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1.
Sci Rep ; 14(1): 15050, 2024 07 01.
Article in English | MEDLINE | ID: mdl-38951205

ABSTRACT

Chalcones are intermediate products in the biosynthesis of flavonoids, which possess a wide range of biological properties, including antimicrobial and anticancer activities. The introduction of a chlorine atom and the glucosyl moiety into their structure may increase their bioavailability, bioactivity, and pharmacological use. The combined chemical and biotechnological methods can be applied to obtain such compounds. Therefore, 2-chloro-2'-hydroxychalcone and 3-chloro-2'-hydroxychalcone were synthesized and biotransformed in cultures of two strains of filamentous fungi, i.e. Isaria fumosorosea KCH J2 and Beauveria bassiana KCH J1.5 to obtain their novel glycosylated derivatives. Pharmacokinetics, drug-likeness, and biological activity of them were predicted using cheminformatics tools. 2-Chloro-2'-hydroxychalcone, 3-chloro-2'-hydroxychalcone, their main glycosylation products, and 2'-hydrochychalcone were screened for antimicrobial activity against several microbial strains. The growth of Escherichia coli 10,536 was completely inhibited by chalcones with a chlorine atom and 3-chlorodihydrochalcone 2'-O-ß-D-(4″-O-methyl)-glucopyranoside. The strain Pseudomonas aeruginosa DSM 939 was the most resistant to the action of the tested compounds. However, chalcone aglycones and glycosides with a chlorine atom almost completely inhibited the growth of bacteria Staphylococcus aureus DSM 799 and yeast Candida albicans DSM 1386. The tested compounds had different effects on lactic acid bacteria depending on the tested species. In general, chlorinated chalcones were more effective in the inhibition of the tested microbial strains than their unchlorinated counterparts and aglycones were a little more effective than their glycosides.


Subject(s)
Anti-Infective Agents , Biotransformation , Chalcones , Chlorine , Microbial Sensitivity Tests , Chalcones/chemistry , Chalcones/pharmacology , Chalcones/chemical synthesis , Chlorine/chemistry , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Anti-Infective Agents/chemical synthesis , Beauveria/metabolism , Fungi/drug effects , Escherichia coli/drug effects , Escherichia coli/growth & development
2.
Appl Microbiol Biotechnol ; 108(1): 408, 2024 Jul 05.
Article in English | MEDLINE | ID: mdl-38967685

ABSTRACT

The simulations and predictions obtained from mathematical models of bioprocesses conducted by microorganisms are not overvalued. Mechanistic models are bringing a better process understanding and the possibility of simulating unmeasurable variables. The Dynamic Energy Budget (DEB) model is an energy balance that can be formulated for any living organism and can be classified as a structured model. In this study, the DEB model was used to describe E. coli growth in a batch reactor in carbon and nitrogen substrate limitation conditions. The DEB model provides a possibility to follow the changes in the microbes' cells including their elemental composition and content of some important cell ingredients in different growth phases in substrate limitation conditions which makes it more informative compared to Monod's model. The model can be used as an optimal choice between Monod-like models and flux-based approaches. KEY POINTS: • The DEB model can be used to catch changes in elemental composition of E. coli • Bacteria batch culture growth phases can be explained by the DEB model • The DEB model is more informative compared to Monod's based models.


Subject(s)
Bioreactors , Carbon , Energy Metabolism , Escherichia coli , Nitrogen , Escherichia coli/growth & development , Escherichia coli/metabolism , Nitrogen/metabolism , Carbon/metabolism , Bioreactors/microbiology , Models, Biological , Culture Media/chemistry , Batch Cell Culture Techniques , Models, Theoretical
3.
Int J Biol Macromol ; 273(Pt 2): 133110, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38876230

ABSTRACT

In food packaging, sodium lignosulfonate nanoparticles (SLS NPs) showed significant antibacterial properties, antioxidant and UV barrier activities. Herein, the SLS NPs were synthesized via a sustainable green method and were added into egg albumin/sodium alginate mixture (EA/SA) to fabricate a safe, edible EA/SA/SNPs food packaging. A composite film EA/SA/SNP was examined microstructurally and physicochemically. The mechanical characteristics, UV protection, water resistance, and the composite film's thermal stability were all enhanced by the inclusion of SLS NPs, and water vapor permeability reduced by 44 %. This composite film exhibited robust antioxidative properties with DPPH and ABTS free radical scavenging rates reaching 76.84 % and 92.56 %, and effective antimicrobial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) with antibacterial rates reaching 98.25 % and 97.13 % for the positively charged nanoparticles interacting with the cell membrane. Freshness tests showed that the EA/SA/SNPs packaging film could delay the quality deterioration of fresh tomatoes. This composite film can slow down spoilage bacteria proliferation and prolongs food's preservation period by eight days at ambient temperature.


