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Bioplastics, synthesized by several microbes, accumulates inside cells under stress conditions as a storage material. Several microbial enzymes play a crucial role in their degradation. This research was carried to test the biodegradability of poly-[beta]-hydroxybutyrate (PHB) utilizing PHB depolymerase, produced by bacteria isolated from sewage waste soil samples. Potent PHB degrader was screened based on the highest zone of hydrolysis followed by PHB depolymerase activity. Soil burial method was employed to check their degradation ability at different incubation periods of 15, 30, and 45 days at 37±2°C, pH 7.0 at 60% moisture with 1% microbial inoculum of Aeromonas caviae Kuk1-(34) (MN414252). Without optimized conditions, 85.76% of the total weight of the PHB film was degraded after 45 days. This degradation was confirmed with Fourier-transform infrared spectroscopy (FTIR) and Scanning electron microscope (SEM) analysis. The presence of bacterial colonies on the surface of the degraded film, along with crest, holes, surface erosion, and roughness, were visible. Media optimization was carried out in statistical mode using Plackett Burman (PB) and Central Composite Design (CCD) of Response Surface Methodology (RSM) by considering ten different factors. Analysis of Variance (ANOVA), Pareto chart, response surface plots, and F-value of 3.82 implies that the above statistical model was significant. The best production of PHB depolymerase enzyme (14.98 U/mL) was observed when strain Kuk1-(34) was grown in a media containing 0.1% PHB, K.sub.2 HPO.sub.4 (1.6 gm/L) at 27 â for seven days. Exploiting these statistically optimized conditions, the culture was found to be a suitable candidate for the management of solid waste, where 94.4% of the total weight of the PHB film was degraded after 45 days of incubation.

Over 30 years, the Gram-positive bacterium Priestia megaterium (previously known as Bacillus megaterium) was systematically developed for biotechnological applications ranging from the production of small molecules like vitamin B12, over polymers like polyhydroxybutyrate (PHB) up to the in vivo and in vitro synthesis of multiple proteins and finally whole-cell applications. Here we describe the use of the natural vitamin B12 (cobalamin) producer P. megaterium for the elucidation of the biosynthetic pathway and the subsequent systematic knowledge-based development for production purposes. The formation of PHB, a natural product of P. megaterium and potential petro-plastic substitute, is covered and discussed. Further important biotechnological characteristics of P. megaterium for recombinant protein production including high protein secretion capacity and simple cultivation on value-added carbon sources are outlined. This includes the advanced system with almost 30 commercially available expression vectors for the intracellular and extracellular production of recombinant proteins at the g/L scale. We also revealed a novel P. megaterium transcription-translation system as a complementary and versatile biotechnological tool kit. As an impressive biotechnology application, the formation of various cytochrome P450 is also critically highlighted. Finally, whole cellular applications in plant protection are completing the overall picture of P. megaterium as a versatile giant cell factory.Key points• The use of Priestia megaterium for the biosynthesis of small molecules and recombinant proteins through to whole-cell applications is reviewed.• P. megaterium can act as a promising alternative host in biotechnological production processes.

Petroleum-derived plastics are linked to a variety of growing environmental issues throughout their lifecycle, including emission of greenhouse gases, accumulation in terrestrial and marine habitats, pollution, among others. There has been a lot of attention over the last decade in industrial and research communities in developing and producing eco-friendly polymers to deal with the current environmental issues. Bioplastics preferably are a fast-developing family of polymeric substances that are frequently promoted as substitutes to petroleum-derived plastics. Polyhydroxyalkanoates (PHAs) have a number of appealing properties that make PHAs a feasible source material for bioplastics, either as a direct replacement of petroleum-derived plastics or as a blend with elements derived from natural origin, fabricated biodegradable polymers, and/or non-biodegradable polymers. Among the most promising PHAs, polyhydroxybutyrates (PHBs) are the most well-known and have a significant potential to replace traditional plastics. These biodegradable plastics decompose faster after decomposing into carbon dioxide, water, and inorganic chemicals. Bioplastics have been extensively utilized in several sectors such as food-processing industry, medical, agriculture, automobile industry, etc. However, it is also associated with disadvantages like high cost, uneconomic feasibility, brittleness, and hydrophilic nature. A variety of tactics have been explored to improve the qualities of bioplastics, with the most prevalent being the development of gas and water barrier properties. The prime objective of this study is to review the current knowledge on PHAs and provide a brief introduction to PHAs, which have drawn attention as a possible potential alternative to conventional plastics due to their biological origin, biocompatibility, and biodegradability, thereby reducing the negative impact of microplastics in the environment. This review may help trigger further scientific interest to thoroughly research on PHAs as a sustainable option to greener bioplastics.

