Effect of plasticizers on the rheological properties of xanthan gum - starch biodegradable films.

The effect of plasticizers, namely glycerol, sorbitol, and citric acid, on the structural and mechanical properties of biodegradable films obtained from xanthan gum (XG) and starch was studied. The plasticizing effect of glycerol, sorbitol, and citric acid on XG-starch films is justified by the destruction of intermolecular contacts between starch and XG macromolecules and the redistribution of hydrogen bonds in the system as a result of the hydrotropic action of plasticizer molecules. The use of glycerol proved to be the most effective for regulating the deformation of films, while the use of sorbitol to preserve strength. The dependence of the film roughness on the type and concentration of plasticizers was characterized. The smallest values of protrusions on the surface of XG-starch films were found in the presence of sorbitol. Considering the effect of the concentration of plasticizers on the stickiness of the surface of XG-starch films and their structural and mechanical properties, 1.5 % concentration of glycerol, sorbitol and citric acid was determined as optimal.

Selective oxidation of glycerol mediated by surface plasmon of gold nanoparticles deposited on titanium dioxide nanowires.

This paper describes an alternative method for the in situ synthesis of gold nanoparticles (AuNPs) with a particle size of less than 3 nm, using nanoreactors formed by reverse micelles of 1,4-bis-(2-ethylhexyl) sulfosuccinate sodium (AOT) and nanoparticle stabilization with l-cysteine, which favor the preparation of nanoparticles with size and shape control, which are homogeneously dispersed (1% by weight) on the support of titanium dioxide nanowires (TNWs). To study the activity and selectivity of the prepared catalyst (AuNPs@TNWs), an aqueous solution of 40 mM glycerol was irradiated with a green laser (λ = 530 nm, power = 100 mW) in the presence of the catalyst and O2 as an oxidant at 22 °C for 6 h, obtaining a glycerol conversion of 86% with a selectivity towards hydroxypyruvic acid (HA) of more than 90%. From the control and reactions, we concluded that the Ti-OH groups promote the glycerol adsorption on the nanowires surface and the surface plasmon of the gold nanoparticles favors the selectivity of the reaction towards the hydroxypyruvic acid.

Glycerol carbonate and solketal carbonate as circular economy bricks for supercapacitors and potassium batteries.

Considering the worldwide market of batteries and supercapacitors, the (partial or total) replacement of conventional fossil-derived carbonates with bio-based ones in electrolyte formulations would allow the production of safer and more sustainable devices. In this work, embracing the 7th principle of green chemistry, glycerol derivatives (namely glycerol carbonate and solketal carbonate) are tested as solvents and additives for electrolyte formulations. Glycerol carbonate is innovatively employed as promising electrolyte solvent for electric double-layer capacitors with excellent performances. On the other hand, a solketal carbonate-laden liquid electrolyte is investigated for potassium-based batteries, showing a rather stable electrochemical behaviour and performance close to those of commercial oil-derived alternatives.

Ozone-Assisted Cu-Based Catalysts for the Efficient Electro-Reforming Glycerol to Formic Acid.

Glycerol electrooxidation reaction (GOR) to produce value-added chemicals, such as formic acid, could make more efficient use of abundant glycerol and meet future demand for formic acid as a fuel for direct or indirect formic acid fuel cells. Non-noble metal Cu-based catalysts have great potential in electro-reforming glycerol to formic acid. However, the high activity, selectivity and stability of Cu based catalysts in GOR cannot be achieved simultaneously. Here, we used ozone-assisted electrocatalyst to convert glycerol to formic acid under alkaline conditions, the onset potential was reduced by 60 mV, the Faraday efficiency (FE) reached 95%. The catalyst has excellent stability within 300 h at the current density of 10 mA cm-2. The electron spin resonance proved that ozone produced superoxide anion during the GOR. In situ Raman spectroscopy, electrochemical studies showed that glycerol can be activated with ozone in GOR, and the C-C bond can be broken to reduce the polymerization of glycerol on the catalyst surface, so as to produce more formic acid at a lower voltage. Moreover, the removal of dissolved O3 from water can be up to 100% after 30 minutes of GOR reaction at a solubility of 50 mg L-1 as measured by UV-VIS spectrophotometry.

Streamlined integrated protein isoelectric focusing using microfluidic paper-based device.

