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Abstract—The review presents publications devoted to a promising biopolymer, poly-3-hydroxybutyrate, and composite materials based on it. The structure and properties of the materials are considered. The main focus is on the biomedical application of these biopolymer materials. The main advantage of poly-3-hydroxybutyrate is its biodegradability in nature and living systems, biocompatibility, hemocompatibility, and thromboresistance. This biopolymer also possesses processability. The products and materials based on it can be processed by solution and melt production methods. The review pays great attention to the production of ultrathin fibers by electrospinning. Materials based on poly-3-hydroxybutyrate even now find wide application in medicine, hygiene, packaging, and ecology. [ABSTRACT FROM AUTHOR]

Abstract Background Microbial biopolymers such as poly-3-hydroxybutyrate (PHB) are emerging as promising alternatives for sustainable production of biodegradable bioplastics. Their promise is heightened by the potential utilisation of photosynthetic organisms, thus exploiting sunlight and carbon dioxide as source of energy and carbon, respectively. The cyanobacterium Synechocystis sp. B12 is an attractive candidate for its superior ability to accumulate high amounts of PHB as well as for its high-light tolerance, which makes it extremely suitable for large-scale cultivation. Beyond its practical applications, B12 serves as an intriguing model for unravelling the molecular mechanisms behind PHB accumulation. Results Through a multifaceted approach, integrating physiological, genomic and transcriptomic analyses, this work identified genes involved in the upregulation of chlorophyll biosynthesis and phycobilisome degradation as the possible candidates providing Synechocystis sp. B12 an advantage in growth under high-light conditions. Gene expression differences in pentose phosphate pathway and acetyl-CoA metabolism were instead recognised as mainly responsible for the increased Synechocystis sp. B12 PHB production during nitrogen starvation. In both response to strong illumination and PHB accumulation, Synechocystis sp. B12 showed a metabolic modulation similar but more pronounced than the reference strain, yielding in better performances. Conclusions Our findings shed light on the molecular mechanisms of PHB biosynthesis, providing valuable insights for optimising the use of Synechocystis in economically viable and sustainable PHB production. In addition, this work supplies crucial knowledge about the metabolic processes involved in production and accumulation of these molecules, which can be seminal for the application to other microorganisms as well.

Poly-3-hydroxybutyrate (P3HB) is a biodegradable polyester produced mainly by bacterial fermentation in an isotactic configuration. Its high crystallinity (about 70%) and brittle behavior have limited the process window and the application of this polymer in different sectors. Atactic poly-3-hydroxybutyrate (a-P3HB) is an amorphous polymer that can be synthesized chemically and blended with the isotactic P3HB to reduce its crystallinity and improve its processability Ring-opening polymerization (ROP) is the most cited synthesis route for this polymer in the literature. In this work, a new synthesis route of a-P3HB by self-polycondensation of racemic ethyl 3-hydroxybutyrate will be demonstrated. Different catalysts were tested regarding their effectiveness, and the reaction parameters were optimized using titanium isopropoxide as the catalyst. The resulting polymers were compared by self-polycondensation for their properties with those of a-P3HB obtained by the ROP and characterized by Fourier transform infrared spectroscopy (FTIR) and gel permeation chromatography (GPC), and the double bond content (DBC) was determined by UV–VIS spectroscopy by using 3-butenoic acid as a standard. Additionally, a life cycle analysis (LCA) of the new method of synthesizing has been carried out to assess the environmental impact of a-P3HB. [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]

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]

The principles of creating biomedical materials based on biopolymers and their compositions for various medical applications are considered. The direction of creating materials with antimicrobial activity is especially emphasized using the example of polyhydroxyalkanoates. These biopolymers and composites based on them are most frequently used in the production of medical devices. Polyhydroxyalkanoates as a class of biopolymers, as well as polyhydroxybutyrate, the representative of this class most suitable for biomedical use, are analyzed in the context of increasing resistance to microorganisms. Progress in this direction achieved in recent years is reported. The influence of the supramolecular and molecular structure of the materials on the ability to biodegrade in the environment and a living organism is considered. The advantage of mixtures of biopolymers for achieving high degradation rates in comparison with the original polymers is noted. Promising antiseptics based on porphyrin metal complexes in combination with biopolymer nonwoven fibrous matrices are demonstrated. Characteristic features of preclinical tests of antiseptic materials are considered. A conclusion is drawn that the structural organization of a polymeric material or composite determines the level of intermolecular interactions during the formation of the material and thereby programs the set of functional properties and mechanism of degradation under the influence of aggressive external factors. [ABSTRACT FROM AUTHOR]

Poly-3-hydroxybutyrate (P3HB) is a biodegradable polyester produced mainly by bacterial fermentation in an isotactic configuration. Its high crystallinity (about 70%) and brittle behavior have limited the process window and the application of this polymer in different sectors. Atactic poly-3-hydroxybutyrate (a-P3HB) is an amorphous polymer that can be synthesized chemically and blended with the isotactic P3HB to reduce its crystallinity and improve its processability Ring-opening polymerization (ROP) is the most cited synthesis route for this polymer in the literature. In this work, a new synthesis route of a-P3HB by self-polycondensation of racemic ethyl 3-hydroxybutyrate will be demonstrated. Different catalysts were tested regarding their effectiveness, and the reaction parameters were optimized using titanium isopropoxide as the catalyst. The resulting polymers were compared by self-polycondensation for their properties with those of a-P3HB obtained by the ROP and characterized by Fourier transform infrared spectroscopy (FTIR) and gel permeation chromatography (GPC), and the double bond content (DBC) was determined by UV–VIS spectroscopy by using 3-butenoic acid as a standard. Additionally, a life cycle analysis (LCA) of the new method of synthesizing has been carried out to assess the environmental impact of a-P3HB.

