Microbial Protein Research Articles

Valorisation of agricultural residues into Thauera sp. Sel9 microbial proteins for aquaculture

Unconventional protein sources are necessary to tackle the increasing demand for food. Microbial proteins (MP) are an alternative source of proteins for feed or food, suitable as feed for aquaculture. Substituting fishmeal with MP obtained from agricultural wastes could reduce the environmental burden of aquaculture and help with waste management. In this study, pure culture MP from the PHA-producer Thauera sp. Sel9 were obtained from agricultural residues (agro-zootechnical digestate and pasta industry leftovers). The produced MP was used in feeding tests with the model fish zebrafish (Danio rerio) to assess potential toxic effects and evaluate overall fish health. The obtained MP was rich in protein (59.5 % w/w over TS) and PHAs (15.0 %) and comprised all fish essential amino acids. The chemical scores and essential amino acid index confirmed the excellent quality of the MP. The feeding tests with 50 % feed substitution with MP resulted in survival rates (80–88 %) comparable to the control group (78 %), with only 100 % MP showing increased mortality. Thauera MP obtained from agricultural residues has the potential to become a partial fishmeal substitute in fish-farming.

Imputing Single-Cell Protein Abundance in Multiplex Tissue Imaging.

Multiplex tissue imaging are a collection of increasingly popular single-cell spatial proteomics and transcriptomics assays for characterizing biological tissues both compositionally and spatially. However, several technical issues limit the utility of multiplex tissue imaging, including the limited number of molecules (proteins and RNAs) that can be assayed, tissue loss, and protein probe failure. In this work, we demonstrate how machine learning methods can address these limitations by imputing protein abundance at the single-cell level using multiplex tissue imaging datasets from a breast cancer cohort. We first compared machine learning methods' strengths and weaknesses for imputing single-cell protein abundance. Machine learning methods used in this work include regularized linear regression, gradient-boosted regression trees, and deep learning autoencoders. We also incorporated cellular spatial information to improve imputation performance. Using machine learning, single-cell protein expression can be imputed with mean absolute error ranging between 0.05-0.3 on a [0,1] scale. Finally, we used imputed data to predict whether single cells were more likely to come from pre-treatment or post-treatment biopsies. Our results demonstrate (1) the feasibility of imputing single-cell abundance levels for many proteins using machine learning; (2) how including cellular spatial information can substantially enhance imputation results; and (3) the use of single-cell protein abundance levels in a use case to demonstrate biological relevance.

Developing fish oil emulsion gel enriched with Lentinula edodes single cell protein and its effect on controlling the growth of Acinetobacter baumannii

In this study, the characterization of fish oil (FO) emulsion gel (EGEL) containing single cell protein (SCP) produced from Lentinula edodes (L. edodes) and its potential inhibition against Acinetobacter baumannii (A. baumannii) were investigated. Oil extracted from the fish liver was emulsified with tween 80 and water, and then gelled using gelatin with the assistance of an ultrasonic homogenizer. The characteristics and surface analysis of SCP-EGEL were examined using FTIR (Fourier-transform infrared spectroscopy) and SEM (Scanning electron microscope). The particle size distribution and zeta potential of SCP-EGEL were measured using a Malvern Zetasizer. When SCP-EGEL was applied to the surface of the medium inoculated with A. baumannii, the inhibition zone (IZ) was 8.2 mm. An expansion of the IZ was observed (10.2 mm) when SCP-EGEL was applied to a fish skin (FS) surface prepared in the shape of a 6-mm diameter disc. In the SEM images, when SCP was added to lipo gel, the gel structure appeared flattened or swollen in some areas. The appearance of SCP cells being covered with gel gave the impression that they have a secondary wall. Therefore, the resulting complex can potentially be used as an additive in animal and human nutrition, in functional food coatings to suppress A. baumannii, and in fish feed to enrich it with protein.

Integrative Transcriptomic and Single-Cell Protein Characterization of Colorectal Carcinoma Delineates Distinct Tumor Immune Microenvironments Associated with Overall Survival.