An innovative integrated paper-based microdevice was developed for protein separation by isoelectric focusing (IEF), allowing for robust design thanks to a 3D-printed holder integrating separation channel, reservoirs, and electrodes. To reach robustness and precision, the optimization focused on the holder geometry, the paper nature, the reservoir design, the IEF medium, and various focusing parameters. A well-established and stable pH gradient was obtained on a glass-fiber paper substrate with simple sponge reservoirs, and the integration of the electrodes in the holder led to a straightforward system. The separation medium composed of water/glycerol (85/15, v/v) allowed for reducing medium evaporation while being an efficient medium for most hydrophobic and hydrophilic proteins, compatible with mass spectrometry detection for further proteomics developments. To our knowledge, this is the first report of the use of glycerol solutions as a separation medium in a paper-based microdevice. Analytical performances regarding pH gradient generation, pI determination, separation efficiency, and resolution were estimated while varying the IEF experimental parameters. The overall process led to an efficient separation within 25 min. Then, this methodology was applied to a sample composed of saliva doped with proteins. A minimal matrix effect was evidenced, underscoring the practical viability of our platform. This low-cost, versatile and robust paper-based IEF microdevice opens the way to various applications, ranging from sample pre-treatment to integration in an overall proteomic-on-a-chip device.

Enhanced cold plasma hydrogenation with glycerol as hydrogen source for production of trans-fat-free margarine.

The quest for better nutritious foods has encouraged novel scientific investigations to find trans-fat reduction methods. This research proposes an innovative approach for the production of healthier trans-fat-free margarine from palm oil by the use of dielectric barrier discharge (DBD) plasma technology with glycerol serving as the principal source of hydrogen. The effectiveness of DBD plasma in hydrogenating palm olein was investigated. By employing a methodical series of experiments and thorough analytical approaches, examination of the saturated fatty acid conversion experienced its iodine value (IV) reduction from 67.16 ± 0.70 to 31.61 ± 1.10 under the optimal process parameters of 1 L min-1 He flow rate, 35 W plasma discharge power, 10 mm gap size, ambient initial temperature, and 12 h reaction time with solid texture. According to the method for producing trans-fat-free margarine in the absence of a catalyst and H2 gas, the hydrogenation rate of the prepared mixture of palm olein-glycerol was remarkably improved; the trans-fat content in the produced product was zero; the efficacy of incorporating cis- and trans-isomerization was lowered, and the method has a promising industrial application prospect.

Efficient Electrocatalytic Oxidation of Glycerol to Formate Coupled with Nitrate Reduction over Cu-doped NiCo Alloy Supported on Nickel Foam.

Electrooxidation of biomass-derived glycerol which is regarded as a main byproduct of industrial biodiesel production, is an innovative strategy to produce value-added chemicals, but currently showcases slow kinetics, limited Faraday efficiency, and unclear catalytic mechanism. Herein, we report high-efficiency electrooxidation of glycerol into formate via a Cu doped NiCo alloy catalyst supported on nickel foam (Cu-NiCo/NF) in a coupled system paired with nitrate reduction. The designed Cu-NiCo/NF delivers only 1.23 V vs. RHE at 10 mA cm-2, and a record Faraday efficiency of formate of 93.8%. The superior performance is ascribed to the rapid generation of NiIII-OOH and CoIII-OOH and favorable coupling of surface *O with reactive intermediates. Using Cu-NiCo/NF as a bifunctional catalyst, the coupled system synchronously produces NH3 and formate, showing 290 mV lower than the coupling of hydrogen evolution reaction, together with excellent long-term stability for up to 144 h. This work lays out new guidelines and reliable strategies from catalyst design to system coupling for biomass-derived electrochemical refinery.

The monotopic quinone reductases from Staphylococcus aureus.

Staphylococcus aureus, a Gram-positive bacterium, is an opportunistic pathogen and one of the most frequent causes for community acquired and nosocomial infections that has become a major public health threat due to the increased incidence of its drug resistance. Although being a prominent pathogen, its energetic metabolism is still underexplored, and its respiratory enzymes have been escaping attention. S. aureus can adapt to different environmental conditions by performing both aerobic and anaerobic respirations, which is particularly important as it frequently colonizes niches with different oxygen concentrations. This adaptability is derived from the composition of its respiratory chain, specifically from the presence of terminal electron acceptor reductases. The plasticity of S. aureus energy metabolism is enlarged by the ten quinone reductases encoded in its genome, eight of them being monotopic proteins. The role of these proteins is critical as they connect the different catabolic pathways to the respiratory chain. In this work, we identify, describe, and revise the monotopic quinone reductases present in S. aureus, providing an integrated view of its respiratory chain.

