Acetic Anhydride Research Articles

Microbial community changes and metabolic pathways analysis during waste activated sludge and meat processing waste anaerobic co-digestion

Waste activated sludge (WAS) and meat processing waste (MPW) were acted as co-substrates in anaerobic co-digestion (AcD), and biochemical methane potential (BMP) test was carried out to investigate the methane production performances. Microbial community structure and metabolic pathways analyses were conducted by 16S rRNA high-throughput sequencing and functional prediction analysis. BMP test results indicated that AcD of 70% WAS+30% MPW and 50% WAS+50% MPW (VS/VS) could significantly improve methane yield to 371.05 mL/g VS and 599.61 mL/g VS, respectively, compared with WAS acting as sole substrate (191.87 mL/g VS). The results of microbial community analysis showed that Syntrophomonas and Petrimonas became the dominant bacteria genera, and Methanomassiliicoccus and Methanobacterium became the dominant archaea genera after MPW addition. 16S functional prediction analysis results indicated that genes expression of key enzymes involved in syntrophic acetate oxidation (SAO), hydrogenotrophic and methylotrophic methanogenesis were up-regulated, and acetoclastic methanogenesis was inhibited after MPW addition. Based on these analyses, it could be inferred that SAO combined with hydrogenotrophic and methylotrophic methanogenesis was the dominant pathway for organics degradation and methane production during AcD. These findings provided systematic insights into the microbial community changes and metabolic pathways during AcD of WAS and MPW.

An Environmentally Friendly Solvent for Cellulose Acetate Production: Ethyl Lactate

The aim of this study was to investigate the possibilities of using biodegradable ethyl lactate instead of acetic acid, the most preferred solvent in industrial production, during cellulose acetate synthesis under certain conditions. In the study, dissolving pulp fibres, which were activated with water and acetic acid respectively, were dispersed in ethyl lactate after pressing and then subjected to acetylation reaction with acetic anhydride catalysed by sulfuric acid. The properties of acetylated cellulose were determined using X-Ray diffraction analysis, Fourier-transform infrared spectroscopy, Differential Scanning Calorimetry and other chemical analysis methods (viscosity, the percentage of bounded acid, chemical resistance). It was determined that it was possible to produce cellulose acetate with a DS value of 2.79 in ethyl lactate at 40°C using dissolving pulp. Fourier-transform infrared spectroscopy studies revealed a characteristic band broadening in the 1739 cm-1 region, indicating the presence of acetyl groups. The results of X-ray diffraction analyses also showed that the crystalline structure of cellulose was completely dispersed and band broadenings on diffractograms occurred. However, the calculation made on X-ray diffractograms also provided interesting findings in terms of crystallite sizes. It was observed that the crystallite sizes of the acetates were higher compared to the dissolving pulp. It was determined that the high crystallinity of the samples posed a problem in terms of processing properties such as dissolution. Among the solvents used, only Dimethylsulfoxide was found to have sufficient dissolving power.

Investigation on synergistic effect of CuO and Cu–O–Ce on efficient oxidation of ethyl acetate over CuO/CeO2 catalysts

Investigation on synergistic effect of CuO and Cu–O–Ce on efficient oxidation of ethyl acetate over CuO/CeO2 catalysts

Adsorption and separation of lead ions in phosphoric acid by co-doped carbon nanotubes with sulfur, oxygen, and manganese

The purpose of this study was to prepare a novel and efficient adsorbent for the removal of trace Pb(II) from phosphoric acid. Manganese oxide/sulfur-oxygen doped carbon nanotube composites (CNTs-S-O-Mn) were prepared from multi-walled carbon nanotubes modified with nitric acid, mercaptoacetic acid, acetic anhydride, concentrated sulfuric acid and potassium permanganate. Characterisation revealed that CNTs-S-O-Mn contained hydroxyl, sulphonate groups, MnO and Mn3O4. At 298 K, 15.362 mg·L-1 initial concentration of lead ions, and 18.4 % phosphoric acid concentration, the adsorption capacity of the composites for lead ions was 38.65 mg·g−1, which was higher than that of the unmodified CNTs, 8.04 mg·g−1. The adsorption kinetic data at 298 K conformed to the quasi-second-order kinetic equation, R2 = 0.997; 298, adsorption isotherms at 308 and 318 K conformed to the Langmuir equation, R2 = 0.990–––0.991. The adsorption capacity increased with decreasing phosphoric acid concentration. In this system, the adsorption is affected by a certain concentration of lead ions. Adsorption occurs via chemisorption and is exothermic. Sulfonic acid groups and hydroxyl groups on manganese oxides play an important role in the adsorption of Pb (II) through surface complexation with Pb (II). These results indicate that the removal of trace Pb (II) from phosphoric acid by modified carbon nanotubes is feasible, revealing a new use of carbon nanotubes for phosphoric acid purification.

