Polyhydroxyalkanoates Research Articles

Understanding the mechanism of microplastic-associated antibiotic resistance genes in aquatic ecosystems: Insights from metagenomic analyses and machine learning

The pervasive presence of microplastics (MPs) in aquatic systems facilitates the transmission of antibiotic resistance genes (ARGs), thereby posing risks to ecosystems and human well-being. However, owing to variations in environmental backgrounds and the limited scope of research subjects, studies on ARGs in MPs lack unified conclusions, particularly regarding whether different types of MPs selectively promote ARG enrichment. Analysing large-scale datasets can better encompass broad spatiotemporal scales and diverse samples, facilitating a more extensive exploration of the complex ecological relationships between MPs and ARGs. The present study integrated existing metagenomic datasets to conduct a comprehensive risk assessment and comparative analysis of resistance groups across various MPs. In addition, we endeavoured to elucidate potential associations between ARGs and bacterial taxa, as well as MP structural features, using machine learning (ML) methods. The findings of our research highlight the pivotal role of MP type in shaping plastispheres, accounting for 9.56 % of the biotic variation (Adonis index) and explaining 18.59 % of the ARG variance. Compared to conventional MPs, biodegradable MPs, such as polyhydroxyalkanoates (PHA) and polylactic acid (PLA), exhibit lower species uniformity and diversity but pose a higher risk of ARG occurrence. These ML approaches effectively forecasted ARG abundance by using the bacterial taxa and molecular structure descriptors (MDs) of MPs (average R2tra = 0.882, R2test = 0.759). Feature analysis showed that MDs associated with lipophilicity, solubility, toxicity, and surface potential significantly influenced the relative abundance of ARGs in the plastispheres. The interpretable multiple linear regression (MLR) model, particularly notable, elucidated a linear relationship between bacterial genera and ARGs, offering promise for identifying potential ARG hosts. This study offers novel insights into ARG dynamics and ecological risks within aquatic plastispheres, highlighting the importance of comprehensive MP monitoring initiatives.

Structure Assessment and Impacts of Lipids' Chemistry on the Structuration of Polyhydroxyalkanoate Biosynthesized by Bacillus licheniformis AZU-A5.

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

Assessing the aerobic/anoxic enrichment efficiency at different C/N ratios: pilot scale polyhydroxyalkanoates production from waste activated sludge

Polyhydroxyalkanoates (PHA) can be produced using fermentation products of an excess sewage sludge fermentation process. An efficient method to enrich a PHA-producing community is an aerobic-feast/anoxic-famine enrichment strategy. The effect of different carbon to nitrogen (C/N) feed ratios of 1, 2 and 3.5 g COD/g N on the process performance was studied. The study was executed on a pilot plant scale using fermented waste activated sludge as the organic carbon source. The system's performance was monitored in terms of removing contaminants, producing PHA, and reducing N2O emissions. The results indicated that a lower C/N ratio results in lower PHA production, with PHA content in the sludge of 20, 24 and 36 % w/w for C/N ratios of 1, 2 and 3.5 g COD/g N, respectively. At the lowest C/N ratio, the highest nitrite accumulation rate (77%), nitrification efficiency (89%) and denitrification efficiency (89%) were observed, but the N2O production was also the highest (0.77 mg N2O-N/L). The long-term comprehensive monitoring carried out in this study revealed high carbon and ammonia removal efficiencies (never below 80%) despite the C/N shifts and high COD and ammonia concentrations. At the same time, the system showed relatively low PHA production and high environmental impact in terms of high gaseous N2O emission. These findings question the sustainability of the aerobic-feast/anoxic-famine enrichment strategy for PHA production in full-scale plants.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate)-based microspheres as a sustained platform for Huperzine A delivery for alzheimer's disease therapy

