Oxygen-limited Conditions Research Articles

Zic-HILIC MS/MS Method for NADomics Provides Novel Insights into Redox Homeostasis in Escherichia coli BL21 Under Microaerobic and Anaerobic Conditions

Background: Nicotinamide adenine dinucleotide (NAD+), its precursors, and its derivatives (collectively NADome) play a crucial role in cellular processes and maintain redox homeostasis. Understanding the dynamics of these metabolic pools and redox reactions can provide valuable insights into metabolic functions, especially cellular regulation and stress response mechanisms. The accurate quantification of these metabolites is challenging due to the interconversion between the redox forms. Methods: Our laboratory previously developed a zwitterionic hydrophilic interaction liquid chromatography (zic-HILIC)–tandem mass spectrometry method for the quantification of five essential pyridine nucleotides, including NAD+ derivatives and it’s reduced forms, with 13C isotope dilution and matrix-matched calibration. In this study, we have improved the performance of the chromatographic method and expanded its scope to twelve analytes for a comprehensive view of NAD+ biosynthesis and utilization. The analytical method was validated and applied to investigate Escherichia coli BL21 under varying oxygen supplies including aerobic, microaerobic, and anaerobic conditions. Conclusions: The intracellular absolute metabolite concentrations ranged over four orders of magnitude with NAD+ as the highest abundant, while its precursors were much less abundant. The composition of the NADome at oxygen-limited conditions aligned more with that in the anaerobic conditions rather than in the aerobic phase. Overall, the NADome was quite homeostatic and E. coli rapidly, but in a minor way, adapted the metabolic activity to the challenging shift in the growth conditions and achieved redox balance. Our findings demonstrate that the zic-HILIC-MS/MS method is sensitive, accurate, robust, and high-throughput, providing valuable insights into NAD+ metabolism and the potential significance of these metabolites in various biological contexts.

Candida glabrata maintains two HAP1 ohnologs, HAP1A and HAP1B, for distinct roles in ergosterol gene regulation to mediate sterol homeostasis under azole and hypoxic conditions.

Invasive and drug-resistant fungal infections pose a significant public health concern. Candida glabrata, a human fungal pathogen, is often difficult to treat due to its intrinsic resistance to azole antifungal drugs and its capacity to develop clinical drug resistance. Therefore, understanding the pathways that facilitate fungal growth and environmental adaptation may lead to novel drug targets and/or more efficacious antifungal therapies. While the mechanisms of azole resistance in Candida species have been extensively studied, the roles of zinc cluster transcription factors, such as Hap1A and Hap1B, in C. glabrata have remained largely unexplored until now. Our research shows that these factors play distinct yet crucial roles in regulating ergosterol homeostasis under azole drug treatment and oxygen-limiting growth conditions. These findings offer new insights into how this pathogen adapts to different environmental conditions and enhances our understanding of factors that alter drug susceptibility and/or resistance.

Effective removal of Pb (II) from wastewater by zinc-iron bimetallic oxide-modified walnut shell biochar: A combined experimental and DFT calculation approach

The modified walnut shell biochar (WBC) was prepared through zinc-iron bimetallic oxide modification (ZF@WBC) at 600 °C under oxygen-limited conditions in this study. Through adsorption experiments, characterization analyses, and density functional theory (DFT) calculations, the adsorption properties of ZF@WBC to Pb (II) were investigated and the mechanism underlying such adsorption was elucidated. Characterization results showed that the surface area (375.9709 m2/g) and total pore volume (0.205319 cm3/g) of ZF@WBC were significantly greater than those of walnut shell biochar. The maximum adsorption capacity of ZF@WBC for Pb (II) was found to be 104.26 mg/g, which is 2.57 times higher than that of WBC according to the adsorption experiments conducted. The observed adsorption behavior followed both the pseudo-second-order (PSO) kinetic model and Langmuir isothermal adsorption model, suggesting that chemisorption plays a major role in the absorption process. Based on SEM, XRD, XPS, FTIR characterizations along with DFT calculations performed in this study, it can be concluded that surface complexation, ion exchange, electrostatic attraction, physical absorption are among the main mechanisms responsible for absorption of Pb (II) by ZF@WBC. Furthermore, even in the presence of interfering ions at different concentrations, ZF@WBC exhibited a removal rate above 70% for Pb (II). Therefore, ZF@WBC has great potential as an effective absorbent for removing Pb (II) from wastewater, while also offering opportunities for biomass waste resource utilization.