Identification and characterization of a potential strain for the production of polyhydroxyalkanoate from glycerol.

While poly (3-hydroxybutyrate) (PHB) holds promise as a bioplastic, its commercial utilization has been hampered by the high cost of raw materials. However, glycerol emerges as a viable feedstock for PHB production, offering a sustainable production approach and substantial cost reduction potential. Glycerol stands out as a promising feedstock for PHB production, offering a pathway toward sustainable manufacturing and considerable cost savings. The identification and characterization of strains capable of converting glycerol into PHB represent a pivotal strategy in advancing PHB production research. In this study, we isolated a strain, Ralstonia sp. RRA (RRA). The strain exhibits remarkable proficiency in synthesizing PHB from glycerol. With glycerol as the carbon source, RRA achieved a specific growth rate of 0.19 h-1, attaining a PHB content of approximately 50% within 30 h. Through third-generation genome and transcriptome sequencing, we elucidated the genome composition and identified a total of eight genes (glpR, glpD, glpS, glpT, glpP, glpQ, glpV, and glpK) involved in the glycerol metabolism pathway. Leveraging these findings, the strain RRA demonstrates significant promise in producing PHB from low-cost renewable carbon sources.

A Histological and Biomechanical Analysis of Human Acellular Dermis (HAD) Created Using a Novel Processing and Preservation Technique.

Background: Large and complex defects requiring reconstruction are challenging for orthopaedic surgeons. The use of human acellular dermal (HAD) matrices to augment large soft tissue defects such as those seen in massive rotator cuff tears, knee extensor mechanism failures and neglected Tendo-Achilles tears has proven to be a valuable tool in surgeons reconstructive armamentarium. Different methods for allograft decellularization and preservation alter the native properties of the scaffold. Traditional processing and preservation methods have shown to have drawbacks that preclude its widespread use. Some of the common issues include inferior biomechanical properties, the risk of rejection, limited customization, difficulty in storing and transporting, the requirement of pre-operative preparation, and last but not the least increased cost. Methods: We describe a novel processing and preservation method utilizing a two-step non-denaturing decellularization method coupled with preservation using a water-sequestering agent (glycerol) to remove immunogenic components while retaining biomechanical properties. The efficiency of this novel process was compared with the traditional freeze-drying method and verified by histological evaluation and biomechanical strength analysis. Results: The absence of cellular components and matrix integrity in hematoxylin and eosin-stained glycerol-preserved HAD (gly-HAD) samples compared to freeze-dried HAD (FD-HAD) demonstrated effective yet gentle decellularization. Biomechanical strength analysis revealed that gly-HADs are stronger with an ultimate tensile load to the failure strength of 210 N compared to FD-HAD (124N). The gly-HADs were found to have an optimal suture-retention strength of 126 N. Finally, sterility testing of the resultant grafts was checked to ensure a sterility assurance level of 10-6 to establish implantability. Conclusion: The novel processing and preservation technique is described in this paper to create a Human Acellular Dermis with higher biomechanical strength and superior histological characteristics. The processing and preservation technique ensured high sterility assurance levels to establish implantability.

Isolation, molecular identification, and genomic analysis of Mangrovibacter phragmitis strain ASIOC01 from activated sludge harboring the bioremediation prowess of glycerol and organic pollutants in high-salinity.

The physiological and genotypic characteristics of Mangrovibacter (MGB) remain largely unexplored, including their distribution and abundance within ecosystems. M. phragmitis (MPH) ASIOC01 was successfully isolated from activated sludge (AS), which was pre-enriched by adding 1,3-dichloro-2-propanol and 3-chloro-1,2-propanediol as carbon sources. The new isolate, MPH ASIOC01, exhibited resilience in a medium containing sodium chloride concentration up to 11% (with optimal growth observed at 3%) and effectively utilizing glycerol as their sole carbon source. However, species delimitation of MGBs remains challenging due to high 16S rRNA sequence similarity (greater than 99% ANI) among different MGBs. In contrast, among the housekeeping gene discrepancies, the tryptophan synthase beta chain gene can serve as a robust marker for fast species delimitation among MGBs. Furthermore, the complete genome of MPH ASIOC01 was fully sequenced and circlized as a single contig using the PacBio HiFi sequencing method. Comparative genomics revealed genes potentially associated with various phenotypic features of MGBs, such as nitrogen-fixing, phosphate-solubilizing, cellulose-digesting, Cr-reducing, and salt tolerance. Computational analysis suggested that MPH ASIOC01 may have undergone horizontal gene transfer events, possibly contributing unique traits such as antibiotic resistance. Finally, our findings also disclosed that the introduction of MPH ASIOC01 into AS can assist in the remediation of wastewater chemical oxygen demand, which was evaluated using gas chromatograph-mass spectrometry. To the best of our knowledge, this study offers the most comprehensive understanding of the phenotypic and genotypic features of MGBs to date.