Subject(s)
Alginates , Anti-Bacterial Agents , Antioxidants , Food Packaging , Lignin , Nanoparticles , Alginates/chemistry , Alginates/pharmacology , Food Packaging/methods , Nanoparticles/chemistry , Antioxidants/pharmacology , Antioxidants/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Lignin/chemistry , Lignin/analogs & derivatives , Lignin/pharmacology , Escherichia coli/drug effects , Escherichia coli/growth & development , Staphylococcus aureus/drug effects , Permeability , Steam
4.
Int J Biol Macromol ; 273(Pt 2): 133091, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38878924

ABSTRACT

The increasing significance of biopolymer-based food packaging can be attributed to its biodegradability and independence from petroleum-derived materials. Concurrently, metal oxide nanoparticles (NPs) have gained prominence as effective antimicrobial agents against both wild-type and antibiotic-resistant microbes. In this study, cerium oxide or ceria, CeO2, nanoparticles with an average diameter of 50 nm were synthesized via a green method utilizing Vibrio sp. VLC cell lysate supernatant. The synthesized CeO2 NPs displayed remarkable antimicrobial properties, inhibiting the growth of Escherichia coli and Staphylococcus aureus by 93.7 % and 98 %, respectively. To enhance the potential of bacterial cellulose (BC) for advanced applications, we developed a BC/xanthan/CeO2 nanocomposite using both ex situ and in situ techniques. The integration of CeO2 NPs within the nanocomposite structure not only improved the inherent properties of BC, but also rendered it suitable for use in active food packaging systems. The nanocomposite exhibited no significant cytotoxicity on the human dermal fibroblast (HDF) cells, confirming its safety. Nanocomposites containing biogenically synthesized CeO2 NPs demonstrated exceptional efficacy for reducing microbial contamination. Bread samples coated with nanocomposite films displayed no signs of microbial growth. These results support the application of BC/xanthan/CeO2 nanocomposites as suitable and effective coating materials for antimicrobial food packaging applications.


Subject(s)
Anti-Bacterial Agents , Cellulose , Cerium , Food Packaging , Nanocomposites , Polysaccharides, Bacterial , Cellulose/chemistry , Cellulose/pharmacology , Food Packaging/methods , Cerium/chemistry , Cerium/pharmacology , Nanocomposites/chemistry , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/pharmacology , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Humans , Staphylococcus aureus/drug effects , Escherichia coli/drug effects , Escherichia coli/growth & development , Microbial Sensitivity Tests , Metal Nanoparticles/chemistry
5.
Int J Mol Sci ; 25(11)2024 May 30.
Article in English | MEDLINE | ID: mdl-38892186

ABSTRACT

Flavonoids are an abundant class of naturally occurring compounds with broad biological activities, but their limited abundance in nature restricts their use in medicines and food additives. Here we present the synthesis and determination of the antibacterial and antioxidant activities of twenty-two structurally related flavonoids (five of which are new) by scientifically validated methods. Flavanones (FV1-FV11) had low inhibitory activity against the bacterial growth of MRSA 97-7. However, FV2 (C5,7,3',4' = OH) and FV6 (C5,7 = OH; C4' = SCH3) had excellent bacterial growth inhibitory activity against Gram-negative E. coli (MIC = 25 µg/mL for both), while Chloramphenicol (MIC = 25 µg/mL) and FV1 (C5,7,3' = OCH3; 4' = OH) showed inhibitory activity against Gram-positive L. monocytogenes (MIC = 25 µg/mL). From the flavone series (FO1-FO11), FO2 (C5,7,3',4' = OH), FO3 (C5,7,4' = OH; 3' = OCH3), and FO5 (C5,7,4' = OH) showed good inhibitory activity against Gram-positive MRSA 97-7 (MIC = 50, 12, and 50 µg/mL, respectively), with FO3 being more active than the positive control Vancomycin (MIC = 25 µg/mL). FO10 (C5,7= OH; 4' = OCH3) showed high inhibitory activity against E. coli and L. monocytogenes (MIC = 25 and 15 µg/mL, respectively). These data add significantly to our knowledge of the structural requirements to combat these human pathogens. The positions and number of hydroxyl groups were key to the antibacterial and antioxidant activities.