Oligomers of poly-3-hydroxybutyrate (PHB) were prepared by aminolysis of high molecular weight PHB with ethylenediamine and 1,4-diaminobutane. Polymer-oligomer blends (10, 30, and 50% content of the oligomers) were prepared as films by solution casting. As the content of oligomers increased, a gradual increase in the hydrophilicity of the polymer surface was observed, resulting reflected in the water contact angle decrease from 84° to 72–76°. In addition, a moderate decrease in elongation at break, Young's modulus, and tensile strength for the blends were observed as more oligomer was added to the film. Finally, the viability of NIH-3T3 mouse fibroblasts was higher compared to intact PHB when growing in non-prepared polymer/oligomer mixtures. These findings confirm the benefits of the introduction of a hydrophilic functionalized oligomer into the PHB matrix in terms of improving the biocompatibility of the resulting polymer/oligomer blends.

Acetate is a low-cost feedstock for the production of different bio-chemicals. Electrochemical reduction of CO2 into acetate and subsequent acetate fermentation is a promising method for transforming CO2 into value-added chemicals. However, the significant inhibitory effect of acetate on microbial growth remains a barrier for acetate-based biorefinery. In this study, the deletion of genes involved in L-leucine degradation was found to be beneficial for the growth of Pseudomonas stutzeri A1501 in acetate. P. stutzeri (Δpst_3217), in which the hydroxymethylglutaryl-CoA lyase catalyzing β-hydroxy-β-methylglutaryl-CoA into acetyl-CoA and acetoacetate was deleted, grew faster than other mutants and exhibited increased tolerance to acetate. Then, the genes phbCAB from Ralstonia eutropha H16 for poly-3-hydroxybutyrate (PHB) biosynthesis were overexpressed in P. stutzeri (∆pst_3217) and the recombinant strain P. stutzeri (∆pst_3217-phbCAB) can accumulate 0.11 g L−1 PHB from commercial acetate. Importantly, P. stutzeri (∆pst_3217-phbCAB) can also use CO2-derived acetate to produce PHB and the accumulated PHB accounted for 5.42% (w/w) of dried cell weight of P. stutzeri (∆pst_3217-phbCAB).

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 article examines the regularities of structure formation of ultrafine fibers based on poly-3-hydroxybutyrat under the influence of technological (electrical conductivity, viscosity), molecular (molecular weight), and external factors (low-molecular and nanodispersed substances of different chemical natures). Systems with polar substances are characterized by the presence of intermolecular interactions and the formation of a more perfect crystalline fiber structure. Changes in technological and molecular characteristics affect the fiber formation process, resulting in alterations in the morphology of the nonwoven fabric, fiber geometry, and supramolecular fiber structure. Polymer molecular weight, electrical conductivity, and solution viscosity influence fiber formation and fiber diameter. The fiber structure is heterogeneous, consisting of both crystalline and non-equilibrium amorphous phases. This article shows that with an increase in the molecular weight and concentration of the polymer, the diameter of the fiber increases. At the same time, the increase in the productivity of the electrospinning process does not affect the fiber geometry. The chemical structure of the solvent and the concentration of polar substances play a decisive role in the formation of fibers of even geometry. As the polarity of the solvent increases, the intermolecular interaction with the polar groups of poly-3-hydroxybutyrate increases. As a result of this interaction, the crystallites are improved, and the amorphous phase of the polymer is compacted. The action of polar molecules on the polymer is similar to the action of polar nanoparticles. They increase crystallinity via a nucleation mechanism. This is significant in the development of matrix-fibrillar systems for drug delivery, bioactive substances, antiseptics, tissue engineering constructs, tissue engineering scaffolds, artificial biodegradable implants, sorbents, and other applications. [ABSTRACT FROM AUTHOR]

Increasing greenhouse gas emissions and decreasing fossil fuel supplies necessitate the development of alternative methods for producing petroleum-based commodities. Plastics are also primarily petroleum-based goods with rising demand, thus there is growing interest in plastic substitutes. Polyhydroxyalkanoates (PHAs) are naturally produced biopolymers that are utilized by microorganisms as a source of energy and carbon storage. Poly-3-hydroxybutyrate (PHB) is a member of the PHA family and is considered the most promising candidate to replace polyethylene (PE). PHB is naturally produced by Cupriavidus necator, but recombinant production has also been recently established. This study is the first to investigate the heterologous production of PHB with recombinant Acetobacterium woodii using CO2 + H2 as a carbon and energy source. The introduction of a synthetic PHB production pathway resulted in the production of 1.23 g/L CDW and 1.9% PHB/cell dry weight (CDW), which corresponds to a production of 23.5 mg/L PHB. PHB quantification was simplified using LipidGreen2 fluorescence measurements. [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]

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]

Due to the current environmental situation, biopolymers are replacing the usual synthetic polymers, and special attention is being paid to poly-3-hydroxybutyrate (PHB), which is a biodegradable polymer of natural origin. In this paper, the rate of biodegradation of films and fibers based on PHB was compared. The influence of exposure to soil on the structure and properties of materials was evaluated using methods of mechanical analysis, the DSC method and FTIR spectroscopy. The results showed rapid decomposition of the fibrous material and also showed how the surface of the material affects the rate of biodegradation and the mechanical properties of the material. It was found that maximum strength decreased by 91% in the fibrous material and by 49% in the film. Additionally, the DSC method showed that the crystallinity of the fiber after exposure to the soil decreased. It was established that the rate of degradation is influenced by different factors, including the surface area of the material and its susceptibility to soil microorganisms. The results obtained are of great importance for planning the structure of features in the manufacture of biopolymer consumer products in areas such as medicine, packaging, filters, protective layers and coatings, etc. Therefore, an understanding of the biodegradation mechanisms of PHB could lead to the development of effective medical devices, packaging materials and different objects with a short working lifespan. [ABSTRACT FROM AUTHOR]

The current study was carried out to evaluate the effects of cartilage-decellularized extracellular matrix (ECM) on Poly (3-hydroxybutyrate) (PHB) electrospun scaffold for cartilage tissue engineering. In this study, PHB scaffolds with three different percentages of ECM (1%, 2%, and 3%) were fabricated. Mechanical, chemical, and physical properties of scaffolds along with their interactions with cells were assessed. Results showed that by adding of ECM to the scaffolds, the Physiomechanical properties of combined scaffolds were modified, compared to the pure PHB scaffold. In the other hand, the biological behavior was increased by increasing the ECM ratio to the PHB. [ABSTRACT FROM AUTHOR]

