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The continuous growth in the biopolymers market underscores the necessity to develop and characterize novel materials to replace conventional polymers. This study evaluated the effect of the processing method on the properties of poly(hydroxybutyrate)/agave fiber (PHB/AF) biocomposites with a coupling agent based on glycidyl methacrylate. The components were first blended using two methods (dry-blending and twin screw-extrusion) before being compression molded. Then, the biocomposites degradation was evaluated via accelerated weathering to determine its effect on the physico-mechanical properties. The results showed that adding AF to PHB increased the porosity (low density), but this effect was decreased by 60% for the extruded compatibilized biocomposites. Consequently, the level of degradation (loss of mechanical properties and dimensional stability) was less affected when a coupling agent was used. However, after weathering, the porosity similarly increased for both uncompatibilized biocomposites. Finally, the tensile, impact, and flexural strength were less affected by weathering when the compatibilizer was mixed via dry-blending. [ABSTRACT FROM AUTHOR]

To tackle the persistent global issue of nonbiodegradable petro‐plastics, this study undertakes the challenge of developing environmentally sustainable and active packaging alternatives. The approach involves preparing composite films by blending poly(hydroxybutyrate) (PHB) and poly(lactic acid) (PLA) while integrating olive oil and zinc oxide to augment their effectiveness. The films are fabricated by mixing different concentrations of ZnO (0.5% to 2.5%) and 10% olive oil into PHB/PLA (70/30) matrix using solvent‐casting. A comprehensive analysis is conducted to assess the properties of prepared films, including Fourier transform infrared spectroscopy, scanning electron microscopy, thermogravimetric analysis, X‐ray diffraction, mechanical testing, water vapor permeability, and UV‐blocking properties. Among the tested samples, film containing 1.5% ZnO (referred to as PPOZ1.5) displays highest tensile strength (30.8 MPa) along with improved water vapor permeability. Consequently, PPOZ1.5 film is selected for further investigations which include DPPH (diphenyl‐picrylhydrazyl) radical scavenging activity, and antimicrobial activity. PPOZ1.5 film exhibits exceptional antioxidant activity (>65%) and significant antimicrobial efficacy against Escherichia coli, Staphylococcus aureus, and Aspergillus niger. Moreover, when utilized to package bread samples, PPOZ1.5 film effectively inhibits microbial growth, ensuring food safety for an extended storage period of more than 12 days, ultimately contributing to the preservation and safety of packaged bakery products. [ABSTRACT FROM AUTHOR]

As part of ongoing efforts to discover novel polyhydroxyalkanoate‐producing bacterial species, we embarked on characterizing the thermotolerant species, Paracoccus kondratievae, for biopolymer synthesis. Using traditional chemical and thermal characterization techniques, we found that P. kondratievae accumulates poly(3‐hydroxybutyrate) (PHB), reaching up to 46.8% of the cell's dry weight after a 24‐h incubation at 42°C. Although P. kondratievae is phylogenetically related to the prototypical polyhydroxyalkanoate producer, Paracoccus denitrificans, we observed significant differences in the PHB production dynamics between these two Paracoccus species. Notably, P. kondratievae can grow and produce PHB at elevated temperatures ranging from 42 to 47°C. Furthermore, P. kondratievae reaches its peak PHB content during the early stationary growth phase, specifically after 24 h of growth in a flask culture. This is then followed by a decline in the later stages of the stationary growth phase. The depolymerization observed in this growth phase is facilitated by the abundant presence of the PhaZ depolymerase enzyme associated with PHB granules. We observed the highest PHB levels when the cells were cultivated in a medium with glycerol as the sole carbon source and a carbon‐to‐nitrogen ratio of 10. Finally, we found that PHB production is induced as an osmotic stress response, similar to other polyhydroxyalkanoate‐producing species. [ABSTRACT FROM AUTHOR]

Polyhydroxybutyrates (PHBs) are biopolymers that are good green alternative for synthetic carbon-based polymers, and are also one of the most researched members of the Polyhydroxyalkanoates (PHA) family. In this study, a gram-positive bacterial strain Bacillus megaterium LSRB 0103 was isolated from Pallikaranai Marshland, Chennai, India. Primary screening using Sudan Black dye revealed the presence of intracellular PHB granules. Minimal Davis Media (MDM) which was used or PHB production gave a yield of 0.7107 g/L. Subsequently, using response surface methodology (RSM), a central composite design (CCD) model was designed for media optimization having cornstarch, urea, and pH as independent variables. The findings of the CCD model were fitted into a second-order polynomial equation. The RSM model predicted the maximum PHB yield of 0.93 g/L, at these independent variable levels, cornstarch, 5 g/L; urea, 2.1 g/L; and pH 7.0; while the experimental PHB yield was 0.94 g/L, with a percentage error of 1.1%. This study is the first-time report of production of PHB by Bacillus megaterium using cornstarch and urea as substrate. [ABSTRACT FROM AUTHOR]

Nanotechnology is one of the newest directions for plant-based therapies. Chronic venous disease often predisposes to long-term and invasive treatment. This research focused on the inclusion of vegetal extracts from Sophorae flos (SE), Calendulae flos (CE), and Ginkgo bilobae folium (GE) in formulations with PHB and PLGA polymers and their physicochemical characterization as a preliminary stage for possible use in the development of a complex therapeutic product. The samples were prepared by an oil–water emulsification and solvent evaporation technique, resulting in suspensions with high spreadability and a pH of 5.5. ATR-FTIR analysis revealed bands for stretching vibrations (O-H, C=O, and C-H in symmetric and asymmetric methyl and methylene) in the same regions as the base components, but switched to high or low wavenumbers and absorbance, highlighting the formation of adducts/complexes between the extracts and polymers. The obtained formulations were in the amorphous phase, as confirmed by XRD analysis. AFM analysis emphasized the morphological peculiarities of the extract–polymer nanoformulations. It could be noticed that, in the case of SE-based formulations, the dominant characteristics for SE-PHB and SE-PLGA composition were the formation of random large (SE-PHB) and smaller uniform (SE-PLGA) particles; further on, these particles tended to aggregate in the case of SE-PHB-PLGA. For the CE- and GE-based formulations, the dominant surface morphology was their porosity, generally with small pores, but larger cavities were observed in some cases (CE- and GE-PHB). The highest roughness values at the (8 µm × 8 μm) scale were found for the following samples and succession: CE-PHB < SE-PLGA < SE-PHB-PLGA. In addition, by thermogravimetric analysis, impregnation in the matrix of compression stockings was evaluated, which varied in the following order: CE-polymer > SE-polymer > GE-polymer. In conclusion, nine vegetal extract–polymer nanoformulations were prepared and preliminarily characterized (by advanced physicochemical methods) as a starting point for further optimization, stability studies, and possible use in complex pharmaceutical products. [ABSTRACT FROM AUTHOR]

Date palm fiber (DPF) holds great potential for composite materials, but its flammability limits its practical applications. In this study, DPF was modified using a pad-drying method to impregnate it with a 5 wt.% solution of ammonium dihydrogen phosphate (ADP). These treated fibers were then utilized to fabricate poly(β-hydroxybutyrate)- (PHB-) based composites. The resulting thermal insulators were comprehensively evaluated for their flammability, physical, mechanical, and thermophysical properties, as well as morphological and thermal stability characteristics. The findings revealed a significant reduction in flame spread and smoke suppression; however, the concentration used is not sufficient to achieve the desired rating grades. The thermal insulation capacity of the modified fiber composites was substantially enhanced, particularly with the 40% PHB/DPF-ADP composite displaying the lowest thermal conductivity at 0.0564 W/m.K. Moreover, the presence of gaps and voids at the interface led to a reduction in tensile strength to 4-7 MPa. Additionally, the modified fiber composites exhibited significantly reduced water absorption (~0.76%), attributed to the formation of a highly water-resistant substance containing a furan compound. This work provides a simple and effective approach for achieving durable flame retardancy and long-term thermal insulation performance, offering promising opportunities for the practical application of biobased PHB composites. [ABSTRACT FROM AUTHOR]

