Transfer Of Substrates Research Articles

Chalcogen Bond-Driven Alkylations: Selenoxide-Pillar[5]arene As A Recyclable Catalyst For Displacement Reactions In Water.

A novel strategy to catalyze alkylation reactions through chalcogen bond interaction using a supramolecular structure is presented herein. Utilizing just 1.0 mol% of selenoxide-pillar[5]arene (P[5]SeO) as the catalyst we achieved efficient catalysis in the cyanation of benzyl bromide in water. Our approach demonstrated high efficiency and effectiveness, with the results supported by designed control experiments and theoretical models, highlighting the catalytic effect of the pillar[5]arene through noncovalent interactions. Quantum-chemical calculations (ωB97X-D/def2-TZVP@SMD) pointed out that the catalyzed cyanation reaction followed an SN2-like mechanism, with energy barriers (ΔH‡) ranging from 16.7 to 18.2 kcal mol-1, exhibiting dissociative character depending on the para-substituent. 1H NMR analysis revealed that P[5]SeO acted as a catalyst through inclusion complex formation, facilitating the transfer of the electrophilic substrate to the aqueous solution for nucleophilic displacement. Our reaction protocol proved applicable to various substrates, including aromatic and alpha-carbonyl derivatives. The use of sodium azide as the nucleophile was also feasible. Importantly, our method allowed scalability, and the catalyst P[5]SeO could be recovered and reused effectively for multiple reaction cycles, showcasing sustainability.

A hybrid TIM complex mediates protein import into hydrogenosomes of Trichomonas vaginalis

BackgroundHydrogenosomes are a specific type of mitochondria that have adapted for life under anaerobiosis. Limited availability of oxygen has resulted in the loss of the membrane-associated respiratory chain, and consequently in the generation of minimal inner membrane potential (Δψ), and inefficient ATP synthesis via substrate-level phosphorylation. The changes in energy metabolism are directly linked with the organelle biogenesis. In mitochondria, proteins are imported across the outer membrane via the Translocase of the Outer Membrane (TOM complex), while two Translocases of the Inner Membrane, TIM22, and TIM23, facilitate import to the inner membrane and matrix. TIM23-mediated steps are entirely dependent on Δψ and ATP hydrolysis, while TIM22 requires only Δψ. The character of the hydrogenosomal inner membrane translocase and the mechanism of translocation is currently unknown.ResultsWe report unprecedented modification of TIM in hydrogenosomes of the human parasite Trichomonas vaginalis (TvTIM). We show that the import of the presequence-containing protein into the hydrogenosomal matrix is mediated by the hybrid TIM22-TIM23 complex that includes three highly divergent core components, TvTim22, TvTim23, and TvTim17-like proteins. The hybrid character of the TvTIM is underlined by the presence of both TvTim22 and TvTim17/23, association with small Tim chaperones (Tim9-10), which in mitochondria are known to facilitate the transfer of substrates to the TIM22 complex, and the coupling with TIM23-specific ATP-dependent presequence translocase-associated motor (PAM). Interactome reconstruction based on co-immunoprecipitation (coIP) and mass spectrometry revealed that hybrid TvTIM is formed with the compositional variations of paralogs. Single-particle electron microscopy for the 132-kDa purified TvTIM revealed the presence of a single ring of small Tims complex, while mitochondrial TIM22 complex bears twin small Tims hexamer. TvTIM is currently the only TIM visualized outside of Opisthokonta, which raised the question of which form is prevailing across eukaryotes. The tight association of the hybrid TvTIM with ADP/ATP carriers (AAC) suggests that AAC may directly supply ATP for the protein import since ATP synthesis is limited in hydrogenosomes.ConclusionsThe hybrid TvTIM in hydrogenosomes represents an original structural solution that evolved for protein import when Δψ is negligible and remarkable example of evolutionary adaptation to an anaerobic lifestyle.

Pickering Emulsions Biocatalysis: Recent Developments and Emerging Trends.

