Search Results (32)

Polyhydroxybutyrate (PHB) is a biopolymer produced by bacteria. This study aimed to implement the production process of experimental medical-grade PHB and to evaluate its morphology and biocompatibility compared to conventional resin-based composites (RBCs). PHB raw material was produced via biological process and then the membrane was generated via electrospinning specifically for this study and imaged with Micro-Computed Tomography (Micro-CT) and scanning electron microscopy (SEM). MTS assay was used to assess the cytotoxicity of PHB compared to other materials. Test groups included two packable resin composites (Point 4-Kerr, G-aenial anterior-GC), two flowable resin composites (Filtek Ultimate Flowable-3M ESPE, Nova Compo HF-Imicryl), a compomer (Nova Compomer-Imicryl), a fissure-sealant (Fissured Nova Plus-Imicryl), and the PHB membrane (Innovaplast Biotechnology Inc., Eskisehir, Turkey). A control group consisting of cells without any test material was also produced. To perform the MTS assay, disc-shaped specimens of the aforementioned materials were prepared and then incubated with mouse fibroblast cells (L929) for 24 and 48 h. Micro-CT and SEM images revealed a homogeneous and fibrillary structure of the PHB. MTS assay revealed the highest cell viability in the PHB, Nova Compomer, and Fissured Nova Plus groups after 24 h. PHB and Nova Compomer showed the highest viability rates at 48 h while other RBCs had rates below 25% (p < 0.05). Considering the cell viability data and its fibrillary structure, from a biological point of view, PHB seems a promising biopolymer that might have applications in the field of dental restorative materials. Full article

In this research work, a main biopolymer group of polyhydroxyalkanoates (PHAs) in the form of polyhydroxybutyrate (PHB) was synthesised by a pure bacterial strain of Azotobacter vinelandii via repeated fed-batch fermentation. An agricultural crop, sugar cane, was used as the sole carbon source. Firstly, batch fermentation was investigated considering variations in incubation times (24 h, 48 h, and 96 h). The highest dry cell weight (DCW) and PHAs of 5.15 ± 0.04 g/L and 4.00 ± 0.04 g/L were obtained after 48 h of incubation time. The optimum time obtained was further varied to investigate the effect of the sugar concentrations in the medium. It was found that bacteria could grow very well and produced the highest DCW and PHAs (11.17 ± 0.15 g/L and 8.77 ± 0.06 g/L) when the culture medium with a 100 g/L sugar concentration was added. Later, repeated fed-batch fermentation was carried out to improve productivity. The results obtained revealed that PHA production was increased in the next cycle of the process. Furthermore, the final productivity (0.104 g/L·h) was increased 1.65-fold compared to the first cycle (0.063 g/L·h). Moreover, the culture strategy showed remarkable results, with reductions in both fermentation time and preparation cost. Full article

Poly-β-hydroxybutyrate (PHB) is a bacterial metabolite produced by bacteria such as Halomonas sp. that serves as a carbon and energy storage compound for bacteria under nutrient-limited conditions. Two experiments were conducted to investigate the effects of dietary supplementation with Halomonas-PHB on hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatu ♂). In experiment I, juvenile groupers were fed basal diets supplemented with 3% Halomonas-PHB (3% HM-PHB) containing 1.4% PHB and 3% Halomonas (3% HM) without PHB, as well as a control diet, for seven weeks. The results showed no significant difference in survival rate, weight gain, and crude fat content between the 3% HM-PHB group and the control group; however, the crude protein of the 3% HM-PHB group was significantly lower than that of the control group. Furthermore, supplementation with 3% HM-PHB increased the fatty acids content in fish muscles, including long-chain unsaturated fatty acids C18:1n9, EPA, and DHA. In experiment II, groupers were fed a basal diet supplemented with 6.5% Halomonas-PHB (6.5% HM-PHB) containing 3% PHB and 6.5% Halomonas (6.5% HM) containing no PHB, as well as a basal diet (Control). After seven weeks of rearing, the fish were challenged with Vibrio anguillarum for 48 h. Although no significant difference in survival rate and growth was observed among different groups, the dietary supplement of 6.5% Halomonas-PHB improved the survival rate of V. anguillarum challenged grouper and significantly increased the gene expressions of catalase (CAT) and superoxide dismutase (SOD) in blood, interleukin 1 (IL1) and interleukin 10 (IL10) in the liver, spleen, head kidney, and blood (p < 0.05). In conclusion, dietary supplementation of Halomonas-PHB had no significantly positive effect on fish growth performance but increased the content of fatty acids, including long-chain unsaturated fatty acids C18:1n9, EPA, and DHA in fish muscle; it also improved the V. anguillarum resistance, possibly through increasing immune-related gene expression in different tissues and organs. Our findings offer compelling evidence that Halomonas-PHB can be utilized as a feed additive in intensive grouper farming to enhance the groupers’ resistance to Vibrio. Full article