Efficient enzymatic synthesis of d-α-tocopherol acetate by Carica papaya lipase-catalyzed acetylation of d-α-tocopherol in a solvent-free system

Enzymatic synthesis of d-α-tocopherol acetate is a promising approach for cost-effective production of liposoluble food and pharmaceutical antioxidant. In this study, we developed an efficient and environmentally friendly process that employed plant-derived Carica papaya lipase (CPL) as biocatalyst for the synthesis of d-α-tocopherol acetate in a solvent-free system. The optimum conditions were: 400 mg/mL d-α-tocopherol, 125 mg/mL CPL (44 U, aw = 0.328), with acetic anhydride as acyl donor, at 60°C and 200 rpm for 30 h. Interestingly, yields of 90.7%, 77.7%, 60.3% and 48.1% of d-α-tocopherol acetate were achieved after the “naturally immobilized” CPL recycled for 4 times, indicating a novel enzyme immobilization mechanism. Moreover, the synthetic product was purified from scale-up reaction, and confirmed by FTIR and GC-MS analysis, providing a total of 97.5% purity (91.1% d-α-tocopherol acetate plus 6.4% d-α-tocopherol). Overall, this work provided an efficient alternative method for green production of d-α-tocopherol acetate probably being applied in food and pharmaceutical industries, and this could also enhance the comprehensive utilization of Carica papaya.

Regulatory mechanisms of biochar alleviating ammonia inhibition to methanogenesis during long-term operation of anaerobic membrane bioreactor treating swine wastewater

Anaerobic Membrane Bioreactor (AnMBR) is regarded as an advanced technology for the anaerobic digestion (AD) of swine wastewater. However, the high ammonia level in swine wastewater is a crucial issue to inhibit methane production. This study aimed to establish a biochar-assisted AnMBR (BC) to mitigate ammonia inhibition during AD of swine wastewater. As the gradual ammonia loading rate (ALR) increased from 3.0 g/L to 6.0 g/L, BC maintained methane yields ranging from 74.9 % to 48.1 %, which were 9.1 % to 33.3 % higher than those of a regular AnMBR (CT). Moreover, the COD removal efficiency steadily surpassed 80 %. Although the increase in ALR had negative effects on both BC and CT, the presence of biochar enhanced the methanogenic activity of bulk sludge, specifically accelerating processes involved in syntrophic acetate oxidation and CO2-reducing methanogenesis under high ALR conditions. Integrated characterizations using PARAFAC-based EPS analysis, INT-ETS, and WO3 probe-based extracellular electron transfer (EET) activities indicated that biochar stimulated the secretion of humic-like and aromatic protein-like compounds in the TB-EPS, which likely established a redox-active matrix with biochar between VFA oxidation bacteria and CO2-reducing methanogens to facilitate EET. This potentially made the syntrophic VFA oxidation with favorably thermodynamic kinetics under high ammonia stress. Furthermore, the analysis of microbial community coupling prediction of functional genes associated with methanogenic pathways revealed that as ALR increased, the presence of biochar re-shaped the microbial communities by enriching electro-active Syntrophomonas and Methanosarcina/Methanospirillum, which mainly triggering EET-based syntrophic methanogenesis by accelerating the intracellular enzymatic transformation from Formylmethanofuran to 5,10-Methylenetetrahydromethanopterin process. We expect the major findings of this study provide new insights to understand the mechanisms of biochar alleviating ammonia inhibition during long-term operation of AnMBR.

A nano-imprinted graphene oxide-cellulose composite as a SERS active substrate.