Background: Industrial biomanufacturing of value-added products using CO2 as a carbon source is considered more sustainable, cost-effective and resource-efficient than using common carbohydrate feedstocks. Cupriavidus necator H16 is a representative H2-oxidizing lithoautotrophic bacterium that can be utilized to valorize CO2 into valuable chemicals and has recently gained much attention as a promising platform host for versatile C1-based biomanufacturing. Since this microbial platform is genetically tractable and has a high-flux carbon storage pathway, it has been engineered to produce a variety of valuable compounds from renewable carbon sources. In this study, the bacterium was engineered to produce resveratrol autotrophically using an artificial phenylpropanoid pathway. Results: The heterologous genes involved in the resveratrol biosynthetic pathway—tyrosine ammonia lyase (TAL), 4-coumaroyl CoA ligase (4CL), and stilbene synthase (STS) —were implemented in C. necator H16. The overexpression of acetyl-CoA carboxylase (ACC), disruption of the PHB synthetic pathway, and an increase in the copy number of STS genes enhanced resveratrol production. In particular, the increased copies of VvSTS derived from Vitis vinifera resulted a 2-fold improvement in resveratrol synthesis from fructose. The final engineered CR-5 strain produced 1.9 mg/L of resveratrol from CO2 and tyrosine via lithoautotrophic fermentation. Conclusions: To the best of our knowledge, this study is the first to describe the valorization of CO2 into polyphenolic compounds by engineering a phenylpropanoid pathway using the lithoautotrophic bacterium C. necator H16, demonstrating the potential of this strain a platform for sustainable chemical production. [ABSTRACT FROM AUTHOR]

Poly-hydroxybutyrate (PHB) is an environmentally friendly alternative for conventional fossil fuel-based plastics that is produced by various microorganisms. Large-scale PHB production is challenging due to the comparatively higher biomanufacturing costs. A PHB overproducer is the haloalkaliphilic bacterium Halomonas campaniensis, which has low nutritional requirements and can grow in cultures with high salt concentrations, rendering it resistant to contamination. Despite its virtues, the metabolic capabilities of H. campaniensis as well as the limitations hindering higher PHB production remain poorly studied. To address this limitation, we present HaloGEM, the first high-quality genome-scale metabolic network reconstruction, which encompasses 888 genes, 1528 reactions (1257 gene-associated), and 1274 metabolites. HaloGEM not only displays excellent agreement with previous growth data and experiments from this study, but it also revealed nitrogen as a limiting nutrient when growing aerobically under high salt concentrations using glucose as carbon source. Among different nitrogen source mixtures for optimal growth, HaloGEM predicted glutamate and arginine as a promising mixture producing increases of 54.2% and 153.4% in the biomass yield and PHB titer, respectively. Furthermore, the model was used to predict genetic interventions for increasing PHB yield, which were consistent with the rationale of previously reported strategies. Overall, the presented reconstruction advances our understanding of the metabolic capabilities of H. campaniensis for rationally engineering this next-generation industrial biotechnology platform. Key points: A comprehensive genome-scale metabolic reconstruction of H. campaniensis was developed. Experiments and simulations predict N limitation in minimal media under aerobiosis. In silico media design increased experimental biomass yield and PHB titer. [ABSTRACT FROM AUTHOR]

Cellulosic fibers obtained from Barley straw were utilized to reinforce PHB. Four different processed fibers were employed as reinforcing material: sawdust (SW), defibered (DFBF), delignified (DBF), and bleached (BBF) fibers. The composite was processed from two different perspectives: a discontinuous (bach) and an intensification process (extrusion). Once processed and transformed into final shape specimens, the materials were characterized by mechanical testing (tensile mode), scanning electron microscopy, and theoretical simulations by finite elements analysis (FEA). In terms of mechanical properties, only the elastic moduli (Et) exhibited results ranging from 37% to 170%, depending on the reinforcement composition. Conversely, strengths at break, under both tensile and bending tests, tended to decrease, indicating poor affinity between the components. Due to the mechanical treatment applied on the fiber, DFBF emerged as the most promising filler, with mechanical properties closest to those of neat PHB. DFBF-based composites were subsequently produced through process intensification using a twin-screw extruder, and molded into flowerpots. Mechanical results showed almost identical properties between the discontinuous and intensification processes. The suitability of the material for agriculture flowerpots was demonstrated through finite analysis simulation (FEA), which revealed that the maximum von Mises stresses (5.38 × 105 N/m2) and deformations (0.048 mm) were well below the limits of the composite materials. [ABSTRACT FROM AUTHOR]

Several composites based on poly(3-hydroxybutyrate) (PHB) and mesoporous SBA-15 silica were prepared by solvent-casting followed by a further stage of compression molding. The thermal stability, phase transitions and crystalline details of these composites were studied, paying special attention to the confinement of the PHB polymeric chains into the mesopores of the silica. For that, differential scanning calorimetry (DSC) and real-time variable-temperature X-ray scattering at small angles (SAXS) were performed. Confinement was stated first by the existence of a small endotherm at temperatures around 20 °C below the main melting or crystallization peak, being later confirmed by a notable discontinuity in the intensity of the main (100) diffraction from the mesoporous silica observed through SAXS experiments, which is related to the change in the scattering contrast before and after the crystallization or melting of the polymer chains. Furthermore, the usual α modification of PHB was developed in all samples. Finally, a preliminary investigation of mechanical and relaxation parameters was carried out through dynamic–mechanical thermal analysis (DMTA). The results show, in the temperature interval analyzed, two relaxations, named α and β (the latest related to the glass transition) in order of decreasing temperatures, in all specimens. The role of silica as a filler is mainly observed at temperatures higher than the glass transition. In such cases, stiffness is dependent on SBA-15 content. [ABSTRACT FROM AUTHOR]

The stringent response exerted by (p)ppGpp and RNA-polymerase binding protein DksA regulates gene expression in diverse bacterial species. To control gene expression (p)ppGpp, synthesized by enzymes RelA and SpoT, interacts with two sites within the RNA polymerase; site 1, located in the interphase between subunits β' and ω (rpoZ), and site 2 located in the secondary channel that is dependent on DksA protein. In Escherichia coli, inactivation of dksA results in a reduced sigma factor RpoS expression. In Azotobacter vinelandii the synthesis of polyhydroxybutyrate (PHB) is under RpoS regulation. In this study, we found that the inactivation of relA or dksA, but not rpoZ, resulted in a negative effect on PHB synthesis. We also found that the dksA, but not the relA mutation reduced both rpoS transcription and RpoS protein levels, implying that (p)ppGpp and DksA control PHB synthesis through different mechanisms. Interestingly, despite expressing rpoS from a constitutive promoter in the dksA mutant, PHB synthesis was not restored to wild type levels. A transcriptomic analysis in the dksA mutant, revealed downregulation of genes encoding enzymes needed for the synthesis of acetyl-CoA, the precursor substrate for PHB synthesis. Together, these data indicate that DksA is required for optimal expression of RpoS which in turn activates transcription of genes for PHB synthesis. Additionally, DksA is required for optimal transcription of genes responsible for the synthesis of precursors for PHB synthesis. [ABSTRACT FROM AUTHOR]

Synthetic polymers have been extensively employed in various sectors of life. They, however, are mostly xenobiotic, toxic and persistent, thus causing certain environmental and health issues. So, the current awareness has raised the demand for eco-friendly biobased polymers. In this context, poly(hydroxybutyrate) (PHB) could be a promising candidate due to its eco-friendly and modifiable nature. The chemo-physical properties of such a biopolyester might be tailored through mediation with certain metallic nanoparticles, thus making its nanobiocomposite with improved mechanical stability. The present work aimed to prepare silver sulfide (Ag2S) nanoparticles (NPs)-mediated PHB-based conductive films and evaluate their structural and functional properties. The Ag2S NPs were prior prepared using the chemical precipitation/reduction approach and then impregnated into PHB-based nanobiocomposite films using the solution casting approach, whereas poly(lactic acid) was used as a reinforcing agent. The UV–visible spectrum of the Ag2S NPs dispersion exhibited a strong absorption peak at 260 nm. The z-average of the Ag2S NPs was recorded as 100 ± 10 nm with a polydispersity index of 0.095 and zeta potential of − 35 ± 5 mV. Fourier transform infrared analysis expressed successful mediation of Ag2S NPs in the biocomposite. The crystallinity of the nanobiocomposite specimen was calculated to be 79.23%, i.e., larger than that of the control PHB/PLA blend film having 53.69%. The prepared nanocomposite films were found electrically conductive with appropriate physicochemical, mechanical and thermal properties, and thus could find possible applications in biosensing and smart packing sectors. [ABSTRACT FROM AUTHOR]