Biocatalysis within biphasic systems is gaining significant attention in the field of synthetic chemistry, primarily for its ability to solve the problem of incompatible solubilities between biocatalysts and organic compounds. By forming an emulsion from these two-phase systems, a larger surface area is created, which greatly improves the mass transfer of substrates to the biocatalysts. Among the various types of emulsions, Pickering emulsions stand out due to their excellent stability, compatibility with biological substances, and the ease with which they can be formed and separated. This makes them ideal for reusing both the emulsifiers and the biocatalysts. This review explores the latest developments in biocatalysis using Pickering emulsions. It covers the structural features, methods of creation, innovations in flow biocatalysis, and the role of interfaces in these processes. Additionally, the challenges and future directions are discussed in combining chemical and biological catalysts within Pickering emulsion frameworks to advance synthetic methodologies.

Transition metals doped Zr2CF2 as promising sensor and adsorbent for NH3

The effective adsorption and treatment of toxic gases have attracted great attention amongst researchers, particularly the use of new two-dimensional materials for achieving the adsorption of toxic gases. We applied the density functional theory to investigate the stability, geometric structural, electrical, and magnetic properties of NH3 molecules adsorbed on pristine, F-vacancy defected, and transition-metals (V, Cr, Mn, Y, Nb, and Mo) doped Zr2CF2. The charge transfer and adsorption energy of the original substrate were low, indicating that the interaction between NH3 and Zr2CF2 was primarily physical adsorption. After introducing F-vacancy and TM-doping, the adsorption stability was enhanced, with the large charge transfer and high adsorption energies indicating that the adsorption of NH3 transitioned to chemisorption. Electronic density of state diagrams suggested that the enhanced interaction was mainly caused by the hybridization of the d orbitals of dopants and p orbitals of the adsorbed NH3 molecules. The adsorption energy of NH3 molecules on Y-doped Zr2CF2 was the highest at a value of −1.425eV, and the material was also explored as an adsorbent for NH3 gas. After adsorbing six NH3 molecules, the Y-doped substrate had an average adsorption energy of −1.350eV and still maintained a good thermal stability at a temperature of 300 K. These results can provide insights into the development of sensors and adsorbents based on MXenes.

Cadmium transport by mammalian ATP-binding cassette transporters.

Cellular responses to toxic metals depend on metal accessibility to intracellular targets, reaching interaction sites, and the intracellular metal concentration, which is mainly determined by uptake pathways, binding/sequestration and efflux pathways. ATP-binding cassette (ABC) transporters are ubiquitous in the human body-usually in epithelia-and are responsible for the transfer of indispensable physiological substrates(e.g. lipids and heme), protection against potentially toxic substances, maintenance of fluid composition, and excretion of metabolic waste products. Derailed regulation and gene variants of ABC transporters culminate in a wide array of pathophysiological disease states, such as oncogenic multidrug resistance or cystic fibrosis. Cadmium (Cd) has no known physiological role in mammalians and poses a health risk due to its release into the environment as a result of industrial activities, and eventually passes into the food chain. Epithelial cells, especially within the liver, lungs, gastrointestinal tract and kidneys, are particularly susceptible to the multifaceted effects of Cd because of the plethora of uptake pathways available. Pertinent to their broad substrate spectra, ABC transporters represent a major cellular efflux pathway for Cd and Cd complexes. In this review, we summarize current knowledge concerning transport of Cd and its complexes (mainly Cd bound to glutathione) by the ABC transporters ABCB1 (P-glycoprotein, MDR1), ABCB6, ABCC1 (multidrug resistance related protein 1, MRP1), ABCC7 (cystic fibrosis transmembrane regulator, CFTR), and ABCG2 (breast cancer related protein, BCRP). Potential detoxification strategies underlying ABC transporter-mediated efflux of Cd and Cd complexesare discussed.

Differentiation of prokaryotic community composition in suspended sediment size-fractionation in a small loess watershed