Enhanced phosphorus adsorption using modified drinking water treatment residues: A comparative analysis of powder and alginate bead forms

This study provides an analysis of the phosphorus adsorption efficacy of three modified drinking water treatment residues (MDWTRs): MDWTR-P (powdered form), MDWTR-D2, and MDWTR-D5 (alginate bead-entrapped forms with bead diameters of 2 mm and 5 mm, respectively). The preparation process involved washing and drying the drinking water treatment residue, followed by grinding and sieving to achieve particle sizes below 90 μm. The residue was then incinerated at 600 °C in oxygen-limited conditions. Subsequently, the MDWTR was formulated into alginate beads by mixing with sodium alginate and FeCl3 solutions, resulting in spherical particles of specified diameters. The evaluation of surface area, pore volume, pore size, and CHN concentration revealed that MDWTR-D5 possesses the largest surface area (284.7 m2 g−1) and highest micropore volume (0.04 cm3 g−1), indicating a greater capacity for adsorption. SEM-EDS analysis demonstrated significant compositional changes post-treatment, particularly elevated phosphorus levels, confirming effective adsorption. Metal content analysis indicated high aluminum levels in MDWTR-P and increased iron content in MDWTR-D5. Toxicity Characteristic Leaching Procedure (TCLP) and in vitro bioaccessibility (IVBA) tests confirmed the non-hazardous nature of all MDWTRs, ensuring their safety for environmental applications. Kinetic analyses using pseudo-first-order, pseudo-second-order, and intraparticle diffusion models highlighted the superior performance of MDWTR-D5, with the highest equilibrium adsorption capacity and initial adsorption rate across all tested concentrations, suggesting both high efficiency and rapid adsorption potential. Further validation using Langmuir and Freundlich isotherms revealed MDWTR-D5's highest monolayer adsorption capacity (22.88 mg g−1) and Freundlich adsorption capacity parameter (6.97 mg g−1). Statistical analysis via one-way ANOVA confirmed significant differences in phosphorus concentrations among the MDWTRs samples (p-value <0.001), consistently underscoring MDWTR-D5's superior adsorption performance. These findings highlight MDWTR-D5's potential as an effective adsorbent for phosphorus removal in wastewater treatment, emphasizing its applicability in environmental remediation strategies.

The Influence of Pyrolysis Temperature and Feedstocks on the Characteristics of Biochar-Derived Dissolved Organic Matter: A Systematic Assessment

Biochar is a carbon-rich product obtained by pyrolyzing biomass under oxygen-limited conditions and has a wide range of potential for environmental applications. In particular, dissolved organic matter (DOM) released from biochar has an important impact on the fate of pollutants. The study aimed to systematically assess how varying pyrolysis temperatures and biomass feedstocks influence the characteristics of biochar-derived DOM. DOM samples were comprehensively characterized utilizing UV-vis spectroscopy and excitation–emission matrix (EEM) fluorescence spectroscopy, coupled with parallel factor (PARAFAC) analysis. The study discovered that pyrolysis temperature significantly affects DOM characteristics more than feedstock type. An increase in pyrolysis temperature correlated with a notable decrease in dissolved organic carbon content, aromaticity, and fluorescence intensity, alongside a marked increase in pH and hydrophilicity. PARAFAC analysis identified three distinct DOM components: two humic-like substances (C1 and C2) and one protein-like substance (C3). The proportion of protein-like substances increased with higher pyrolysis temperatures, while the humic-like substances’ proportion declined. The compositional shifts in DOM with pyrolysis temperature may significantly influence its environmental behavior and functionality. Further research is necessary to explore the long-term environmental impact and potential applications of biochar-derived DOM.

Iodate reduction by marine aerobic bacteria.