In this study PLA and PHB were melt-blended with/without maleic anhydride (MA) as a compatibilizer and melt-spun into a monofilament. The aquatic biodegradation of PLA/PHB fibers was tracked regarding the PLA and PHB components and the total carbon in the blend. Pure PLA fiber was recalcitrant to aquatic biodegradation in the present conditions, whereas PHB was readily degradable, as expected. The 75/25% and 50/50% PLA/PHB fibers showed zero biodegradation. The 25/75% PLA/PHB blend showed only an 11% final biodegradation extent and a 13% PHB biodegradation extent. It is shown that an unintended consequence of blending PLA with PHB is that after biodegradation, micro/nano plastic pollution is evolved. MA increased the miscibility, which further decreased the PHB biodegradation. The lower biodegradation extent with MA indicates that increased miscibility and more intimate coverage of the biodegradable polymer with the unbiodegradable polymer may have a negative effect on the biodegradation of biodegradable polymers. [ABSTRACT FROM AUTHOR]

The grating structure and periodic assembly in D -(−)-poly(3-hydroxybutyrate) (PHB), or simply poly(3-hydroxybutyrate), with diluent-modulated optical regularity are analyzed using microscopy techniques coupled with a unique 3D interior dissection technique. The top-surface valley band is a result of impingement of branching crystals evolving from normal-oriented main lamellar stalks, and the top-surface band is a result of the protruding lamellar stalks. The 3D dissection shows direct experimental proof of discontinuous interfaces between the successive bands, which are created by periodic impingement of neighboring lamellar bundles growing in the normal direction from the substrate. The iridescence phenomenon from the PHB periodic bands can be attributed to the grating-assembly structures with cross-bar pitches tuned to a suitable range by combination of diluents and temperature of growth, whose function resembles those seen in nature's iridescent crystals. [ABSTRACT FROM AUTHOR]

Controlled thermal degradation of high-molecular-weight poly-3-hydroxybutyrate (PHB) was studied in dimethylformamide (DMF), bromoform, tetrabromoethane, and trichloropropane. Samples with molecular weights of 1.6 kDa and higher were obtained. PHB degradation in pure DMF and in trichloropropane resulted in the formation of C-terminal crotonic acid residues at the sites of polymer chain scission. In the case of bromoform and tetrabromoethane, 3-bromobutyric acid residues were formed (61–68% at 100 °C and 29–54% at 145 °C), along with crotonic acid residues. This indicates the possibility of halogen capture from solvent molecules at the moment of chain scission. Presumably, the reaction follows the E1 mechanism involving the formation of an intermediate carbocation. The results show the possibility of using thermochemical degradation of biodegradable polyesters in solutions for obtaining their oligomeric bromo-substituted derivatives. [ABSTRACT FROM AUTHOR]

Azospirillum brasilense Sp7 produces PHB, which is covered by granule-associated proteins (GAPs). Phasins are the main GAPs. Previous studies have shown phasins can regulate PHB synthesis. When A. brasilense grows under stress conditions, it uses sigma factors to transcribe genes for survival. One of these factors is the σ24 factor. This study determined the possible interaction between phasins and the σ24 factor or phasin-σ24 factor complex and DNA. Three-dimensional structures of phasins and σ24 factor structures were predicted using the I-TASSER and SWISS-Model servers, respectively. Subsequently, a molecular docking between phasins and the σ24 factor was performed using the ClusPro 2.0 server, followed by molecular docking between protein complexes and DNA using the HDOCK server. Evaluation of the types of ligand–receptor interactions was performed using the BIOVIA Discovery Visualizer for three-dimensional diagrams, as well as the LigPlot server to obtain bi-dimensional diagrams. The results showed the phasins (Pha4Abs7 or Pha5Abs7)-σ24 factor complex was bound near the −35 box of the promoter region of the phaC gene. However, in the individual interaction of PhaP5Abs7 and the σ24 factor, with DNA, both proteins were bound to the −35 box. This did not occur with PhaP4Abs7, which was bound to the −10 box. This change could affect the transcription level of the phaC gene and possibly affect PHB synthesis. [ABSTRACT FROM AUTHOR]

Background: Polyhydroxybutyrate is a biopolymer produced by bacteria and archaea under nitrogen-limiting conditions. PHB is an essential polymer in the bioplastic sector because of its biodegradability, eco-friendliness, and adaptability. The characterization of PHB is a multifaceted process for studying the structure and its properties. This entire aspect can assure the long-term viability and performance attributes of the PHB. The characteristics of PHB extracted from the halophile Brachybacterium paraconglomeratum were investigated with the objective of making films for application in healthcare. Results: This was the first characterization study on PHB produced by a rare halophile, Brachybacterium paraconglomeratum (MTCC 13074). In this study, the strain produced 2.72 g/l of PHB for.5.1 g/l of biomass under optimal conditions. Methods are described for the determination of the physicochemical properties of PHB. The prominent functional groups CH3 and C = O were observed by FT-IR and the actual chemical structure of the PHB was deduced by NMR. GCMS detects the confirmation of four methyl ester derivatives of the extracted PHB in the sample. Mass spectrometry revealed the molecular weight of methyl 3-hydroxybutyric acid (3HB) present in the extract. The air-dried PHB films were exposed to TGA, DSC and a universal testing machine to determine the thermal profile and mechanical stability. Additionally, the essential property of biopolymers like viscosity was also assessed for the extracted PHB. Conclusions: The current study demonstrated the consistency and quality of B. paraconglomeratum PHB. Therefore, Brachybacterium sps are also a considerable source of PHB with desired characteristics for industrial production. [ABSTRACT FROM AUTHOR]

Background: Polyhydroxybutyrate is a biopolymer produced by bacteria and archaea under nitrogen-limiting conditions. PHB is an essential polymer in the bioplastic sector because of its biodegradability, eco-friendliness, and adaptability. The characterization of PHB is a multifaceted process for studying the structure and its properties. This entire aspect can assure the long-term viability and performance attributes of the PHB. The characteristics of PHB extracted from the halophile Brachybacterium paraconglomeratum were investigated with the objective of making films for application in healthcare. Results: This was the first characterization study on PHB produced by a rare halophile, Brachybacterium paraconglomeratum (MTCC 13074). In this study, the strain produced 2.72 g/l of PHB for.5.1 g/l of biomass under optimal conditions. Methods are described for the determination of the physicochemical properties of PHB. The prominent functional groups CH3 and C = O were observed by FT-IR and the actual chemical structure of the PHB was deduced by NMR. GCMS detects the confirmation of four methyl ester derivatives of the extracted PHB in the sample. Mass spectrometry revealed the molecular weight of methyl 3-hydroxybutyric acid (3HB) present in the extract. The air-dried PHB films were exposed to TGA, DSC and a universal testing machine to determine the thermal profile and mechanical stability. Additionally, the essential property of biopolymers like viscosity was also assessed for the extracted PHB. Conclusions: The current study demonstrated the consistency and quality of B. paraconglomeratum PHB. Therefore, Brachybacterium sps are also a considerable source of PHB with desired characteristics for industrial production. [ABSTRACT FROM AUTHOR]