The transfer of particle-bound prokaryotes and substrates from slopes to rivers during soil erosion is a crucial dynamic process that greatly influences terrestrial biological and geochemical cycles. Particle size strongly affects the transport distance of suspended sediments and likely induces prokaryote and substrate variations along the course of the river. Previous studies have mainly focused on the spatial redistribution of soil prokaryotes, whereas the variation in prokaryotic diversity and community composition among different particle size-fractions in suspended sediments remain unclear. In this study, suspended sediments were collected at four plots (upstream, middle stream, downstream, and dam) along a river that runs through a valley. Sampled sediments were size-fractionated into > 63 μm and < 63 μm particles (referred to as small and large particle fractions hereafter), to identify the corresponding prokaryotic community composition, interactions, and functions. The richness index (Chao1 and observed species) and the diversity index (Shannon) of sediment-associated prokaryotic communities significantly decreased from upstream towards dam plots, except in the small particle fraction in downstream and dam plots. The highest β-diversity index value was found for the large particle fractions, while the geospatial contribution to β-diversity varied. The concentration of Proteobacteria in downstream (41.39 %) and dam (34.86 %) plots was lower than that in the upstream (47.65 %) and middle stream (45.90 %) plots. In contrast, Cyanobacteria (4.89 % and 6.75 %) and Verrucomicrobiota (5.98 % and 11.00 %) concentrations were greater in the downstream and dam plots, respectively, than those in the upstream (0.93 % and 2.43 %) and middle stream (3.37 % and 3.11 %) plots, respectively. Additionally, the concentrations of Verrucomicrobiota and Bacteroidota were significantly higher in the small particle fractions (2.26–8.12 % and 4.33–7.72 %) than those in the large particle fractions (2.03–5.94 % and 3.70–7.11 %). Compared to the other plots, upstream plots had a larger clustered network and a greater number of co-occurrences within the prokaryotic community in the bulk sediment and in the small particle fraction, whereas more complex interactions among prokaryotic communities were found in the large particle fraction from the dam plots. Soil organic carbon (SOC), total nitrogen (TN), and Olsen phosphorus were the three most influential factors, explaining 67.41 % of the variation in prokaryotic community. Proteobacteria and Myxococcota correlated negatively with SOC, TN, and Olsen P but positively with C = O. Conversely, Cyanobacteria and Verrucomicrobiota correlated positively with SOC, TN, and Olsen P but negatively with C = O. Functional groups linked to the biogeochemical cycling of carbon (methanol oxidation and methylotrophy) were found in high concentrations, whereas those linked to nitrogen (nitrogen and nitrate respiration) were found in low concentrations in upstream and middle stream plots. Reassembly of the sediment physicochemical characteristics among geospatial plots along the transport transect significantly altered the prokaryotic community composition and metabolic functional groups.

Solute Carrier Transporters in Synovial Membrane and Hoffa's Pad of Patients with Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a complex autoimmune disease that leads to joint destruction. A number of immune cells that affect joint tissues are involved in the pathogenesis of this disease. This leads to the synthesis of many pro-inflammatory mediators. The transport of drugs, as well as many cytokines involved in the development of inflammation in RA patients, is mediated by membrane transporters. Membrane transporters are proteins that mediate the transfer of substrates across biological membranes. But to date there are no studies examining the expression of solute carrier (SLC) transporters in joint tissues. The aim of the study was to evaluate the expression of individual SLC family transporters in the synovial membranes (SMs) and infrapatellar fat pad (Hoffa's pad) of RA patients. The study included 20 patients with rheumatoid arthritis and 20 with osteoarthritis as the control group who were undergoing joint replacement surgery as a normal part of clinical care. In the SM and Hoffa's pad of RA patients the following 17 membrane transporters were defined at relevant expression levels for SLC transporter superfamily: SLC15A2, SLC16A3, SLC19A1, SLC2A9, SLC22A1, SLC22A3, SLC22A4, SLC22A5, SLC22A18, SLC33A1, SLC47A1, SLC51A, SLC7A5, SLC7A6, SLC01C1, SLC02B1, SLC04A1. The confirmed expression of these transporters in the SMs as well as Hoffa's pad of patients with RA and OA, and the differences in their expression between these groups, suggests the involvement of SLC transporters in both the maintenance of homeostasis under physiological conditions in the tissues of the joints, as well as in the inflammatory process in RA.