Electrospun fabrics have unique properties due to their uniform morphology and high surface area to volume ratio. Ultrathin nonwoven fabrics are produced for many applications: biomedical, nanosensors, tissue engineering and filtration systems. In this work, nonwoven polylactide, polylactide/natural rubber, poly-3-hydroxybutyrate, and poly-3-hydroxybutyrate/nitrile butadiene rubber fabrics were prepared by electrospinning methods. The obtained fabric samples were used as substrates for the growth of winter wheat seeds "Yubileinaya 100" (Triticum aestivum L.). The stimulating effect of polymer substrates on seed germination and plant growth was shown. The structure and properties of nonwoven agromaterials were controlled by differential scanning calorimetry, IR-spectroscopy, and optical microscopy. The mechanical properties of the obtained fabrics before and after their utilization as substrates were studied. After the wheat growing experiment, the degree of crystallinity of PHB and PHB/NBR samples decreased by 12% and they completely lost their mechanical properties. It is shown that the main factors providing the efficiency of seed growth technology on polymer substrates are the chemical nature and structure of the biodegradable matrix. [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.

Vibrio alginolyticus LHF01 was engineered to efficiently produce poly-3-hydroxybutyrate (PHB) from starch in this study. Firstly, the ability of Vibrio alginolyticus LHF01 to directly accumulate PHB using soluble starch as the carbon source was explored, and the highest PHB titer of 2.06 g/L was obtained in 18 h shake flask cultivation. Then, with the analysis of genomic information of V. alginolyticus LHF01, the PHB synthesis operon and amylase genes were identified. Subsequently, the effects of overexpressing PHB synthesis operon and amylase on PHB production were studied. Especially, with the co-expression of PHB synthesis operon and amylase, the starch consumption rate was improved and the PHB titer was more than doubled. The addition of 20 g/L insoluble corn starch could be exhausted in 6-7 h cultivation, and the PHB titer was 4.32 g/L. To the best of our knowledge, V. alginolyticus was firstly engineered to produce PHB with the direct utilization of starch, and this stain can be considered as a novel host to produce PHB using starch as the raw material.

The effect of the hemin (Hmi) on the structure and properties of nanocomposite electrospun materials based on poly-3-hydroxybutyrate (PHB) is discussed in the article. The additive significantly affected the morphology of fibers allowed to produce more elastic material and provided high antimicrobial activity. The article considers also the impact of the hemin on the biocompatibility of the nonwoven material based on PHB and the prospects for wound healing. [ABSTRACT FROM AUTHOR]

Polyhydroxyalkanoates (PHAs) including poly-3-hydroxybutyrate (P3HB) as secondary metabolisms were in vitro produced by the edible basidiomycetous mushroom Astraeus odoratus during its growth on malt agar extract. Various carbon and nitrogen sources containing cellulose, glucose, glycerol, rice straw powder, soybean meal and peptone were investigated for the growth of basidiomycetous mushrooms. During cultivation, the A. odoratus culture exudated the considerably extracellular fluid up to approx. 2.3 ml on 2% malt extract agar plate within 7 days. The chemical compounds of the exudated fluid were further investigated by Fourier-transform infrared spectroscopy (FTIR) and gas chromatography/mass spectrometry (GC/MS); and its morphology of the lyophilized sample was observed by scanning electron microscope (SEM). FTIR results showed the characteristic bands of OH at 3445 cm−1, CH/CH2/symmetric CH3 (stretch) at 2923 and 2852 cm−1, C=O at 1730 cm−1, asymmetric CH3 (bend) at 1454 and 1414 cm−1, C−O of COO– at 1396 cm−1 and C–O–C at 1223, 1160, 1116, 1058 and 1019 cm−1 which were similar to the absorptive characteristics of P3HB. Methyl ester derivatives of GC/MS results identified 7 compounds including: 3-hydroxybutanoic (monomer of PHB), aminobenzoic, salicylic, hexadecenoic, octadecadienoic, octadecenoic and octadecanoic acids. SEM images revealed a fibriform and porous materials. Hence, the occurrence of PHAs was first described in a basidiomycetous mushroom A. odoratus. Thus, PHAs could be found not only in bacteria and but also in basidiomycetous mushroom, which can be promising target for bioplastics and green environmental studies. [ABSTRACT FROM AUTHOR]

Blend nanofibers composed of synthetic and natural polymers with carbon nanomaterial, have a great potential for bone tissue engineering. In this study, the electrospun nanocomposite scaffolds based on polyhydroxybutyrate(PHB)-Starch-multiwalled carbon nanotubes (MWCNTs) were fabricated with different concentrations of MWCNTs including 0.5, 0.75 and 1 wt%. The synthesized scaffolds were characterized in terms of morphology, porosity, thermal and mechanical properties, biodegradation, bioactivity, and cell behavior. The effect of the developed structures on MG63 cells was determined by real-time PCR quantification of collagen type I, osteocalcin, osteopontin and osteonectin genes. Our results showed that the scaffold containing 1 wt% MWCNTs presented the lowest fiber diameter (124 ± 44 nm) with a porosity percentage above 80 % and the highest tensile strength (24.37 ± 0.22 MPa). The addition of MWCNTs has a positive effect on surface roughness and hydrophilicity. The formation of calcium phosphate sediments on the surface of the scaffolds after immersion in SBF is observed by SEM and verified by EDS and XRD analysis.MG63 cells were well cultured on the scaffold containing MWCNTs and presented more cell viability, ALP secretion, calcium deposition and gene expression compared to the scaffolds without MWCNTs. The PHB-starch-1wt.%MWCNTs scaffold can be considerable for studies of supplemental bone tissue engineering applications. [ABSTRACT FROM AUTHOR]