In this work, we have studied the potential application for 3D-printing of a polymer made from combining a biodegradable and biocompatible polymer (i.e., polyhydroxybutyrate, PHB) with natural bio-based fiber (i.e., cellulose). To this end, a masterbatch at 15 wt.% in filler content was prepared by melt-blending, and then this system was "diluted" with pure PHB in a second extrusion phase in order to produce filaments at 1.5 and 3 wt.% of cellulose. For comparison, a filament made of 100% virgin PHB pellets was prepared under the same conditions. All the systems were then processed in the 3D-printer apparatus, and specimens were mainly characterized by static (tensile and flexural testing) and dynamic mechanical analysis. Thermogravimetric analysis, differential scanning calorimetry, spectroscopic measurements, and morphological aspects of PHB polymer and composites were also discussed. The results showed a significant negative impact of the process on the mechanical properties of the basic PHB with a reduction in both tensile and flexural mechanical properties. The PHB–cellulose composites showed a good dispersion filler in the matrix but a poor interfacial adhesion between the two phases. Furthermore, the cellulose had no effect on the melting behavior and the crystallinity of the polymer. The addition of cellulose improved the thermal stability of the polymer and minimized the negative impact of extrusion. The mechanical performance of the composites was found to be higher compared to the corresponding (processed) polymer. [ABSTRACT FROM AUTHOR]

Simple Summary: Trehalose is an easily and economically obtainable non-reducing disaccharide, readily taken up by living organisms upon external application. Arthrospira platensis grew photoautotrophically under nitrogen-deficient conditions supplied with the appropriate trehalose, promoting glycogen and PHB production. This is supported by a notable increase in the upregulation of glgA and phaC expression, along with the induction of enzyme activity, including glycogen synthase encoded by glgA and PHA synthase encoded by phaC. The present study offers new insights into the contribution of exogenous trehalose in the regulation of glycogen and PHB production in A. platensis. Glycogen and poly-3-hydroxybutyrate (PHB) are excellent biopolymer products from cyanobacteria. In this study, we demonstrate that nitrogen metabolism is positively influenced by the exogenous application of trehalose (Tre) in Arthrospira platensis under nitrogen-deprived (−N) conditions. Cells were cultivated photoautotrophically for 5 days under −N conditions, with or without the addition of exogenous Tre. The results revealed that biomass and chlorophyll-a content of A. platensis experienced enhancement with the addition of 0.003 M and 0.03 M Tre in the −N medium after one day, indicating relief from growth inhibition caused by nitrogen deprivation. The highest glycogen content (54.09 ± 1.6% (w/w) DW) was observed in cells grown for 2 days under the −N + 0.003 M Tre condition (p < 0.05), while the highest PHB content (15.2 ± 0.2% (w/w) DW) was observed in cells grown for 3 days under the −N + 0.03 M Tre condition (p < 0.05). The RT-PCR analysis showed a significant increase in glgA and phaC transcript levels, representing approximately 1.2- and 1.3-fold increases, respectively, in A. platensis grown under −N + 0.003 M Tre and −N + 0.03 M Tre conditions. This was accompanied by the induction of enzyme activities, including glycogen synthase and PHA synthase with maximal values of 89.15 and 0.68 µmol min−1 mg−1 protein, respectively. The chemical structure identification of glycogen and PHB from A. platensis was confirmed by FTIR and NMR analysis. This research represents the first study examining the performance of trehalose in promoting glycogen and PHB production in cyanobacteria under nitrogen-deprived conditions. [ABSTRACT FROM AUTHOR]

Under normal growth (NORMAL) condition with adequate supply of phosphorus and nitrogen, the wellstudied cyanobacterium Synechocystis wild type (WT) produced a high biomass level but subtle contents of the two biopolymers: poly-3-hydroxybutyrate (PHB) and glycogen. In contrast, under nitrogen deprivation, growth of WT was terminated, while the accumulations of PHB and glycogen were increased. Thus, a strain and a cultivation strategy that simultaneously allows both biomass production and the increased storages of the two biopolymers are desirable. Here, under phosphorus deprivation (-P), WT grew and still produced biomass at 850 mg/l (accounting for 62% of the biomass level obtained under NORMAL cultivation) with slightly increased contents of PHB and glycogen at 0.5% and 5.5% (w/w DW), respectively. Under -P, the △G mutant with the inactivated glycogen synthesis produced biomass at 751 mg/l and enhanced PHB accumulation to 4.9% (w/w DW), corresponding to PHB production of 36 mg/l. The △HP mutant with the inactivation of both PHB and hydrogen-gas syntheses under -P produced biomass at 713 mg/l and increased glycogen storage to 14.6% (w/w DW), corresponding to glycogen production of 118 mg/l. The significantly reduced levels of both chlorophyll and phycobilisomes (the main photosynthetic protein complex) found in △G and △HP mutants under -P suggested that photosynthetic proteins were degraded into amino acids which might subsequently be used for PHB and glycogen syntheses. The results indicated that the metabolic engineering combined with -P cultivation concurrently allowed cell growth and increased the production of two biopolymers in the studied Synechocystis. [ABSTRACT FROM AUTHOR]

The marine diatom Phaeodactylum tricornutum is an emerging host for metabolic engineering, but little is known about how introduced pathways are integrated into the existing metabolic framework of the host or influence transgene expression. In this study, we expressed the heterologous poly-3-hydroxybutyrate (PHB) pathway using episomal expression, which draws on the precursor acetyl coenzyme-A (AcCoA). By experimentally perturbing cultivation conditions, we gained insight into the regulation of the endogenous metabolism in transgenic lines under various environmental scenarios, as well as on alterations in AcCoA flux within the host cell. Biosynthesis of PHB led to distinct shifts in the metabolome of the host, and further analysis revealed a condition-dependent relationship between endogenous and transgenic metabolic pathways. Under N limitation, which induced a significant increase in neutral lipid content, both metabolic and transcriptomic data suggest that AcCoA was preferably shunted into the endogenous pathway for lipid biosynthesis over the transgenic PHB pathway. In contrast, supply of organic carbon in the form of glycerol supported both fatty acid and PHB biosynthesis, suggesting cross-talk between cytosolic and plastidial AcCoA precursors. This is the first study to investigate the transcriptomic and metabolomic response of diatom cell lines expressing a heterologous multi-gene pathway under different environmental conditions, providing useful insights for future engineering attempts for pathways based on the precursor AcCoA. Key points: • PHB expression had minimal effects on transcription of adjacent pathways. • N limitation favoured native lipid rather than transgenic PHB synthesis. • Glycerol addition allowed simultaneous lipid and PHB accumulation. [ABSTRACT FROM AUTHOR]

The ability of materials to adhere bacteria on their surface is one of the most important aspects of their development and application in bioengineering. In this work, the effect of the properties of films and electrospun scaffolds made of composite materials based on biosynthetic poly(3-hydroxybutyrate) (PHB) with the addition of magnetite nanoparticles (MNP) and their complex with graphene oxide (MNP/GO) on the adhesion of E. coli and L. fermentum under the influence of a low-frequency magnetic field and without it was investigated. The physicochemical properties (crystallinity; surface hydrophilicity) of the materials were investigated by X-ray structural analysis, differential scanning calorimetry and "drop deposition" methods, and their surface topography was studied by scanning electron and atomic force microscopy. Crystal violet staining made it possible to reveal differences in the surface charge value and to study the adhesion of bacteria to it. It was shown that the differences in physicochemical properties of materials and the manifestation of magnetoactive properties of materials have a multidirectional effect on the adhesion of model microorganisms. Compared to pure PHB, the adhesion of E. coli to PHB-MNP/GO, and for L. fermentum to both composite materials, was higher. In the magnetic field, the adhesion of E. coli increased markedly compared to PHB-MNP/GO, whereas the effect on the adhesion of L. fermentum was reversed and was only evident in samples with PHB-MNP. Thus, the resultant factors enhancing and impairing the substrate binding of Gram-negative E. coli and Gram-positive L. fermentum turned out to be multidirectional, as they probably have different sensitivity to them. The results obtained will allow for the development of materials with externally controlled adhesion of bacteria to them for biotechnology and medicine. [ABSTRACT FROM AUTHOR]