Subject(s)
Anti-Bacterial Agents , Antioxidants , Flavonoids , Microbial Sensitivity Tests , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/chemical synthesis , Antioxidants/pharmacology , Antioxidants/chemistry , Antioxidants/chemical synthesis , Flavonoids/pharmacology , Flavonoids/chemistry , Escherichia coli/drug effects , Escherichia coli/growth & development , Listeria monocytogenes/drug effects , Listeria monocytogenes/growth & development , Flavanones/pharmacology , Flavanones/chemistry , Methicillin-Resistant Staphylococcus aureus/drug effects
6.
Biotechnol J ; 19(6): e2400140, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38896410

ABSTRACT

Artificial Intelligence (AI) technology is spearheading a new industrial revolution, which provides ample opportunities for the transformational development of traditional fermentation processes. During plasmid fermentation, traditional subjective process control leads to highly unstable plasmid yields. In this study, a multi-parameter correlation analysis was first performed to discover a dynamic metabolic balance among the oxygen uptake rate, temperature, and plasmid yield, whilst revealing the heating rate and timing as the most important optimization factor for balanced cell growth and plasmid production. Then, based on the acquired on-line parameters as well as outputs of kinetic models constructed for describing process dynamics of biomass concentration, plasmid yield, and substrate concentration, a machine learning (ML) model with Random Forest (RF) as the best machine learning algorithm was established to predict the optimal heating strategy. Finally, the highest plasmid yield and specific productivity of 1167.74 mg L-1 and 8.87 mg L-1/OD600 were achieved with the optimal heating strategy predicted by the RF model in the 50 L bioreactor, respectively, which was 71% and 21% higher than those obtained in the control cultures where a traditional one-step temperature upshift strategy was applied. In addition, this study transformed empirical fermentation process optimization into a more efficient and rational self-optimization method. The methodology employed in this study is equally applicable to predict the regulation of process dynamics for other products, thereby facilitating the potential for furthering the intelligent automation of fermentation processes.


Subject(s)
Bioreactors , Escherichia coli , Fermentation , Machine Learning , Plasmids , Plasmids/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Escherichia coli/growth & development , Bioreactors/microbiology , Batch Cell Culture Techniques/methods , Biomass
7.
Food Microbiol ; 122: 104561, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38839224

ABSTRACT

The current study aimed to determine if the 3D-printing speed and temperature would impact the transferability of foodborne pathogens from the stainless-steel (SS) food cartridge to the 3D-printed food ink. Staphylococcus aureus and Escherichia coli were inoculated onto the interior surface of the SS food cartridges. Subsequently, a model food ink was extruded with a recommended macronutrient contribution of 55.8, 23.7, and 20.5% of carbohydrates, proteins, and fat, respectively. The impact of 3D-printing temperatures and speeds on transfer rates was analysed using a Two-Way ANOVA. S. aureus was transferred more from the cartridge to the food ink with a population of 3.39, 2.98, and 3.09 log CFU/g compared to 2.03, 2.06, and 2.00 log CFU/g for E. coli at 2000, 3000, and 4000 mm/s printing speed, respectively, at 25 °C. A Kruskal-Wallis Test was employed to investigate the effect of different speeds and temperatures on the transferability of S. aureus and E. coli. Speed was the main factor affecting S. aureus transferability, while temperature (25 and 50 °C) had the greatest impact on E. coli transferability. This research seeks to advance the understanding of 3D-printing parameters in pathogen transferability and help the food industry move towards this technology's quick and safe adoption.


Subject(s)
Escherichia coli , Food Microbiology , Printing, Three-Dimensional , Staphylococcus aureus , Temperature , Staphylococcus aureus/growth & development , Escherichia coli/growth & development , Stainless Steel , Food Handling/instrumentation , Food Handling/methods , Food Contamination/analysis , Colony Count, Microbial
8.
ISME J ; 18(1)2024 Jan 08.
Article in English | MEDLINE | ID: mdl-38874171

ABSTRACT

Although enteric bacteria normally reside within the animal intestine, the ability to persist extraintestinally is an essential part of their overall lifestyle, and it might contribute to transmission between hosts. Despite this potential importance, few genetic determinants of extraintestinal growth and survival have been identified, even for the best-studied model, Escherichia coli. In this work, we thus used a genome-wide library of barcoded transposon insertions to systematically identify functional clusters of genes that are crucial for E. coli fitness in lake water. Our results revealed that inactivation of pathways involved in maintaining outer membrane integrity, nucleotide biosynthesis, and chemotaxis negatively affected E. coli growth or survival in this extraintestinal environment. In contrast, inactivation of another group of genes apparently benefited E. coli growth or persistence in filtered lake water, resulting in higher abundance of these mutants. This group included rpoS, which encodes the general stress response sigma factor, as well as genes encoding several other global transcriptional regulators and RNA chaperones, along with several poorly annotated genes. Based on this co-enrichment, we identified these gene products as novel positive regulators of RpoS activity. We further observed that, despite their enhanced growth, E. coli mutants with inactive RpoS had reduced viability in lake water, and they were not enriched in the presence of the autochthonous microbiota. This highlights the duality of the general stress response pathway for E. coli growth outside the host.