Huperzine A (HupA) is used in Alzheimer's disease (AD) therapy for its effective inhibition of acetylcholinesterase (AChE) and enhancement of cholinergic neuronal function. However, direct oral administration and injection of HupA cause side effects like nausea, anorexia, and rapid metabolism. Using a tripolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate (PBVHx), from the polyhydroxyalkanoate (PHA) family synthesized via synthetic biology, we present a novel AD therapy strategy with peritoneally administered PBVHx microspheres loaded with HupA (HupA-PBVHxMs). This approach extends HupA's metabolic duration in the blood and brain, enhancing AChE inhibition efficacy. Uniformly sized HupA-PBVHxMs, created using microfluidics and rotary evaporation, show up to 70.4 % drug encapsulation efficiency, sustained HupA release for 40 days, reduced neurotoxicity from Aβ25–35, and maintained in vivo HupA supply and AChE inhibition for over 20 days. In cognitive tests, HupA-PBVHxMs improved function in AD mice. Thus, PBVHx microspheres with slower HupA release and lower biotoxicity offer a superior platform for sustained AChE inhibitor release, outperforming commercial PLGA microspheres.

Optimization and prediction of additively manufactured PLA-PHA biodegradable polymer blend using TOPSIS and GA-ANN

Recent years have seen the proliferation of fused deposition modeling (FDM) as a means of manufacturing biodegradable products, for different applications such as rigid packaging, agricultural and biomedical. Blends of Polyhydroxyalkanoates (PHA) and polylactic acid (PLA) have been investigated to ascertain their prospective applications through FDM. This paper includes three parameters that affect the build process: layer height, nozzle temperature, and flow rate. 3D printed PLA/PHA can be characterized mechanically, and process parameters can be optimized to maximize design functionality. The experimental setup utilized a Taguchi L9 design, and TOSPIS was employed to optimize the output results. Using TOPSIS analysis, 0.2 mm layer thickness, 195 °C nozzle temperature, and 100 % flow rate were found to be the most optimal initiation parameters. The Taguchi analysis was used to analyze the output responses, and it was found that layer height had the greatest influence on mechanical properties, followed by flow rate and nozzle temperature. The percentage elongation at break has been improved significantly by adding PHA i.e., 170 % compared to PLA (5–10 %). This paper presents a framework for in-depth mechanical characterization of PLA-PHA 3D-printed parts, along with methods for optimizing process parameters to achieve optimal performance, as well as tools for modeling output responses using GA-ANN with an accuracy of 95 %.

Recovery and fractionation of volatile fatty acids from fermented solutions by electrodialysis: electrochemical characterization of anion-exchange membranes

Electrodialysis can be used to recover charged precursors, such as volatile fatty acids (VFAs), of the biopolymers polyhydroxyalkanoates (PHAs). In general, all VFAs are recovered in a receiver solution, despite their advantageous partial fractionation. Herein, the separation of acetic, propionic, butyric, and valeric acids by electrodialysis was evaluated using three anion-exchange membranes (namely, Ralex AMH-PES, Fumasep FAS-PET-130 and PC200D) applying cell voltages from 0V to 2.88V. The mass transfer mechanisms were evaluated by linear sweep voltammetry and chronopotentiometry. Fumasep showed a slightly greater VFAs fractionation capacity than Ralex, while it was much greater for PC200D due to the presence of tertiary amines in its fixed functional groups. Overall, increasing the operating voltage and/or time reduced the degree of VFAs fractionation with all membranes. The higher percent extraction values and greater VFAs fractionation degrees obtained with the PC200D membrane could enhance PHAs storage efficiency.

Novel genomic and phenotypic traits of polyhydroxyalkanoate-producing bacterium ZZQ-149, the type strain of Halomonas qinghailakensis

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

Multiscale modeling and experimental study of molecular weight distribution and monomeric ratio in PHA production

While plastics have brought considerable convenience to our daily lives, most of them are still made up of petroleum-based materials. The plastic production industry accounts for a substantial portion of greenhouse gas emissions, and these products cause serious environmental pollution due to their poor degradability. Hence, bio-derived polymers have gained much research interest to substitute the existing plastic materials. Among them, polyhydroxyalkanoates (PHAs) are regarded as the most promising candidate owing to their biocompatible and biodegradable nature. In particular, poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB) is now in the limelight for its unique mechanical properties. However, understanding the sophisticated mechanism and structure is quite challenging, posing obstacles to its successful commercialization. In response to this difficulty, we devised a multiscale model using the kinetic Monte Carlo (kMC) algorithm to enable capturing the extensive characteristics of PHA, including molecular weight distribution and monomeric ratio. Considering the different time and length scales of the overall process, the model is developed with two layers for the fermentation and polymerization reactions, and they are successfully integrated. The results not only validate the experimental results but also showcase a wide range of PHA information that cannot be measured through instrumental analyses.