The degradation pattern of bacterial poly-3-hydroxybutyrate (PHB) in dimethylformamide (DMF) and dioxane solutions at 100 °C assisted by ethylenediamine, 1,4-diaminobutane and monoaminoethanol was studied. When diamines were introduced into the PHB solution in DMF in the amount of 1 mol of the reagent to 5 or 10 mol of PHB monomers, a rapid decrease in the molecular weight of the polymer was observed. The initial value of the weight average molecular weight (Mw) 840 kDa had decreased by 20–30 times within the first 10–20 min of the experiment, followed by its gradual decrease to several thousand Da. When a similar molar quantity of aminoethanol was added, the molecular weight decreased slower. PHB had been degrading much slower in the dioxane solution than in DMF. By varying the number of reagents, it was possible to reach stabilization of the Mw at 1000–3000 Da when using diamines and 8000–20,000 Da using aminoethanol. 1H NMR analysis of the oligomers revealed of amino and amido groups forming in their structure. From the opposite end of the polymer chain, residues of 3-hydroxybutyric, crotonic and isocrotonic acids were formed during degradation. Differential scanning calorimetry indicated that after oligomerization there was a decrease in the melting point from 178 °C to 140–170 °C depending on the decrease in the molecular weight. The method proposed can be used for obtaining aminated PHB oligomers. [ABSTRACT FROM AUTHOR]

Volatile fatty acids (VFAs) derived from biomass are considered to be economical and environmentally friendly feedstocks for microbial fermentation. Converting VFAs to polyhydroxyalkanoate (PHA) could reduce the substrate cost and provide an economically viable route for the commercialization of PHA. The halophilic bacteria Salinivibrio spp. TGB4 and TGB19, newly isolated from salt fields, were found to accumulate poly-3-hydroxybutyrate (PHB) using acetate or butyrate as the substrate. Both strains exhibited considerable cell growth (OD 600 of ~8) even at acetate concentration of 100 g/L. In shake flask cultures, TGB4 produced PHB titers of 0.90 and 1.34 g/L, while TGB19 produced PHB titers of 0.25 and 2.53 g/L with acetate and butyrate, respectively. When acetate and butyrate were both applied, PHB production was significantly increased, and the PHB titer of TGB4 and TGB19 reached 6.14 and 6.84 g/L, respectively. After optimizing the culture medium, TGB19 produced 8.42 g/L PHB, corresponding to 88.55 wt% of cell dry weight. During fed-batch cultivation, TGB19 produced a PHB titer of 53.23 g/L. This is the highest reported PHB titer using acetate and butyrate by pure microbial cultures and would provide promising hosts for the industrial production of PHA from VFAs. • Salinivibrio spp. TGB4 and TGB19 were newly isolated to produce PHB. • Both strains utilized VFAs but not carbohydrates for PHB production. • The acetate tolerance level was as high as 100 g/L. • The highest PHB titer of 53.23 g/L using VFAs was achieved. [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.

The article examines the regularities of structure formation of ultrafine fibers based on poly-3-hydroxybutyrat under the influence of technological (electrical conductivity, viscosity), molecular (molecular weight), and external factors (low-molecular and nanodispersed substances of different chemical natures). Systems with polar substances are characterized by the presence of intermolecular interactions and the formation of a more perfect crystalline fiber structure. Changes in technological and molecular characteristics affect the fiber formation process, resulting in alterations in the morphology of the nonwoven fabric, fiber geometry, and supramolecular fiber structure. Polymer molecular weight, electrical conductivity, and solution viscosity influence fiber formation and fiber diameter. The fiber structure is heterogeneous, consisting of both crystalline and non-equilibrium amorphous phases. This article shows that with an increase in the molecular weight and concentration of the polymer, the diameter of the fiber increases. At the same time, the increase in the productivity of the electrospinning process does not affect the fiber geometry. The chemical structure of the solvent and the concentration of polar substances play a decisive role in the formation of fibers of even geometry. As the polarity of the solvent increases, the intermolecular interaction with the polar groups of poly-3-hydroxybutyrate increases. As a result of this interaction, the crystallites are improved, and the amorphous phase of the polymer is compacted. The action of polar molecules on the polymer is similar to the action of polar nanoparticles. They increase crystallinity via a nucleation mechanism. This is significant in the development of matrix-fibrillar systems for drug delivery, bioactive substances, antiseptics, tissue engineering constructs, tissue engineering scaffolds, artificial biodegradable implants, sorbents, and other applications.

Increasing greenhouse gas emissions and decreasing fossil fuel supplies necessitate the development of alternative methods for producing petroleum-based commodities. Plastics are also primarily petroleum-based goods with rising demand, thus there is growing interest in plastic substitutes. Polyhydroxyalkanoates (PHAs) are naturally produced biopolymers that are utilized by microorganisms as a source of energy and carbon storage. Poly-3-hydroxybutyrate (PHB) is a member of the PHA family and is considered the most promising candidate to replace polyethylene (PE). PHB is naturally produced by Cupriavidus necator, but recombinant production has also been recently established. This study is the first to investigate the heterologous production of PHB with recombinant Acetobacterium woodii using CO2 + H2 as a carbon and energy source. The introduction of a synthetic PHB production pathway resulted in the production of 1.23 g/L CDW and 1.9% PHB/cell dry weight (CDW), which corresponds to a production of 23.5 mg/L PHB. PHB quantification was simplified using LipidGreen2 fluorescence measurements.