Subject(s)
Escherichia coli , Lakes , Escherichia coli/genetics , Escherichia coli/growth & development , Lakes/microbiology , Sigma Factor/genetics , Sigma Factor/metabolism , Genome, Bacterial , DNA Transposable Elements , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Gene Expression Regulation, Bacterial , Water Microbiology
9.
J Biomed Mater Res B Appl Biomater ; 112(7): e35442, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38923117

ABSTRACT

The development of drug-resistant microorganisms is taking a heavy toll on the biomedical world. Clinical infections are costly and becoming increasingly dangerous as bacteria that once responded to standard antibiotic treatment are developing resistance mechanisms that require innovative treatment strategies. Nitric oxide (NO) is a gaseous molecule produced endogenously that has shown potent antibacterial capabilities in numerous research studies. Its multimechanistic antibacterial methods prevent the development of resistance and have shown potential as an alternative to antibiotics. However, there has yet to be a direct comparison study evaluating the antibacterial properties of NO against antibiotic susceptible and antibiotic-resistant clinically isolated bacterial strains. Herein, standardized lab and clinically isolated drug-resistant bacterial strains are compared side-by-side for growth and viability following treatment with NO released from S-nitrosoglutathione (GSNO), an NO donor molecule. Evaluation of growth kinetics revealed complete killing of E. coli lab and clinical strains at 17.5 mM GSNO, though 15 mM displayed >50% killing and significantly reduced metabolic activity, with greater dose dependence for membrane permeability. Clinical P. aeruginosa showed greater susceptibility to GSNO during growth curve studies, but metabolic activity and membrane permeability demonstrated similar effects for 12.5 mM GSNO treatment of lab and clinical strains. MRSA lab and clinical strains exhibited total killing at 17.5 mM treatment, though metabolic activity was decreased, and membrane permeation began at 12.5 mM for both strains. Lastly, both S. epidermidis strains were killed by 15 mM GSNO, with sensitivities in metabolic activity and membrane permeability at 12.5 mM GSNO. The mirrored antibacterial effects seen by the lab and clinical strains of two Gram-negative and two Gram-positive bacteria reveal the translational success of NO as an antibacterial therapy and potential alternative to standard antibiotic treatment.


Subject(s)
Anti-Bacterial Agents , Escherichia coli , Nitric Oxide , Nitric Oxide/pharmacology , Nitric Oxide/chemistry , Nitric Oxide/metabolism , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Escherichia coli/drug effects , Escherichia coli/growth & development , Humans , S-Nitrosoglutathione/pharmacology , S-Nitrosoglutathione/chemistry , Nitric Oxide Donors/pharmacology , Nitric Oxide Donors/chemistry , Drug Resistance, Bacterial/drug effects , Microbial Sensitivity Tests , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/growth & development
11.
Environ Microbiol Rep ; 16(3): e13308, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38924372

ABSTRACT

Microorganisms, including potential pathogens, can colonise plastic surfaces in aquatic environments. This study investigates the colonisation of plastic pellets by Escherichia coli (E. coli) as a proxy for faecal pathogens in aquatic environments. Plastic pellets from a polluted beach were placed in seawater aquaria spiked with E. coli. Diverse bacteria, primarily from the Proteobacteria phylum, rapidly colonised the pellets within 24 h, with notable species known for plastic or hydrocarbon degradation. Over 26 days, biofilms formed on the plastic surfaces, reaching bacterial populations of up to 6.8·105 gene copies (gc) of the 16S rRNA mm-2. E. coli, was detected in the pellets for up to 7 days using culture methods, exhibiting varying attachment densities regardless of source or environmental factors. The study highlights plastic biofilms as reservoirs for E. coli, contributing to the survival and persistence of faecal bacteria in aquatic systems. These findings deepen our understanding of the risks associated with plastic pollution in marine settings, offering insights into the behaviour of faecal indicators and their implications for water quality assessments, while providing valuable information on potential pathogen dissemination within plastic-associated microbial communities.