Iodate reductase (Idr) gene cluster (idrABP1P2 ) is involved in bacterial iodate (IO3 -) respiration under anaerobic conditions. Putative idr gene clusters are present in both anaerobic and aerobic bacteria; however, the specific physiological roles of idr genes in aerobic bacteria remain unclear. Therefore, in this study, three marine aerobic bacteria with putative idr gene clusters (Roseovarius azorensis, Notoacmeibacter marinus, and Aliiroseovarius sediminilitoris) were grown in the presence of iodate to determine whether they can reduce iodate to iodide (I-). All tested bacteria almost completely reduced 2 mM iodate under static conditions but only reduced 0.1-0.5 mM iodate under shaking conditions. Moreover, the washed cell suspension of R. azorensis reduced iodate only when the cells were pre-grown statically in the presence of iodate. Transcriptional analysis revealed that the expression levels of idrA, idrB, idrP1 , and idrP2 genes were upregulated in R. azorensis when the cells were grown statically in the presence of iodate. Specifically, idrA expression was induced by 0.1 μM iodate and was up to 14-fold higher compared to that of the non-iodate control. These results suggest that marine aerobic bacteria reduce iodate under oxygen-limited conditions, and that this capacity is induced by environmentally relevant levels of iodate in seawater. Our results suggest that marine aerobic bacteria contribute to iodide production in marine surface waters, thereby affecting the global iodine cycling and ozone budget.

Bioupcycling Methane and CO2 for Succinate Production by an Engineered Type I Methanotrophic Bacterium.

Methane, a byproduct of agricultural activities, has shown potential as a nonedible substrate for biomanufacturing. The production of succinate by a methanotrophic bacterium utilizing methane presents an innovative route for the sustainable synthesis of chemicals. In this study, Methylotuvimicrobium buryatense 5GB1S was genetically modified through the reconstruction of an artificial serine cycle to enable the bioconversion of both methane and CO2 into succinate. The 13C labeling analysis confirmed the CO2 fixing in M. buryatense 5GB1S, leading to a 46% improvement in carbon conversion efficiency and a 107% increase in succinate production compared to the wild-type strain. The transcriptome data on carbon metabolisms was assessed to guide future optimizations for strengthening the overall carbon flux from methane to succinate. Finally, the maximum succinate titer of 299.36 mg/L was achieved under oxygen-limited conditions in 3 L bioreactors, which resulted in the volumetric productivity of 199.60 mg/L/day, representing a 23-fold enhancement compared to the wild-type strain. This study offers a new strategy for upcycling greenhouse gases into succinate in a sustainable manner through methanotrophic-based biomanufacturing.

Physiological Metabolic Analysis of VB12 Accumulation in Ensifer adhaerens Casida A Enhanced by Oxygen Limitation.

Cobalamin (VB12) is in enormous demand across the fields of medicine, food, and feed additives. However, the oxygen supply plays a critical role in VB12 biosynthesis by Ensifer adhaerens Casida A and has been identified as a bottleneck for economical substrate consumption. This study elucidates the relationship between oxygen limitation and VB12 accumulation with transcriptomic and metabolomic analyses. Under oxygen limitation, E. adhaerens enhances oxygen transport and storage by increasing expression of flavin hemoglobin (Hmp), which was up-regulated 6-fold at 24h of oxygen restriction compared to the oxygen restriction of 4h (p<0.01). Because of the cofactor of Hmp is heme, the demand for heme increases, leading to the upregulation of genes in the heme biosynthesis pathway. Similarly, genes involved in biosynthesis of its precursor, 5-ALA, were upregulated as well. 5-ALA is also a direct precursor of VB12, further leading to the upregulation of genes in the VB12 biosynthesis pathway. This process initiates biosynthesis and accumulation of VB12. As VB12 and heme biosynthesis progresses, genes associated with the biosynthesis and transportation pathways of compounds related to their biosynthesis were likewise upregulated, including genes involved in S-adenosyl methionine (SAM) biosynthesis, and the transport of Fe2+ and Co2+. Additionally, amino acids and organic acids associated with biosynthesis were also extensively consumed, such as methionine, which is used for synthesizing SAM, decreased by 310% after 24 h of oxygen limitation compared to 20% dissolved oxygen (p<0.05). At the same time, genes related to growth-associated metabolic pathways, such as pentose phosphate pathway (PPP), were significantly downregulated. Therefore, the potential mechanism by which E. adhaerens accumulates VB12 under oxygen-limited conditions by enhancing Hmp expression, which facilitates the porphyrin metabolic pathway and promotes VB12 biosynthesis. This research provides valuable insights for increasing VB12 production through metabolic engineering and process optimization.