Graphene coating reduces the heat transfer performance of water vapor condensation on copper surfaces: A molecular simulation study

Water vapor condensation is a key process for many applications. Existing studies in water vapor heterogeneous condensation have used different methods to modify the hydrophobicity of the water vapor condensation surface. One way to modify the surface properties is to use graphene coating. On a macroscale, surface modification using graphene coating can improve water vapor condensation heat transfer of various substrates. However, on the molecular scale, its effect is poorly understood. This work investigated the effect of graphene coating on water vapor condensation using molecular dynamics simulations (MDS). We examined the water vapor condensation on bare copper surfaces considering the effects of initial temperature difference, water model, and surface size, parameters that have not been investigated by previous studies employing MDS for the same water-surface configuration. One water model was then used to simulate the condensation on a copper surface with and without graphene coating. We then investigated the effect of graphene defect, the energy and vibration of graphene atoms, and the interaction between graphene and copper layers. The surface size notably influenced the condensation rate and heat transfer performance. The condensation rate and heat transfer performance were significantly reduced when the copper surface was coated by graphene. The number of water molecules condensed was 1253 molecules/ns on the bare copper surface, compared to 587 molecules/ns on the graphene-coated copper surface. Moreover, the water molecules condensed on the graphene-coated copper surface tended to return to the bulk vapor phase. Other important results are also provided. This study gives an insight into the water vapor condensation on graphene-coated copper surfaces, useful to pursuit the design and optimization of graphene-coated copper surfaces for applications that need efficient water vapor condensation, such as for industrial applications, like thermal, chemical, and nuclear.

Equine maternal aging affects the metabolomic profile of oocytes and follicular cells during different maturation time points.

Introduction: Oocyte quality and fertility decline with advanced maternal age. During maturation within the ovarian follicle, the oocyte relies on the associated somatic cells, specifically cumulus and granulosa cells, to acquire essential components for developmental capacity. Methods: A nontargeted metabolomics approach was used to investigate the effects of mare age on different cell types within the dominant, follicular-phase follicle at three time points during maturation. Metabolomic analyses from single oocytes and associated cumulus and granulosa cells allowed correlations of metabolite abundance among cell types. Results and Discussion: Overall, many of the age-related changes in metabolite abundance point to Impaired mitochondrial metabolic function and oxidative stress in oocytes and follicular cells. Supporting findings include a higher abundance of glutamic acid and triglycerides and lower abundance of ceramides in oocytes and somatic follicular cells from old than young mares. Lower abundance of alanine in all follicular cell types from old mares, suggests limited anaerobic energy metabolism. The results also indicate impaired transfer of carbohydrate and free fatty acid substrates from cumulus cells to the oocytes of old mares, potentially related to disruption of transzonal projections between the cell types. The identification of age-associated alterations in the abundance of specific metabolites and their correlations among cells contribute to our understanding of follicular dysfunction with maternal aging.

Investigating the effect of anode materials on the performance and microbial community in an integrated chamber-free microbial fuel cell

Conventionally, there are two basic configurations for microbial fuel cell (MFC), one with anode and cathode chambers, as known as dual-chamber MFC, one with only anode chamber and a air–cathode, called single-chamber MFC. However, electrode materials' complex configuration and high resistance limit their practical applications in power generation and wastewater purification. To this end, we constructed three MFCs using carbon felt, Ni/Ti, and Ag-Ni/Ti as anodes based on our previously constructed integrated chamber-free MFC (iMFC). After 21 days of operation, the Ag-Ni/Ti-MFC performed best in bioelectricity output and removal of target pollutants. The bioelectricity output parameters were an open-circuit voltage of 0.773 V, a maximum current density of 552.1429 mA/m2, and a power density of 426.8064 mW/m2. Furthermore, the MFC achieved removals of 67.90% for the chemical oxygen demand (COD), 55.20% for total nitrogen (TN), and 61.83% for ammonia nitrogen (NH3-N). Introducing silver nanoparticles to the transmembrane and outer membrane boosted the charge extraction efficiency in the MFCs. Moreover, 16S rDNA analysis indicated that typical electricity-producing bacteria, including Comamonas, Paraclostridium, and Asaccharospora, had a higher relative abundance in the Ag-Ni/Ti-MFC anode than in other anodes. Overall, the surface structure of different anode materials plays a critical role in bioelectricity generation and wastewater purification using iMFC by affecting the mass transfer of substrates and metabolites.

Engineering BioBricks for Deoxysugar Biosynthesis and Generation of New Tetracenomycins.