Subject(s)
Biofilms , Escherichia coli , Plastics , Seawater , Biofilms/growth & development , Escherichia coli/genetics , Escherichia coli/physiology , Escherichia coli/isolation & purification , Escherichia coli/growth & development , Seawater/microbiology , RNA, Ribosomal, 16S/genetics , Water Microbiology
12.
Molecules ; 29(12)2024 Jun 15.
Article in English | MEDLINE | ID: mdl-38930918

ABSTRACT

PURPOSE OF THE STUDY: the creation of a dextran coating on cerium oxide crystals using different ratios of cerium and dextran to synthesize nanocomposites, and the selection of the best nanocomposite to develop a nanodrug that accelerates quality wound healing with a new type of antimicrobial effect. MATERIALS AND METHODS: Nanocomposites were synthesized using cerium nitrate and dextran polysaccharide (6000 Da) at four different initial ratios of Ce(NO3)3x6H2O to dextran (by weight)-1:0.5 (Ce0.5D); 1:1 (Ce1D); 1:2 (Ce2D); and 1:3 (Ce3D). A series of physicochemical experiments were performed to characterize the created nanocomposites: UV-spectroscopy; X-ray phase analysis; transmission electron microscopy; dynamic light scattering and IR-spectroscopy. The biomedical effects of nanocomposites were studied on human fibroblast cell culture with an evaluation of their effect on the metabolic and proliferative activity of cells using an MTT test and direct cell counting. Antimicrobial activity was studied by mass spectrometry using gas chromatography-mass spectrometry against E. coli after 24 h and 48 h of co-incubation. RESULTS: According to the physicochemical studies, nanocrystals less than 5 nm in size with diffraction peaks characteristic of cerium dioxide were identified in all synthesized nanocomposites. With increasing polysaccharide concentration, the particle size of cerium dioxide decreased, and the smallest nanoparticles (<2 nm) were in Ce2D and Ce3D composites. The results of cell experiments showed a high level of safety of dextran nanoceria, while the absence of cytotoxicity (100% cell survival rate) was established for Ce2D and C3D sols. At a nanoceria concentration of 10-2 M, the proliferative activity of fibroblasts was statistically significantly enhanced only when co-cultured with Ce2D, but decreased with Ce3D. The metabolic activity of fibroblasts after 72 h of co-cultivation with nano composites increased with increasing dextran concentration, and the highest level was registered in Ce3D; from the dextran group, differences were registered in Ce2D and Ce3D sols. As a result of the microbiological study, the best antimicrobial activity (bacteriostatic effect) was found for Ce0.5D and Ce2D, which significantly inhibited the multiplication of E. coli after 24 h by an average of 22-27%, and after 48 h, all nanocomposites suppressed the multiplication of E. coli by 58-77%, which was the most pronounced for Ce0.5D, Ce1D, and Ce2D. CONCLUSIONS: The necessary physical characteristics of nanoceria-dextran nanocomposites that provide the best wound healing biological effects were determined. Ce2D at a concentration of 10-3 M, which stimulates cell proliferation and metabolism up to 2.5 times and allows a reduction in the rate of microorganism multiplication by three to four times, was selected for subsequent nanodrug creation.


Subject(s)
Cerium , Dextrans , Escherichia coli , Fibroblasts , Nanocomposites , Wound Healing , Cerium/chemistry , Cerium/pharmacology , Dextrans/chemistry , Dextrans/pharmacology , Nanocomposites/chemistry , Humans , Wound Healing/drug effects , Escherichia coli/drug effects , Escherichia coli/growth & development , Fibroblasts/drug effects , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/chemical synthesis , Cell Proliferation/drug effects , Microbial Sensitivity Tests , Cell Line
13.
Molecules ; 29(12)2024 Jun 19.
Article in English | MEDLINE | ID: mdl-38930981

ABSTRACT

ZnO nanorod nonwoven fabrics (ZNRN) were developed through hydrothermal synthesis to facilitate the prevention of the transmission of respiratory pathogens. The superhydrophobicity and antibacterial properties of ZNRN were improved through the response surface methodology. The synthesized material exhibited significant water repellency, indicated by a water contact angle of 163.9°, and thus demonstrated antibacterial rates of 91.8% for Escherichia coli (E. coli) and 79.75% for Staphylococcus aureus (S. aureus). This indicated that E. coli with thinner peptidoglycan may be more easily killed than S. aureus. This study identified significant effects of synthesis conditions on the antibacterial effectiveness, with comprehensive multivariate analyses elucidating the underlying correlations. In addition, the ZnO nanorod structure of ZNRN was characterized through SEM and XRD analyses. It endows the properties of superhydrophobicity (thus preventing bacteria from adhering to the ZNRN surface) and antibacterial capacity (thus damaging cells through the puncturing of these nanorods). Consequently, the alignment of two such features is desired to help support the development of personal protective equipment, which assists in avoiding the spread of respiratory infections.