The aim of the presented work was to functionalize a blend based on polyhydroxyalkanoate (PHA): poly(hydroxybutyrate (PHB) with poly(lactic acid) (PLA) and a mixture of three selected herb extracts, namely, Hypericum L., Urtica L. and Chelidonium L., (E), zinc oxide (ZnO) and a combined system (EZnO), produced via extrusion. Before processing with bioresin, the natural modifiers were characterized using thermal analysis, FTIR and antimicrobial tests. The results revealed interactions between the extracts and the filler, leading to higher thermal stability in EZnO than when using E alone. Moreover, the mixture of extracts exhibited antimicrobial properties toward both Gram-negative (S. aureus) as well as Gram-positive bacteria (E. coli). Modified regranulates were transformed into films by cast extrusion. The influence of the additives on thermal (DSC, TGA and OIT), mechanical, barrier (WVTR and OTR), morphological (FTIR) and optical properties was investigated. The EZnO additive had the highest impact on the mechanical, barrier (OTR and WVTR) and optical properties of the bioresin. The microbial test results revealed that PHA-EZnO exhibited higher activity than PHA-ZnO and PHA-E and also reduced the number of S. aureus, E. coli and C. albicans cells. The findings confirmed the synergistic effect between the additive components. Modified polyester films did not eliminate the phi6 bacteriophage particles completely, but they did decrease their number, confirming moderate antiviral effectiveness. Full article

Environmental pollution caused by petroleum-derived plastics continues to increase annually. Consequently, current research is interested in the search for eco-friendly bacterial polymers. The importance of Bacillus bacteria as producers of polyhydroxyalkanoates (PHAs) has been recognized because of their physiological and genetic qualities. In this study, twenty strains of Bacillus genus PHA producers were isolated. Production was initially evaluated qualitatively to screen the strains, and subsequently, the strain B12 or Bacillus sp. 12GS, with the highest production, was selected through liquid fermentation. Biochemical and molecular identification revealed it as a novel isolate of Bacillus cereus. Production optimization was carried out using the Taguchi methodology, determining the optimal parameters as 30 °C, pH 8, 150 rpm, and 4% inoculum, resulting in 87% and 1.91 g/L of polyhydroxybutyrate (PHB). Kinetic studies demonstrated a higher production within 48 h. The produced biopolymer was analyzed using Fourier-transform infrared spectroscopy (FTIR), confirming the production of short-chain-length (scl) polyhydroxyalkanoate, named PHB, and differential scanning calorimetry (DSC) analysis revealed thermal properties, making it a promising material for various applications. The novel B. cereus isolate exhibited a high %PHB, emphasizing the importance of bioprospecting, study, and characterization for strains with biotechnological potential. Full article