Tetracenomycins and elloramycins are polyketide natural products produced by several actinomycetes that exhibit antibacterial and anticancer activities. They inhibit ribosomal translation by binding in the polypeptide exit channel of the large ribosomal subunit. The tetracenomycins and elloramycins are typified by a shared oxidatively modified linear decaketide core, yet they are distinguished by the extent of O-methylation and the presence of a 2',3',4'-tri-O-methyl-α-l-rhamnose appended at the 8-position of elloramycin. The transfer of the TDP-l-rhamnose donor to the 8-demethyl-tetracenomycin C aglycone acceptor is catalyzed by the promiscuous glycosyltransferase ElmGT. ElmGT exhibits remarkable flexibility toward transfer of many TDP-deoxysugar substrates to 8-demethyltetracenomycin C, including TDP-2,6-dideoxysugars, TDP-2,3,6-trideoxysugars, and methyl-branched deoxysugars in both d- and l-configurations. Previously, we developed an improved host, Streptomyces coelicolor M1146::cos16F4iE, which is a stable integrant harboring the required genes for 8-demethyltetracenomycin C biosynthesis and expression of ElmGT. In this work, we developed BioBricks gene cassettes for the metabolic engineering of deoxysugar biosynthesis in Streptomyces spp. As a proof of concept, we used the BioBricks expression platform to engineer biosynthesis for d-configured TDP-deoxysugars, including known compounds 8-O-d-glucosyl-tetracenomycin C, 8-O-d-olivosyl-tetracenomycin C, 8-O-d-mycarosyl-tetracenomycin C, and 8-O-d-digitoxosyl-tetracenomycin C. In addition, we generated four new tetracenomycins including one modified with a ketosugar, 8-O-4'-keto-d-digitoxosyl-tetracenomycin C, and three modified with 6-deoxysugars, including 8-O-d-fucosyl-tetracenomycin C, 8-O-d-allosyl-tetracenomycin C, and 8-O-d-quinovosyl-tetracenomycin C. Our work demonstrates the feasibility of BioBricks cloning, with the ability to recycle intermediate constructs, for the rapid assembly of diverse carbohydrate pathways and glycodiversification of a variety of natural products.

MOFs-derived CuO–Fe3O4@C with abundant oxygen vacancies and strong Cu–Fe interaction for deep mineralization of bisphenol A

A novel CuO–Fe3O4 encapsulated in the carbon framework with abundant oxygen vacancies (CuO–Fe3O4@C) was successfully prepared by thermal conversion of Cu(OAc)2/Fe-metal organic framework. The as-prepared catalyst exhibited excellent peroxymonosulfate (PMS) activation performance, good recyclability and fast magnetic separation. Under optimal conditions, the added BPA (60 mg/L) could be completely removed by CuO–Fe3O4@C/PMS system within 15 min with the degradation rate constant (k) of 0.32 min−1, being 10.3 and 246.2 times that in CuO/PMS (0.031min−1) and Fe3O4/PMS (0.0013 min−1) system. A deep mineralization rate of BPA (＞80%) was achieved within 60 min. The results demonstrated the synergistic effect of bimetallic clusters, oxygen vacancies and carbon framework was a key benefit for the exposure of more active sites, the electron donor capacity and the mass transfer of substrates, thereby promoting the decomposition of BPA. Capture experiments and EPR indicated that 1O2 was the predominant reactive oxygen species (ROSs). The degradation routes of BPA and the activation mechanism of PMS were proposed. This study offers an opportunity to develop promising MOFs-derived hybrid catalysts with tailored structures and properties for the practical application of SR-AOPs.

Genetic design of co-expressing a novel aconitase with cis-aconitate decarboxylase and chaperone GroELS for high-level itaconic acid production

Itaconic acid (IA) is regarded as one of the 12 important building blocks as recognized by US Department of Energy. Thus, the production of IA is crucial and urgent. In this study, we attempted to explore a new aconitase from an acidophilic E. coli Nissle 1917, denoted as ENAcnA, and was compared to Corynebacterium glutamicum for the first time. The native ENAcnA has 10 amino acid mutations and the supposed critical mutations at N397S, Q409L, S522G, and G826E in the aconitase family domain contributed to the high hydratase activity at the low pH condition. Co-expression of ENAcnA and cis-aconitate decarboxylase from Aspergillus terreus in the recombinant E. coli BL21(DE3) resulted in IA titer and productivity of 80.4 g/L and 10.1 g/L/h, respectively. Integration of GroELS chaperon to chromosome of BL21(DE3) for generating BD::7G strain even enhanced IA titer by 13 %. The cold treatment of whole-cell biocatalyst substantially assisted the mass transfer of substrate through the cell membrane, and consequently achieved the maximum IA productivity. The discovery of ENAcnA, chaperon assistance in host, and cold treatment of whole cell biocatalysts contributed to high-level IA production up to 95.5 g/L for the industrial scale.