Subject(s)
Anti-Bacterial Agents , Escherichia coli , Hydrophobic and Hydrophilic Interactions , Nanotubes , Staphylococcus aureus , Zinc Oxide , Zinc Oxide/chemistry , Zinc Oxide/pharmacology , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Nanotubes/chemistry , Escherichia coli/drug effects , Escherichia coli/growth & development , Staphylococcus aureus/drug effects , Staphylococcus aureus/growth & development , Textiles/microbiology , Microbial Sensitivity Tests , Surface Properties
14.
Proc Natl Acad Sci U S A ; 121(23): e2315850121, 2024 Jun 04.
Article in English | MEDLINE | ID: mdl-38814871

ABSTRACT

Rosettes are self-organizing, circular multicellular communities that initiate developmental processes, like organogenesis and embryogenesis, in complex organisms. Their formation results from the active repositioning of adhered sister cells and is thought to distinguish multicellular organisms from unicellular ones. Though common in eukaryotes, this multicellular behavior has not been reported in bacteria. In this study, we found that Escherichia coli forms rosettes by active sister-cell repositioning. After division, sister cells "fold" to actively align at the 2- and 4-cell stages of clonal division, thereby producing rosettes with characteristic quatrefoil configuration. Analysis revealed that folding follows an angular random walk, composed of ~1 µm strokes and directional randomization. We further showed that this motion was produced by the flagellum, the extracellular tail whose rotation generates swimming motility. Rosette formation was found to require de novo flagella synthesis suggesting it must balance the opposing forces of Ag43 adhesion and flagellar propulsion. We went on to show that proper rosette formation was required for subsequent morphogenesis of multicellular chains, rpoS gene expression, and formation of hydrostatic clonal-chain biofilms. Moreover, we found self-folding rosette-like communities in the standard motility assay, indicating that this behavior may be a general response to hydrostatic environments in E. coli. These findings establish self-organization of clonal rosettes by a prokaryote and have implications for evolutionary biology, synthetic biology, and medical microbiology.


Subject(s)
Escherichia coli , Flagella , Escherichia coli/metabolism , Escherichia coli/genetics , Escherichia coli/growth & development , Flagella/metabolism , Cell Division , Escherichia coli Proteins/metabolism , Escherichia coli Proteins/genetics
15.
mBio ; 15(6): e0079024, 2024 Jun 12.
Article in English | MEDLINE | ID: mdl-38742872

ABSTRACT

Loss of the Escherichia coli inner membrane protein YhcB results in pleomorphic cell morphology and clear growth defects. Prior work suggested that YhcB was directly involved in cell division or peptidoglycan assembly. We found that loss of YhcB is detrimental in genetic backgrounds in which lipopolysaccharide (LPS) or glycerophospholipid (GPL) synthesis is altered. The growth defect of ΔyhcB could be rescued through inactivation of the Mla pathway, a system responsible for the retrograde transport of GPLs that are mislocalized to the outer leaflet of the outer membrane. Interestingly, this rescue was dependent upon the outer membrane phospholipase PldA that cleaves GPLs at the bacterial surface. Since the freed fatty acids resulting from PldA activity serve as a signal to the cell to increase LPS synthesis, this result suggested that outer membrane lipids are imbalanced in ΔyhcB. Mutations that arose in ΔyhcB populations during two independent suppressor screens were in genes encoding subunits of the acetyl coenzyme A carboxylase complex, which initiates fatty acid biosynthesis (FAB). These mutations fully restored cell morphology and reduced GPL levels, which were increased compared to wild-type bacteria. Growth of ΔyhcB with the FAB-targeting antibiotic cerulenin also increased cellular fitness. Furthermore, genetic manipulation of FAB and lipid biosynthesis showed that decreasing FAB rescued ΔyhcB filamentation, whereas increasing LPS alone could not. Altogether, these results suggest that YhcB may play a pivotal role in regulating FAB and, in turn, impact cell envelope assembly and cell division.IMPORTANCESynthesis of the Gram-negative cell envelope is a dynamic and complex process that entails careful coordination of many biosynthetic pathways. The inner and outer membranes are composed of molecules that are energy intensive to synthesize, and, accordingly, these synthetic pathways are under tight regulation. The robust nature of the Gram-negative outer membrane renders it naturally impermeable to many antibiotics and therefore a target of interest for antimicrobial design. Our data indicate that when the inner membrane protein YhcB is absent in Escherichia coli, the pathway for generating fatty acid substrates needed for all membrane lipid synthesis is dysregulated which leads to increased membrane material. These findings suggest a potentially novel regulatory mechanism for controlling the rate of fatty acid biosynthesis.