Fast-track adaptive laboratory evolution of Cupriavidus necator H16 with divalent metal cations.

Microbial strain improvement through adaptive laboratory evolution (ALE) has been a key strategy in biotechnology for enhancing desired phenotypic traits. In this Biotech Method paper, we present an accelerated ALE (aALE) workflow and its successful implementation in evolving Cupriavidus necator H16 for enhanced tolerance toward elevated glycerol concentrations. The method involves the deliberate induction of genetic diversity through controlled exposure to divalent metal cations, enabling the rapid identification of improved variants. Through this approach, we observed the emergence of robust variants capable of growing in high glycerol concentration environments, demonstrating the efficacy of our aALE workflow. When cultivated in 10% v/v glycerol, the adapted variant Mn-C2-B11, selected through aALE, achieved a final OD600 value of 56.0 and a dry cell weight of 15.2 g L-1, compared to the wild type (WT) strain's final OD600 of 39.1 and dry cell weight of 8.4 g L-1. At an even higher glycerol concentration of 15% v/v, Mn-C2-B11 reached a final OD600 of 48.9 and a dry cell weight of 12.7 g L-1, in contrast to the WT strain's final OD600 of 9.0 and dry cell weight of 3.1 g L-1. Higher glycerol consumption by Mn-C2-B11 was also confirmed by high-performance liquid chromatography (HPLC) analysis. This adapted variant consumed 34.5 times more glycerol compared to the WT strain at 10% v/v glycerol. Our method offers several advantages over other reported ALE approaches, including its independence from genetically modified strains, specialized genetic tools, and potentially carcinogenic DNA-modifying agents. By utilizing divalent metal cations as mutagens, we offer a safer, more efficient, and cost-effective alternative for expansion of genetic diversity. With its ability to foster rapid microbial evolution, aALE serves as a valuable addition to the ALE toolbox, holding significant promise for the advancement of microbial strain engineering and bioprocess optimization.

Ensuring optimal lipid oxidation during physical activity in overweight and obese patients using a capillary glycerol sensor.

AIM: To describe lipid oxidation during physical activity (PA) in overweight and obese patients using a real-time capillary glycerol sensor and to propose a personalized strategy to optimize lipolysis. METHODS: Healthy adult volunteers with a BMI >25 kg/m2 were recruited. All participants performed 27 sessions of 30-min PA covering all possible combinations of exercises (low, moderate, high intensities) and pre-exercise meals (high-carbohydrate, high-fat, or fasting) with 3 replicates. Glycerol measurements were performed five times during each session: at T0, T20, T30, T45, and T60. The evolution of capillary glycerolemia during PA was modeled by a nonlinear mixed-effects model. We developed a personalized PA recommendation based on the first 2 glycerolemia measurements that indicates whether a change in activity type is preferable. RESULTS: Thirteen patients were included in the study and 1232 capillary glycerolemia measurements were performed. Hyperglycemic and hyperlipidemic meals were associated with lower baseline glycerolemia (p < 0.0001 for both) compared to fasting. In contrast, the type of meal was not significantly associated with PA-induced lipolysis (p = 0.52 and p = 0.76 for high-carbohydrate and hyperlipidemic meal respectively). Compared with exercises done at low and moderate intensities, high intensity exercises were associated with a higher PA-induced lipolysis (p = 0.0002). Specific strength exercises were associated with a lower PA-induced lipolysis (p < 0.0001). The 2 initial glycerolemia measurements appeared to be a pertinent criterion which help the personalization of PA programs for such individuals (c-index 0.67). CONCLUSION: The inter-individual lipolysis responses to PA and the availability of a real-time capillary blood glycerol sensor may enable the personalization of PA programs, thus promoting optimal lipolysis in subjects with overweight and obesity.

Engineering Lewis-Acid Defects on ZnO Quantum Dots by Trace Transition-Metal Single Atoms for High Glycerol-to-Glycerol Carbonate Conversion.