Colorectal carcinoma (CRC) is a heterogeneous group of tumors with varying therapeutic response and prognosis, and evidence suggests the tumor immune microenvironment (TIME) plays a pivotal role. Using advanced molecular and spatial biology technologies, we aimed to evaluate the TIME in patients with CRC to determine whether specific alterations in the immune composition correlated with prognosis. We identified primary and metastatic tumor samples from 31 consented patients, which were profiled with whole-exome sequencing and bulk RNA-seq. Immune cell deconvolution followed by gene set enrichment analysis and unsupervised clustering was performed. A subset of tumors underwent in situ analysis of the TIME spatial composition at single-cell resolution through Imaging Mass Mass Cytometry. Gene set enrichment analysis revealed two distinct groups of advanced CRC, one with an immune activated phenotype and the other with a suppressed immune microenvironment. The activated TIME phenotype contained increased Th1 cells, activated dendritic cells, tertiary lymphoid structures, and higher counts of CD8+ T cells whereas the inactive or suppressed TIME contained increased macrophages and a higher M2/M1 ratio. Our findings were further supported by RNA-seq data analysis from the TCGA CRC database, in which unsupervised clustering also identified two separate groups. The immunosuppressed CRC TIME had a lower overall survival probability (HR 1.66, p=0.007). This study supports the pertinent role of the CRC immune microenvironment in tumor progression and patient prognosis. We characterized the immune cell composition to better understand the complexity and vital role that immune activity states of the TIME play in determining patient outcome.

Standard Operating Procedure for the determination of trace elements in hydrothermal fluids by Inductively Coupled Plasma Mass Spectrometry (ICP-MS).

This standard operating procedure (SOP) validates an inductively coupled plasma mass spectrometry (ICP-MS) procedure for the determination of trace elements in hydrothermal fluids. Hydrothermal fluids are aqueous solutions with a wide range of temperature, salinity, pH and trace elements that can be used by a set of microbial proteins containing redox-sensitive transition metals as their catalytic core. Due to the high variability of these samples, we have developed this protocol taking into account the special features of the matrices analyzed. An ICP-MS 7900 Agilent system was used. Calibration curves are linear in the 0.01 to 100 μg/L concentration range.

Imidazo[1,2-a]pyridine-appended chalcone and Schiff base conjugates: Synthetic, spectrophotometric, biological, and computational aspects

Herein we describe 2-chloroimidazo[1,2-a]pyridine-appended Schiff base and chalcone conjugates, exploring their synthetic, spectrophotometric, biological, and computational aspects. UV–visible study revealed that scaffolds VIc and VIIIa exhibit bathochromic and hypsochromic shifts respectively compared to their analogs, due to auxochrome effect. The molecular docking simulations were performed to investigate the binding interactions of motifs with three cancerous target proteins (4HJO, 1M14, and 1M17) and four microbial target proteins. The compound VIIIb exhibited significant anti-lung cancer activity (GI50: 22.3) against the A-459 cell line. Moreover, the synthesized compounds exhibited auspicious antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Klebsiella pneumoniae, with MIC ranging from 64 to 175 µg/mL. Additionally, in-silico target prediction analysis suggested a considerable likelihood (26.7%) of acting on kinases for compounds VIIIa and VIIIc. In-silico toxicity scrutiny anticipated all compounds as non-fatal and less toxic as they belong to toxicity class 4 (300 < LD50 ≤ 2000). Further structural insights into the synthesized compounds were gained through optimized geometries, FMOs, MEP plots, and global reactivity descriptors (µ, ɳ, S, and ω), aiding in understanding their structural features, potential reactivity, and toxicity. Overall, these results highlight the potential of the prepared moieties as hopeful intrants for further development.

Soybean molasses can be used as a substitute for corn in grazing beef cattle supplements during the rainy season.

