sequences revealed high level of diversity, with a significant proportion of putative PhaC genes exhibiting low sequence identity with each other and PhaC gene in the database. Notably, the sequence variation observed within the same PhaC gene classes suggests the potential presence of previously unidentified PhaC gene variants. Conclusions: Overall, this research improves our understanding of the diversity of PHA-producing taxa and PhaC genes in municipal solid waste environments, providing opportunities for sustainable PHA production and waste management strategies. However, additional studies, including the isolation and characterization of specific strains, are necessary to confirm the PHA production capabilities of these strains and explore their biotechnological potential. [ABSTRACT FROM AUTHOR]

The search for alternatives to petrochemical plastics has intensified, with increasing attention being directed toward bio-based polymers (bioplastics), which are considered healthier and more environmentally friendly options. In this review, a comprehensive overview of polyhydroxyalkanoates (PHAs) is provided, including their characterization, applications, and the mechanisms underlying their biosynthesis. PHAs are natural polyesters produced by a wide range of prokaryotic and some eukaryotic organisms, positioning them as a significant and widely studied type of bioplastic. Various strategies for the production of PHAs from agroindustrial waste, such as cacao shells, cheese whey, wine, wood, and beet molasses, are reviewed, emphasizing their potential as sustainable feedstocks. Industrial production processes for PHAs, including the complexities associated with extraction and purification, are also examined. Although the use of waste materials offers promise in reducing costs and environmental impact, challenges remain in optimizing these processes to enhance efficiency and cost-effectiveness. The need for continued research and development to improve the sustainability and economic viability of PHA production is emphasized, positioning PHAs as a viable and eco-friendly alternative to conventional petroleum-based plastics. [ABSTRACT FROM AUTHOR]

Abstract Background Polyhydroxyalkanoates (PHAs) are naturally produced biopolymers with significant scientific and biotechnological potential. This study aimed to investigate the diversity of the PHA-producing bacterial community and PhaC genes in soil samples collected from a municipal solid waste disposal site known as “Qoshe” in Addis Ababa, Ethiopia, using a shotgun metagenomics approach. The SqueezeMeta pipeline was used to analyze the microbial community in the waste samples. A CD search against the TIGRFAM protein family database was performed to identify the complete-length multidomain sequences of PhaC genes and classify them into their respective classes. Statistical analysis and data visualization were performed using RStudio with R version 4.2.3. Results The findings of this study suggest that known and unknown taxa likely contribute to the phaC genes of municipal solid waste. Taxonomic profiling of the metagenomic data revealed that the majority of the PHA-producing taxa belonged to the phylum Proteobacteria (80%), followed by Actinomycetota (16.5%). Furthermore, this study identified Thiomonas and unclassified Mycobacterium as the main contributors to class I PhaC genes. Class II PhaC genes are predominantly associated with the Pseudomonadaceae family, followed by unclassified Hyphomicrobials and Acidimicrobiales. Class III PhaC genes are abundantly related to the Methylococcaceae family, specifically the Methylocaldum genus. The analysis of PhaC gene sequences revealed high level of diversity, with a significant proportion of putative PhaC genes exhibiting low sequence identity with each other and PhaC gene in the database. Notably, the sequence variation observed within the same PhaC gene classes suggests the potential presence of previously unidentified PhaC gene variants. Conclusions Overall, this research improves our understanding of the diversity of PHA-producing taxa and PhaC genes in municipal solid waste environments, providing opportunities for sustainable PHA production and waste management strategies. However, additional studies, including the isolation and characterization of specific strains, are necessary to confirm the PHA production capabilities of these strains and explore their biotechnological potential.

Polyhydroxyalkanoate (PHA) is a type of polymer that is biodegradable, biocompatible, and eco-friendly. In this research, certain bacteria from textile waste, such as curing and bleaching, leather, and sizing waste, were found to produce PHA and remove color from textile dyes. Three types of bacteria were identified as Bacillus cereus SH2, Priestia megaterium SH3, and Bacillus cereus SH4 through genetic sequencing. These bacteria were grown in a medium with Disperse Red 73 to assess their ability to remove color and produce PHAs. The presence of PHA in the extracted biopolymer was confirmed using Fourier transform infrared spectroscopy (FTIR). Among the bacteria, SH4 was the most efficient at producing PHA (49.09 %), with a concentration of 0.54 gL-1. SH2 showed significant color removal ability (98.91 %), indicating its potential in the treatment of dye wastewaters. The isotherm, kinetics, and thermodynamics of the process were also investigated. The findings indicated that the absorption kinetics and isotherms were most accurately described by the pseudo-first-order and Langmuir models, respectively. The thermodynamic analysis of the adsorption process revealed that the adsorption occurred spontaneously and was endothermic in nature. This is the first report of the newly isolated strains from textile waste and their ability in degradation of Disperse Red 73 and produce PHA. [ABSTRACT FROM AUTHOR]