Efficient conversion of biomass wastes into valuable chemicals has been regarded as a sustainable approach for green and circular economy. Herein, a highly efficient catalytic conversion of glycerol (Gly) into glycerol carbonate (GlyC) by carbonylation with the commercially available urea is presented using low-cost transition metal single atoms supported on zinc oxide quantum dots (M1-ZnO QDs) as a catalyst without using any solvent. A facile one-step wet chemical synthesis allows various types of metal single atoms to simultaneously dope and introduce Lewis-acid defects in the ZnO QD structure. It is found that doping with a trace amount of isolated metal atoms greatly boosts the catalytic activity with Gly conversion of 90.7%, GlyC selectivity of 100.0%, and GlyC yield of 90.6%. Congruential results from both Density Functional Theory (DFT) and in situ Diffuse Reflectance Infrared Fourier Transform Spectroscopy (in situ DRIFTS) studies reveal that the superior catalytic performance can be attributed to the enriched Lewis acid sites that endow optimal adsorption, formation of the intermediate for coupling between urea and Gly, and desorption of GlyC. Moreover, the tiny size of ZnO QDs efficiently promotes the accessibility of these active sites to the reactants.

H-NS involved in positive regulation of glycerol dehydratase gene expression in Klebsiella pneumoniae 2e.

Glycerol dehydratase is the key and rate-limiting enzyme in the 1,3-propanediol synthesis pathway of Klebsiella pneumoniae, which determined the producing rate and yield of 1,3-propanediol. However, the expression regulation mechanism of glycerol dehydratase gene dhaB remains poorly unknown. In this study, a histone-like nucleoid-structuring (H-NS) protein was identified and characterized as the positive transcription regulator for dhaB expression in K. pneumoniae 2e, which exhibited high tolerance against crude glycerol in our previous study. Deletion of hns gene significantly decreased the transcription level of dhaB in K. pneumoniae 2e, which led to a remarkable defect on strain growth, glycerol dehydratase activity, and 3-hydroxypropanal production during glycerol fermentation. The transcription level of dhaB was significantly up-regulated in crude glycerol relative to pure glycerol, while the inactivation of H-NS resulted in more negative effect for transcription level of dhaB in the former. Though the H-NS expression level was almost comparable in both substrates, its multimer state was reduced in crude glycerol relative to pure glycerol, suggesting that the oligomerization state of H-NS might have contributed for positive regulation of dhaB expression. Furthermore, electrophoretic mobility shift and DNase I footprinting assays showed that H-NS could directly bind to the upstream promoter region of dhaB by recognizing the AT-rich region. These findings provided new insight into the transcriptional regulation mechanism of H-NS for glycerol dehydratase expression in K. pneumoniae, which might offer new target for engineering bacteria to industrially produce 1,3-propanediol.IMPORTANCEThe biological production of 1,3-propanediol from glycerol by microbial fermentation shows great promising prospect on industrial application. Glycerol dehydratase catalyzes the penultimate step in glycerol metabolism and is regarded as one of the key and rate-limiting enzymes for 1,3-propanediol production. H-NS was reported as a pleiotropic modulator with negative effects on gene expression in most studies. Here, we reported for the first time that the expression of glycerol dehydratase gene is positively regulated by the H-NS. The results provide insight into a novel molecular mechanism of H-NS for positive regulation of glycerol dehydratase gene expression in K. pneumoniae, which holds promising potential for facilitating construction of engineering highly efficient 1,3-propanediol-producing strains.

Development of New Edible Biodegradable Films Containing Camu-Camu and Agro-Industry Residue.

The use of edible films has garnered significant interest in the food and environmental sectors due to their potential to prevent food deterioration and their biodegradability. This study aimed to develop and characterize edible films based on camu-camu residue, gelatin, and glycerol, evaluating their solubility, thermal, degradability, antioxidant, and water vapor permeability properties of the gelatin matrix. This is the first study incorporating camu-camu into a gelatin and glycerol matrix. The films produced with camu-camu residue were manageable and soluble, with some non-soluble residues, providing a shiny and well-presented appearance. In the biodegradation results, samples 3 and 4 appeared to degrade the most, being two of the three most affected samples in the triplicate. The films showed degradation modifications from the third day of the experiment. In the germination and plant growth analysis, sample 4 exhibited satisfactory development compared to the other samples, emerging as the sample with the best overall result in the analyses, attributed to a 13.84 cm increase in the growth of the upper part of the seedling. These results indicate that the produced materials have potential for food packaging applications.