Soybean molasses (SBMO) is a byproduct derived from the production of soy protein concentrate, obtained through solubilization in water and alcohol. The utilization of SBMO as an animal feed ingredient shows promising potential, primarily due to its low cost and as a potential energy concentrate. This study aimed to assess the intake, digestibility, ruminal parameters (pH and ruminal ammonia - NH3), nitrogen retention (NR) and microbial protein synthesis in grazing beef cattle supplemented with SBMO as a substitute for corn during the rainy season. Five Nellore (10-month-old) bulls with an average initial weight of 246 ± 11.2kg were utilized in a 5 × 5 Latin square design. The animals were housed in five paddocks, each consisting of 0.34ha of Marandu grass (Urochloa brizantha). Five isonitrogenous protein-energy supplements (300g crude protein [CP]/kg supplement) were formulated, with SBMO replacing corn at varying levels (0, 0.25, 0.50, 0.75, or 1.00g-1g). The supplements were provided daily at a quantity of 2.0kg-1 animal. The inclusion of SBMO at any level of corn substitution did not significantly affect the intake of pasture dry matter or total dry matter (P > 0.10). Likewise, the intake of CP and, consequently, the ruminal concentration of NH3 did not differ among the SBMO levels. Increasing the inclusion of SBMO did not have a significant impact on NR (P > 0.10), indicating that animals receiving supplements containing 100% SBMO as a substitute for corn may perform similarly to animals receiving supplements with 100% corn (0% SBMO). Soybean molasses represents a viable alternative energy source for grazing beef cattle during the rainy season and can entirely replace corn without adversely affecting animal nutritional performance.

Contextual AI models for single-cell protein biology

Understanding protein function and developing molecular therapies require deciphering the cell types in which proteins act as well as the interactions between proteins. However, modeling protein interactions across biological contexts remains challenging for existing algorithms. Here we introduce PINNACLE, a geometric deep learning approach that generates context-aware protein representations. Leveraging a multiorgan single-cell atlas, PINNACLE learns on contextualized protein interaction networks to produce 394,760 protein representations from 156 cell type contexts across 24 tissues. PINNACLE’s embedding space reflects cellular and tissue organization, enabling zero-shot retrieval of the tissue hierarchy. Pretrained protein representations can be adapted for downstream tasks: enhancing 3D structure-based representations for resolving immuno-oncological protein interactions, and investigating drugs’ effects across cell types. PINNACLE outperforms state-of-the-art models in nominating therapeutic targets for rheumatoid arthritis and inflammatory bowel diseases and pinpoints cell type contexts with higher predictive capability than context-free models. PINNACLE’s ability to adjust its outputs on the basis of the context in which it operates paves the way for large-scale context-specific predictions in biology.

Nitrate recovery from groundwater and simultaneous upcycling into single-cell protein using a novel hybrid biological-inorganic system

Nitrate pollution in groundwater is a serious problem worldwide, as its concentration in many areas exceeds the WHO-defined drinking water standard (50 mg/L). Hydrogen-oxidizing bacteria (HOB) are a group of microorganisms capable of producing single-cell protein (SCP) using hydrogen and oxygen. Furthermore, HOB can utilize various nitrogen sources, including nitrate. This study developed a novel hybrid biological-inorganic (HBI) system that coupled a new submersible water electrolysis system driven by renewable electricity with HOB fermentation for in-situ nitrate recovery from polluted groundwater and simultaneously upcycling it together with CO2 into single-cell protein. The performance of the novel HBI system was first evaluated in terms of bacterial growth and nitrate removal efficiency. With 5 V voltage applied and the initial nitrate concentration of 100 mg/L, the nitrate removal efficiency of 85.52 % and raw of 47.71 % (with a broad amino acid spectrum) were obtained. Besides, the HBI system was affected by the applied voltages and initial nitrogen concentrations. The water electrolysis with 3 and 4 V cannot provide sufficient H2 for HOB and the removal of nitrate was 57.12 % and 59.22 % at 180 h, while it reached 65.14 % and 65.42 % at 5 and 6 V, respectively. The nitrate removal efficiency reached 58.40 % and 50.72 % within 180 h with 200 and 300 mg/L initial nitrate concentrations, respectively. Moreover, a larger anion exchange membrane area promoted nitrate removal. The monitored of the determination of different forms of nitrogen indicated that around 60 % of the recovered nitrate was assimilated into cells, and 40 % was bio-converted to N2. The results demonstrate a potentially sustainable method for remediating nitrate contaminant in groundwater, upcycling waste nitrogen, CO2 sequestration and valorization of renewable electricity into food or feed.

Lupin Seed Supplementation as a Functional Feed Additive: In Vitro Ruminal Gas, Methane and Carbon Dioxide Production, Fermentation Kinetics, and Nutrient Degradability.