An Azorhizobium caulinodans phaC mutant (OPS0865) unable to make poly-3-hydroxybutyrate (PHB), grows poorly on many carbon sources and cannot fix nitrogen in laboratory culture. However, when inoculated onto its host plant, Sesbania rostrata, the phaC mutant consistently fixed nitrogen. Upon reisolation from S. rostrata root nodules, a suppressor strain (OPS0921) was isolated that has significantly improved growth on a variety of carbon sources and also fixes nitrogen in laboratory culture. The suppressor retains the original mutation and is unable to synthesize PHB. Genome sequencing revealed a suppressor transition mutation, G to A (position 357,354), 13 bases upstream of the ATG start codon of phaR in its putative ribosome binding site (RBS). PhaR is the global regulator of PHB synthesis but also has other roles in regulation within the cell. In comparison with the wild type, translation from the phaR native RBS is increased approximately sixfold in the phaC mutant background, suggesting that the level of PhaR is controlled by PHB. Translation from the phaR mutated RBS (RBS*) of the suppressor mutant strain (OPS0921) is locked at a low basal rate and unaffected by the phaC mutation, suggesting that RBS* renders the level of PhaR insensitive to regulation by PHB. In the original phaC mutant (OPS0865), the lack of nitrogen fixation and poor growth on many carbon sources is likely to be due to increased levels of PhaR causing dysregulation of its complex regulon, because PHB formation, per se, is not required for effective nitrogen fixation in A. caulinodans.[Graphic: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Thermoplastic film materials with a structure of pseudo-interpenetrating polymer networks (pseudo-IPNs) were created on the basis of natural semi-crystalline poly-3-hydroxybutyrate (PHB) and natural rubber (NR), the content of the NR was varied from 2 to 40 wt.%. Using the methods of Fourier Transform Infra-Red (FTIR) spectroscopy, scanning electron microscopy (SEM), thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), structureproperties relationships were studied for PHB/NR thermoplastic pseudo-IPNs at different ratios of the components. Significant non-additive increase in mechanical characteristics compared to the individual PHB was found for PHB/NR pseudo-IPNs samples with a low content (up to 15 wt.%) of NR. For the sample with a content of 5 wt.% NR, the greatest increase (~18.4%) in strength at break was fixed (σb ≈ 22.5 MPa); for a sample with a content of 10 wt.% NR, the largest increase (~ 215%) in elongation at break (εb ≈ 48.8%) was observed. By FTIR spectroscopy, it was found that certain changes in the crystal structure of the PHB matrix occurred in the samples of pseudo-IPNs at increasing the NR content; however, no changes in the chemical structure of the components were detected. The SEM method revealed spherical domains of the dispersed elastomeric NR phase distributed in the PHB matrix in the samples of PHB/NR pseudo-IPNs. These observations evidence the microphase separation of the system components due to their thermodynamic incompatibility. It is worth to note that the sizes of the domains of the dispersed elastomeric phase naturally increased from ~10-20 μm for pseudo-IPNs PHB/NR = 85/15 wt.%, up to ~ 100 μm for PHB/NR = 60/40 wt.%. Using TGA method, it is established that PHB/NR pseudo-IPNs samples are characterized by high resistance to thermo-oxidative destruction, which non-additively increases with increasing the NR content. Destruction of PHB/NR pseudo-IPNs samples occur in two main stages: at I stage (Td ~ 240-285 °С) the main weight loss of the samples is occurred due to the destruction of macromolecules of the PHB matrix; at II stage (Td ~ 320-380 °С) the oxidative destruction of NR component takes place. Calculations have shown that with increasing the NR content, the maximum rate of thermo-oxidative destruction of pseudo-IPNs samples decreases compared to the individual PHB by 5-52 % with increasing the NR content from 2 to 40 wt.%. Therefore, it means that their resistance to thermo-oxidative destruction increases. The results obtained by DSC method have shown that the introduction of NR and changes of the components ratio significantly and nonadditively affect all the thermophysical characteristics of the samples studied. This indicates a significant restructuring of the microphase (amorphous and crystalline) structure of pseudo-IPNs synthesized due to the interpenetration of the components into the microphases of each other and the formation of mixed PHB/NR microphases with different ratios of the components. [ABSTRACT FROM AUTHOR]

With the threat of 'white pollution', the large‐scale production and application of biodegradable plastics have become crucial. Poly‐3‐hydroxybutyrate (PHB) is the simplest and most used member of the polyhydroxyalkanoate family, with a uniform C4 structural unit (C4H6O2). To reduce the impact of PHB on waste treatment, various recycling strategies have been developed for the tailored conversion of PHB to targeted chemicals and fuels. Given the special structural unit, PHB and its plastic forms have the potential to serve as a sustainable feedstock for producing a series of value‐added chemicals and fuels, including (i) crotonic acid via pyrolysis, (ii) (R)‐β‐hydroxybutyric acid via hydrolysis, (iii) propylene via depolymerization and decarboxylation, (iv) liquid fuels via deoxygenation and oligomerization, (v) n‐biobutanol via catalytic hydrogenolysis, (vi) methyl crotonate via thermolysis and esterification, (vii) (R)‐3‐hydroxybutyrate methyl ester via catalytic methanolysis, and (viii) β‐aminobutyric acid via ammonolysis. This review article provides a systematic review focusing on the reaction pathways, primary applications and market potential of the target products, and the pros and cons of current bio‐refineries in comparison with conventional petroleum refineries. The thermochemical processes developed for PHB degradation and polypropylene (a typical petroleum‐derived plastic to be replaced by PHB) are also comprehensively evaluated to reveal the future opportunities and challenges of PHB's broad‐scale production and utilization. PHB‐derived chemicals and fuels have a mature market already, and it is confirmed that the tailored valorization of PHB is a promising pathway to expand the market and gain more environmental and energy benefits. © 2022 Society of Chemical Industry and John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The theoretical prediction of the mechanical behavior of fibrous materials for medicine in different environments is of great scientific interest. The deformation mechanisms under loading of biopolymer electrospun fibrous materials differ significantly in dependence on the phase state of the medium, namely, liquid state, gaseous state, or at their interface. In this paper, the influence of various mediums on the mechanical behavior of fibrous materials based on poly‐3‐hydroxybutyrate under tension is examined. The oxidative stress (ozone treatment) and the incorporation of low concentrations of modifying additives (complexes of tetraphenyl‐porphyrin with Zn) are proposed as the most effective methods of directed control of stress‐related characteristics without a negative effect on the functional properties of medical materials. [ABSTRACT FROM AUTHOR]

Halophilic Halomonas bluephagenesis, a natural producer of poly-3-hydroxybutyrate (PHB), was metabolically engineered to synthesize ethylene glycol and glycolate from xylose. Xylose utilization was achieved by overexpressing either the xylonate pathway or the ribulose-1-phosphate pathway. The key genes encoding for xylonate dehydratase and 2-keto-3-deoxy-xylonate aldolase in the xylonate pathway were screened. With further overexpressing aldehyde reductase gene yjgB, ethylene glycol accumulation was improved to 0.91 g/L, accompanied with 1.48 g/L of PHB accumulation. The disruption of native glycolate oxidase was found to be essential for glycolate production, and the defective recombinant strain produced 0.80 g/L glycolate with 1.14 g/L PHB in shake flask cultures. These results indicated that H. bluephagenesis has the potential to produce diverse metabolic chemicals from xylose., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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.