An Ultrasonic Target Detection System Based on Piezoelectric Micromachined Ultrasonic Transducers

In this paper, an ultrasonic target detection system based on Piezoelectric Micromachined Ultrasonic Transducers (PMUTs) is proposed, which consists of the PMUTs based ultrasonic sensor and the sensor system. Two pieces of 3 × 3 PMUTs arrays with the resonant frequency of 115 kHz are used as transmitter and receiver of the PMUTs-based ultrasonic sensor. Then, the sensor system can calculate the target's position through the signal received by the above receiver. The static and dynamic performance of the proposed prototype system are characterized on black, white, and transparent targets. The experiment results demonstrated that the proposed system can detect targets of different colors, transparencies, and motion states. In the static experiments, the static location errors of the proposed system in the range of 200 mm to 320 mm are 0.51 mm, 0.50 mm and 0.53 mm, whereas the errors of a commercial laser sensor are 2.89 mm, 0.62 mm, and N\A. In the dynamic experiments, the experimental materials are the targets with thicknesses of 1 mm, 1.5 mm, 2 mm and 2.5 mm, respectively. The proposed system can detect the above targets with a maximum detection error of 4.00%. Meanwhile, the minimum resolution of the proposed system is about 0.5 mm. Finally, in the comprehensive experiments, the proposed system successfully guides a robotic manipulator to realize the detecting, grasping, and moving of a transparent target with 1 mm. This ultrasonic target detection system has demonstrated a cost-effective method to detect targets, especially transparent targets, which can be widely used in the detection and transfer of glass substrates in automated production lines.

A novel inverse membrane bioreactor for efficient bioconversion from methane gas to liquid methanol using a microbial gas-phase reaction

BackgroundMethane (CH4), as one of the major energy sources, easily escapes from the supply chain into the atmosphere, because it exists in a gaseous state under ambient conditions. Compared to carbon dioxide (CO2), CH4 is 25 times more potent at trapping radiation; thus, the emission of CH4 to the atmosphere causes severe global warming and climate change. To mitigate CH4 emissions and utilize them effectively, the direct biological conversion of CH4 into liquid fuels, such as methanol (CH3OH), using methanotrophs is a promising strategy. However, supplying biocatalysts in an aqueous medium with CH4 involves high energy consumption due to vigorous agitation and/or bubbling, which is a serious concern in methanotrophic processes, because the aqueous phase causes a very large barrier to the delivery of slightly soluble gases.ResultsAn inverse membrane bioreactor (IMBR), which combines the advantages of gas-phase bioreactors and membrane bioreactors, was designed and constructed for the bioconversion of CH4 into CH3OH in this study. In contrast to the conventional membrane bioreactor with bacterial cells that are immersed in an aqueous phase, the filtered cells were placed to face a gas phase in the IMBR to supply CH4 directly from the gas phase to bacterial cells. Methylococcus capsulatus (Bath), a representative methanotroph, was used to demonstrate the bioconversion of CH4 to CH3OH in the IMBR. Cyclopropanol was supplied from the aqueous phase as a selective inhibitor of methanol dehydrogenase, preventing further CH3OH oxidation. Sodium formate was added as an electron donor to generate NADH, which is necessary for CH3OH production. After optimizing the inlet concentration of CH4, the mass of cells, the cyclopropanol concentration, and the gas flow rate, continuous CH3OH production can be achieved over 72 h with productivity at 0.88 mmol L−1 h−1 in the IMBR, achieving a longer operation period and higher productivity than those using other types of membrane bioreactors reported in the literature.ConclusionsThe IMBR can facilitate the development of gas-to-liquid (GTL) technologies via microbial processes, allowing highly efficient mass transfer of substrates from the gas phase to microbial cells in the gas phase and having the supplement of soluble chemicals convenient.