The inevitable enteric gas emission from ruminants is considered a modern-day problem from an environmental perspective. Addressing this problem requires nutritional approaches such as the use of phytogenic additives in ruminant diets. In this regard, lupin seed (LS) can be a useful additive due to its phytochemical constituents. Therefore, this study investigated the effects of lupin (Lupinus angustifolius) seed supplementation as a functional and sustainable feed additive in sheep diet (50:50 concentrate-to-forage ratio) on in vitro gas production (GP; mL/g DM), methane (CH4; mL/g DM) and carbon dioxide (CO2; mL/g DM) emissions, fermentation parameters, and nutrient degradability (g/kg DM incubated). Gas production and CH4 were measured per gram of incubated dry matter (DM), degradable DM (dDM), degradable neutral detergent fiber (dNDF), and degradable acid detergent fiber (dADF). Lupin seeds were included at 0 (control), 0.5, 1.0, 1.5, and 2% of the diet. The seeds contained 3.27% essential oils (DM basis), with eucalyptol as the main phytochemical. The highest GP per gram of DM and dDM was observed (p < 0.01) with 2.0% LS supplementation level. While 1.0% LS had the highest GP per gram of dNDF, 0.5% lupin diet had the highest GP per gram of dADF. Asymptotic GP and CH4 emissions linearly and quadratically increased (p < 0.01) with increasing LS level, while lag time decreased. Despite increased CH4 production, the proportion of CH4 in total biogas was lower (p = 0.008) for LS treatments than the control, with the 0.5% LS showing the lowest CH4 proportion. Production of CO2 increased with lupin seed treatments, with 0.5% LS producing the highest proportion (p = 0.027). Degradability of DM, NDF, and ADF was greater (p < 0.01) for the high LS supplementation level, while 0.5% supplementation level decreased ADF degradability. Total short-chain fatty acids, acetic acid, and propionic acid increased (p < 0.05) with LS supplementation level, leading to a reduced acetate:propionate ratio. Rumen pH decreased (p = 0.036) with LS supplementation, while ammonia-N decreased (p = 0.045) and estimated metabolizable energy increased (p < 0.001) linearly. Calculated microbial protein synthesis (p = 0.005) and gas yield (p = 0.047) increased with LS supplementation level. LS supplementation at 2.0% of diet (DM basis) increased GP and CH4 emission (mL/g DM) and enhanced nutrient degradability, suggesting its potential use as a functional feed additive for ruminants when supplemented at a 2.0% level into diet.

Effects of corn grain processing and phosphorus content in calf starters on intake, growth performance, nutrient digestibility, blood metabolites, and urinary purine derivatives

Corn grain with a high phosphorus (P) content (mainly in the form of phytate-P) may need to be processed to improve the digestibility of nutrients for young calves. Processing corn grains can improve the accessibility of phytate-P to the rumen enzymes and increase the bioavailability of P, which benefits the growth and development of calves. The objective of this study was to investigate the effects of feeding starter diets with steam-flaked corn (SFC) compared with ground corn (GC) with 2 P contents of 0.4% and 0.7% DM basis on intake, growth performance, nutrient digestibility, blood metabolites and urinary purine derivatives in dairy calves. A total of 48 female Holstein dairy calves (3 d old; average initial weight 39.7 ± 3.9 kg) were randomly assigned to a 2 × 2 factorial arrangement of treatments (12 calves/treatment) in a randomized complete block design. The treatment groups were: 1) a starter diet of GC with 0.4% P (GC-0.4P); 2) a starter diet of GC with 0.7% P (GC-0.7P); 3) a starter diet of SFC with 0.4% P (SFC-0.4P); 4) a starter diet of SFC with 0.7% P (SFC-0.7P). Calves received 6 L/d of transition milk on d 2-3 and 5 L/d of whole milk on d 4-30, which was increased to 7 L/d on d 31-45, then decreased to 5 L/d on d 46-60 and reduced to a single feeding of 2 L on d 61-62. All calves had free access to starter feed and water. All calves were weaned on d 63 and remained in the study until d 83. Rumen fluid samples were collected on d 38 (pre-weaning) and d 76 (post-weaning). Blood samples were collected on d 40 and 80 and urine samples were collected on 4 consecutive days from d 79 to 82 to analyze urinary excretion of PD. The phytate-P content ranged from 0.23 to 0.17 for GC and SFC, respectively. In particular, the interaction between corn processing method and P content showed that the SFC-0.7P diets had a greater intake of starter feed during the pre- and post-weaning periods compared with the other experimental groups. In addition, calves fed the SFC-0.7P diet had greater average daily gain, body weight, withers height at weaning, better organic matter digestibility, higher blood β-hydroxybutyrate levels and higher microbial protein synthesis compared with all other groups. Feeding the SFC diet also resulted in improved feed efficiency, improved P digestibility and a tendency toward a lower rumen pH, albeit with a tendency toward an increase in blood glucose concentration during the pre-weaning period. In addition, the inclusion of 0.7% P to the starter diet resulted in increased fiber digestibility and a slight improvement in growth performance, which was particularly evident in hip height. Overall, the inclusion of SFC in the calf starter diet, especially in combination with a 0.7% DM basis P supplement, improved growth performance and nutrient utilization in dairy calves compared with GC.