Changes of glycolysis and gluconeogenesis key enzymes in the muscle of the shrimp Penaeus vannamei in response to hypoxia and reoxygenation

Although the shrimp Penaeus vannamei can tolerate oxygen-limited conditions, the organisms may suffer detrimental effects in several processes, including growth, reproduction, immune responses, and others. During hypoxia, the shrimp makes metabolic adjustments in energy use and glucose homeostasis. Glucose is hydrolyzed during glycolysis and synthesized from non-glycosidic metabolites in gluconeogenesis. Although these pathways share many enzymes, the processes are not exactly the reverse of each other. We studied the first and last irreversible steps of glycolysis and gluconeogenesis catalyzed by hexokinase/glucose-6-phosphatase (HK/G6Pase) and pyruvate kinase/pyruvate carboxylase/phosphoenolpyruvate carboxykinase (PK/PC/PEPCK), respectively, in muscle under hypoxia and hypoxia followed by reoxygenation to gain insights about the coordinated metabolic responses that maintain glucose balance during oxygen stress. For the first group, HK gene expression was induced in hypoxia while G6Pase was reduced in hypoxia and reoxygenation, whereas for enzymes activity, HK was maintained and G6Pase was only detected in reoxygenation. For the second group, gene expression increased in PK in hypoxia, in PC in reoxygenation and in both cases, the enzymatic activities were maintained. Mitochondrial PEPCK expression increased during hypoxia and cytosolic PEPCK decreased, while total PEPCK activity was maintained. Intracellular lactate decreased during hypoxia while glucose levels were maintained. Altogether the results show that in limited oxygen, the expression of glycolytic enzymes-genes are induced, the gluconeogenic counterparts are active, although with more varied responses to oxygen limited conditions, and intracellular glucose levels are maintained.

Properties of Biochar Prepared by Solar Pyrolysis and Its Adsorption of Cu&amp;lt;sup&amp;gt;2+&amp;lt;/sup&amp;gt; in Water

This study investigates the potential of biochar produced via a solar pyrolysis system and its effectiveness in removing copper (Cu&amp;lt;sup&amp;gt;2+&amp;lt;/sup&amp;gt;) ions from water, presenting a sustainable and energy-efficient method for biochar production and biomass recycling. Two common agricultural and livestock wastes, corn straw and cow dung, were used as raw materials to produce biochar. These materials underwent solar pyrolysis under limited oxygen conditions to produce biochar, which was then compared to biochar produced via traditional pyrolysis. The comparison involved elemental analyses, infrared spectroscopy, scanning electron microscopy, and specific surface area and pore size analysis to highlight differences in their physical and chemical properties. Adsorption experiments were conducted to evaluate the adsorptive capacity of biochar for copper ions (Cu&amp;lt;sup&amp;gt;2+&amp;lt;/sup&amp;gt;) from water, determining the optimal pH conditions and underlying adsorption mechanisms. The findings reveal that biochar produced through solar pyrolysis exhibits similar properties and Cu&amp;lt;sup&amp;gt;2+&amp;lt;/sup&amp;gt; adsorption capacities to those prepared by traditional methods. Specifically, cow dung biochar demonstrated a higher adsorption capacity for Cu&amp;lt;sup&amp;gt;2+&amp;lt;/sup&amp;gt; compared to corn straw biochar. The Cu&amp;lt;sup&amp;gt;2+&amp;lt;/sup&amp;gt; adsorption by corn straw biochar followed the Langmuir isothermal adsorption model and pseudo-second-order kinetic equation, whereas cow dung biochar conformed to the Freundlich isothermal adsorption model and pseudo-second-order kinetic equation. By demonstrating the comparable efficacy of solar pyrolysis biochar in heavy metal adsorption, this study highlights its potential for sustainable environmental remediation and biomass utilization.