A novel PEC and ECL bifunctional aptasensor based on V2CTx MXene-derived MOF embedded with silver nanoparticles for selectively aptasensing miRNA-126.

A novel photoelectrochemical (PEC) and electrochemiluminescence (ECL) bifunctional aptasensor has been established for the detection of miRNA-126 using V2CTx MXene-derived porphyrin-based metal-organic framework embedded with Ag nanoparticles (Ag NPs) (denoted as AgNPs@V-PMOF) as a robust bioplatform. Due to the presence of V nodes in V2CTx MXene nanosheets, V-based MOF was prepared using tetrakis(4-carboxyphenyl)porphyrin as ligand, followed by the incorporation of Ag+ ions to form the AgNPs@V-PMOF Schottky heterojunction. Benefiting from the fast electron transfer of the V2CTx substrate and well-matched band-edge energy level of the photosensitive Ag NPs and V-PMOF, the constructed AgNPs@V-PMOF Schottky heterojunction exhibited the promoted transfer of the photogenerated carriers, showing superior PEC and ECL performances. Moreover, a large number of the complementary DNA strand of miRNA-126 can be immobilized over AgNPs@V-PMOF in view of the combined interaction of π-π stacking, van der Waals force, and Ag-N coordination between AgNPs@V-PMOF. Consequently, the developed AgNPs@V-PMOF-based aptasensor illustrated extremely low detection limits of 0.78 and 0.53 fM within a wide range from 1.0 fM to 1.0 nM of miRNA-126 detected by PEC and ECL techniques, respectively, superior to most reported miRNA aptasensors. Also, the provided bifunctional aptasensor demonstrated high selectivity, good stability, fine reproducibility, and acceptable regenerability, as well as promising potential for the analysis of miRNA-126 from living cancer cells. This work puts forward the development of aptasensors for the early and accurate diagnosis of cancer markers and extends the application of MOF in the biosensing field.

LBMON299 Aldosterone Production Is Regulated By Gap Junctions In Human Adrenal Cells

Abstract Aldosterone plays a critical role in blood pressure regulation via the renin-angiotensin-aldosterone system. Autonomous production of aldosterone, primary aldosteronism (PA), is the commonest curable cause of hypertension worldwide. Two cell types in the adrenal cortex produce aldosterone: zona glomerulosa (ZG) cells and aldosterone producing nodules (APN), and in cells of aldosterone producing adenomas (APA). Adrenal cortex cells undergo phenotypic metamorphosis as they migrate from the subcapsular region, centripetally, towards the corticomedullary junction, changing from aldosterone- to cortisol- producing cells as they go. Gap junctions (GJs) are specialised channels of cytoplasmic communication between cells, permitting the regulated transfer of substrates. Loss-of-function mutations in the CADM1 gene, which affects GJ protein CX43, have been found in APAs, implicating CX43 in aldosterone regulation. This study aims to demonstrate the dysregulation of aldosterone production by CX43 inhibition in human adrenal cell line, H295R, and elucidate GJ distribution in human adrenal cortex. Methodology H295R cells were treated with 1×10-8M angiotensin II and connexin mimetic GJ inhibitor, GAP27, at 0µM, 25µM, 83µM and 250µM. Immunofluorescence (IF) was used to identify the abundance of CX43 in different cell types of the adrenal cortex, performed on ex-vivo para-APA adrenal tissue, using antibodies against CX43 (GJA1), colocalised with markers of aldosterone-producing cells (aldosterone synthase, CYP11B2), cells of the zona glomerulosa (ZG) (VSNL1, and DAB2) and zona fasciculata (ZF) (CYP7A1). Results Inhibition of CX43 with Gap27 results in a dose-dependent increase in CYP11B2 mRNA expression and aldosterone production. At the highest concentration of 250µM GAP27, CYP11B2 mRNA expression increased by 133-fold (p=&amp;lt;0. 00001) and aldosterone production by 29-fold (p=0. 0006). IF studies demonstrate a crescendo gradient of CX43 in a centripetal direction from cells of APN to ZF. The abundance was lowest in aldosterone-producing APN cells, moderate in non-aldosterone-producing ZG cells and highest in cortisol-producing ZF cells. GJ distribution was pan-membranous in ZF, but, where present, mostly cytoplasmic in APN. Evidence of recent active communication, in the form of annular gap junctions (AJG), was present in all three cell types. Conclusion The CX43 inhibition-induced upregulation of CYP11B2 expression and aldosterone synthesis indicates that it has an important regulatory role in physiological aldosterone production via CYP11B2 expression. The distribution of CX43 in adrenal tissue suggests its transport substrate not only regulates aldosterone synthesis, but also in adrenal cortex cell phenotypic metamorphosis/migration. Future work will investigate the substrate of CX43 in adrenal cells and its possible effect on adrenal cell phenotype. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Enzyme-Immobilized Metal-Organic Frameworks: From Preparation to Application.