Contextual AI models for single-cell protein biology.

Understanding protein function and developing molecular therapies require deciphering the cell types in which proteins act as well as the interactions between proteins. However, modeling protein interactions across biological contexts remains challenging for existing algorithms. Here, we introduce Pinnacle, a geometric deep learning approach that generates context-aware protein representations. Leveraging a multi-organ single-cell atlas, Pinnacle learns on contextualized protein interaction networks to produce 394,760 protein representations from 156 cell type contexts across 24 tissues. Pinnacle's embedding space reflects cellular and tissue organization, enabling zero-shot retrieval of the tissue hierarchy. Pretrained protein representations can be adapted for downstream tasks: enhancing 3D structure-based representations for resolving immuno-oncological protein interactions, and investigating drugs' effects across cell types. Pinnacle outperforms state-of-the-art models in nominating therapeutic targets for rheumatoid arthritis and inflammatory bowel diseases, and pinpoints cell type contexts with higher predictive capability than context-free models. Pinnacle's ability to adjust its outputs based on the context in which it operates paves way for large-scale context-specific predictions in biology.

From yeast screening for suitability as single cell protein to fed-batch cultures.

Fed-batch cultures have rarely been used in single cell protein (SCP) research. This work evaluated multiple yeast species for suitability as SCP cultivated using glucose- and sucrose-based substrate and performed in-depth studies of fed-batch SCP cultivation kinetics for selected yeasts, including determination of specific crude nitrogen-to-protein conversion factors. SCP was cultivated using fully synthetic media in flask batch or bioreactor fed-batch cultures. Crude nitrogen and nucleic acid content were determined using the Dumas method and fluorescence assay kits, respectively. C. utilis compared favorably to other yeasts in flask batch cultures in terms of process yield (0.52 ± 0.01 gxgs-1) and crude nitrogen content (10.0 ± 0.5 and 9.9 ± 0.5%CDW for glucose and sucrose, respectively). This is the first time biomass composition data was reported for SCP cultivated in fed-batch mode. C. utilis crude nitrogen content was consistent across the tested conditions (protein content stabilized around 50%CDW in fed-batch), while that of the benchmark yeast S. cerevisiae was higher in batch cultures and at the beginning of fed-batch relative to the end (protein content decreased over time and stabilized around 43%CDW). Total nucleic acid content of the yeasts was similar (6.8%CDW and 6.3%CDW, for C. utilis and S. cerevisiae, respectively), with crude nitrogen-to-protein conversion factors of 4.97 and 5.80. This study demonstrated the suitability of C. utilis as SCP, notably the robustness of its crude nitrogen content (as an indicator of protein content) across batch and fed-batch conditions, compared to that of the benchmark yeast S. cerevisiae.

THE EFFECTS OF SINGLE CELL PROTEIN ON AL-AWASSI LAMBS IN IRAQ

The effects of single cell protein (SCP) were studied in Al-Awassi lambs in Iraq. Four groups of lambs received 0%, 5%, 10% and 15% of (SCP) as a replacer for a soybean. The influence of (SCP) on the performance and carcass data of lambs and their blood pictures were studied. The total uric acid, blood urea and the histopathological changes were also investigated. The results - showed” that ‘there were no significant (P&gt;0.05) differences ‘among the initial, slaughtered, total weights gained and chilled carcass weights. The wholesale cuts showed. no significant (P&gt;0.05) differences. The eye rib area, fat thickness of. longissmus doris muscle, average percentage of organs and offal dressing percentage (calculated in live and empty body weight basis) were similar in all groups examined. the blood and serum parameters did not differ significantly (P&gt;0.05) among the experimental groups.