Electrospinning of biomimetic materials is of particular interest due to the possibility of producing flexible layers with highly developed surfaces from a wide range of polymers. Additionally, electrospinning is characterized by a high simplicity of implementation and the ability to modify the produced fibrous materials, which resemble structures found in living organisms. This study explores new electrospun materials based on polyhydroxyalkanoates, specifically poly-3-hydroxybutyrate, modified with chlorophyll derivatives. The research investigates the impact of chlorophyll derivatives on the morphology, supramolecular structure, and key properties of nonwoven materials. The obtained results are of interest for the development of new flexible materials with low concentrations of chlorophyll derivatives. [ABSTRACT FROM AUTHOR]

This work focuses on the obtaining and the bactericidal properties study, in vitro, of hybrid films as potential coating materials to inhibit bacteria proliferation. In consequence, hybrid films from nickel oxide (NiO) and nickel ferrite (NiFe2O4) nanoparticles (NPs) embedded in poly-3-hydroxybutyrate (P3HB) were obtained by the solvent casting method. P3HB@NiO and P3HB@NiFe2O4 hybrid films and P3HB film were characterized by X-ray diffraction (XRD), Raman scattering, and scanning electron microscopy (SEM). The XRD of the hybrid films showed that NiO and NiFe2O4 NPs incorporated in the P3HB conserved their nanometric size, and by Energy-dispersive X-ray spectroscopy (EDS) were observed that NPs are homogeneously distributed in the films. The bactericidal effect of the obtained films was evaluated in vitro from the broth surface method against two opportunistic and nosocomial pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. The results showed that P3HB film, P3HB@NiO, and P3HB@NiFe2O4 hybrid films reduced 90%, 98%, and 97% of the growth of S. aureus, respectively. For P. aeruginosa, their growth was reduced by 90%, 94%, and 96%, respectively. In addition, the cytotoxic effect of NiO and NiFe2O4 NPs, as well as P3HB film, and P3HB@NiO, and P3HB@NiFe2O4 hybrid films was evaluated using human skin cells; keratinocytes and fibroblast, being the NPs less cytotoxic than films. Although P3HB is known as a biocompatible polymer, here is demonstrated that in our work conditions, their films have bactericidal properties and are cytotoxic to keratinocytes and fibroblasts, the first barrier of the human skin. However, the P3HB@NiO and P3HB@NiFe2O4 hybrid films synergize the bactericidal effect between the P3HB and the NPs. On the other hand, the NPs decrease the P3HB cytotoxicity to keratinocytes. The methodology used in this work is particularly suitable for producing hybrid films with antibacterial activity against Gram-positive and Gram-negative bacterial strains. [ABSTRACT FROM AUTHOR]

Predictability and robustness are challenges for bioproduction because of the unstable intracellular synthetic activities. With the deeper understanding of the gene expression process, fine-tuning has become a meaningful tool for biosynthesis optimization. This study characterized several gene expression elements and constructed a multiple inducible system that responds to ten different small chemical inducers in halophile bacterium Halomonas bluephagenesis. Genome insertion of regulators was conducted for the purpose of gene cluster stabilization and regulatory plasmid simplification. Additionally, dynamic ranges of the multiple inducible systems were tuned by promoter sequence mutations to achieve diverse scopes for high-resolution gene expression control. The multiple inducible system was successfully employed to precisely control chromoprotein expression, lycopene and poly-3-hydroxybutyrate (PHB) biosynthesis, resulting in colorful bacterial pictures, optimized cell growth, lycopene and PHB accumulation. This study demonstrates a desirable approach for fine-tuning of rational and efficient gene expressions, displaying the significance for metabolic pathway optimization. • Gene expression elements and multiple inducible systems were constructed in Halomonas bluephagenesis. • Multiple inducible systems with diverse scopes were achieved for high-resolution gene expression control. • Chromoprotein expression, lycopene and (PHB) biosynthesis were tuned with multiple inducible systems. [ABSTRACT FROM AUTHOR]

Biocomposite development, as a sustainable alternative to fossil-derived materials with diverse industrial applications, requires expediting the design process and reducing production costs. Simulation methods offer a solution to these challenges. The main aspects to consider in simulating composite materials successfully include accurately representing microstructure geometry, carefully selecting mesh elements, establishing appropriate boundary conditions representing system forces, utilizing an efficient numerical method to accelerate simulations, and incorporating statistical tools like experimental designs and re-regression models. This study proposes a comprehensive methodology encompassing these aspects. We present the simulation using a numerical homogenization technique based on FEM to analyze the mechanical behavior of a composite material of a polyhydroxybutyrate (PHB) biodegradable matrix reinforced with cylindrical inclusions of flax and kenab. Here, the representative volume element (RVE) considered the geometry, and the numerical homogenization method (NHM) calculated the macro-mechanical behavior of composites. The results were validated using the asymptotic homogenization method (AHM) and experimental data, with error estimations of 0.0019% and 7%, respectively. This model is valuable for predicting longitudinal and transverse elastic moduli, shear modulus, and Poisson's coefficient, emphasizing its significance in composite materials research. [ABSTRACT FROM AUTHOR]

Summary: Plant‐based co‐production of polyhydroxyalkanoates (PHAs) and seed oil has the potential to create a viable domestic source of feedstocks for renewable fuels and plastics. PHAs, a class of biodegradable polyesters, can replace conventional plastics in many applications while providing full degradation in all biologically active environments. Here we report the production of the PHA poly[(R)‐3‐hydroxybutyrate] (PHB) in the seed cytosol of the emerging bioenergy crop Camelina sativa engineered with a bacterial PHB biosynthetic pathway. Two approaches were used: cytosolic localization of all three enzymes of the PHB pathway in the seed, or localization of the first two enzymes of the pathway in the cytosol and anchoring of the third enzyme required for polymerization to the cytosolic face of the endoplasmic reticulum (ER). The ER‐targeted approach was found to provide more stable polymer production with PHB levels up to 10.2% of the mature seed weight achieved in seeds with good viability. These results mark a significant step forward towards engineering lines for commercial use. Plant‐based PHA production would enable a direct link between low‐cost large‐scale agricultural production of biodegradable polymers and seed oil with the global plastics and renewable fuels markets. [ABSTRACT FROM AUTHOR]

The aim of this study was to utilize cassava pulp to prepare biocomposites comprising microcrystalline cellulose from cassava pulp (CP-MCC) as a filler and polyhydroxybutyrate (PHB) synthesized in-house by Cupriavidus necator strain A-04. The CP-MCC was extracted from fresh cassava pulp. Next, the CP-MCC surface was modified with butyryl chloride (esterified to CP-MCC butyrate) to improve dissolution and compatibility with the PHB. FTIR results confirmed that the esterified CP-MCC butyrate had aliphatic chains replacing the hydroxyl groups; this substitution increased the solubilities in acetone, chloroform, and tetrahydrofuran. Biocomposite films were prepared by varying the composition of esterified CP-MCC butyrate as a filler in the PHB matrix at 0, 5, 10, 15, 20 and 100 wt%. The results for the 95:5 and 90:10 CP-MCC butyrate biocomposite films showed that esterification led to improvements in the thermal properties and increased tensile strengths and elongations at break. All prepared biocomposite films maintained full biodegradability. [ABSTRACT FROM AUTHOR]