Cellulose is a sustainable material capable of forming optically active nanoarrays on its surface. We created a composite of cellulose acetate (CA) and graphene oxide (GO), by mixing GO (0.1 mg mL-1) into CA. This was then imprinted with nanoscale surface features that form Bragg-like modes in resonance with the excitation laser when a thin layer of silver is vapor deposited onto the surface of the substrate. The addition of GO leads to improved surface-enhanced Raman scattering (SERS) signal strengths, obtaining an average SERS signal increase of 1.4-fold following the inclusion of GO. The combination of photonic and electromagnetic effects with charge transfer-based processes that support the SERS chemical mechanism and the possible presence of electromagnetic hot spots from the roughened surface results in an enhanced SERS signal strength when GO is added. This work shows the potential for nanoimprinted graphene oxide/cellulose acetate composites as flexible sensor platforms to detect target molecules.

One-pot synthesis of 4,5,6-triacetoxyisoindole-1,3‑dione derivatives by hydride transferring agent in Ac2O: Explanation of the regioselectivity in the molecule by theoretical calculations

Isoindole-1,3‑dione derivatives containing an epoxy alcohol unit were synthesized. Epoxy alcohols were converted to the corresponding 1,2,3-triacetate derivatives in a one-pot manner using the NaBH4/Ac20 system for the first time and in high yield. C-3 selective ring-opening products were obtained with good yield and with stereospecificity with the NaBH4/Ac2O system at 139–140 °C. The exact configuration of 2-methyl-1,3-dioxooctahydro-1H-isoindole-4,5,6-triyltriacetate was determined by X-ray diffraction analysis which was obtained from the reaction of epoxy alcohol by hydride transferring agent in acetic anhydride. On the other hand, theoretical computations were carried out to explain the regioselectivity in the ring opening reaction of epoxy alcohols. The results showed that the ring-opening reaction of epoxy alcohol proceeds in a thermodynamically controlled manner and regioselectivity occurs depending on the stability of the products.

Effect of Cannabis sativa L. extracts, phytocannabinoids and their acetylated derivates on the SHSY-5Y neuroblastoma cells’ viability and caspases 3/7 activation

BackgroundThere is a need for novel treatments for neuroblastoma, despite the emergence of new biological and immune treatments, since refractory pediatric neuroblastoma is still a medical challenge. Phyto cannabinoids and their hemisynthetic derivatives have shown evidence supporting their anticancer potential. The aim of this research was to examine Phytocannabinoids or hemisynthetic cannabinoids, which reduce the SHSY-5Y, neuroblastoma cell line’s viability.MethodsHexane and acetyl acetate extracts were produced starting with Cannabis sativa L. as raw material, then, 9-tetrahidrocannabinol, its acid counterpart and CBN were isolated. In addition, acetylated derivatives of THC and CBN were synthesized. The identification and purity of the chemicals was determined by High Performance Liquid Chromatography and 1H y 13C Magnetic Nuclear Resonance. Then, the capacity to affect the viability of SHSY-5Y, a neuroblastoma cell line, was examined using the resazurin method. Finally, to gain insight into the mechanism of action of the extracts, phytocannabinoids and acetylated derivatives on the examined cells, a caspase 3/7 determination was performed on cells exposed to these compounds.ResultsThe structure and purity of the isolated compounds was demonstrated. The extracts, the phytocannabinoids and their acetylated counterparts inhibited the viability of the SHSY 5Y cells, being CBN the most potent of all the tested molecules with an inhibitory concentration of 50 percent of 9.5 µM.ConclusionEach of the evaluated molecules exhibited the capacity to activate caspases 3/7, indicating that at least in part, the cytotoxicity of the tested phytocannabinoids and their hemi-synthetic derivatives is mediated by apoptosis.

Effects of twelve organic solvents on process hazards and reaction selectivity of m-xylene nitration: Combined calorimetric technique and DFT calculations

Selection of appropriate solvents is of great significance for achieving efficient and safe m-xylene nitration. In this study, the exothermic behavior of m-xylene nitration under twelve organic solvents were studied by reaction calorimetry. The process hazards were assessed by calculation of thermal safety parameters. The reaction selectivity was quantified. The anhydrides exhibited the prominent promotion as water absorbent and active reagent in nitration, followed by selected aliphatic hydrocarbons, chlorinated hydrocarbons. While acetic acid, propionic acid and ethyl acetate had negative effects. The maximum yield of 95.6 % was achieved in nitric acid-acetic anhydride (HNO3-Ac2O) system. The thermal weight loss of the products appeared at around 150−230 °C by thermogravimetry experiments. The activation energy was determined to be 192.9 kJ/mol through adiabatic kinetic calculations. Furthermore, the solvent effects were analyzed by calculating the solvent polarity and its interaction energy with HNO3. The different action mechanisms of Ac2O and Pc2O on NO2+ generation were revealed. The density functional theory calculations were in agreement with the experimental results. These findings can provide guidance for understanding of the solvent effect on the reaction hazards and selectivity of m-xylene nitration, and provide experimental and theoretical basis for solvent screening for the nitration of similar aromatic compounds.