Effect of mouthwash containing poly l-Lysine and glycerol monolaurate on oral Helicobacter pylori relating to biofilm eradication, anti-adhesion, and pro-inflammatory cytokine suppression.

Background/purpose: Helicobacter pylori has been found to be related to periodontitis, and the oral cavity has been considered a reservoir for H. pylori gastritis infection. Thus, this study evaluated the effect of mouthwash containing poly l-Lysine and glycerol monolaurate on inhibiting H. pylori growth, biofilm formation, cell cytotoxicity, adhesion ability, cagA mRNA expression, and pro-inflammatory cytokines stimulated by H. pylori. Materials and methods: Nineteen H. pylori strains were isolated from the oral cavity. The effectiveness of mouthwash containing poly l-Lysine and glycerol monolaurate was examined for its ability to inhibit H. pylori growth and biofilm formation and was tested for cell viability in oral epithelial cells (H357), gastric adenocarcinoma cells (AGS), and periodontal ligament cells (PDL). Additionally, the mouthwash was tested for reducing cagA mRNA expression, adhesion ability to H357 and AGS cells, and pro-inflammatory cytokines stimulated with H. pylori in AGS and PDL cells. Results: The mouthwash containing poly l-Lysine and glycerol monolaurate could eradicate the biofilm by 14.9-19.9% after incubation at 5 min, and cell viability revealed 77.2, 79.8, and 100.0% for AGS, H357, and PDL cells, respectively. Moreover, the mouthwash containing poly l-Lysine and glycerol monolaurate could down-regulate cagA mRNA expression, reduce adhesion of H. pylori by approximately 9.5-47.8% for H357 cells and 24.5-62.9% for AGS cells, and decrease pro-inflammatory cytokines, especially interleukin-8, stimulated with H. pylori. Conclusion: Mouthwash containing poly l-Lysine and glycerol monolaurate could inhibit H. pylori growth and reduce their virulence expression. The mouthwash also revealed low cytotoxicity to oral and gastric cells.

Glycerol-slaved 1H-1H NMR cross-relaxation in quasi-native lysozyme.

1H-1H nuclear cross-relaxation experiments have been carried out with lysozyme in variable glycerol viscosity to study intramolecular motion, self-diffusion, and isotropic rigid-body rotational tumbling at 298 K, pH 3.8. Dynamics of intramolecular 1H-1H cross-relaxation rates, the increase in internuclear spatial distances, and lateral and rotational diffusion coefficients all show fractional viscosity dependence with a power law exponent κ in the 0.17-0.83 range. The diffusion coefficient of glycerol Ds with the bulk viscosity itself is non-Stokesian, having a fractional viscosity dependence on the medium viscosity (Ds âˆ¼ Î·-κ, κ ≈ 0.71). The concurrence and close similarity of the fractional viscosity dependence of glycerol diffusion on the one hand, and diffusion and intramolecular cross-relaxation rates of the protein on the other lead to infer that relaxation of glycerol slaves protein relaxations. Glycerol-transformed native lysozyme to a quasi-native state does not affect the conclusion that both global and internal fluctuations are slaved to glycerol relaxation.

Start-up of glycerol-driven denitrifying phosphorus removal from wastewater: The effects of the microaerobic environment.

Glycerol, an abundant by-product of biodiesel production, represented a promising carbon source for enhancing nutrient removal from low C/N ratio wastewater. This study discovered a novel approach to initiate glycerol-driven denitrifying phosphorus removal (DPR) in situ by creating a short-term microaerobic environment within the aerobic zone. This approach facilitated the in-situ conversion of glycerol, which was subsequently utilized by denitrifying phosphate accumulating organisms (DPAOs) for DPR. The feasibility and stability of glycerol-driven DPR were validated in a continuous-flow pilot-scale reactor. Anaerobic phosphorus release increased from 1.0 mg/L/h to 2.5 mg/L/h, with fermentation bacteria and related functional genes showing significant increases. The stable stage exhibited 92.8% phosphorus removal efficiency and 55.5% DPR percentage. The microaerobic environment enhanced fermentation bacteria enrichment, crucial for glycerol-driven DPR stability. The collaborative interaction between fermentation bacteria and phosphate accumulating organisms (PAOs) played a key role in sustaining glycerol-driven DPR stability. These findings provide a robust theoretical foundation for applying glycerol-driven DPR in established wastewater treatment plants.