Cyanobacteria are photosynthetic microorganisms with significant biotechnological potential owing to their ability to produce valuable biopolymers such as polyhydroxybutyrate (PHB). This study focuses on PHB production in the cyanobacteria, Limnospira fusiformis NRMCF6962 and Spirulina major NRMCF6963, under nitrogen-limited conditions. Qualitative and quantitative analyses confirmed PHB accumulation in L. fusiformis and its absence in S. major . Whole-genome sequencing of L. fusiformis revealed 5,177 coding sequences. Comparative genomic analysis of 26 cyanobacterial strains belonging to genera of Spirulina, Arthrospira, and Limnospira revealed significant variations in gene content, particularly within PHB biosynthesis pathways. Functional annotation revealed that PHB metabolism is absent in S. major , which instead relies on alternative stress-response pathways including chlorophyll degradation, selenocysteine metabolism and xanthine metabolism. Phylogenomic and metabolic pathway analyses suggests an evolutionary adaptation among the three genera of Oscillatoriales, with S. major as a divergent species from PHB-producing descendants. Orthologous Average Nucleotide Identity and digital DNA-DNA hybridization validated the taxonomic distinction between two major clades within strains. Metabolic insights revealed the critical role of PHB and alternative metabolic pathways in cyanobacteria for stress adaptation. This research advances the understanding of PHB metabolism and evolutionary mechanisms in cyanobacteria, underscoring the potential for cyanobacterial strain improvement and scale-up in bioprocess industry.

Poly–hydroxybutyrate (PHB) is a potential source of biodegradable plastics that are environmentally friendly due to their complete degradation to water and carbon dioxide. This study aimed to investigate PHB production in the cyanobacterium Synechocystis sp. PCC6714 MTₐ24 in an outdoor bioreactor using urban wastewater as a sole nutrient source. The culture was grown in a thin-layer raceway pond with a working volume of 100 L, reaching a biomass density of up to 3.5 g L.sup.-1 of cell dry weight (CDW). The maximum PHB content was found under nutrient-limiting conditions in the late stationary phase, reaching 23.7 ± 2.2% PHB per CDW. These data are one of the highest reported for photosynthetic production of PHB by cyanobacteria, moreover using urban wastewater in pilot-scale cultivation which multiplies the potential of sustainable cultivation approaches. Contamination by grazers (Poterioochromonas malhamensis) was managed by culturing Synechocystis in a highly alkaline environment (pH about 10.5) which did not significantly affect the culture growth. Furthermore, the strain MTₐ24 showed significant wastewater nutrient remediation removing about 72% of nitrogen and 67% of phosphorus. These trials demonstrate that the photosynthetic production of PHB by Synechocystis sp. PCC6714 MTₐ24 in the outdoor thin-layer bioreactor using urban wastewater and ambient carbon dioxide. It shows a promising approach for the cost-effective and sustainable production of biodegradable carbon-negative plastics.

Plastic pollution is fueling the grave environmental threats currently facing humans, the animal kingdom, and the planet. The pursuit of renewable resourced biodegradable materials commenced in the 1970s with the need for carbon neutral fully sustainable products driving important progress in recent years. The development of bioplastic materials is highlighted as imperative to the solutions to our global environment challenges and to the restoration of the wellbeing of our planet. Bio-based plastics are becoming increasingly sustainable and are expected to substitute fossil-based plastics. Bioplastics currently include both, nondegradable and biodegradable compositions, depending on factors including the origins of production and post-use management and conditions. Among the most promising materials being developed and evaluated is polyhydroxybutyrate (PHB), a microbial bioprocessed polyester belonging to the polyhydroxyalkanoate (PHA) family. This biocompatible and non-toxic polymer is biosynthesized and accumulated by a number of specialized bacterial strains. The favorable mechanical properties and amenability to biodegradation when exposed to certain active biological environments, earmark PHB as a high potential replacement for petrochemical based polymers such as ubiquitous high density polyethylene (HDPE). To date, high production costs, minimal yields, production technology complexities, and difficulties relating to downstream processing are limiting factors for its progression and expansion in the marketplace. This review examines the chemical, mechanical, thermal, and crystalline characteristics of PHB, as well as various fermentation processing factors which influence the properties of PHB materials.