Abstract Blood vessels are highly dynamic and complex structures with a variety of physiological functions, including the transport of oxygen, nutrients, and metabolic wastes. Their normal functioning involves the close and coordinated cooperation of a variety of cells. However, adverse internal and external environmental factors can lead to vascular damage and the induction of various vascular diseases, including atherosclerosis and thrombosis. This can have serious consequences for patients, and there is an urgent need for innovative techniques to repair damaged blood vessels. Polyesters have been extensively researched and used in the treatment of vascular disease and repair of blood vessels due to their excellent mechanical properties, adjustable biodegradation time, and excellent biocompatibility. Given the high complexity of vascular tissues, it is still challenging to optimize the utilization of polyesters for repairing damaged blood vessels. Nevertheless, they have considerable potential for vascular tissue engineering in a range of applications. This summary reviews the physicochemical properties of polyhydroxyalkanoate (PHA), polycaprolactone (PCL), poly-lactic acid (PLA), and poly(lactide-co-glycolide) (PLGA), focusing on their unique applications in vascular tissue engineering. Polyesters can be prepared not only as 3D scaffolds to repair damage as an alternative to vascular grafts, but also in various forms such as microspheres, fibrous membranes, and nanoparticles to deliver drugs or bioactive ingredients to damaged vessels. Finally, it is anticipated that further developments in polyesters will occur in the near future, with the potential to facilitate the wider application of these materials in vascular tissue engineering.

Androgenetic alopecia (AGA), the most prevalent clinical hair loss, lacks safe and effective treatments due to downregulated angiogenic genes and insufficient vascularization in the perifollicular microenvironment of the bald scalp in AGA patients. In this study, a hyaluronic acid (HA) based hydrogel-formed microneedle (MN) was designed, referred to as V-R-MNs, which was simultaneously loaded with vascular endothelial growth factor (VEGF) and the novel hair loss drug Ritlecitinib, the latter is encapsulated in slowly biodegradable polyhydroxyalkanoates (PHAs) nanoparticles (R-PHA NPs) for minimally invasive AGA treatment. The integration of HA based hydrogel alongside PHA nanoparticles significantly bolstered the mechanical characteristics of microneedles and enhanced skin penetration efficiency. Due to the biosafety, mechanical strength, and controlled degradation properties of HA hydrogel formed microneedles, V-R-MNs can effectively penetrate the skin's stratum corneum, facilitating the direct delivery of VEGF and Ritlecitinib in a minimally invasive, painless and long-term sustained release manner. V-R-MNs not only promoted angiogenesis and improve the immune microenvironment around the hair follicle to promote the proliferation and development of hair follicle cells, but also the application of MNs to the skin to produce certain mechanical stimulation could also promote angiogenesis. In comparison to the clinical drug minoxidil for AGA treatment, the hair regeneration effect of V-R-MN in AGA model mice is characterized by a rapid onset of the anagen phase, improved hair quality, and greater coverage. This introduces a new, clinically safer, and more efficient strategy for AGA treatment, and serving as a reference for the treatment of other related diseases.

Abstract Background Plastic is widely utilized in packaging, frameworks, and as coverings material. Its overconsumption and slow degradation, pose threats to ecosystems due to its toxic effects. While polyhydroxyalkanoates (PHA) offer a sustainable alternative to petroleum-based plastics, their production costs present significant obstacles to global adoption. On the other side, a multitude of household and industrial activities generate substantial volumes of wastewater containing both organic and inorganic contaminants. This not only poses a threat to ecosystems but also presents opportunities to get benefits from the circular economy. Main body of abstract Production of bioplastics may be improved by using the nutrients and minerals in wastewater as a feedstock for microbial fermentation. Strategies like feast-famine culture, mixed-consortia culture, and integrated processes have been developed for PHA production from highly polluted wastewater with high organic loads. Various process parameters like organic loading rate, organic content (volatile fatty acids), dissolved oxygen, operating pH, and temperature also have critical roles in PHA accumulation in microbial biomass. Research advances are also going on in downstream and recovery of PHA utilizing a combination of physical and chemical (halogenated solvents, surfactants, green solvents) methods. This review highlights recent developments in upcycling wastewater resources into PHA, encompassing various production strategies, downstream processing methodologies, and techno-economic analyses. Short conclusion Organic carbon and nitrogen present in wastewater offer a promising, cost-effective source for producing bioplastic. Previous attempts have focused on enhancing productivity through optimizing culture systems and growth conditions. However, despite technological progress, significant challenges persist, such as low productivity, intricate downstream processing, scalability issues, and the properties of resulting PHA. Graphical abstract