THR USE OF DIFFERENT LEVELS OF SINGLE CELL PROTEIN IN BROILER RATIONS

The present work was carried out in poultry experiment farm of Animal Resources Dept.., College of Agric., Baghdad University, Iraq. Five different levels of single cell protein (0, 5, 10, 15, 20%) replaced to soybean meal in the diets of broiler chick and 600 one day old chicks (Hubberd). The birds were randomly distributed among five treatments (two replicate for each treatment.), 60 chicks per replicate. The results showed that. the diet containing 10% single cell protein was the best diet among the other treatments. The average body weight in six weeks old were 1081.5, 1097, 1171, 1095, 332 gr for the treatments used respectively.

ADTnorm: Robust Integration of Single-cell Protein Measurement across CITE-seq Datasets.

CITE-seq enables paired measurement of surface protein and mRNA expression in single cells using antibodies conjugated to oligonucleotide tags. Due to the high copy number of surface protein molecules, sequencing antibody-derived tags (ADTs) allows for robust protein detection, improving cell-type identification. However, variability in antibody staining leads to batch effects in the ADT expression, obscuring biological variation, reducing interpretability, and obstructing cross-study analyses. Here, we present ADTnorm (https://github.com/yezhengSTAT/ADTnorm), a normalization and integration method designed explicitly for ADT abundance. Benchmarking against 14 existing scaling and normalization methods, we show that ADTnorm accurately aligns populations with negative- and positive-expression of surface protein markers across 13 public datasets, effectively removing technical variation across batches and improving cell-type separation. ADTnorm enables efficient integration of public CITE-seq datasets, each with unique experimental designs, paving the way for atlas-level analyses. Beyond normalization, ADTnorm includes built-in utilities to aid in automated threshold-gating as well as assessment of antibody staining quality for titration optimization and antibody panel selection. Applying ADTnorm to a published COVID-19 CITE-seq dataset allowed for identifying previously undetected disease-associated markers, illustrating a broad utility in biological applications.

Characteristics of Neonatal Sepsis and Predictive Values of Polyfunctional Assessment of Umbilical Cord Neutrophils Based on Single Cell Proteomic Secretion.

The early diagnosis of neonatal sepsis is crucial as it remains a prevalent cause of neonatal mortality. In this study, we conducted an analysis on the clinical data and detection indicators of 22 cases with sepsis and 62 cases without sepsis among neonates. Our findings indicate that the clinical signs observed in neonates with sepsis lack specificity. In addition, the commonly used clinical inflammatory indicators (such as leukocyte count, neutrophil-to-lymphocyte ratio [NLR], C-reactive protein [CRP], procalcitonin) exhibit limited sensitivity and specificity. Furthermore, the current clinical measures lack the assessment of inflammatory factors. Therefore, in order to enhance the accuracy of early sepsis diagnosis in neonates, we have employed a novel microfluidic-based single-cell technology platform for the analysis of 32 cytokines secreted by neutrophils at the individual cell level under various toxin stimulation conditions. We have further investigated and compared the disparities in single-cell protein secretomics between umbilical cord blood neutrophils and healthy adult peripheral neutrophils within an in vitro sepsis model. Our findings indicate that in a resting state UCB neutrophils exhibited lower polyfunctionality compared with healthy adult blood neutrophils, and notable variations in cytokine secretion profiles were detected between the two groups. However, the polyfunctionality of UCB neutrophils significantly increased and surpassed that of healthy adult neutrophils when exposed to alpha-hemolysin or lipopolysaccharide. UCB neutrophilssecreted awiderangeof chemokines and inflammatory factors, among which GM-CSF andIL-18were the most significant. Furthermore, we initially categorized the functional subgroups of neutrophils by considering the secretion of five primary cytokines by neutrophils (GM-CSF, IL-18, IL-8, MIP-1β, and MIF). The currentstudy, for the first time,examinedin detail the heterogeneity of protein secretion and thefunctional diversity of UCB neutrophils stimulated by different antigens. Moreover, new insight into neonatal sepsis, early diagnosis, and wider clinical applications of UCB neutrophils are provided by these data.