Evolution and Competitive Struggles of Lactiplantibacillus plantarum under Different Oxygen Contents.

Lactiplantibacillus (Lb.) plantarum is known as a benign bacterium found in various habitats, including the intestines of animals and fermented foods. Since animal intestines lack oxygen, while fermented foods provide a limited or more oxygen environment, this study aimed to investigate whether there were genetic differences in the growth of Lb. plantarum under aerobic vs. anaerobic conditions. Genomic analysis of Lb. plantarum obtained from five sources-animals, dairy products, fermented meat, fermented vegetables, and humans-was conducted. The analysis included not only an examination of oxygen-utilizing genes but also a comparative pan-genomic analysis to investigate evolutionary relationships between genomes. The ancestral gene analysis of the evolutionary pathway classified Lb. plantarum into groups A and B, with group A further subdivided into A1 and A2. It was confirmed that group A1 does not possess the narGHIJ operon, which is necessary for energy production under limited oxygen conditions. Additionally, it was found that group A1 has experienced more gene acquisition and loss compared to groups A2 and B. Despite an initial assumption that there would be genetic distinctions based on the origin (aerobic or anaerobic conditions), it was observed that such differentiation could not be attributed to the origin. However, the evolutionary process indicated that the loss of genes related to nitrate metabolism was essential in anaerobic or limited oxygen conditions, contrary to the initial hypothesis.

Improving the transfer of dissolved oxygen in a biological Fe2+ oxidation process using a venturi jet as an intensive aeration system

The low bio-production of Fe3+ as a leaching agent is one of the main limitations to implementing industrial bio-processes at feasibility conditions. The main limitation of the bio-oxidation process of Fe2+ is the low oxygen transfer to the aqueous phase because of the low oxygen solubility. This study assesses the effectiveness of the venturi jet as an innovative and intensive aeration device for overcoming the oxygen limitation in a continuous ferrous oxidation process in a fixed-bed reactor with immobilized acidithiobacillus ferrooxidans, in contrast to the conventional diffuser aeration device. Firstly, the influence of the airflow and the influence of the medium concentration were determined for the following parameters for both aeration devices; Volumetric mass transfer coefficient (kLa), Standard Oxygen Transfer Rate (SOTR), Standard Aeration Efficiency (SAE), and Standard Oxygen Transfer Efficiency (SOTE). Then, both aeration devices were compared in a continuous bio-oxidation process in an up-flow packed bio-reactor (UFPB). The system performance was assessed by monitoring temperature, pH, oxidation-reduction potential, and dissolved oxygen concentration for 69 days. Findings displayed that when aerating with the diffuser, the ferrous oxidation rate was restricted by the low dissolved oxygen availability, being about 1 ppm (1 mg L−1). Under these oxygen-limiting conditions, the maximum ferrous (Fe2+) oxidation rate was 9.09 g L−1 h−1. However, when aerating with the venturi jet, the dissolved oxygen concentration increased up to 2.70 mg L−1, achieving a maximum of 29.11 g L−1 h−1. So, this study has demonstrated that the change in the aeration device has resulted in an improvement in the process, achieving a 3.5-fold increase in the oxidation rate. Furthermore, the venturi jet offered additional advantages over the diffuser, such as requiring less power to deliver the same amount of air, being unaffected by jarosite precipitates, and not requiring a supply of compressed air.

Development of a Robust Saccharomyces cerevisiae Strain for Efficient Co-Fermentation of Mixed Sugars and Enhanced Inhibitor Tolerance through Protoplast Fusion.