Subject(s)
Escherichia coli Proteins , Escherichia coli , Fatty Acids , Escherichia coli/genetics , Escherichia coli/metabolism , Escherichia coli/growth & development , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Fatty Acids/metabolism , Fatty Acids/biosynthesis , Glycerophospholipids/metabolism , Lipopolysaccharides/biosynthesis , Membrane Proteins/genetics , Membrane Proteins/metabolism
16.
World J Microbiol Biotechnol ; 40(7): 213, 2024 May 24.
Article in English | MEDLINE | ID: mdl-38789629

ABSTRACT

Co-fermentation performed by Saccharomyces cerevisiae and Escherichia coli or other microbes has been widely used in industrial fermentation. Meanwhile, the co-cultured microbes might regulate each other's metabolisms or cell behaviors including oxidative stress tolerance through secreting molecules. Here, results based on the co-culture system of S. cerevisiae and E. coli suggested the promoting effect of E. coli on the oxidative stress tolerance of S. cerevisiae cells. The co-cultured E. coli could enhance S. cerevisiae cell viability through improving its membrane stability and reducing the oxidized lipid level. Meanwhile, promoting effect of the co-cultured supernatant on the oxidative stress tolerance of S. cerevisiae illustrated by the supernatant substitution strategy suggested that secreted compounds contained in the co-cultured supernatant contributed to the higher oxidative stress tolerance of S. cerevisiae. The potential key regulatory metabolite (i.e., hexadecanoic acid) with high content difference between co-cultured supernatant and the pure-cultured S. cerevisiae supernatant was discovered by GC-MS-based metabolomics strategy. And exogenous addition of hexadecanoic acid did suggest its contribution to higher oxidative stress tolerance of S. cerevisiae. Results presented here would contribute to the understanding of the microbial interactions and provide the foundation for improving the efficiency of co-fermentation performed by S. cerevisiae and E. coli.


Subject(s)
Coculture Techniques , Escherichia coli , Fermentation , Oxidative Stress , Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolism , Escherichia coli/metabolism , Escherichia coli/growth & development , Metabolomics , Microbial Viability , Gas Chromatography-Mass Spectrometry
17.
NPJ Syst Biol Appl ; 10(1): 61, 2024 May 29.
Article in English | MEDLINE | ID: mdl-38811603

ABSTRACT

Under ideal conditions, Escherichia coli cells divide after adding a fixed cell size, a strategy known as the adder. This concept applies to various microbes and is often explained as the division that occurs after a certain number of stages, associated with the accumulation of precursor proteins at a rate proportional to cell size. However, under poor media conditions, E. coli cells exhibit a different size regulation. They are smaller and follow a sizer-like division strategy where the added size is inversely proportional to the size at birth. We explore three potential causes for this deviation: degradation of the precursor protein and two models where the propensity for accumulation depends on the cell size: a nonlinear accumulation rate, and accumulation starting at a threshold size termed the commitment size. These models fit the mean trends but predict different distributions given the birth size. To quantify the precision of the models to explain the data, we used the Akaike information criterion and compared them to open datasets of slow-growing E. coli cells in different media. We found that none of the models alone can consistently explain the data. However, the degradation model better explains the division strategy when cells are larger, whereas size-related models (power-law and commitment size) account for smaller cells. Our methodology proposes a data-based method in which different mechanisms can be tested systematically.


Subject(s)
Escherichia coli , Models, Biological , Escherichia coli/growth & development , Cell Division/physiology , Cell Size , Escherichia coli Proteins/metabolism
18.
World J Microbiol Biotechnol ; 40(7): 203, 2024 May 16.
Article in English | MEDLINE | ID: mdl-38753033

ABSTRACT

The viable but non-culturable (VBNC) state is considered a survival strategy employed by bacteria to endure stressful conditions, allowing them to stay alive. Bacteria in this state remain unnoticed in live cell counts as they cannot proliferate in standard culture media. VBNC cells pose a significant health risk because they retain their virulence and can revive when conditions normalize. Hence, it is crucial to develop fast, reliable, and cost-effective methods to detect bacteria in the VBNC state, particularly in the context of public health, food safety, and microbial control assessments. This research examined the biomolecular changes in Escherichia coli W3110 induced into the VBNC state in artificial seawater under three different stress conditions (temperature, metal, and antibiotic). Initially, confirmation of VBNC cells under various stresses was done using fluorescence microscopy and plate counts. Subsequently, lipid peroxidation was assessed through the TBARS assay, revealing a notable increase in peroxidation end-products in VBNC cells compared to controls. ATR-FTIR spectroscopy and chemomometrics were employed to analyze biomolecular changes, uncovering significant spectral differences in RNA, protein, and nucleic acid concentrations in VBNC cells compared to controls. Notably, RNA levels increased, while protein and nucleic acid amounts decreased. ROC analyses identified the 995 cm- 1 RNA band as a consistent marker across all studied stress conditions, suggesting its potential as a robust biomarker for detecting cells induced into the VBNC state under various stressors.