A rod-shaped, motile, Gram-negative bacterial strain named DM-R-R2A-13T was isolated from the plant Cannabis sativa L. 'Cheungsam'. The phylogenetic analysis of the 16S rRNA gene sequence revealed that strain DM-R-R2A-13T belongs to the family Oxalobacteraceae and is closely related to members of the genus Massilia, with Massilia flava (97.58% sequence similarity) and Massilia armeniaca (97.37% sequence similarity) being the closest members. The digital DNA-DNA hybridization (dDDH) values between strain DM-R-R2A-13T and Massilia flava CGMCC 1.10685T and Massilia armeniaca ZMN-3Twere 22.2% and 23.3%, while the average nucleotide identity (ANI) values were 78.85% and 79.63%, respectively. The DNA G+C content was measured to be 64.6 mol%. Moreover, the bacterium was found to contain polyhydroxyalkanoate (PHA) granules based on transmission electron microscopy, indicating its potential to produce bioplastic. Genome annotation revealed the presence of PHA synthase genes (phaC, phaR, phaP, and phaZ), and the biopolymer was identified as poly-3-hydroxybutyrate (PHB) based on nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopy (FTIR) analyses. Using maltose as a carbon source, the strain produced PHB of up to 58.06% of its dry cell weight. Based on the phenotypic, chemotaxonomic, and phylogenetic characteristics, it has been determined that DM-R-R2A-13T represents a novel species belonging to the genus Massilia. As such, the name Massilia endophytica sp. nov. is proposed for this newly identified species. The type strain is DM-R-R2A-13T (= KCTC 92072T = GDMCC 1.2920T). [ABSTRACT FROM AUTHOR]

Simulation is used by scientists to imitate a real-life experimental setup in order to save time, costs and effort. Geant4, a toolkit based on the Monte Carlo method, has been widely used in investigating the radiation-shielding properties of different materials. In many recent studies, researchers have focused on polymers and their shielding capabilities. Poly(lactic acid) (PLA) is a widely used biopolymer in many applications due to its excellent mechanical properties. However, it has limitations related to its degree of crystallinity and molecular characteristics, which could be improved through blending with other biodegradable polymers such as poly(hydroxybutyrate) (PHB). Previous published studies have shown that the mechanical properties of such blends can be improved further. In this work, the effect of blending PHB with PLA on the photon- and neutron-shielding capabilities will be investigated using Geant4 over a wide energy range, as well as the effect of doping those blends with metal oxides. The results show that the shielding properties of the polymers are affected by blending with other polymers and by doping the polymer blends with different metal oxides, and they confirm that Geant4 is a very reliable tool that can simulate any material's shielding properties against photons and neutrons. [ABSTRACT FROM AUTHOR]

The mechanical behavior of polymer materials is heavily influenced by a phenomenon known as crazing. Crazing is a precursor to damage and leads to the formation of cracks as it grows in both thickness and tip size. The current research employs an in situ SEM method to investigate the initiation and progression of crazing in all-biopolymeric blends based on Polyhydroxyalkanoates (PHAs). To this end, two chemically different grades of PHA, namely poly(hydroxybutyrate) (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBHV), were melt-blended with polybutyrate adipate terephthalate (PBAT). The obtained morphologies of blends, the droplet/fibrillar matrix, were highly influenced by the plasticity of the matrices as well as the content of the minor phase. Increasing the concentration of PBAT from 15 to 30 wt.% resulted in the brittle to ductile transition. It changed the mechanism of plastic deformation from single craze-cracking to homogeneous and heterogeneous intensified crazing for PHB and PHBHV matrices, respectively. Homogeneous tensile crazes formed perpendicularly to the draw direction at the initial stages of deformation, transformed into shear crazes characterized by oblique edge propagation for the PHBHV/PBAT blend. Such angled crazes suggested that the displacement might be caused by shear localized deformation. The crazes' strength and the time to failure increased with the minor phase fibers. These fibers, aligned with the tensile direction and spanning the width of the crazes, were in the order of a few micrometers in diameter depending on the concentration. The network of fibrillar PBAT provided additional integrity for larger plastic deformation values. This study elucidates the mechanism of crazing in PHA blends and provides strategies for controlling it. [ABSTRACT FROM AUTHOR]

A rod-shaped, motile, Gram-negative bacterial strain named DM-R-R2A-13T was isolated from the plant Cannabis sativa L. 'Cheungsam'. The phylogenetic analysis of the 16S rRNA gene sequence revealed that strain DM-R-R2A-13T belongs to the family Oxalobacteraceae and is closely related to members of the genus Massilia, with Massilia flava (97.58% sequence similarity) and Massilia armeniaca (97.37% sequence similarity) being the closest members. The digital DNA-DNA hybridization (dDDH) values between strain DM-R-R2A-13T and Massilia flava CGMCC 1.10685T and Massilia armeniaca ZMN-3Twere 22.2% and 23.3%, while the average nucleotide identity (ANI) values were 78.85% and 79.63%, respectively. The DNA G+C content was measured to be 64.6 mol%. Moreover, the bacterium was found to contain polyhydroxyalkanoate (PHA) granules based on transmission electron microscopy, indicating its potential to produce bioplastic. Genome annotation revealed the presence of PHA synthase genes (phaC, phaR, phaP, and phaZ), and the biopolymer was identified as poly-3-hydroxybutyrate (PHB) based on nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopy (FTIR) analyses. Using maltose as a carbon source, the strain produced PHB of up to 58.06% of its dry cell weight. Based on the phenotypic, chemotaxonomic, and phylogenetic characteristics, it has been determined that DM-R-R2A-13T represents a novel species belonging to the genus Massilia. As such, the name Massilia endophytica sp. nov. is proposed for this newly identified species. The type strain is DM-R-R2A-13T (= KCTC 92072T = GDMCC 1.2920T). [ABSTRACT FROM AUTHOR]

This study developed a technical task associated with the formation of welded joints based on biodegradable polymers and their subsequent physicochemical characterization. The primary objective was to establish the effect of the welding process and modification of natural poly(3-hydroxybutyrate) (PHB) with N,N-dibutylundecenoylamide (DBUA) as a plasticizing agent on the structure and properties of PHB-based biopolymer materials as well as the process and structure of welded joints formation using ultrasonic welding technique. The weldability of biodegradable layers based on PHB and PHB/DBUA mixture was ultrasonically welded and optimized using a standard Branson press-type installation. The effect of the DBUA plasticizer and welding process on the structure of PHB-based biodegradable material was investigated using scanning electron microscopy, X-ray diffraction, FT-IR spectroscopy, differential scanning calorimetry, and thermomechanical analysis. The results confirmed that the DBUA acted as an effective plasticizer of PHB, contributing to lower crystallinity of the PHB/DBUA mixture (63%) in relation to the crystallinity degree of pure PHB film (69%). Ultrasonic welding resulted in an additional increase (approximately 8.5%) in the degree of crystallinity in the PHB/DBUA in relation to the initial PHB/DBUA mixture. The significant shift toward lower temperatures of the crystallization and melting peaks of PHB modified with DBUA were observed using DSC concerning pure PHB. The melt crystallization process of PHB was affected by welding treatment, and a shift toward higher temperature was observed compared with the unwelded PHB/DBUA sample. The butt-welded joints of biodegradable PHB/DBUA materials made using the ultrasonic method tested for tensile strength have damaged the area immediately outside the joining surface. [ABSTRACT FROM AUTHOR]

In order to create new biodegradable nanocomposites for biomedicine, packaging, and environmentally effective adsorbents, ultra-thin composite fibers consisting of poly(3-hydroxybutyrate) (PHB) and graphene oxide (GO) were obtained by electrospinning. Comprehensive studies of ultrathin fibers combining thermal characteristics, dynamic electron paramagnetic resonance (ESR) probe measurements, and scanning electron microscopy (SEM) were carried out. It is shown that at the addition of 0.05, 0.1, 0.3, and 1% OG, the morphology and geometry of the fibers and their thermal and dynamic characteristics depend on the composite content. The features of the crystalline and amorphous structure of the PHB fibers were investigated by the ESR and DSC methods. For all compositions of PHB/GO, a nonlinear dependence of the correlation time of molecular mobility TEMPO probe (τ) and enthalpy of biopolyether melting (ΔH) is observed. The influence of external factors on the structural-dynamic properties of the composite fiber, such as hydrothermal exposure of samples in aqueous medium at 70 °C and ozonolysis, leads to extreme dependencies of τ and ΔH, which reflect two processes affecting the structure in opposite ways. The plasticizing effect of water leads to thermal destruction of the orientation of the pass-through chains in the amorphous regions of PHB and a subsequent decrease in the crystalline phase, and the aggregation of GO nanoplates into associates, reducing the number of GO-macromolecule contacts, thus increasing segmental mobility, as confirmed by decreasing τ values. The obtained PHB/GO fibrillar composites should find application in the future for the creation of new therapeutic and packaging systems with improved biocompatibility and high-barrier properties. [ABSTRACT FROM AUTHOR]