In recent years, polyhydroxyalkanoates (PHAs) have gained notoriety because of their desirable properties that include proven biodegradability, biocompatibility, and thermal stability, which make them suitable alternatives to fossil-based polymers. However, the widespread use of PHAs is still challenging because of their production costs, which are greatly associated with the cultivation medium used for bacterial cultivation. In Portugal, one-quarter of the forest area is covered by Eucalyptus globulus wood, making its residues a cheap, abundant, and sustainable potential carbon source for biotechnological uses. In this work, eucalyptus bark was used as the sole feedstock for PHA production in a circular bioeconomic approach. Eucalyptus bark hydrolysate was obtained after enzymatic saccharification using Cellic^® CTec3, resulting in a sugar-rich solution containing glucose and xylose. Although with differing performances, several bacteria were able to grow and produce PHA with distinct compositions, using the enzymatic hydrolysate as the sole carbon source. Pseudomonas citronellolis NRRL B-2504 achieved a high cellular growth rate in bioreactor assays (24.4 ± 0.15 g/L) but presented a low accumulation of a medium-chain-length PHA (mcl-PHA) comprising the monomers hydroxydecanoate (HD, 65%), hydroxydodecanoate (HDd, 25%), and hydroxytetradecanoate (HTd, 14%). Burkholderia thailandensis E264, on the other hand, reached a lower cellular growth rate (8.87 ± 0.34 g/L) but showed a higher biopolymer accumulation, with a polyhydroxybutyrate (PHB) content in the cells of 12.3 wt.%. The new isolate, Pseudomonas sp., revealed that under nitrogen availability, it was able to reach a higher accumulation of the homopolymer PHB (31 wt.%). These results, although preliminary, demonstrate the suitability of eucalyptus bark’s enzymatic hydrolysate as a feedstock for PHA production, thus offering an exciting avenue for achieving sustainable and environmentally responsible plastic products from an undervalued forestry waste. Full article

This study evaluated the feasibility of contextually producing hydrogen, microbial proteins, and polyhydroxybutyrate (PHB) using a mixed culture of purple phototrophic bacteria biomass under photo fermentative conditions. To this end, three consecutive batch tests were conducted to analyze the biomass growth curve and to explore the potential for optimizing the production process. Experimental findings indicated that inoculating reactors with microorganisms from the exponential growth phase reduced the duration of the process. Furthermore, the most effective approach for simultaneous hydrogen production and the valorization of microbial biomass was found when conducting the process during the exponential growth phase of the biomass. At this stage, achieved after 3 days of fermentation, the productivities of hydrogen, PHB, and microbial proteins were measured at 63.63 L/m³ d, 0.049 kg/m³ d, and 0.045 kg/m³ d, respectively. The biomass composition comprised a total intracellular compound percentage of 56%, with 27% representing PHB and 29% representing proteins. Under these conditions, the estimated daily revenue was maximized, amounting to 0.6 $/m³ d. Full article

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. Full article

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. Full article

Microalgae can treat waste streams containing elevated levels of organic carbon and nitrogen. This process can be economically attractive if high value products are created simultaneously from the relatively low-cost waste stream. Co-production of two high value microalgal products, phycocyanin and polyhydroxybutyrate (PHB), was investigated using non-axenic Arthrospira platensis MUR126 and supplemental organic carbon (acetate, oxalate, glycerol and combinations). All supplemented cultures had higher biomass yield (g/L) than photoautotrophic control. All cultures produced PHB (3.6–7.8% w/w), except the control and those fed oxalate. Supplemented cultures showed a two to three-fold increase in phycocyanin content over the eight-day cultivation. Results indicate co-production of phycocyanin and PHB is possible in A. platensis, using mixed-waste organic carbon. However, supplementation resulted in growth of extremophile bacteria, particularly in cultures fed glycerol, and this had a negative impact on culture health. Refinement of the carbon dosing rate is required to minimise impacts of native bacterial contamination. Full article