Waste paper holds great potential as a substrate for the microbial production of bioplastic (Poly-3-hydroxybutyrate (PHB)). This study aimed to produce PHB by utilizing office paper as a substrate using Cupriavidus necator through batch and fed-batch simultaneous saccharification and fermentation (SSF) approach. For the batch experiment, different loadings of shredded office paper (3, 5 and 10%) with two different pretreatments H2O2 (OPH) and H2O2 and Triton X-100 (OPTH) were carried out. For the fed-batch experiment, paper loading started with 3% and two more additions were made at 36 and 84 h. Both experiments were conducted at 30 °C, 200 rpm and pH 7 using 55.5 FPU/g of cellulase and 37.5 CBU/g of β-glucosidase with a fixed amount of nitrogen source. High PHB yield was observed with OPH in all loadings, though the OPHT showed a better hydrolysis. Maximum PHB yield (4.27 g/L) was achieved with 10% OP on the sixth day of fermentation in batch SSF. Whereas, maximum PHB yield (4.19 g/L) was obtained within a shorter time (66 h)with OPH in the fed-batch experiment. The extracted PHB showed well-matched characteristic features to the standard PHB. Finally, this study proves the feasibility of employing the SSF process for PHB production using waste paper as an alternative approach to overcome the shortcoming of the separate hydrolysis and fermentation (SHF) process. [ABSTRACT FROM AUTHOR]

Dental implant success is threatened by peri-implantitis, an inflammation leading to implant failure. Conventional treatments struggle with the intricate microbial and host factors involved. Antibacterial membranes, acting as barriers and delivering antimicrobials, may offer a promising solution. Thus, this study highlights the potential of developing antibacterial membranes of poly-3-hydroxybutyrate and silver nanoparticles (Ag Nps) to address peri-implantitis challenges, discussing design and efficacy against potential pathogens. Electrospun membranes composed of PHB microfibers and Ag Nps were synthesized in a blend of DMF/chloroform at three different concentrations. Various studies were conducted on the characterization and antimicrobial activity of the membranes. The synthesized Ag Nps ranged from 4 to 8 nm in size. Furthermore, Young's modulus decreased, reducing from 13.308 MPa in PHB membranes without Ag Nps to 0.983 MPa in PHB membranes containing higher concentrations of Ag Nps. This demonstrates that adding Ag Nps results in a less stiff membrane. An increase in elongation at break was noted with the rise in Ag Nps concentration, from 23.597 % in PHB membranes to 60.136 % in PHB membranes loaded with Ag Nps. The antibiotic and antibiofilm activity of the membranes were evaluated against Pseudomonas aeruginosa , Staphylococcus aureus , Streptococcus mutans , and Candida albicans. The results indicated that all PHB membranes containing Ag Nps exhibited potent antibacterial activity by inhibiting the growth of biofilms and planktonic bacteria. However, inhibition of C. albicans occurred only with the PHB-Ag Nps C membrane. These findings emphasize the versatility and potential of Ag Nps-incorporated membranes as a multifunctional approach for preventing and addressing microbial infections associated with peri-implantitis. The combination of antibacterial and antibiofilm properties in these membranes holds promise for improving the management and treatment of peri-implantitis-related complications. • The study involved synthesizing silver nanoparticles (Ag Nps) into the Poly-3-hydroxybutyrate (PHB) solution. • PHB solution containing silver nanoparticles was electrospun to develop scaffolds for tissue regeneration. • The antimicrobial activity of the PHB/Ag Nps scaffolds was evaluated over four different microorganisms. • The scaffolds loaded with Ag Nps showed a strong antibacterial activity over the different strains evaluated • Notably, inhibiting the growth of C. albicans required higher concentrations of Ag Nps loaded in scaffolds than the bacteria. [ABSTRACT FROM AUTHOR]

Due to the current environmental situation, biopolymers are replacing the usual synthetic polymers, and special attention is being paid to poly-3-hydroxybutyrate (PHB), which is a biodegradable polymer of natural origin. In this paper, the rate of biodegradation of films and fibers based on PHB was compared. The influence of exposure to soil on the structure and properties of materials was evaluated using methods of mechanical analysis, the DSC method and FTIR spectroscopy. The results showed rapid decomposition of the fibrous material and also showed how the surface of the material affects the rate of biodegradation and the mechanical properties of the material. It was found that maximum strength decreased by 91% in the fibrous material and by 49% in the film. Additionally, the DSC method showed that the crystallinity of the fiber after exposure to the soil decreased. It was established that the rate of degradation is influenced by different factors, including the surface area of the material and its susceptibility to soil microorganisms. The results obtained are of great importance for planning the structure of features in the manufacture of biopolymer consumer products in areas such as medicine, packaging, filters, protective layers and coatings, etc. Therefore, an understanding of the biodegradation mechanisms of PHB could lead to the development of effective medical devices, packaging materials and different objects with a short working lifespan.

Electrospun fabrics have unique properties due to their uniform morphology and high surface area to volume ratio. Ultrathin nonwoven fabrics are produced for many applications: biomedical, nanosensors, tissue engineering and filtration systems. In this work, nonwoven polylactide, polylactide/natural rubber, poly-3-hydroxybutyrate, and poly-3-hydroxybutyrate/nitrile butadiene rubber fabrics were prepared by electrospinning methods. The obtained fabric samples were used as substrates for the growth of winter wheat seeds “Yubileinaya 100” (Triticum aestivum L.). The stimulating effect of polymer substrates on seed germination and plant growth was shown. The structure and properties of nonwoven agromaterials were controlled by differential scanning calorimetry, IR-spectroscopy, and optical microscopy. The mechanical properties of the obtained fabrics before and after their utilization as substrates were studied. After the wheat growing experiment, the degree of crystallinity of PHB and PHB/NBR samples decreased by 12% and they completely lost their mechanical properties. It is shown that the main factors providing the efficiency of seed growth technology on polymer substrates are the chemical nature and structure of the biodegradable matrix.