The economical and efficient commercial production of second-generation bioethanol requires fermentation microorganisms capable of entirely and rapidly utilizing all sugars in lignocellulosic hydrolysates. In this study, we developed a recombinant Saccharomyces cerevisiae strain, BLH510, through protoplast fusion and metabolic engineering to enhance its ability to co-ferment glucose, xylose, cellobiose, and xylooligosaccharides while tolerating various inhibitors commonly found in lignocellulosic hydrolysates. The parental strains, LF1 and BLN26, were selected for their superior glucose/xylose co-fermentation capabilities and inhibitor tolerance, respectively. The fusion strain BLH510 demonstrated efficient utilization of mixed sugars and high ethanol yield under oxygen-limited conditions. Under low inoculum conditions, strain BLH510 could completely consume all four kinds of sugars in the medium within 84 h. The fermentation produced 33.96 g/L ethanol, achieving 84.3% of the theoretical ethanol yield. Despite the challenging presence of mixed inhibitors, BLH510 successfully metabolized all four sugars above after 120 h of fermentation, producing approximately 30 g/L ethanol and reaching 83% of the theoretical yield. Also, strain BLH510 exhibited increased intracellular trehalose content, particularly under conditions with mixed inhibitors, where the intracellular trehalose reached 239.3 mg/g yeast biomass. This elevated trehalose content contributes to the enhanced stress tolerance of BLH510. The study also optimized conditions for protoplast preparation and fusion, balancing high preparation efficiency and satisfactory regeneration efficiency. The results indicate that BLH510 is a promising candidate for industrial second-generation bioethanol production from lignocellulosic biomass, offering improved performance under challenging fermentation conditions. Our work demonstrates the potential of combining protoplast fusion and metabolic engineering to develop superior S. cerevisiae strains for lignocellulosic bioethanol production. This approach can also be extended to develop robust microbial platforms for producing a wide array of lignocellulosic biomass-based biochemicals.

Oxygen regulation of microbial communities and chemical compounds in cigar tobacco curing.

Curing is a critical process that determines the sensory quality of cigars. The impact of oxygen on cigar curing and the mechanisms by which it regulates microbial changes affecting cigar quality are not well understood. In this study, we selected handmade cigars from the same batch and conducted curing experiments in environments with varying oxygen concentrations (equivalent to 0.1%, 6-12, and 15% of atmospheric oxygen concentration). We collected samples over 60 days and analyzed the distribution of microbial communities using high-throughput sequencing. Combined with the analysis of total sugars, proteins, flavor substances, and other chemical compounds, we elucidated how different oxygen concentrations affect the cigar curing process, influence microbial community succession, and ultimately impact cigar quality. Our results revealed significant differences in bacterial community composition under different oxygen conditions. Under aerobic conditions, Cyanobacteria were the dominant bacteria, while under oxygen-limited conditions, Staphylococcus and Corynebacterium predominated. As oxygen concentration decreased, so did the richness and diversity of the bacterial community. Conversely, oxygen concentration had a lesser impact on fungi; Aspergillus was the dominant genus in all samples. We also found that Enterococcus showed a positive correlation with aspartic acid, alanine, and 4-aminobutyric acid and a negative correlation with cysteine. Cigars cured at 15% oxygen concentration for 60 days exhibited optimal quality, particularly in terms of flavor richness and sweetness. These findings suggest that oxygen concentration can alter cigar quality by regulating aerobic and anaerobic microbial community succession. The relationship between specific microbial communities and flavor compounds also provides a theoretical reference for developing artificial control technologies in the cigar curing process.

Methane-Driven Perchlorate Reduction by a Microbial Consortium.

The phenomenon of methane oxidation linked to perchlorate reduction has been reported in multiple studies; yet, the underlying microbial mechanisms remain unclear. Here, we enriched suspended cultures by performing methane-driven perchlorate reduction under oxygen-limiting conditions in a membrane bioreactor (MBR). Batch test results proved that perchlorate reduction was coupled to methane oxidation, in which acetate was predicted as the potential intermediate and oxygen played an essential role in activating methane. By combining DNA-based stable isotope probing incubation and high-throughput sequencing analyses of 16S rRNA gene and functional genes (pmoA, pcrA, and narG), we found that synergistic interactions between aerobic methanotrophs (Methylococcus and Methylocystis) and perchlorate-reducing bacteria (PRB; Denitratisoma and Dechloromonas) played active roles in mediating methane-driven perchlorate reduction. This partnership was further demonstrated by coculture experiments in which the aerobic methanotroph could produce acetate to support PRB to complete perchlorate reduction. Our findings advance the understanding of the methane-driven perchlorate reduction process and have implications for similar microbial consortia linking methane and chlorine biogeochemical cycles in natural environments.