Adsorption simulation of 2,4-D pesticide on novel zinc-based 2-amino-4-(1H-1,2,4-triazole-4-yl)benzoic acid coordination complexes using machine learning approach.

The capacity of zinc-based 2-amino-4-(1H-1,2,4-triazole-4-yl)benzoic acid coordination complex (Zn(NH2-TBA)2) and modified Zn(NH-TBA)2COMe complex for removal of 2,4-dichlorophenoxyacetic acid (2,4-D) from aqueous solutions was investigated through adsorption modeling and artificial intelligence tools. Analyzing the adsorption characteristics of pesticides helps in studying the groundwater pollution by pesticides in agriculture area.In this study, Zn(NH2-TBA)2 was synthesized using Schiff base and its surface was modified using acetic anhydride group and their physical characteristics were identified using proton NMR, FTIR, and XRD. NMR results showed maximum modification yield obtained was 65% after 5days. The porous structure and surface area monitored using nitrogen isotherm and BET surface area analysis presented relatively less surface area and porosity after modification. Adsorption modelling indicated that Toth model with a maximum adsorption capacity of 150.8mg/g and 100.7mg/g represents the homogenous adsorption systems which satisfy both low- and high-end boundary of adsorbate concentration in all settings according to the optimum point, while the kinetics and rate of 2,4-D adsorption follow the pseudo-first-order kinetic model in all situations. Artificial neural network (ANN), support vector regression, and particle swarm optimized least squares-support vector regression (PSO-LSSVR) were used for the optimization and modelling of adsorbent mass, adsorbate concentration, contact time, and temperature to develop predictive equations for the simulation of the adsorption efficiency of 2,4-D pesticide. The obtained results exhibited the better performance of ANN and PSO-LSSVR for prediction of adsorption results. The mean square error values of ANN (0.001, 0.012) and PSO-LSSVR (0.121, 0.105) were obtained for Zn(NH2-TBA)2 and Zn(NH-TBA)2COMe, respectively, while their respective coefficient of determination (R2) obtained were 0.999 and 0.988 for ANN and 0.980 and 0.825 for PSO-LSSVR. The study specified that machine learning predictive behavior performed better for Zn(NH2-TBA)2 compared to Zn(NH-TBA)2COMe that is also supported by theoretical kinetics and isotherm models. The research concludes that artificial intelligence models are the most efficient tools for studying the predictive behavior of adsorption data.

Insights into the Mechanism of Catalytic Activity of Plasmodium Parasite Malate-Quinone Oxidoreductase.

Plasmodium malate-quinone oxidoreductase (MQO) is a membrane flavoprotein catalyzing the oxidation of malate to oxaloacetate and the reduction of quinone to quinol. Recently, using a yeast expression system, we demonstrated that MQO, expressed in place of mitochondrial malate dehydrogenase (MDH), contributes to the TCA cycle and the electron transport chain in mitochondria, making MQO attractive as a promising drug target in Plasmodium malaria parasites, which lack mitochondrial MDH. However, there is little information on the structure of MQO and its catalytic mechanism, information that will be required to develop novel drugs. Here, we investigated the catalytic site of P. falciparum MQO (PfMQO) using our yeast expression system. We generated a model structure for PfMQO with the AI tool AlphaFold and used protein footprinting by acetylation with acetic anhydride to analyze the surface topology of the model, confirming the computational prediction to be reasonably accurate. Moreover, a putative catalytic site, which includes a possible flavin-binding site, was identified by this combination of protein footprinting and structural prediction model. This active site was analyzed by site-directed mutagenesis. By measuring enzyme activity and protein expression levels in the PfMQO mutants, we showed that several residues at the active site are essential for enzyme function. In addition, a single substitution mutation near the catalytic site resulted in enhanced sensitivity to ferulenol, an inhibitor of PfMQO that competes with malate for binding to the enzyme. This strongly supports the notion that the substrate binds to the proposed catalytic site. Then, the location of the catalytic site was demonstrated by structural comparison with a homologous enzyme. Finally, we used our results to propose a mechanism for the catalytic activity of MQO by reference to the mechanism of action of structurally or functionally homologous enzymes.