Enhancing nitrogen removal by simultaneous nitritation and denitritation in a multi-cycle SBR with supplementation of solid carbon sources

Simultaneous nitritation and denitritation have the potential to significantly improve nitrogen removal in sewage treatment processes. However, their application in low-strength sewage treatment systems presents challenges. This study explored the impact of four solid carbon sources (SCSs) on N-removal via nitrite in a multi-cycle SBR with biocarriers. Results showed that both N-removal efficiencies and nitrite accumulation rates increased with higher COD/N ratios, indicating that high COD/N ratios can improve the competition between denitrifiers and nitrite-oxidizing bacteria for nitrite, leading to stable simultaneous nitritation and denitritation. The supplementation of SCSs further enhanced this high-efficiency N-removal process, with polybutylene succinate (PBS) and polycaprolactone (PCL) showing greater increases in N-removal via nitrite than poly-hydroxybutyrate (PHB) and poly-hydroxyalkanoate (PHA). Moreover, PBS showed the most significant increase in denitrification efficiency in anoxic conditions, while PHA was the most effective external SCS at a moderate level of dissolved oxygen. These findings suggest that the incorporation of external SCSs can facilitate the simultaneous nitritation and denitrification process in multi-cycle SBRs, underscoring the importance of selecting an appropriate SCS for optimizing nitrogen removal in sewage treatment projects.

Effect of culture history and carbon sources on polyhydroxyalkanoates production in activated sludge systems

ABSTRACT Polyhydroxyalkanoates (PHAs) are important and completely biodegradable alternatives to regular plastics, and they can be produced by activated sludge systems during wastewater treatment. Wastewaters with high organic content are being used for PHA production, which is an important resource recovery option. In this context, the effect of sludge retention time and different carbon sources, such as acetate, peptone-mixture and industrial wastewater (containing acetic acid (AA), lactic acid (LA) and propionic acid (PA)), on PHA storage was investigated. Oxygen utilisation rate (OUR) profiles were generated in respirometric tests and were evaluated by activated sludge modelling. Results showed that high storage (AA: 70%; LA: 49%; PA: 60% and industrial wastewater: 52%) was achievable in the feast phase even when the biomass was fed with a high organic acid content substrate to which it is not acclimated.

Polyhydroxyalkanoates production from cheese whey under near-seawater salinity conditions

Treating saline streams presents considerable challenges due to their adverse effects on conventional biological processes, thereby leading to increased expenses in managing those side streams. With this in consideration, this study explores into the potential for valorizing fermented cheese whey (CW), a by-product of the dairy industry, into polyhydroxyalkanoates (PHA) using mixed microbial cultures (MMC) under conditions of near-seawater salinity (30 gNaCl/L). The selection of a PHA-accumulating MMC was successfully achieved using a sequential batch reactor operated under a feast and famine regime, with a hydraulic retention time of 14.5 h, a variable solids retention time of 3 and 4.5 days, and an organic loading rate (OLR) of 60 Cmmol/(L d). The selected culture demonstrated efficient PHA production rates and yields, maintaining robust performance even under high salinity conditions. During PHA accumulation, a maximum PHA content in biomass of 56.4 % wt. was achieved for a copolymer P(3HB-co-3HHx) with a 3HHx content of 7 %. Additionally, to asses the capacity of the culture to produce polymers with different compositions, valeric acid was supplemented to the real fermented feedstock which resulted in the production of terpolymers P(3HB-co-3HV-co-3HHx) with varied monomeric content and a higher maximum PHA content of 62 % wt. Additionally, this study highlights the potential utilization of seawater as alternative to freshwater for PHA production, thereby enhancing circular economy principles and promoting environmental sustainability.