Background: Polyhydroxyalkanoates (PHAs) are optimal potential materials for industrial and medical uses, characterized by exceptional sustainability, biodegradability, and biocompatibility. These are primarily from various bacteria and archaea. Bacterial strains with effective PHA formation capabilities and minimal production cost form the foundation for PHA production. Detailed genomic analysis of these PHA-generating bacteria is vital to understand their PHA production pathways and enhance their synthesis capability. Results: ZZQ-149, a halophilic, PHA-producing bacterium, was isolated from the sediment of China's Qinghai Lake. Here, we decoded the full genome of ZZQ-149 using Single Molecule Real Time (SMRT) technology based on PacBio RS II platform, coupled with Illumina sequencing platforms. Physiological, chemotaxonomic traits, and phylogenetic analysis based on 16 S rRNA gene and single copy core genes of ninety-nine Halomonas type strains identified ZZQ-149 as the type strain of Halomonas qinghailakensis. Furthermore, a low average nucleotide identity (ANI, < 95%) delineated the genetic differences between ZZQ-149 and other Halomonas species. The ZZQ-149 genome, with a DNA G + C content of 52%, comprises a chromosome (3, 798, 069 bps) and a plasmid (6, 107 bps). The latter encodes the toxin-antitoxin system, BrnT/BrnA. Through comprehensive genome sequencing and analysis, we identified multiple PHA-synthesizing enzymes and an unprecedented combination of eight PHA-synthesizing pathways in ZZQ-149. Conclusions: Being a halophilic, PHA-producing bacterium, ZZQ-149 exhibits potential as a high PHA producer for engineered bacteria via genome editing while ensuring low-cost PHA production through continuous, unsterilized fermentation. [ABSTRACT FROM AUTHOR]

In the current study, the ability of an indigenous marine Actinomycete Nocardiopsis dassonvillei (NCIM 5124) to degrade poly(3-hydroxybutyrate)-PHB was examined. From the whole genome sequencing data of the organism, information regarding the PHB depolymerase gene and amino acid sequence (Accession number: MCK9871921.1) was retrieved. In silico studies indicated the presence of a signal peptide characteristic of extracellular enzymes. ProtParam tool predicted that the protein had a molecular mass of 42.46 kDa with an isoelectric point of 4.51. Aliphatic and instability index values suggested that the protein was stable and the observed GARVY value indicated its hydrophilic nature. 3D structure prediction and multiple sequence alignments revealed the presence of Type I catalytic domain [including the oxyanion histidine towards the N terminal, the catalytic triad with serine (as a part of GLSAG pentapeptide), aspartate and histidine], substrate binding and linker domain. The organism was able to grow on PHB in solid and liquid media and effectively degrade it. Maximum enzyme activity (1.8 U/mL/min) was observed after 5 d of incubation in Bushnell Hass Medium containing 0.1% PHB, 1.5% sodium chloride, at 30 °C, pH 7.5 with agitation at 130 rpm. Application of the organism in disintegrating films of PHB and its copolymers was successfully demonstrated on the basis of weight loss and scanning electron microscope analysis. To the best of our knowledge, this is the first report on production of PHB depolymerase with high efficiency by N. dassonvillei, an organism that holds promise in degrading PHB-derived waste material. [ABSTRACT FROM AUTHOR]