The influence of elastomers—natural rubber (NR) and synthetic ethylene-propylene rubber (EPR)—on the morphology and physical and mechanical characteristics of composite materials based on polylactide (PLA) and poly-3-hydroxybutyrate (PHB), respectively, is studied. Differential scanning calorimetry shows differences in the patterns of changes in the thermophysical characteristics of the PLA—NR (with a rubber content of 5 to 15 wt %)—and PHB—synthetic EPR (10–30 wt %)—compositions. The melting point and the degree of crystallinity increase in the case of PLA and decrease in the experiment with PHB. The mechanical characteristics of the investigated compositions have similar dependences: with an increase in the content of rubbers, the relative elongation increases and the tensile strength and elastic modulus decrease. [ABSTRACT FROM AUTHOR]

Abstract Waste plastic and methane emissions are two anthropogenic by-products exacerbating environmental pollution. Methane-oxidizing bacteria (methanotrophs) hold the key to solving these problems simultaneously by utilising otherwise wasted methane gas as carbon source and accumulating the carbon as poly-3-hydroxybutyrate, a biodegradable plastic polymer. Here we present the isolation and characterisation of two novel Methylocystis strains with the ability to produce up to 55.7 ± 1.9% poly-3-hydroxybutyrate of cell dry weight when grown on methane from different waste sources such as landfill and anaerobic digester gas. Methylocystis rosea BRCS1 isolated from a recreational lake and Methylocystis parvus BRCS2 isolated from a bog were whole genome sequenced using PacBio and Illumina genome sequencing technologies. In addition to potassium nitrate, these strains were also shown to grow on ammonium chloride, glutamine and ornithine as nitrogen source. Growth of Methylocystis parvus BRCS2 on Nitrate Mineral Salt (NMS) media with 0.1% methanol vapor as carbon source was demonstrated. The genetic tractability by conjugation was also determined with conjugation efficiencies up to 2.8 × 10–2 and 1.8 × 10–2 for Methylocystis rosea BRCS1 and Methylocystis parvus BRCS2 respectively using a plasmid with ColE1 origin of replication. Finally, we show that Methylocystis species can produce considerable amounts of poly-3-hydroxybutyrate on waste methane sources without impaired growth, a proof of concept which opens doors to their use in integrated bio-facilities like landfills and anaerobic digesters.

Designing laboratory-scale degradation experiments for polymer-based biomaterials is crucial for the development of safe and functional implants, in particular regarding high molecular weight polyesters. Within this work, we compared accelerated degradation of solvent cast HMW-PLLA and HMW-P3HB films at 55 °C (16 weeks) with enzyme-associated (proteinase K and lipases) real-time degradation at 37 °C (108 weeks). During real-time degradation, PLLA showed mass loss up to 83%, in contrast to accelerated conditions, where no changes occurred. Moreover, we observed wave-shape development of crystallinity for PLLA and PHB for both degradation conditions applied, whereby PLLA χ value nearly doubled to up to 75%. These results were used to develop a correlation model based on molecular weight decrease and were furthermore discussed in light of a detailed literature review. In summary, real-time in vitro studies could be adapted to accelerated protocols providing the same limiting conditions such as molecular weight and initial crystallinity are given.

The aim of this study was to investigate the biocompatibility and osteoinductive properties of fibrous scaffolds implanted in the bone tissue of white rats in vivo. The effect of various concentrations of conductive PANi (2 and 3 wt%) and rGO (0.2 and 1 wt%) in the PHB scaffolds on the reparative processes of bone callus formation in white rats was studied. On the 28th day after implantation, a histomorphological study of the preparations of the transverse section of the diaphysis of the femur of the implantation area was performed. The results of implantation tests in the femur revealed that all the studied scaffolds are biocompatible and the most pronounced stimulating effect on bone formation was observed for hybrid PHB scaffolds doped with 3 wt% PANi and 0.2 wt% rGO compared with pure PHB scaffolds, thus determining further prospects to study the osteoinductive features of hybrid PHB scaffolds.

Over the past decade, formic acid and acetic acid have gained increasing attention as alternative feedstocks for poly-3-hydroxybutyrate (PHB) production as these potentially CO 2 -derived molecules are naturally assimilated by Cupriavidus necator. Both organic acids were individually evaluated in fed-batch fermentations at bioreactor scale. Acetic acid was revealed as the most promising carbon source yielding 42.3 g L−1 PHB, whereas no significant amount of PHB was produced from formic acid. Hence, acetic acid was further used as the substrate during process intensification. Key performance characteristics, including process stability, PHB titer, and productivity were optimized by introducing NH 4 -acetate as the nitrogen source, extending the growth phase, and implementing a repeated fed-batch procedure, respectively. These advanced fermentation strategies resulted in the establishment of a stable fermentation process reaching 58.5 g L−1 PHB, while doubling the productivity to 0.93 g L−1 h−1 PHB. [Display omitted] • Acetic acid was revealed as most promising CO 2 -derived feedstock over formic acid. • Ammonium acetate as nitrogen source enables stable pH-static feeding of acetic acid. • Extension of the growth phase by linear feeding increases final PHB concentration. • Repeated fed-batch strategy enhances PHB productivity. • 58.5 g L−1 PHB was produced with a productivity of 0.93 g L−1 h−1 PHB. [ABSTRACT FROM AUTHOR]