Subject(s)
Biomarkers , Escherichia coli , Lipid Peroxidation , Microbial Viability , Escherichia coli/growth & development , Escherichia coli/genetics , Escherichia coli/metabolism , Spectroscopy, Fourier Transform Infrared/methods , Anti-Bacterial Agents/pharmacology , Stress, Physiological , Seawater/microbiology , Seawater/chemistry , Temperature , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Culture Media/chemistry
19.
Colloids Surf B Biointerfaces ; 240: 113997, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38815309

ABSTRACT

In this paper, a simple, bottom up, bioinspired technique is proposed for the synthesis of highly stable colloids of silica supported spherical silver nanoparticles (SiO2@Ag) that act as efficient catalytic and antimicrobial coatings for an organic substrate, filter paper. The core - shell structure and the highly branched dendritic polymer, poly(ethylene)imine, enabled the precise control of growth rate and morphology of silica and silver nanoparticles. The polymer also enabled the deposition of these nanoparticles onto an organic substrate, filter paper, through immersion by modifying its surface. The catalytic and antibacterial properties of these samples were assessed. The results obtained from this analysis showed a complete degradation of an aqueous pollutant, 4-nitrophenol, for 6 successive catalytic cycles without intermediate purification steps. Furthermore, the polymeric silica-silver suspension proved to express antibacterial activity against both Gram-positive and Gram-negative bacteria (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa). The antibacterial properties were evaluated according to the disk diffusion method, whereas the Minimum Inhibitory Concentration was also determined. The samples were examined by Scanning Electron Microscopy, Transmission Electron Microscopy, X-ray diffraction analysis, z-potential analysis, Fourier Transform Infrared Spectroscopy and Ultraviolet-visible Spectroscopy.


Subject(s)
Anti-Bacterial Agents , Colloids , Microbial Sensitivity Tests , Silicon Dioxide , Silver , Silver/chemistry , Silver/pharmacology , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/chemical synthesis , Silicon Dioxide/chemistry , Silicon Dioxide/pharmacology , Catalysis , Colloids/chemistry , Metal Nanoparticles/chemistry , Polymers/chemistry , Polymers/pharmacology , Polymers/chemical synthesis , Escherichia coli/drug effects , Escherichia coli/growth & development , Paper , Staphylococcus aureus/drug effects , Pseudomonas aeruginosa/drug effects , Surface Properties , Particle Size , Nitrophenols/chemistry
20.
Int J Biol Macromol ; 272(Pt 1): 132631, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38810852

ABSTRACT

A new series of chromone and furochromone-based sulfonamide Schiff's base derivatives 3-12 were synthesized and evaluated for their antimicrobial activity against S. aureus, E. coli, C. albicans, and A. niger using agar diffusion method. Compound 3a demonstrated potent antimicrobial activities with MIC values of 9.76 and 19.53 µg/mL against S. aureus, E. coli and C. albicans, which is 2-fold and 4-fold more potent than neomycin (MIC = 19.53, 39.06 µg/mL respectively). To improve the effectiveness of 3a, it was encapsulated into chitosan nanoparticles (CS-3aNPs). The CS-3aNPs size was 32.01 nm, as observed by transmission electron microscope (TEM) images and the zeta potential value was 14.1 ± 3.07 mV. Encapsulation efficiency (EE) and loading capacity (LC) were 91.5 % and 1.6 %, respectively as indicated by spectral analysis. The CS-3aNPs extremely inhibited bacterial growth utilizing the colony-forming units (CFU). The ability of CS-3aNPs to protect skin wounds was evaluated in vivo. CS-3aNPs showed complete wound re-epithelialization, hyperplasia of the epidermis, well-organized granulation tissue formation, and reduced signs of wound infection, as seen through histological assessment which showed minimal inflammatory cells in comparison with untreated wound. Overall, these findings suggest that CS-3aNPs has a positive impact on protecting skin wounds from infection due to their antimicrobial activity.


Subject(s)
Chitosan , Chromones , Microbial Sensitivity Tests , Nanoparticles , Sulfonamides , Wound Healing , Chitosan/chemistry , Chitosan/pharmacology , Nanoparticles/chemistry , Wound Healing/drug effects , Chromones/chemistry , Chromones/pharmacology , Animals , Sulfonamides/pharmacology , Sulfonamides/chemistry , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Staphylococcus aureus/drug effects , Candida albicans/drug effects , Mice , Escherichia coli/drug effects , Escherichia coli/growth & development , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Bacteria/drug effects , Bacteria/growth & development
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