Methanotrophs are bacteria that consume methane (CH4) as their sole carbon and energy source. These microorganisms play a crucial role in the carbon cycle by metabolizing CH4 (the greenhouse gas), into cellular biomass and carbon dioxide (CO2). Polyhydroxyalkanoates (PHAs) are biopolymers produced by various microorganisms, including methanotrophs. PHA production using methanotrophs is a promising strategy to address growing concerns regarding plastic pollution and the need for sustainable, biodegradable materials. Various factors, including nutrient availability, environmental conditions, and metabolic engineering strategies, influence methanotrophic production. Nutrient limitations, particularly those of nitrogen or phosphorus, enhance PHA production by methanotrophs. Metabolic engineering approaches, such as the overexpression of key enzymes involved in PHA biosynthesis or the disruption of competing pathways, can also enhance PHA yields by methanotrophs. Overall, PHA production by methanotrophs represents a sustainable and versatile approach for developing biomedical materials with numerous potential applications. Additionally, alternative feedstocks, such as industrial waste streams or byproducts can be explored to improve the economic feasibility of PHA production. This review briefly describes the potential of methanotrophs to produce PHAs, with recent updates and perspectives. [ABSTRACT FROM AUTHOR]

Polyhydroxyalkanoates(PHAs) are considered a good alternative to petroleum-based plastics because of their good biodegradability and biocompatibility. The synthesis of PHAs using activated sludge can not only solve the problem of the high cost of pure cultures but also improve the utilization value of activated sludge. In this study, sludge activity recovery experiments were firstly conducted and the effects of different initial sludge concentrations on the activated sludge PHAs synthesis system were further investigated. the initial sludge concentrations were 1#SBR (2800 ± 50) mg/L, 2#SBR (4200 ± 50) mg/L, and 3#SBR (5500 ± 50) mg/L. The results showed that the activity, sedimentation performance and PHAs synthesis capacity of activated sludge were enhanced after the sludge activity recovery experiment. At the initial sludge concentration of 4200 mg/L, the activated sludge PHAs synthesis system was operated stably and the synthesis efficiency of PHAs was enhanced. In contrast, at the initial sludge concentration of 2800 and 5500 mg/L, the steady state of the activated sludge PHAs synthesis system was damaged to different degrees at different times, and the synthesis efficiency of PHAs was greatly reduced. [ABSTRACT FROM AUTHOR]

The Bacillus megaterium LVN01 species native to Colombia has demonstrated the ability to metabolize different coproducts or industrial waste (such as fique juice, cane molasses, and residual glycerol) and accumulate polyhydroxybutyrate (PHB), giving it potential in the bioplastics industry. In this research, the potential of liquid digestate as a carbon source for the production of PHA polymers in fermentation processes with this bacterial strain was evaluated. Favorably, it was found that B. megaterium utilizes the nutrients from this residual substrate to multiply appropriately and efficiently synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Bench-scale aerobic batch fermentation, under the operational conditions of this research [volume: 3 L; temperature: 30.8 °C; agitation: 400 rpm; pH: 7.0 ± 0.2; dissolved oxygen: 100% saturation; antifoam: 10% (v/v)], generated maximum values of dry cell weight (DCW) (0.56 g cell L−1) at 60 h, while the maximum PHBV yield (360 mg PHBV L−1) occurred at 16 h, which is very favorable for sustainable degradable bioplastics production. Additionally, GC–MS and NMR analyses confirmed that the PHBV copolymer synthesized by B. megaterium is made up of the monomers 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). Furthermore, the thermal properties determined by TGA (Tonset = 283.1 °C; Tendset = 296.98 °C; Td = 290.114 °C) and DSC (Tm = °C 155.7 °C; ΔHf = 19.80 J g−1; Xcr = 18.17%) indicate that it is a thermally stable biopolymer with low percentages of crystallinity, providing flexibility that facilitates molding, adaptation, and application in various industrial sectors. [ABSTRACT FROM AUTHOR]