As a kind of ubiquitous emerging pollutant, microplastics (MPs) are persistent in the environment and have a large impact on the ecosystem. Fortunately, some microorganisms in the natural environment can degrade these persistent MPs without creating secondary pollution. In this study, 11 different MPs were selected as carbon sources to screen the microorganisms for degradable MPs and explore the possible mechanism of degradation. After repeated domestication, a relatively stable microbial community was obtained after approximately 30 days later. At this time, the biomass of the medium ranged from 88 to 699 mg/L. The growth of bacteria with different MPs ranged from 0.030 to 0.090 optical density (OD) 600 of the first generation to 0.009–0.081 OD 600 of the third generation. The weight loss method was used to determine the biodegradation ratios of different MPs. The mass losses of polyhydroxybutyrate (PHB), polyethylene (PE), and polyhydroxyalkanoate (PHA) were relatively large, at 13.4%, 13.0%, and 12.7%, respectively; these figures for polyvinyl chloride (PVC) and polystyrene (PS) were relatively slight, 8.90% and 9.10%, respectively. The degradation half-life (t_1/2) of 11 kinds of MPs ranges from 67 to 116 days. Among the mixed strains, Pseudomonas sp., Pandoraea sp., and Dyella sp. grew well. The possible degradation mechanism is that such microbial aggregates can adhere to the surface of MPs and form complex biofilms, secrete extracellular and intracellular enzymes, etc., break the hydrolyzable chemical bonds or ends of molecular chains by attacking the plastic molecular chains, and produce monomers, dimers, and other oligomers, leading to the reduction of the molecular weight of the plastic itself. Full article

The aim of this work was to study the valorization of argan seed pulp, a waste material obtained from argan oil extraction, for the biosynthesis of polyhydroxybutyrate (PHB). A new species that showed the metabolic capacity for the conversion of argan waste into the bio-based polymer was isolated from an argan crop located in Teroudant, a southwestern region of Morocco, where the arid soil is exploited for goat grazing. The PHB accumulation efficiency of this new species was compared to the previously identified species 1B belonging to the genus Sphingomonas, and results were reported as dry cell weight residual biomass and PHB final yield measured. Temperature, incubation time, pH, NaCl concentration, nitrogen sources, residue concentrations, and culture medium volumes were analyzed with the aim of obtaining a maximum accumulation of PHB. UV-visible spectrophotometry and FTIR analysis confirmed that PHB was present in the material extracted from the bacterial culture. The results of this wide investigation indicated that the new isolated species 2D1 had a higher efficiency in PHB production compared to the previously identified strain 1B, which was isolated from a contaminated argan soil in Teroudant. PHB final yield of the two bacterial species, i.e., the new isolated and 1B, cultivated under optimal culture conditions, in 500 mL MSM enriched with 3% argan waste, were 21.40% (5.91 ± 0.16 g/L) and 8.16% (1.92 ± 0.23 g/L), respectively. For the new isolated strain, the result of the UV-visible spectrum indicates the absorbance at 248 nm, while the FTIR spectrum showed peaks at 1726 cm⁻¹ and 1270 cm⁻¹: these characteristic peaks indicated the presence of PHB in the extract. The data from the species 1B UV-visible and FTIR spectra were previously reported and were used in this study for a correlation analysis. Furthermore, additional peaks, uncharacteristic of standard PHB, suggest the presence of impurities (e.g., cell debris, solvent residues, biomass residues) that persisted after extraction. Therefore, a further enhancement of the sample purification during extraction is recommended for more accuracy in the chemical characterization. If 470,000 tons of argan fruit waste can be produced annually, and 3% of waste is consumed in 500 mL culture by 2D1 to produce 5.91 g/L (21.40%) of the bio-based polymer PHB, it can be estimated that the amount of putative PHB that can be extracted annually from the total argan fruit waste is about 2300 tons. Full article