When applying electron or gamma irradiation to poly-3-hydroxybutyrate (P3HB), main chain scissions are the dominant material reactions. Though propositions have been made that crosslinking in the amorphous phase of P3HB occurs under irradiation, a conclusive method to achieve controlled additive free irradiation crosslinking has not been shown and no mechanism has been derived to the best of our knowledge. By applying irradiation in a molten state at 195 °C and doses above 200 kGy, we were able to initiate crosslink reactions and achieved gel formation of up to 16%. The gel dose Dgel was determined to be 200 kGy and a range of the G values, the number of scissions and crosslinks for 100 eV energy deposition, is given. Rheology measurements, as well as size exclusion chromatography (SEC), showed indications for branching at doses from 100 to 250 kGy. Thermal analysis showed the development of a bimodal peak with a decrease in the peak melt temperature and an increase in peak width. In combination with an increase in the thermal degradation temperature for a dose of 200 kGy compared to 100 kGy, thermal analysis also showed phenomena attributed to branching and crosslinking. [ABSTRACT FROM AUTHOR]

The production of biomass and biopolyesters (polyhydroxyalkanoates) by the marine bacterium Saccharophagus degradans from: glucose, starch, glucosamine, and xylose were studied. The effect of yeast extract, trace elements, vitamins, and initial concentration of each carbon source on biomass production was evaluated. The addition of yeast extract, together with trace elements, increased the biomass production by 5- fold, obtaining the maximum values when using an initial substrate concentration of 20 g/L for: starch, 8.61; glucosamine, 7.06; and xylose, 4.47 (g dry cell weight/ L). From glucose at 40 g/L the maximum biomass production was 9.07 g/L. Next, the biopolymer production under nutrient-limiting conditions (by N, P, S and Mg) was investigated, as well as the ability of S. degradans to produce copolymers from precursors. The biopolyester produced in all cases was poly(3-hydroxybutyrate) (PHB). Different accumulation percentages (g PHB/100 g biomass) were achieved depending on the carbon source used: glucose, 27.72; starch, 18.11; glucosamine 3.33; there was not biopolymer accumulated from xylose. Finally, the PHB produced from each carbohydrate was characterized, presenting molecular weights between 53 and 58 KDa, and fusion temperatures between 164 and 174°C. [ABSTRACT FROM AUTHOR]

A new class of bio-composites was developed by utilizing four kinds of lignocellulosic biomass fiber (bagasse, bamboo, rice husk, and rice straw) as filling fibers. Poly-β-hydroxybutyrate (PHB) and poly(butylene succinate) (PBS) in a mixture ratio of 7:3 were used as matrix materials with hot-press molding. The performance of the resulting composites was evaluated by compositional analyses, mechanical analysis, Fourier transform infrared (FTIR) spectroscopy, thermogravimetry, and morphological analysis. The interfacial adhesion, thermal stability, and comprehensive mechanical properties of the alkali treated bamboo/PHB/PBS composite were highest among the four bio-composites. The bending strength, tensile strength, and impact strength for alkali treated bamboo/PHB/PBS composite was 19.82 MPa, 12.97 MPa, and 4.30 kJ/m², respectively. The thermal stability for NaOH modified bamboo/PHB/PBS composite was slightly superior to the other three composites, with the initial pyrolysis temperature of 248 °C, moderate pyrolysis speed, and the amount of pyrolysis residue (5.81%). The results showed the suitability of biomass fiber and biodegradable polymer for producing environmentally friendly composite materials. [ABSTRACT FROM AUTHOR]

Poly-3-hydroxybutyrate (P(3HB)), a member of the polyhydroxyalkanoate (PHA) family, is a biodegradable polyester with diverse industrial applications. NADPH-dependent acetoacetyl-CoA reductase (pha B) is the enzyme which plays an essential role in P(3HB) synthesis by catalyzing the conversion of the intermediates. The expression of pha B enzyme using the recombinant Escherichia coli BL-21(DE3) and the purification of the synthesized enzyme were studied. The pET-B3 plasmid harbouring the pha B gene derived from Ralstonia eutropha H16, was driven by the lac promoter in E. coli BL-21(DE3). The enzyme was expressed with different induction time, temperatures and cell age. Results showed that the cell age of 4 h, induction time of 12 h at 37°C were identified as the optimal conditions for the enzyme reductase expression. A specific activity of 0.151 U mg−1 protein and total protein concentration of 0.518 mg mg−1 of dry cell weight (DCW) were attained. Affinity chromatography was performed to purify the His-tagged pha B enzyme, in which enhanced the specific activity (14.44 U mg−1) and purification fold (38-fold), despite relative low yield (44.6%) of the enzyme was obtained. The purified pha B showed an optimal enzyme activity at 30°C and pH 8.0. The findings provide an alternative for the synthesis of the reductase enzyme which can be used in the industrial-scale production of the biodegradable polymers. • The His-tagged pha B was successfully expressed in recombinant Escherichia coli BL-21(DE3). • Affinity chromatography was applied for the purification of His-tagged pha B enzyme. • Specific activity and purification fold of purified pha B enzyme was notably enhanced. [ABSTRACT FROM AUTHOR]

Bacteria frequently store excess carbon in hydrophobic granules of polyhydroxybutyrate (PHB) that in some growth conditions can occupy most of the cytoplasmic space. Different types of proteins associate to the surface of the granules, mainly enzymes involved in the synthesis and utilization of the reserve polymer and a diverse group of proteins known as phasins. Phasins have different functions, among which are regulating the size and number of the granules, modulating the activity of the granule‐associated enzymes and helping in the distribution of the granules inside the cell. Caulobacter crescentus is an oligotrophic bacterium that shows several morphological and regulatory traits that allow it to grow in very nutrient‐diluted environments. Under these conditions, storage compounds should be particularly relevant for survival. In this work, we show an initial proteomic characterization of the PHB granules and describe a new type of phasin (PhaH) characterized by the presence of an N‐terminal hydrophobic helix followed by a helix‐hairpin‐helix (HhH) domain. The hydrophobic helix is required for maximal PHB accumulation and maintenance during the stationary phase while the HhH domain is involved in determining the size of the PHB granules and their distribution in the cell. [ABSTRACT FROM AUTHOR]

The development of biopolymer scaffolds for intestine regeneration is one of the most actively developing areas in tissue engineering. However, intestinal regenerative processes after scaffold implantation depend on the activity of the intestinal microbial community that is in close symbiosis with intestinal epithelial cells. In this work, we study the impact of different scaffolds based on biocompatible poly(3-hydroxybutyrate) (PHB) and alginate (ALG) as well as PHB/ALG scaffolds seeded with probiotic bacteria on the composition of gut microbiota of Wistar rats. Implantation of PHB/ALG scaffolds on the large-intestine wall to close its injury showed that alpha diversity of the gut microbiota was not reduced in rats implanted with different PHB/ALG scaffolds except for the PHB/ALG scaffolds with the inclusion of Lactobacillus spheres (PHB/ALG-L). The composition of the gut microbiota of rats implanted with PHB/ALG scaffolds with probiotic bacteria or in simultaneous use of an antimicrobial agent (PHB/ALG-AB) differed significantly from other experimental groups. All rats with implanted scaffolds demonstrated shifts in the composition of the gut microbiota by individual operational taxonomic units. The PHB/ALG-AB construct led to increased abundance of butyrate-producing bacteria: Ileibacterium sp. dominated in rats with implanted PHB/ALG-L and Lactobacillus sp. and Bifidobacterium sp. dominated in the control group. In addition, the PHB/ALG scaffolds had a favourable effect on the growth of commensal bacteria. Thus, the effect of implantation of the PHB/ALG scaffold compared to other scaffolds on the composition of the gut microbiota was closest to the control variant, which may demonstrate the biocompatibility of this device with the microbiota. [ABSTRACT FROM AUTHOR]