A newly isolated strain for poly(3-hydroxybutyrate) production under anaerobic conditions and the key enzyme analysis

Polyhydroxyalkanoates (PHAs) are biodegradable polyesters with great possibility to replace petroleum-based plastics, but the high costs of production associated with energy inputs have been an obstacle to their commercial application. The production of PHAs under anaerobic environments without aeration appears to be more attractive but has not been adequately investigated. A strain capable of accumulating poly(3-hydroxybutyrate) (PHB) under anaerobic conditions was isolated. The strain anaerobically accumulated PHB reached 38.7 % of cell dry weight (CDW), which was 1.9 times higher than under aerobic conditions. It was also found that the concentrations of key enzymes for PHB synthesis were higher under anaerobic conditions than under aerobic conditions. The highest PHB accumulation was 77.9 % obtained at oxygen-limited conditions and the highest PHB yield was 0.13 g PHB/g carbon obtained at anaerobic conditions. It suggests that the conditions should be well controlled according to the purpose of PHB production. This study offers a new alternative of PHAs formation.

Investigation of characteristics and application of food waste-derived bone char on plant growth

Waste bone, a by-product originating from animal husbandry and food industry, primarily ends up in landfills and may cause soil pollution if no treatment method is applied. In this article, pork bone char (PBc), a porous carbonaceous material, was manufactured by pyrolyzing waste pork bones collected from food waste sources. The investigation covered temperatures from 400 to 800 °C in limited oxygen conditions. The prepared PBc samples were characterized and applied as an alternative source of phosphorus fertilizer by evaluating the growth of Ipomoea aquatica planted on the PBc/soil mixture and assessing the properties of SPBc.As a result, the PBc materials exhibit a clear hydroxyapatite mineral structure along with an abundance of micro-mesoporous features. The SPBc samples demonstrate a significant change in phosphorus concentration (up to 1.6 %) compared to PBc non-amended soil, and the growth of the Ipomoea aquatica plant differed significantly from the plant not fertilized by PBc. The results after four weeks indicate that samples added with PBc synthesized at temperatures of 500, 600, and 700 °C promoted its vigorous growth corresponding to the ratio of Ca/P of hydroxyapatite. The specific surface area, pore volume, and average pore size by various pyrolysis temperatures can impact the release of phosphorus from the PBc into the soil, significantly influencing the growth of Ipomoea aquatica. The achieved outcome not only presents a promising substance for reducing reliance on conventional phosphorus fertilizers derived from apatite minerals but also establishes a means to pursue the objectives of sustainable development and a circular economy.

Development of Foam-Free Biosurfactant Production Processes Using Bacillus licheniformis

Microbial biosurfactants are considered environmentally friendly alternatives to synthetic surfactants in numerous applications. One of the main bottlenecks to their widespread use is the lack of effective processes for their production on an industrial scale. Biosurfactant production using conventional aerated bioreactors results in excessive foaming due to the combination of air injection and their tensioactive properties. A not widely explored approach to overcome this problem is the development of foam-free production processes, which require the identification and characterization of appropriate microorganisms. Bacillus licheniformis EL3 was evaluated for biosurfactant production under oxygen-limiting conditions in a bioreactor, using a mineral medium containing glucose as a carbon source and NaNO3 and NH4Cl as nitrogen sources. After optimizing the operational conditions, glucose concentration, and inoculum strategy, B. licheniformis EL3 produced 75 ± 3 mg biosurfactant/L in 43 h. The purified biosurfactant exhibited exceptional surface active properties, with minimum surface tension values (29 mN/m) and a critical micelle concentration (27 mg/L) similar to those achieved with commercial surfactin. Furthermore, biosurfactant yield per substrate (YP/S = 0.007 g biosurfactant/g glucose) was similar to the figures reported for Bacillus subtilis strains grown in similar conditions, whereas biosurfactant yield per biomass (YP/X = 0.755 g biosurfactant/g biomass) and specific biosurfactant productivity (qBS = 0.018 g biosurfactant/(g biomass × h)) were almost three times higher when compared to previous reports. The results obtained indicate that B. licheniformis EL3 is a promising candidate for the development of foam-free biosurfactant production processes at an industrial scale.