Biotreatment of Industrial Pollutant (Carbon Dioxide) and its Role in Bioplastics Production

Polyhydroxyalkanoates (PHAs) are a class of biopolymers with characteristics resembling those of petrochemical-based plastics that have gained recent attention and a growing amount of research effort due to their environmental friendliness. Numerous bacteria, including Alcaligenes spp., Pseudomonas spp., Staphylococcus spp., and algae create (PHAs) in the presence of carbon and limiting nutrients like nitrogen, and these biopolymers can successfully replace industrial plastics. Therefore, the goal of this study is to employ carbon dioxide, an industrial pollutant emitted into the environment, as a cheap carbon source to lower production costs and remove pollution at the same time. Only three of the nine bacterial isolates utilized were able to synthesize the polymer in the presence of CO2, and the best isolate was belonging to the genus Alcaligenes after 48 hours of incubation at 30°C and pH 7, which are the optimal conditions for polymer synthesis. Bacterial growth resulted in the production of 5.2gm/l of PHA and 6.2gm/l of biomass under these conditions.

Evaluation of fouling phenomena and cation-exchange membrane cleaning in Donnan dialysis for separation of ammonium from fermented solutions rich in volatile fatty acids

Polyhydroxyalkanoates (PHA) are promising biobased and biodegradable alternatives to the petroleum-based plastics. For their production, controlling cell growth is crucial and can be achieved by limiting nutrients, such as ammonium. The use of Donnan dialysis (DD) has been considered for this purpose. However, the effects of long–term DD fermentative operations need to be evaluated since fouling and sorption phenomena can affect the process performance. In this study, DD operations were performed over 10 consecutive batches to separate ammonium from a real fermented stream derived from an acidogenic fermentation process using cation-exchange membranes with distinct properties (a homogenous Fumasep FKS–PET-130 and a heterogenous Ralex CMH-PES) and NaCl or HCl aqueous receiver solutions. A cleaning procedure with acidic and alkaline solutions was proposed for the Ralex. It was verified that sorption and fouling phenomena occurred on/inside both membranes using a NaCl solution as receiver. This occurred more intensely with the Ralex, thus reducing the ammonium percent extraction from 55% (first batch) to approximately 40% in the subsequent batches. On the other hand, Fumasep exhibited higher percent extractions, and its values remained constant at around 65% along the batches when only rinsed with distilled water. The proposed cleaning procedure for Ralex effectively regenerated this membrane and increased the subsequent ammonium percent extractions. However, the acidic and/or alkaline solutions formed cavities on/in the membrane. The use of a HCl solution as receiver led to a lower reduction in the percent extraction throughout the batches when compared to NaCl, but the acidic receiver solution also damaged the membrane. It is suggested the use of the Fumasep membrane for the application evaluated herein or that a milder cleaning procedure is developed and applied for the Ralex membrane.

Promotion of polyhydroxyalkanoates-producing granular sludge formation by lactic acid using anaerobic dynamic feeding process

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

Halorubrum miltondacostae sp. nov., a potential polyhydroxyalkanoate producer isolated from an inland solar saltern in Rio Maior, Portugal

One hundred and sixty-three extreme halophiles were recovered from a single sample collected from an inland solar saltern in Rio Maior. Based on random amplified polymorphic DNA (RAPD) profiles and partial 16S rRNA gene sequencing 125 isolates were identified as members of the Archaea domain within the genus Halorubrum. Two strains, RMP-11T and RMP-47, showed 99.1 % sequence similarity with the species Halorubrum californiense based on phylogenetic analysis of the 16S rRNA gene sequence. However, phylogenetic analysis based on five housekeeping genes, atpB, EF-2, glnA, ppsA and rpoB’, showed Halorubrum coriense as the closest related species with 96.7 % similarity. The average nucleotide identity (ANI) of strains RMP-11T, RMP-47 and species Hrr. coriense were within the range of 90.0–90.5 %, supporting that strains RMP-11T and RMP-47 represent a novel species of the genus Halorubrum. These strains formed red-pigmented colonies that were able to grow in a temperature range of 25–50 °C. Polyhydroxyalkanoate (PHA) granules were detected in both strains. The polar lipid profile was identical to the neutrophilic species of the genus Halorubrum. The Rio Maior sample from which both strains were isolated was metagenome sequenced. We identified five metagenome-assembled genomes representing novel Halorubrum species but distinct from the species represented by strains RMP-11T and RMP-47. Based on phylogenetic, phylogenomic, comparative genomics, physiological and chemotaxonomic parameters, we describe a new species of the genus Halorubrum represented by strains RMP-11T (=CECT 30760T = DSM 115521T) and RMP-47 (=CECT 30761 = DSM 115541) for which we propose the name Halorubrum miltondacostae sp. nov.