As a class of widely used biocatalysts, enzymes possess advantages including high catalytic efficiency, strong specificity and mild reaction condition. However, most free enzymes have high requirements on the reaction environment and are easy to deactivate. Immobilization of enzymes on nanomaterial-based substrates is a good way to solve this problem. Metal-organic framework (MOFs), with ultra-high specific surface area and adjustable porosity, can provide a large space to carry enzymes. And the tightly surrounded protective layer of MOFs can stabilize the enzyme structure to a great extent. In addition, the unique porous network structure enables selective mass transfer of substrates and facilitates catalytic processes. Therefore, these enzyme-immobilized MOFs have been widely used in various research fields, such as molecule/biomolecule sensing and imaging, disease treatment, energy and environment protection. In this review, the preparation strategies and applications of enzyme-immobilized MOFs are illustrated and the prospects and current challenges are discussed.

Effect of surfactants on the production and biofunction of Tremella fuciformis polysaccharide through submerged fermentation

Tremella fuciformis polysaccharide (TFPS) have been widely used in the fields of food and medicine due to their multiple functions. submerged cultivation of mushroom is viewed as a promising alternative for efficient production of TFPS. Here, span 80 promoting the transfer of substrates and TFPS production were studied. The TFPS yield with addition of 0.3 % (v/v) span 80 in the culture reached 10.56 ± 0.53 g/L, which was 22.51 % higher than that of the control group. In a 7.5 L fermenter with glucose fed-batch strategy, the TFPS yield reached 25.94 ± 0.53 g/L. Span 80 improved the permeability of cell membrane and facilitated the secretion of TFPS. Furthermore, the glucuronic acid content of TFPS from Span 80 containing medium increased to 15.44 ± 0.15 %, and exhibited stronger antioxidant activity against DPPH• and •OH radicals. This study provided further evidence for elucidating the promoting effect of span 80 in submerged fermentation and guidance for the production of TFPS from T. fuciformis spore.

Insight Into Distinct Functional Roles of the Flagellar ATPase Complex for Flagellar Assembly in Salmonella.

Most motile bacteria utilize the flagellar type III secretion system (fT3SS) to construct the flagellum, which is a supramolecular motility machine consisting of basal body rings and an axial structure. Each axial protein is translocated via the fT3SS across the cytoplasmic membrane, diffuses down the central channel of the growing flagellar structure and assembles at the distal end. The fT3SS consists of a transmembrane export complex and a cytoplasmic ATPase ring complex with a stoichiometry of 12 FliH, 6 FliI and 1 FliJ. This complex is structurally similar to the cytoplasmic part of the FOF1 ATP synthase. The export complex requires the FliH12-FliI6-FliJ1 ring complex to serve as an active protein transporter. The FliI6 ring has six catalytic sites and hydrolyzes ATP at an interface between FliI subunits. FliJ binds to the center of the FliI6 ring and acts as the central stalk to activate the export complex. The FliH dimer binds to the N-terminal domain of each of the six FliI subunits and anchors the FliI6-FliJ1 ring to the base of the flagellum. In addition, FliI exists as a hetero-trimer with the FliH dimer in the cytoplasm. The rapid association-dissociation cycle of this hetero-trimer with the docking platform of the export complex promotes sequential transfer of export substrates from the cytoplasm to the export gate for high-speed protein transport. In this article, we review our current understanding of multiple roles played by the flagellar cytoplasmic ATPase complex during efficient flagellar assembly.