The haloarchaeon Haloferax mediterranei synthesizes poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) under unfavorable nutritional conditions without the addition of any precursor to the culture, which is an advantage compared to other microbial counterparts able to synthesize polyhydroxyalkanoates (PHA). PHBV is a biodegradable polymer showing physiochemical properties of biotechnological and biomedical interest and can be used as an alternative to plastics made from chemical synthesis (which are not environmentally friendly). The versatile metabolism of H. mediterranei makes the use of waste as a carbon source for cellular growth and PHA synthesis possible. In this work, cellular growth and the production and characterization of PHBV using two different types of confectionery waste were analyzed and compared with cellular growth and PHBV synthesis in a standard culture media with glucose of analytical grade as a carbon source. The PHBV granules produced were analyzed by TEM and the biopolymer was isolated and characterized by GC-MS, FTIR NMR, and DSC. The results reveal that H. mediterranei can use these two residues (R1 and R2) for pure PHBV production, achieving 0.256 and 0.983 g PHBV/L, respectively, which are among the highest yields so far described using for the first-time waste from the candy industry. Thus, a circular economy-based process has been designed to optimize the upscaling of PHBV production by using haloarchaea as cell factories and valorizing confectionery waste. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Petroleum Research & Studies is the property of Republic of Iraq Ministry of Higher Education & Scientific Research (MOHESR) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Increasing human population demands more usage of consumable items thus causes more destruction to the environment by developing advanced materials and technologies. Synthetic plastic is one of the most used material in daily life which results in accumulation of non-degradable waste material that stays in the environment for thousands of years and creates plastic pollution. Many species on the earth suffer from pollution caused by plastic and it is one of the leading causes of global warming. For sustainable development, we need to protect our environment from plastic pollution by making use of biodegradable materials. In the past few years, biodegradable materials with plastic like properties have gained much attention. They have been made from biomass such as sugarcane, corn, wheat, rice, banana peels, etc. Such desirable properties are also present in PHAs (polyhydroxyalkanoates) synthesized by microorganisms. Several types of PHAs have been discovered from bacteria and used as a bioplastic candidate. However, there are major challenges also such as high production cost and expensive raw material which limits its commercial applications. Modern tools and strategies have allowed precise metabolic engineering of various microorganisms to produce PHAs using highly efficient microbial cell factories. This review article focuses on the research done on the microbial bioplastic production and its various applications. We have also discussed the future perspectives of bioplastic for sustainable development. [ABSTRACT FROM AUTHOR]

Copyright of Plastics Science & Technology / Suliao Ke-Ji is the property of Plastics Science & Technology Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Polyhydroxyalkanoates (PHAs) are promising biodegradable polymers known for their biodegradability and eco-friendly properties. Recent studies indicate that PHAs can reduce the environmental impact by up to 50% compared to petroleum-based plastics. This comprehensive review evaluates the application of PHAs in sustainable food packaging, covering over 100 studies published between 2018 and 2023. The review highlights advancements in PHA production, with a focus on submerged and solid-state fermentation methods, achieving up to a 60% improvement in production efficiency through optimized culture selection. Sustainable extraction and purification methods have been identified, reducing energy consumption by 30%. Blending PHAs with other biodegradable polymers like polylactic acid, starch, and cellulose enhances material performance, with up to a 40% improvement in mechanical properties. The incorporation of antimicrobial agents and essential oils has been shown to extend the shelf life by 25% while maintaining food safety standards. This review underscores the potential of active PHA-based packaging in improving the barrier properties by 35% when combined with coatings, positioning PHA as a key material for the future of environmentally responsible and safe food packaging. [ABSTRACT FROM AUTHOR]

High polyhydroxyalkanoate (PHA) yield from selected substrates associated with minimal accumulation of residual biomass can improve the process economy. In this study, different carbon (glucose, sucrose, glycerol, and acetic acid) and nitrogen (NH4Cl and urea) sources were screened for PHA production by Cupriavidus necator. The effects of incubation time, nitrogen, and phosphate concentration on biomass growth and PHA production were co-optimized through response surface methodology (RSM) and genetic algorithm-optimized artificial neural network (GA-ANN). Sucrose and urea were found to offer significantly better (p <0.001) biomass (1.468 ± 0.007 g l-1) and PHA (0.924 ± 0.02 g l-1) yield when compared with other carbon and nitrogen sources. Though the performance of both the models remains similar for biomass (R2 = 0.97–0.98), GA-ANN (with six neurones in a hidden layer) seems exceptionally better in predicting PHA yield (R2 = 0.97) when compared to the polynomial model (R2 = 0.92). The maximum PHA concentration of 2.69 g l-1 was predicted by the ANN model at an incubation time of 62.80 h with 2.0 g l-1 of nitrogen and 4.0 g l-1 of phosphate concentration. The multi-composite desirability using the GA-ANN model projected a better polymer-to-biomass ratio compared to the polynomial model. The inclusion of a cost-benefit analysis framework may be warranted before recommending the optimal conditions obtained through multivariate regression and GA-ANN models. [ABSTRACT FROM AUTHOR]