ENTEROBACTER CLOACAE LIPOPOLYSACCHARIDE EXPORT SYSTEM PROTEIN (LPTC) GENE EXPRESSION VARIATION VIA EXPOSE TO BIOSYNTHESIZED ZINC OXIDE NANOPARTICLES

Synthetic nanoparticles (NPs) have become more widely used in the disinfection and health sectors owing to their ability to effectively penetrate biological systems. Recent research has demonstrated the ability of microbial proteins and enzymes to function as reducing agents throughout the NP-production process, providing a different option to chemical and physical techniques. This method not only saves money and is effective, but also has the least possible environmental impact. This work used Lactobacillus spp., as the reducing and capping agent to create zinc oxide nanoparticles, (ZnO NPs). Numerous analytical techniques were employed to examine the generated ZnO NPs; these techniques include ultraviolet–visible spectroscopy (UV–vis), scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FT-IR). As an opportunistic pathogenic bacterium, Enterobacter cloaca possesses several virulence characteristics that enable it to infiltrate target tissues. However, bacteria can become resistant to antibiotics, which in turn makes treating bacterial infections becomes more difficult. With the development of science at the biotechnology level, using zinc oxide ZnO NPs on isolates of Enterobacter cloacae, we conducted research in the promising field of nanotechnology. The impact of three different concentrations of ZnO NPs on the gene expression of Enterobacter cloaca lipopolysaccharide transport (Lpt) proteins in isolates obtained from some patients suffering of various infections was investigated. Bacteria with the highest multidrug resistance were selected for further analysis. Expression of LptC gene was reduced significantly at concentration of 125 mg/mL (P &lt; 0.01) for all the isolate that treated with ZnO NPs.

Atom-economical synthesis, antimicrobial assessment, and molecular dynamics of indole-derived peptidomimetics for improved therapeutic insights

The emergence of multi-drug resistant strains spanning bacteria, fungi, viruses, and parasites poses a severe threat to public health and the economy. Tackling antimicrobial resistance requires swiftly identifying new agents with unique mechanisms. Multicomponent reactions offer a vital method to produce diverse analogs in various heterocyclic compounds, with broad medicinal applications. This study synthesized a range of peptidomimetics using isocyanide-based multicomponent Ugi reactions. These analogs were screened for antimicrobial efficacy against both Gram-negative (Escherichia coli, Pseudomonas aeruginosa) and Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis, Staphylococcus aureus IGMS-01), with ampicillin as a reference antibiotic, and for antifungal activity against Candida albicans, using fluconazole as a reference. The compounds showed moderate but significant efficacy across all tests. Docking analysis with microbial proteins revealed notable binding affinities for peptidomimetics 5h (N-(2-(1H-indol-3-yl)ethyl)-N-(1-(4-chlorophenyl)-2-(cyclohexylamino)-2-oxoethyl)stearamide) and 5i (N-(2-(1H-indol-3-yl)ethyl)-N-(1-(4-chlorophenyl)-2-(cyclohexylamino)-2-oxoethyl)oleamide). Molecular dynamics simulations confirmed stable interactions between the ligands and proteins. Furthermore, the long alkyl chain analogs demonstrated the ability to form organo/hydrogels, facilitating the incorporation of silver sulfadiazine for controlled antimicrobial release. This innovative approach enhances drug dispersion within the gel structure, effectively inhibiting microbial growth. These findings highlight the potential of these adaptable materials to advance drug development efforts in the foreseeable future.

Applications of cold plasma technique to enhance the safety and quality of different food products

Cold plasma (CP) has emerged as a novel technology for food processing. Its uses have been expanded into the food industry globally as an effective non-thermal processing technique with various applications. CP treatment uses reactive species for microbial inactivation, mycotoxin degradation, enzyme inactivation, protein modification and hydrogenation. CP treatment improves the physiochemical properties of different food products and the antimicrobial properties of packaging films. This review article demonstrates the applications of CP treatment in different food industries. It also explains the mechanism of microbial reduction in food components by using CP. The plasma source and impact of CP on food quality and safety have been discussed. CP is a key technology to increase the shelf life of food products while enhancing their safety with minimal impact on their bioactive compounds.