Polyhydroxyalkanoate Production by Methanotrophs: Recent Updates and Perspectives.

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

Partial least squares regression based rapid quantification of intracellular biopolymers from a Sudan black absorption assay

Rapid quantitation of intracellular polyhydroxyalkanoate (PHA) is essential for monitoring and possibly intervening in ongoing bioprocesses. The traditional multistep protocol involving cell separation, lysis, organic solvent extraction, and purification of intracellular polymers is time intensive. In this study, indirect Sudan black (ISB) adsorption data combined with culture optical density and intracellular protein measurements were used to develop partial least squares (PLS) regression models to predict the intracellular PHA concentration in Cupriavidus necator. ISB analysis supported the hypothesized partitioning of Sudan black into residual biomass, PHA, and abiotic compartments. The PLS regression model developed with autoscaled data offered excellent fit on cross-validation (R2CV of 0.987) and independent prediction (R2P of 0.994) sets. The method limit of detection for the PLS model (0.036 g l−1) was reasonably low to track biopolymer accumulation in microbial culture, even in early growth phase. The conservative estimate suggests a significantly shorter processing time with the machine learning-based approach (190 min) than with the traditional protocol (940 min) while retaining analytical interchangeability.

Embracing Nature's Clockwork: Crafting Plastics for Degradation in Plant Agricultural Systems.

In the 21st century, global agriculture confronts the urgent challenge of increasing food production by 70% by 2050 while simultaneously addressing environmental and health concerns. Plastics, integral to agricultural innovation, present sustainability challenges due to their non-biodegradable nature and contribution to pollution. This perspective examines the transition to bioplastics, emphasizing their bio-based origin and their crucial characteristic of being readily biodegradable in the soil. Key bioplastics such as poly(lactic acid) (PLA), polyhydroxyalkanoates (PHAs), and biomass-derived polymers are discussed, particularly regarding the microplastic generation in soil resulting from their use in specific applications like mulch films, delivery systems, and soil conditioners. Embracing bioplastics signifies a significant step forward in achieving sustainable agriculture and addressing plastic waste. However, it is highlighted that while some bioplastics can be recovered and recycled, special applications where the plastic is in intimate contact with soil pose challenges for recovery. In these cases, that represent more than the 50% of plastics used in agriculture, meticulous design for biodegradation in soil synchronized with agricultural cycles is necessary. This approach ensures minimal environmental impact and promotes a circular approach to plastic use in agriculture.

Production of Polyhydroxybutyrate by halotolerant Halomonas cerina YK44 using sugarcane molasses and soybean flour in tap water

As environmental pollution intensifies, the interest in bioplastics is growing. The bioplastic polyhydroxyalkanoates (PHAs), which are produced and degraded by microorganisms, have received considerable attention. However, the production cost of PHA is still high, and several ways to increase economy of PHA production have been studied. Therefore, as one way of solution, Halomonas species were screened and evaluated with cheap substrates such as molasses and soybean flour. Among tested strains, Halomonas cerina YK44 was selected and used for polyhydroxybutyrate (PHB) production with molasses and soybean flour together, whose combination was not evaluated well before, in tap water. The medium composition optimization showed maximum PHB production at 4 % sugarcane molasses, 2 % NaCl, 0.05 % soybean flour, and pH 8 in tap water (9.2 g/L DCW, 7.3 g/L PHB, and 79.7 % PHB contents). However, cell growth of halotolerant H. cerina YK44 was disturbed by 0.2 % furfural, which existed in biomass based sugars as inhibitors. Physical and thermal analyses revealed that PHB film started from sugarcane molasses and soybean flour was no different from that initiated from simple sugars (Tm was 175.8 °C and 176.2 °C, PDI was 1.29, and 1.31, respectively).

Hints from nature for a PHA circular economy: Carbon synthesis and sharing by Pseudomonas solani GK13

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