Polyhydroxyalkanoates (PHA) are popular biopolymers due to their potential use as biodegradable thermoplastics. In this study, three aerobic sequencing batch reactors were operated identically except for their temperatures, which were set at 15 °C, 35 °C, and 48 °C. The reactors were subjected to a feast-famine feeding regime, where carbon sources are supplied intermittently, to enrich PHAaccumulating microbial consortia. The biomass was sampled for 16S rRNA gene amplicon sequencing of both DNA (during the enrichment phase) and cDNA (during the enrichment and accumulation phases). All temperatures yielded highly enriched PHA-accumulating consortia. Thermophilic communities were significantly less diverse than those at low or mesophilic temperatures. In particular, Thauera was highly adaptable, abundant, and active at all temperatures. Low temperatures resulted in reduced PHA production rates and yields. Analysis of the microbial community revealed a collapse of community diversity during low-temperature PHA accumulation, suggesting that the substrate dosing strategy was unsuccessful at low temperatures. This points to future possibilities for optimizing low-temperature PHA accumulation. [ABSTRACT FROM AUTHOR]

The present study examines the use of waste cooking oil (WCO) as a substrate for medium-chain-length polyhydroxyalkanoates (mcl-PHA) production by Pseudomonas rhizophila S211. The genome analysis revealed that the S211 strain has a mcl-PHA cluster (phaC1ZC2DFI) encoding two class II PHA synthases (PhaC1 and PhaC2) separated by a PHA depolymerase (PhaZ), a transcriptional activator (PhaD) and two phasin-like proteins (PhaFI). Genomic annotation also identified a gene encoding family I.3 lipase that was able to hydrolyze plant oils and generate fatty acids as favorable carbon sources for cell growth and PHA synthesis via β-oxidation pathway. Using a three-variable Doehlert experimental design, the optimum conditions for mcl-PHA accumulation were achieved in 10% of WCO-based medium with an inoculum size of 10% and an incubation period of 48 h at 30 °C. The experimental yield of PHA from WCO was 1.8 g/L close to the predicted yield of 1.68 ± 0.14 g/L. Moreover, 1H nuclear magnetic resonance spectroscopy analysis confirmed the extracted mcl-PHA. Overall, this study describes P. rhizophila as a cell factory for biosynthesis of biodegradable plastics and proposes green and efficient approach to cooking oil waste management by decreasing the cost of mcl-PHA production, which can help reduce the dependence on petroleum-based plastics. [ABSTRACT FROM AUTHOR]

Lignocellulose is a major biopolymer in plant biomass with a complex structure and composition. It consists of a significant amount of high molecular aromatic compounds, particularly vanillin, syringeal, ferulic acid, and muconic acid, that could be converted into intracellular metabolites such as polyhydroxyalkanoates (PHA) and hydroxybutyrate (PHB), a key component of bioplastic production. Several pre-treatment methods were utilized to release monosaccharides, which are the precursors of the relevant pathway. The consolidated bioprocessing of lignocellulose-capable microbes for biomass depolymerization was discussed in this study. Carbon can be stored in a variety of forms, including PHAs, PHBs, wax esters, and triacylglycerides. From a biotechnology standpoint, these compounds are quite adaptable due to their precursors' utilization of hydrogen energy. This study lays the groundwork for the idea of lignocellulose valorization into value-added products through several significant dominant pathways. [ABSTRACT FROM AUTHOR]

Biodegradable biobased polymers derived from biomass (such as plant, animal, marine, or forestry material) show promise in replacing conventional petrochemical polymers. Research and development have been conducted for decades on potential biodegradable biobased polymers such as polylactic acid (PLA), polyhydroxyalkanoates (PHAs), and succinate polymers. These materials have been evaluated for practicality, cost, and production capabilities as limiting factors in commercialization; however, challenges, such as the environmental limitations on the biodegradation rates for biodegradable biobased polymer, need to be addressed. This review provides a history and overview of the current development in the synthesis process and properties of biodegradable biobased polymers, along with a techno-commercial analysis and discussion on the environmental impacts of biodegradable biobased polymers. Specifically, the techno-commercial analysis focuses on the commercial potential, financial assessment, and life-cycle assessment of these materials, as well as government initiatives to facilitate the transition towards biodegradable biobased polymers. Lastly, the environmental assessment focuses on the current challenges with biodegradation and methods of improving the recycling process and reusability of biodegradable biobased polymers. [ABSTRACT FROM AUTHOR]