The valorization of wood waste as a sustainable bacterial feedstock for the production of Polyhydroxybutyrate (PHB) is explored in this study, aiming to provide an environmentally friendly alternative to conventional plastics. Wood waste, treated with 4% sulfuric acid, served as the carbon source for isolating bacteria from Jalandhar waste streams, with the strain Klebsiella sp. MK3 identified as the most effective in PHB production after 16s rRNA sequencing. Analytical methods including the Molisch test, DNS, and sugar utilization tests confirmed sugar presence and consumption by the bacterial isolate. Media optimization using Design Expert 12.0 utilized a quadratic model, achieving a robust fit with an R² value of 98.6%. Optimization via Plackett-Burman design and response surface methodology enhanced PHB yield to 4.37 mg/mL, a significant increase over previous benchmarks. This yield was achieved under optimal conditions of 1.7% carbon concentration, 0.105% nitrogen concentration, and a constant temperature of 37 °C. Qualitative analysis of PHB by UV-Vis spectroscopy, FTIR, and NMR confirmed its purity and composition. The study highlights the potential of wood waste and wastewater as substrates for cost-effective PHB production, with significant applications in packaging, agriculture, medicine, and more, thus promoting reduced reliance on non-renewable resources and advancing sustainability goals.

In order to make bioplastics accessible for a wider spectrum of applications, ready-to-use plastic material formulations should be available with tailored properties. Ideally, these kinds of materials should also be “home-compostable” to simplify their organic recycling. Therefore, materials based on PLA (polylactid acid) and PHB (polyhydroxybutyrate) blends are presented which contain suitable additives, and some of them contain also thermoplastic starch as a filler, which decreases the price of the final compound. They are intended for various applications, as documented by products made out of them. The produced materials are fully biodegradable under industrial composting conditions. Surprisingly, some of the materials, even those which contain more PLA than PHB, are also fully biodegradable under home-composting conditions within a period of about six months. Experiments made under laboratory conditions were supported with data obtained from a kitchen waste pilot composter and from municipal composting plant experiments. Material properties, environmental conditions, and microbiology data were recorded during some of these experiments to document the biodegradation process and changes on the surface and inside the materials on a molecular level.

Plastic pollution is a worldwide concern causing the death of animals (mainly aquatic fauna) and environmental deterioration. Plastic recycling is, in most cases, difficult or even impossible. For this reason, new research lines are emerging to identify highly biodegradable bioplastics or plastic formulations that are more environmentally friendly than current ones. In this context, microbes, capable of synthesizing bioplastics, were revealed to be good models to design strategies in which microorganisms can be used as cell factories. Recently, special interest has been paid to haloarchaea due to the capability of some species to produce significant concentrations of polyhydroxyalkanoate (PHA), polyhydroxybutyrate (PHB), and polyhydroxyvalerate (PHV) when growing under a specific nutritional status. The growth of those microorganisms at the pilot or industrial scale offers several advantages compared to that of other microbes that are bioplastic producers. This review summarizes the state of the art of bioplastic production and the most recent findings regarding the production of bioplastics by halophilic microorganisms with special emphasis on haloarchaea. Some protocols to produce/analyze bioplastics are highlighted here to shed light on the potential use of haloarchaea at the industrial scale to produce valuable products, thus minimizing environmental pollution by plastics made from petroleum.

Background Plastic pollution is a significant environmental problem caused by its high resistance to degradation. One potential solution is polyhydroxybutyrate (PHB), a microbial biodegradable polymer. Mexico has great uncovered microbial diversity with high potential for biotechnological applications. The best polymer producers tend to be isolated from environments that require survival adaptations from microorganisms, the high-producing Bacillus cereus strain saba.zh comes from refinery wastewater, the costs of production have been a limiting factor for biopolymer production, and one of the focuses of interest has been finding novel strains with better production or singular traits that help in industrial processes. Results The isolates were taxonomically classified as Bacillus cereus MSF4 and Bacillus inaquosorum MSD1 from Mina, Nuevo Leon; B. cereus S07C; and Paenibacillis dendritiformis from the active volcano “El Chichonal” on Chiapas. The strains had growth temperatures ranging from 35 to 50 °C and pH tolerance values ranging from 3 to 9. The best PHB-producing strain, B. cereus MSF4 , produced 0.43 g/kg PHB on orange peels, followed by B. inaquosorum MSD1 at 0.40 g/kg, B. cereus S07C at 0.23 g/kg and P. dendritiformis at 0.26 g/kg. Conclusions The findings of this study affirm the potential of the Mexican isolated strains as PHB-producing organisms, enabling further studies to test their viability as industrial producers. The ability of P. dendritiformis and B. inaquosorum to synthetize PHB was also confirmed by the observations made providing novel evidence to consider these species as potential producers.