In recent years, there has been a growing interest in bio-based degradable plastics as an alternative to synthetic plastic. Polyhyroxybutyrate (PHB) is a macromolecule produced by bacteria as a part of their metabolism. Bacteria accumulate them as reserve materials when growing under different stress conditions. PHBs can be selected as alternatives for the production of biodegradable plastics because of their fast degradation properties when exposed to natural environmental conditions. Hence, the present study was undertaken in order to isolate the potential PHB-producing bacteria isolated from the municipal solid waste landfill site soil samples collected from the Ha’il region of Saudi Arabia to assess the production of PHB using agro-residues as a carbon source and to evaluate the growth of PHB production. In order to screen the isolates for producing PHB, a dye-based procedure was initially employed. Based on the 16S rRNA analysis of the isolates, Bacillus flexus (B. flexus) accumulated the highest amount of PHB of all the isolates. By using a UV–Vis spectrophotometer and Fourier-transform infrared spectrophotometer (FT-IR), in which a sharp absorption band at 1721.93 cm⁻¹ (C=O stretching of ester), 1273.23 cm⁻¹ (–CH group), multiple bands between 1000 and 1300 cm⁻¹ (stretching of the C–O bond), 2939.53 cm⁻¹ (–CH₃ stretching), 2880.39 cm⁻¹ (–CH₂ stretching) and 3510.02 cm⁻¹ (terminal –OH group), the extracted polymer was characterized and confirmed its structure as PHB. The highest PHB production by B. flexus was obtained after 48 h of incubation (3.9 g/L) at pH 7.0 (3.7 g/L), 35 °C (3.5 g/L) with glucose (4.1 g/L) and peptone (3.4 g/L) as carbon and nitrogen sources, respectively. As a result of the use of various cheap agricultural wastes, such as rice bran, barley bran, wheat bran, orange peel and banana peel as carbon sources, the strain was found to be capable of accumulating PHB. Using response surface methodology (RSM) for optimization of PHB synthesis using a Box–Behnken design (BBD) proved to be highly effective in increasing the polymer yield of the synthesis. With the optimum conditions obtained from RSM, PHB content can be increased by approximately 1.3-fold when compared to an unoptimized medium, resulting in a significant reduction in production costs. Thus, isolate B. flexus is a highly promising candidate for the production of industrial-size quantities of PHB from agricultural wastes and is capable of removing the environmental concerns associated with synthetic plastics from the industrial production process. Moreover, the successful production of bioplastics using a microbial culture provides a promising avenue for the large-scale production of biodegradable and renewable plastics with potential applications in various industries, including packaging, agriculture and medicine. Full article

Polyhydroxybutyrate (PHB) is a biodegradable bioplastic with potential applications as an alternative to petroleum-based plastics. However, efficient PHB production remains difficult. The main cost of PHB production is attributed to carbon sources; hence, finding inexpensive sources is important. Galactose is a possible substrate for polyhydroxyalkanoate production as it is abundant in marine environments. Marine bacteria that produce PHB from galactose could be an effective resource that can be used for efficient PHB production. In this study, to identify a galactose utilizing PHB producer, we examined 16 Halomonas strains. We demonstrated that Halomonas cerina (Halomonas sp. YK44) has the highest growth and PHB production using a culture media containing 2% galactose, final 4% NaCl, and 0.1% yeast extract. These culture conditions yielded 8.98 g/L PHB (78.1% PHB content (w/w)). When galactose-containing red algae (Eucheuma spinosum) hydrolysates were used as a carbon source, 5.2 g/L PHB was produced with 1.425% galactose after treatment with activated carbon. Since high salt conditions can be used to avoid sterilization, we examined whether Halomonas sp. YK44 could produce PHB in non-sterilized conditions. Culture media in these conditions yielded 72.41% PHB content. Thus, Halomonas sp. YK44 is robust against contamination, allowing for long-term culture and economical PHB production. Full article

Bioplastics, which are plastic materials produced from renewable bio-based feedstocks, have been investigated for their potential as an attractive alternative to petroleum-based plastics. Despite the harmful effects of plastic accumulation in the environment, bioplastic production is still underdeveloped. Recent advances in strain development, genome sequencing, and editing technologies have accelerated research efforts toward bioplastic production and helped to advance its goal of replacing conventional plastics. In this review, we highlight bioengineering approaches, new advancements, and related challenges in the bioproduction and biodegradation of plastics. We cover different types of polymers, including polylactic acid (PLA) and polyhydroxyalkanoates (PHAs and PHBs) produced by bacterial, microalgal, and plant species naturally as well as through genetic engineering. Moreover, we provide detailed information on pathways that produce PHAs and PHBs in bacteria. Lastly, we present the prospect of using large-scale genome engineering to enhance strains and develop microalgae as a sustainable production platform. Full article