Oxygen-to-ammonium-nitrogen ratio as an indicator for oxygen supply management in microoxic bioanodic ammonium oxidation

Microbial electrolysis cells (MECs) have been proven effective for oxidizing ammonium (NH4+), where the anode acts as an electron acceptor, reducing the energy input by substituting oxygen (O2). However, O2 has been proved to be essential for achieving high removal rates MECs. Thus, precise control of oxygen supply is crucial for optimizing treatment performance and minimizing energy consumption. Unlike previous studies focusing on dissolved oxygen (DO) levels, this study introduces the O2/NH4+-N ratio as a novel control parameter for balancing oxidation rates and the selectivity of NH4+ oxidation towards dinitrogen gas (N2) under limited oxygen condition. Our results demonstrated that the O2/NH4+-N ratio is a more relevant oxygen supply indicator compared to DO level. Oxygen served as a more favorable electron acceptor than the electrode, increasing NH4+ oxidation rates but also resulting in more oxidized products such as nitrate (NO3−). Additionally, nitrous oxide (N2O) and N2 production were higher with the electrode as the electron acceptor compared to oxygen alone. An O2/NH4+-N ratio of 0.5 was found to be optimal, achieving a balance between product selectivity for N2 (51.4 % ± 4.5 %) and oxidation rates (344.6 ± 14.7 mg-N/L*d), with the columbic efficiency of 30.7 % ± 2.0 %. Microbial community analysis revealed that nitrifiers and denitrifiers were the primary bacteria involved, with oxygen promoting the growth of nitrite-oxidizing bacteria, thus facilitating complete NH4+ oxidation to NO3−. Our study provides new insights and guidelines on the appropriate oxygen dosage, offering strategies into optimizing operational conditions for NH4+ removal using MECs.

Synergism of floated paperboard sludge cake /sewage sludge for maximizing biomethane yield and biochar recovery from digestate: A step towards circular economy

Anaerobic digestion of floated paperboard sludge (PS) cake suffers from volatile fatty acids (VFAs) accumulation, nutrient unbalanced condition, and generation of digestate with a risk of secondary pollution. To overcome these drawbacks, sewage sludge (SS) was added to PS cake for biogas recovery improvement under a co-digestion process followed by the thermal treatment of solid fraction of digestate for biochar production. Batch experimental assays were conducted at different SS:PS mixing ratios of 70:30, 50:50, 30:70, and 20:80 (w/w), and their anaerobic co-digestion performances were compared to the mono-digestion systems at 35 ± 0.2 °C for 45 days. The highest methane yield (MY) of 241.68 ± 14.81 mL/g CODremoved was obtained at the optimum SS:PS ratio of 50:50 (w/w). This experimental condition was accompanied by protein, carbohydrate, and VFA conversion efficiencies of 47.3 ± 3.2%, 46.8 ± 3.2%, and 56.3 ± 3.8%, respectively. The synergistic effect of SS and PS cake encouraged the dominance of Bacteroidota (23.19%), Proteobacteria (49.65%), Patescibacteria (8.12%), and Acidovorax (12.60%) responsible for hydrolyzing the complex organic compounds and converting the VFAs into biomethane. Further, the solid fraction of digestate was subjected to thermal treatment at a temperature of 500 °C for 2.0 h, under an oxygen-limited condition. The obtained biochar had a yield of 0.48 g/g dry digestate, and its oxygen-to-carbon (O/C), carbon-to-nitrogen (C/N), and carbon-to-phosphorous (C/P) ratios were 0.55, 10.23, and 16.42, respectively. A combined anaerobic co-digestion/pyrolysis system (capacity 50 m3/d) was designed based on the COD mass balance experimental data and biogenic CO2 market price of 22 USD/ton. This project could earn profits from biogas (12,565 USD/yr), biochar (6641 USD/yr), carbon credit (8014 USD/yr), and COD shadow price (6932 USD/yr). The proposed project could maintain a payback period of 6.60 yr. However, further studies are required to determine the associated life cycle cost model that is useful to validate the batch experiment assumptions.