In this work we investigated tenogenic differentiation of adipose-derived mesenchymal stem cells (AdMSCs), which were seeded onto silk fibroin/poly-3-hydroxybutyrate (SF/P3HB) scaffolds with aligned topography, and high mechanical strength. The electrospinning process was optimized by using the response surface method (RSM) and SF/P3HB nanofibrous matrices with a total polymer concentration of 5% (SF: PHB = 3: 1), flow rate 1 mL/h, collector rotation speed 2000 rpm, applied voltage 14 kV, and collector distance 25 cm were obtained. The average fiber diameter was 699 ± 203 nm and 80% of the nanofibers were aligned within the ±15o range. SF reinforcement reduced the crystallinity of P3HB, and the elastic modulus was found to be 197.0 ± 7.7 MPa. The scaffolds showed bacteriostatic effect. A 21-day of cell culture study was performed with rat rAdMSCs in the absence and presence of tenogenic differentiation factor-5 (GDF-5). The results demonstrated that SF/P3HB scaffolds allow the cells to proliferate and differentiate to the tenocytes. However, no significant effect of GDF-5 on the differentiation of cells was observed. These findings indicated that our aligned SF/P3HB scaffolds have a significant potential to be used for tendon tissue engineering. [ABSTRACT FROM AUTHOR]

Poly-3-hydroxybutyrate (PHB) is an environmentally friendly polymer and can be produced in Escherichia coli cells after overexpression of the heterologous gene cluster phaCAB. The biosynthesis of the outer membrane (OM) consumes many nutrients and influences cell morphology. Here, we engineered the OM by disrupting all gene clusters relevant to the polysaccharide portion of lipopolysaccharide (LPS), colanic acid (CA), flagella, and/or fimbria in E. coli W3110. All these disruptions benefited PHB production. Especially, disrupting all these OM components increased the PHB content to 83.0 wt% (PHB content percentage of dry cell weight), while the wild-type control produced only 1.5 wt% PHB. The increase was mainly due to the LPS truncation to Kdo2 (3-deoxy-D-manno-octulosonic acid)-lipid A, which resulted in 82.0 wt% PHB with a 25-fold larger cell volume, and disrupting CA resulted in 57.8 wt% PHB. In addition, disrupting LPS facilitated advantageous fermentation features, including 69.1% less acetate, a 550% higher percentage of autoaggregated cells among the total culture cells, 69.1% less biofilm, and a higher broken cell ratio. Further detailed mechanism investigations showed that disrupting LPS caused global changes in envelope and cellular metabolism: (i) a sharp decrease in flagella, fimbria, and secretions; (ii) more elastic cells; (iii) much greater carbon flux toward acetyl coenzyme A (acetyl-CoA) and supply of cofactors, including NADP, NAD, and ATP; and (iv) a decrease in by-product acids but increase in g-aminobutyric acid by activating s E factor. Disrupting CA, flagella, and fimbria also improved the levels of acetyl-CoA and cofactors. The results indicate that engineering the OM is an effective strategy to enhance PHB production and highlight the applicability of OM engineering to increase microbial cell factory performance. [ABSTRACT FROM AUTHOR]

Efficient design for bone tissue engineering requires an understanding of the appropriate selection of biomimetic natural or synthetic materials and scalable fabrication technologies. In this research, poly (3-hydroxybutyrate) (PHB) and starch (5-15 wt%) as biological macromolecules were used to fabricate novel biomimetic scaffolds by electrospinning method. SEM results of electrospun scaffolds revealed bead-free nanofibers and three-dimensional homogenous structures with highly interconnected pores. Results of FTIR and Raman demonstrated that there were hydrogen bonds between the two polymers. The tensile strength of scaffolds was significantly improved by adding starch up to 10 wt%, from 3.05 to 15.54 MPa. In vitro degradation and hydrophilicity of the scaffolds were improved with the presence of starch. The viability and proliferation of MG-63 cells and alkaline phosphatase (ALP) activity were remarkably increased in the PHB-starch scaffolds compared to the PHB and control samples. The mineralization and calcium deposition of MG-63 cells were confirmed by alizarin red staining. It is concluded that PHB/starch electrospun scaffold could be a good candidate for bone tissue engineering applications. • PHB-starch scaffolds were made using electrospinning method. • The hydrophilicity and biodegradation of the scaffolds were increased by adding starch. • Tensile strength of the scaffolds was increased in the presence of 10 wt% starch. • Cell viability was significantly increased by adding starch to the scaffolds. • PHB-starch electrospun scaffolds could create a favorable microenvironment for bone tissue engineering applications. [ABSTRACT FROM AUTHOR]

The effect of the hemin (Hmi) on the structure and properties of nanocomposite electrospun materials based on poly-3-hydroxybutyrate (PHB) is discussed in the article. The additive significantly affected the morphology of fibers allowed to produce more elastic material and provided high antimicrobial activity. The article considers also the impact of the hemin on the biocompatibility of the nonwoven material based on PHB and the prospects for wound healing.

The degradation pattern of bacterial poly-3-hydroxybutyrate (PHB) in dimethylformamide (DMF) and dioxane solutions at 100 °C assisted by ethylenediamine, 1,4-diaminobutane and monoaminoethanol was studied. When diamines were introduced into the PHB solution in DMF in the amount of 1 mol of the reagent to 5 or 10 mol of PHB monomers, a rapid decrease in the molecular weight of the polymer was observed. The initial value of the weight average molecular weight (Mw) 840 kDa had decreased by 20–30 times within the first 10–20 min of the experiment, followed by its gradual decrease to several thousand Da. When a similar molar quantity of aminoethanol was added, the molecular weight decreased slower. PHB had been degrading much slower in the dioxane solution than in DMF. By varying the number of reagents, it was possible to reach stabilization of the Mw at 1000–3000 Da when using diamines and 8000–20,000 Da using aminoethanol. 1H NMR analysis of the oligomers revealed of amino and amido groups forming in their structure. From the opposite end of the polymer chain, residues of 3-hydroxybutyric, crotonic and isocrotonic acids were formed during degradation. Differential scanning calorimetry indicated that after oligomerization there was a decrease in the melting point from 178 °C to 140–170 °C depending on the decrease in the molecular weight. The method proposed can be used for obtaining aminated PHB oligomers.