An efficient methodological approach for synthesis of selenopyridines: generation, reactions, anticancer activity, EGFR inhibitory activity and molecular docking studies.

In the present work, we successfully synthesized Se-alkyl selenopyridines 1 and 3, selenopheno[2,3-b]pyridine 2, and bis-selenopyridine 4 derivatives using an eco-friendly method by utilizing NaHSe instead of toxic hydrogen selenide. The effect of the temperature on the reaction was screening at various temperatures. The regiospecific reaction of selenopyridine 1 with bromine afforded an unexpected product 4,6-diamino-5-bromo-2-[(cyanomethyl)selenyl]-pyridine-3-carbonitrile (5), which was cyclized to selenopheno[2,3-b]pyridine (7) by refluxing in the presence of TEA. While its treatment with thiophenol and/or p-chlorothiophenol gave 8a, b. On the other hand, its reaction with aminothiophenol afforded 2-(benzo[d]-thiazol-2-yl)-5-bromoselenopheno[2,3-b]pyridine-3,4,6-triamine (9). Also, N-(2-cyano-4-methyl-5H-1-seleno-3,5,8-triazaacenaphthylen-7-yl)acetamide (11) and a novel series of selenoazo dyes 12a-d were synthesized by treatment of selenopheno[2,3-b]pyridine 2 with acetic anhydride and/or diazonium chlorides of aromatic amines, respectively. Then, we ascertained the potential activity of synthesized compounds against highly metastatic prostate cancer cells (PC-3) and osteosarcoma cells (MG-63) and found that 12a, 12b, 12c, and 12d were more cytotoxic than doxorubicin in both tested cell lines, showing nearly the same anticancer activity with IC50 values ranging from 2.59 ± 0.02µM to 3.93 ± 0.23µM. Mechanistically, the most potent compounds 12a and 12b proved to be potent EGFR inhibitors with IC50 values of 0.301 and 0.123µM, respectively, compared to lapatinib as a positive reference (IC50 = 0.049µM). Moreover, the docking results are in good agreement with the anticancer activity as well as the EGFR inhibitory activity, suggesting these two compounds as promising EGFR anticancer candidates.

Magnetite particles accelerate methanogenic degradation of highly concentrated acetic acid in anaerobic digestion process

The anaerobic digestion (AD) process has become significant for its capability to convert organic wastewater into biogas, a valuable energy source. Excessive acetic acid accumulation in the anaerobic digester can inhibit methanogens, ultimately leading to the deterioration of process performance. Herein, the effect of magnetite particles (MP) as an enhancer on the methanogenic degradation of highly-concentrated acetate (6 g COD/L) was examined through long-term sequential AD batch tests. Bioreactors with (AM) and without (AO) MP were compared. AO experienced inhibition and its methane production rate (qm) converged to 0.45 L CH4/g VSS/d after 10 sequential batches (AO10, the 10th batch in a series of the sequential batch tests conducted using bioreactors without MP addition). In contrast, AM achieved 3–425% higher qm through the sequential batches, indicating that MP could counteract the inhibition caused by the highly-concentrated acetate. MP addition to inhibited bioreactors (AO10) successfully restored them, achieving qm of 1.53 L CH4/g VSS/d, 3.4 times increase from AO10 after 8 days lag time, validating its potential as a recovery strategy for inhibited digesters with acetate accumulation. AM exhibited higher microbial populations (1.8–3.8 times) and intracellular activity (9.3 times) compared to AO. MP enriched Methanosaeta, Peptoclostridium, Paraclostridium, OPB41, and genes related to direct interspecies electron transfer and acetate oxidation, potentially driving the improvement of qm through MP-mediated methanogenesis. These findings demonstrated the potential of MP supplementation as an effective strategy to accelerate acetate-utilizing methanogenesis and restore an inhibited anaerobic digester with high acetate accumulation.