Background: Poly-β-hydroxybutyrate (PHB), produced by a variety of microbial organisms, is a good substitute for petrochemically derived plastics due to its excellent properties such as biocompatibility and biodegradability. The high cost of PHB production is a huge barrier for application and popularization of such bioplastics. Thus, the reduction of the cost is of great interest. Using low-cost substrates for PHB production is an efficient and feasible means to reduce manufacturing costs, and the construction of microbial cell factories is also a potential way to reduce the cost. Results: In this study, an engineered Sphingomonas sanxanigenens strain to produce PHB by blocking the biosynthetic pathway of exopolysaccharide was constructed, and the resulting strain was named NXdE. NXdE could produce 9.24 ± 0.11 g/L PHB with a content of 84.0% cell dry weight (CDW) using glucose as a sole carbon source, which was significantly increased by 76.3% compared with the original strain NX02. Subsequently, the PHB yield of NXdE under the co-substrate with different proportions of glucose and xylose was also investigated, and results showed that the addition of xylose would reduce the PHB production. Hence, the Dahms pathway, which directly converted D-xylose into pyruvate in four sequential enzymatic steps, was enhanced by overexpressing the genes xylB, xylC, and kdpgA encoding xylose dehydrogenase, gluconolactonase, and aldolase in different combinations. The final strain NX02 (ΔssB, pBTxylBxylCkdpgA) (named NXdE II) could successfully co-utilize glucose and xylose from corn straw total hydrolysate (CSTH) to produce 21.49 ± 0.67 g/L PHB with a content of 91.2% CDW, representing a 4.10-fold increase compared to the original strain NX02. Conclusion: The engineered strain NXdE II could co-utilize glucose and xylose from corn straw hydrolysate, and had a significant increase not only in cell growth but also in PHB yield and content. This work provided a new host strain and strategy for utilization of lignocellulosic biomass such as corn straw to produce intracellular products like PHB. [ABSTRACT FROM AUTHOR]

Poly-3-hydroxybutyrate (PHB) is a bio-based and biodegradable polymer produced by microbial fermentation from wide variety of feedstocks. Methanotrophic organism Methylosinus trichosporium has been reported for the production of PHB using alternate feedstock like methane which is economical and abundantly available. PHB was produced from methane by cultivating M. trichosporium in a bioreactor with continuous cell recycle system. Different gas sparging strategies were evaluated for impact on cell biomass and PHB production. The fermentation was conducted in different modes including batch and continuous culture with and without membrane recycle. Using membrane based recycle system the biomass increased from 1.5 to maximum of 7.31 g DCW/L in 27 days. Although PHB content was comparable in both batch mode and membrane based recycle system, a 4.9 fold increase in biomass production enhanced the PHB titer by 6.1 fold as compared to batch culture. Continuous cell recycle processes provide an opportunity to increase the competitiveness of gas fermentation based processes for methanotrophs based PHB production. [ABSTRACT FROM AUTHOR]

Biopolymers incorporating biologically active compounds are promising for applications in regenerative medicine and controlled drug delivery. We have studied the interaction of the neurotransmitter acetylcholine (ACh) with biocompatible polymers, polylactide (PLA), poly-3-hydroxybutyrate (PHB), and poly-ε-caprolactone (PCL). Analysis of particle sizes and zeta potentials in solutions in chloroform by dynamic light scattering showed that the addition of 1 mmol L−1 of ACh to polymer solutions led to a decrease in the size of globules and a decrease in the zeta potential, which indicated a polymer—ACh interaction. Individual PLA, PCL, PHB, and ACh dissolved in chloroform accelerated the radical decomposition of hydroperoxides, and synergism in the generation of radicals was observed in a mixture of polymers with ACh. [ABSTRACT FROM AUTHOR]

α-tricalcium (α-TCP) phosphate is widely used as an osteoinductive biocompatible material, serving as an alternative to synthetic porous bone materials. The objective of this study is to obtain a highly filled fibrous nonwoven material composed of poly-3-hydroxybutyrate (PHB) and α-TCP and to investigate the morphology, structure, and properties of the composite obtained by the electrospinning method (ES). The addition of α-TCP had a significant effect on the supramolecular structure of the material, allowing it to control the crystallinity of the material, which was accompanied by changes in mechanical properties, FTIR spectra, and XRD curves. The obtained results open the way to the creation of new osteoconductive materials with a controlled release of the source of calcium into the living organism. [ABSTRACT FROM AUTHOR]

Gram-negative Azospirillum brasilense accumulates approximately 80% of polyhydroxybutyrate (PHB) as dry cell weight. For this reason, this bacterium has been characterized as one of the main microorganisms that produce PHB. PHB is synthesized inside bacteria by the polymerization of 3-hydroxybutyrate monomers. In this review, we are focusing on the analysis of the PHB production by A. brasilense in order to understand the metabolism during PHB accumulation. First, the carbon and nitrogen sources used to improve PHB accumulation are discussed. A. brasilense accumulates more PHB when it is grown on a minimal medium containing a high C/N ratio, mainly from malate and ammonia chloride, respectively. The metabolic pathways to accumulate and mobilize PHB in A. brasilense are mentioned and compared with those of other microorganisms. Next, we summarize the available information to understand the role of the genes involved in the regulation of PHB metabolism as well as the role of PHB in the physiology of Azospirillum. Finally, we made a comparison between the properties of PHB and polypropylene, and we discussed some applications of PHB in biomedical and commercial areas. [ABSTRACT FROM AUTHOR]

Nature, especially in hot environments, triggers the phase change of the abundant metabolites in plants from solid to liquid as a survival mechanism. Therefore, taking cues from nature, these metabolites could have potential as green solvents for the dissolution of biopolymers. Because of their green credentials, plant-derived aromatic monoterpenoids have gained considerable attention for the extraction of bioactive compounds to replace toxic halogenated solvents. In this work, we report for the first time the fast dissolution of high concentration poly(3-hydroxybutyrate) (PHB) using carvacrol (5-isopropyl-2-methylphenol) – a plant metabolite – as a solvent. Remarkably, 20 wt% PHB was completely dissolved within 30 min at 100 °C, whereas a 10 wt% PHB dissolved in 21 min, and even faster dissolution for 1 wt%, 3 wt% and 5 wt% PHB. The carvacrol/PHB solution exhibited stability characterized by rheology measurements and the atomistic molecular dynamics (MD) simulations elucidate the mechanism behind carvacrol's rapid dissolution of PHB. Most importantly, the carvacrol solvent can be fully recovered by adding anti-solvents. Furthermore, the resultant highly viscous carvacrol/PHB solution could find applications in coating formulations, adhesives, and consumer care additives. [ABSTRACT FROM AUTHOR]

This work appraises the prospect of utilising groundnut shell hydrolysate as a feedstock used for PHB biosynthesis by Azotobacter chroococcum MTCC 3853 under SMF conditions. Sugar reduction: untreated and pretreated 20% H2SO4 (39.46 g/l and 62.96 g/l, respectively), untreated and enzymatic hydrolysis (142.35 mg/g and 568.94 mg/g). The RSM-CCD optimization method was used to generate augment PHB biosynthesis from groundnut shell hydrolysate (30 g/l), ammonium sulphate (1.5 g/l), ammonium chloride (1.5 g/l), peptone (1.5 g/l), pH 7, 30 °C, and a 48 h incubation time. The most convincing factors (p < 0.0001), coefficient R2 values of biomass 0.9110 and PHB yield 0.9261, PHB production, highest biomass (17.23 g/l), PHB Yield(11.46 g/l), and 66.51 (wt% DCW) values were recorded. The control (untreated GN) PHB yield value of 2.86 g/l increased up to fourfold in pretreated GN. TGA results in a melting range in the peak perceived at 270.55 °C and a DSC peak range of 172.17 °C, correspondingly. According to the results, it furnishes an efficient agricultural waste executive approach by diminishing the production expenditure. It reinforces the production of PHB, thereby shrinking our reliance on fossil fuel-based plastics. [ABSTRACT FROM AUTHOR]