Metal-modified CeO2 nanobelts for catalytic oxidation of ethyl acetate: Roles of surface lattice oxygen and hydrolysis behaviors

Based on the comparison of three counterparts [Mg-, Al- and Cu-modified CeO2 nanobelts], it is illustrated that the low-temperature hydrolysis of ethyl acetate (EA) rapidly occurs and the removal of absorbed hydrolyzates (mainly acetates) is considered the rate-determining step. As the temperature increases, the escape of surface lattice oxygen (Os-latt) breaks through the shield of hydrolyzates and promotes the deep oxidation of intermediate products. Additionally, the activated Os-latt also facilitates the C-O bond cleavage in EA by nucleophilic attack. Due to the Os-latt escape, surface metal ions achieve more unsaturated coordinations to absorb the CH3COO– species, leading to competitive adsorption between O2 and hydrolyzates. The Cu modification strengthens the coordination adsorption of CH3COO– species, and the Al modification exhibits a low Os-latt mobility. These factors all contribute to a strong adsorption or accumulation of surface hydrolyzates. Therefore, a favorable ester hydrolysis-oxidation process over CeO2-based catalysts depends not only on excellent Os-latt mobility but also the suitable adsorption capacity of hydrolyzates, such as Mg modification.

Hemisynthesis of Pentacyclic Triterpenoids from Diospyros foxworthyi with In vitro and In silico Anti-malarial Evaluation

Abstract: A total of twelve pentacyclic triterpenoid derivatives based on betulin (1) and lupeol (2) scaffolds isolated from Diospyros foxworthyi were hemisynthesized by acylation or acetylation reactions with appropriate acid chloride or acetic anhydride. The structures of the hemisynthesised compounds were characterised by means of FT-IR, 1D- and 2D-NMR, as well as HRMS analysis. These compounds were assayed for in vitro anti-malarial studies by inhibition of β-hematin formation assay with chloroquine as a positive control. Compounds 1d and 2f showed the strongest potential as β-hematin formation inhibitors with IC50 values of 6.66 ± 1.36 and 11.89 ± 0.15 μM, respectively, compared with the positive control (chloroquine; IC50 = 37.50 ± 0.60 μM). In silico molecular docking simulations were performed using AutoDock Vina for compounds 1d and 2f to investigate the binding interactions and free energy of binding (FEB) with the hemozoin supercell crystal structure (CCDC number: XETXUP01). The findings revealed several hydrophobic interaction modes between the 1d, 2f and hemozoin, with calculated FEBs of -8.4 ± 0.2 and -8.9 ± 0.0 kcal mol-1, indicating strong and favourable interactions.

In-situ acetylation followed by liquid-liquid extraction and gas chromatography – mass spectrometry for the determination of bromophenols in urine

A novel and simple method combining in-situ acetylation, liquid-liquid extraction and gas chromatography-mass spectrometry (GC-MS) has been developed for the quantification of 10 bromophenols in urine, used as biomarkers of exposure to polybrominated diphenyl ethers. The analytical process involves an enzymatic hydrolysis of the bromophenol glucuronide fraction followed by an aqueous derivatization of the phenol group with acetic anhydride. A subsequent liquid-liquid extraction of the sample with hexane allows the injection of the organic layer, using a programmed temperature vaporizer, into a gas chromatograph coupled to a single quadrupole mass spectrometer. Quantification is performed by the standard addition method. Limits of detection are in the pg mL−1 range. Trueness, assessed in terms of percentages of recovery, varies between 100 % and 118 % in synthetic urine and between 79 % and 117 % in human urine. Precision, assessed at two different levels, 0.25 ng mL−1 and 2.5 ng mL−1, shows values of relative standard deviation below 14 % both in intra- and inter-day studies for both matrices. The method has been applied to the analysis of seven urine samples, measuring concentrations higher than the LOQ in three of them. These levels are in agreement with others found in literature, but they have been obtained by applying a much simpler and faster protocol. In addition, the replacement of silylating reagents by acetic anhydride, to derivatize the phenol moiety, provides a greener alternative to other GC-MS procedures published up to date.

Lysine acetylation of Hsp16.3: Effect on its structure, chaperone function and influence towards the growth of Mycobacterium tuberculosis

Hsp16.3 plays a vital role in the slow growth of Mycobacterium tuberculosis via its chaperone function. Many secretory proteins, including Hsp16.3 undergo acetylation in vivo. Seven lysine (K) residues (K64, K78, K85, K114, K119, K132 and K136) in Hsp16.3 are acetylated inside pathogen. However, how lysine acetylation affects its structure, chaperone function and pathogen's growth is still elusive. We examined these aspects by executing in vitro chemical acetylation (acetic anhydride modification) and by utilizing a lysine acetylation mimic mutant (Hsp16.3-K64Q/K78Q/K85Q/K114Q/K119Q/K132Q/K136Q). Far- and near-UV CD measurements revealed that the chemically acetylated proteins(s) and acetylation mimic mutant has altered secondary and tertiary structure than unacetylated/wild-type protein. The chemical modification and acetylation mimic mutation also disrupted the oligomeric assembly, increased surface hydrophobicity and reduced stability of Hsp16.3, as revealed by GF-HPLC, 4,4′-dianilino-1,1′-binaphthyl-5,5′-disulfonic acid binding and urea denaturation experiments, respectively. These structural changes collectively led to an enhancement in chaperone function (aggregation and thermal inactivation prevention ability) of Hsp16.3. Moreover, when the H37Rv strain expressed the acetylation mimic mutant protein, its growth was slower in comparison to the strain expressing the wild-type/unacetylated Hsp16.3. Altogether, these findings indicated that lysine acetylation improves the chaperone function of Hsp16.3 which may influence pathogen's growth in host environment.

Metagenomes from microbial populations beneath a chromium waste tip give insight into the mechanism of Cr (VI) reduction

Dumped Chromium Ore Processing Residue (COPR) at legacy sites poses a threat to health through leaching of toxic Cr(VI) into groundwater. Previous work implicates microbial activity in reducing Cr(VI) to less mobile and toxic Cr(III), but the mechanism has not been explored. To address this question a combined metagenomic and geochemical study was undertaken. Soil samples from below the COPR waste were used to establish anaerobic microcosms which were challenged with Cr(VI), with or without acetate as an electron donor, and incubated for 70 days. Cr was rapidly reduced in both systems, which also reduced nitrate, nitrite then sulfate, but this sequence was accelerated in the acetate amended microcosms. 16S rRNA gene sequencing revealed that the original soil sample was diverse but both microcosm systems became less diverse by the end of the experiment. A high proportion of 16S rRNA gene reads and metagenome-assembled genomes (MAGs) with high completeness could not be taxonomically classified, highlighting the distinctiveness of these alkaline Cr impacted systems. Examination of the coding capacity revealed widespread capability for metal tolerance and Fe uptake and storage, and both populations possessed metabolic capability to degrade a wide range of organic molecules. The relative abundance of genes for fatty acid degradation was 4× higher in the unamended compared to the acetate amended system, whereas the capacity for dissimilatory sulfate metabolism was 3× higher in the acetate amended system. We demonstrate that naturally occurring in situ bacterial populations have the metabolic capability to couple acetate oxidation to sequential reduction of electron acceptors which can reduce Cr(VI) to less mobile and toxic Cr(III), and that microbially produced sulfide may be important in reductive precipitation of chromate. This capability could be harnessed to create a Cr(VI) trap-zone beneath COPR tips without the need to disturb the waste.

Time-resolved spectroscopic investigation for the practical application of a photocatalytic air purifier

The photocatalytic efficiency for removing volatile organic compounds (VOCs) is significantly influenced by operational parameters like humidity and flow velocity, exhibiting notable and inconsistent fluctuations in both lab-scale and large-scale demonstrations. In this study, operando spectroscopy and isotope analysis were employed to investigate the correlation between humidity levels and degradation of gaseous acetaldehyde using TiO2 photocatalysts, aiming to demonstrate the scaling-up of photocatalytic air purifier. It was observed that rate constants for the mineralization of acetaldehyde rapidly decreased by 30% as relative humidity increased from 25% to 80% in the flow system (with an air velocity, v = 0.78 m/s). However, batch system showed smaller change with only a 10% reduction of the rate constant. Humidity fluctuations were more pronounced under high-speed conditions and were amplified in air purifier (v = 3.8 m/s). Time-resolved operando spectroscopy using an 13C isotope of acetaldehyde revealed that humidity’s distinct role in dark adsorption and photocatalytic reactions. Water was found to inhibit the formation of crotonaldehyde during aldol condensation reaction in dark condition. Moreover, water suppressed photocatalytic mineralization by inhibiting acetate oxidation to formate. These findings provide valuable insights for improving realistic air purification processes, underscoring the importance of identifying key intermediates and controlling humidity to enhance the selectivity of gaseous pollutant oxidation reactions.