Polymer-enhanced peroxidase activity of ceria nanozyme for highly sensitive detection of alkaline phosphatase.

Nanoceria have demonstrated a wide array of catalytic activity similar to natural enzymes, holding considerable significance in the colorimetric detection of alkaline phosphatase (ALP), which is a biomarker of various biological disorders. However, the issues of physiological stability and formation of protein corona, which are strongly related to their surface chemistry, limit their practical application. In this work, CeO2 nanoparticles characterized by enhanced dimensional uniformity and specific surface area were synthesized, followed by encapsulation with various polymers to further increase catalytic activity and physiological stability. Notably, the CeO2 nanoparticles encapsulated within each polymer exhibited improved catalytic characteristics, with PAA-capped CeO2 exhibiting the highest performance. We further demonstrated that the PAA-CeO2 obtained with enhanced catalytic activity was attributed to an increase in surface negative charge. PAA-CeO2 enabled the quantitative assessment of AA activity within a wide concentration range of 10 to 60 µM, with a detection limit of 0.111 µM. Similarly, it allowed for the evaluation of alkaline phosphatase activity throughout a broad range of 10 to 80 U/L, with a detection limit of 0.12 U/L. These detection limits provided adequate sensitivity for the practical detection of ALP in human serum.

Enhanced osteogenic and bactericidal performance of premixed calcium phosphate cement with photocrosslinked alginate thin film.

The increasing prevalence of implant-associated infections (IAI) in orthopedics remains a public health challenge. Calcium phosphates (CaPs) are critical biomaterials in dental treatments and bone regeneration. It is highly desirable to endow CaPs with antibacterial properties. To achieve this purpose, we developed a photocrosslinked methacrylated alginate co-calcium phosphate cement (PMA-co-PCPC) with antibacterial properties, using α-tricalcium phosphate (α-TCP) powders with 16% amorphous contents as solid phase, liquid phases containing CuCl2 and SrCl2 as an inhibitor, and CaCl2 as an activator to construct PCPC. When CaCl2 started to activate the hydration reaction, Sr2+ or Cu2+ ions were exchanged with Ca2+, and α-TCP dissolution was restarted and gradually hydrated to form calcium-deficient hydroxyapatite (CDHA). PMA was added to crosslink with Cu/Sr ions and form gel-layer-wrapped hydrated CDHA. This study explored the binding mechanism of PMA and PCPC and the ion release rule of Ca2+ → Sr2+/Cu2+, optimized the construction of several antibacterial PMA-co-PCPC materials, and analyzed the physical, chemical, and biological properties. Because of the combined effect of Cu and Sr ions, the scaffold exhibited a potential antibacterial activity, promoting bone formation and vascular regeneration. This work provides a basis for designing antibacterial calcium phosphate biomaterials with controllable treatment, which is an important characteristic for preventing IAI of biomaterials.

Rapid detection of mexX in Pseudomonas aeruginosa based on CRISPR-Cas13a coupled with recombinase polymerase amplification.

The principal pathogen responsible for chronic urinary tract infections, immunocompromised hosts, and cystic fibrosis patients is Pseudomonas aeruginosa, which is difficult to eradicate. Due to the extensive use of antibiotics, multidrug-resistant P. aeruginosa has evolved, complicating clinical therapy. Therefore, a rapid and efficient approach for detecting P. aeruginosa strains and their resistance genes is necessary for early clinical diagnosis and appropriate treatment. This study combines recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats-association protein 13a (CRISPR-Cas13a) to establish a one-tube and two-step reaction systems for detecting the mexX gene in P. aeruginosa. The test times for one-tube and two-step RPA-Cas13a methods were 5 and 40 min (including a 30 min RPA amplification reaction), respectively. Both methods outperform Quantitative Real-time Polymerase Chain Reactions (qRT-PCR) and traditional PCR. The limit of detection (LoD) of P. aeruginosa genome in one-tube and two-step RPA-Cas13a is 10 aM and 1 aM, respectively. Meanwhile, the designed primers have a high specificity for P. aeruginosa mexX gene. These two methods were also verified with actual samples isolated from industrial settings and demonstrated great accuracy. Furthermore, the results of the two-step RPA-Cas13a assay could also be visualized using a commercial lateral flow dipstick with a LoD of 10 fM, which is a useful adjunt to the gold-standard qRT-PCR assay in field detection. Taken together, the procedure developed in this study using RPA and CRISPR-Cas13a provides a simple and fast way for detecting resistance genes.

Selecting Suitable Near-Native Lignins for Research.

There are several methods to isolate near-native lignins, including milled-wood lignin, enzymatic lignin, cellulolytic enzyme lignin, and enzymatic mild-acidolysis lignin. Which one is the most representative of the native lignin? Herein, near-native lignins were isolated from different plant groups and structurally analyzed to determine how well these lignins represented their native lignin counterparts. Analytical methods were applied to understand the molecular weight, monomer composition, and distribution of interunit linkages in the structure of the lignins. The results indicated that either enzymatic lignin or cellulolytic enzyme lignin may be used to represent native lignin in softwoods and hardwoods. None of the lignins, however, appeared to represent native lignins in grasses (monocot plants) because of substantial syringyl/guaiacyl differences. Complicating the understanding of grass lignin structure, large amounts of hydroxycinnamates acylate their polysaccharides and, when released, are often conflated with actual lignin monomers.

Antimicrobial Peptide-Inspired Design of Amino-Modified Lignin with Improved Antimicrobial Activities.

A major challenge to make use of lignin as an antimicrobial material is the weak antimicrobial activity of industrial lignin. Inspired by the antimicrobial mechanism of actions of antimicrobial peptides, alkyldiamines were employed as lysine mimics for lignin modifications. Accordingly, aminoalkyl-modified lignins with different degrees of substitution of amino groups and different hydrophobicity were synthesized. The chemical structure, properties, and antimicrobial activities of the as-prepared aminoalkyl lignins were thoroughly characterized with state-of-the-art technologies. The results indicated that aminobutyl lignin showed enhanced antimicrobial activity against S. aureus and E. coli and performed even better than copper ions. The antimicrobial mechanism of action of the as-prepared aminobutyl lignin was similar to that of polylysine, which damaged the cell membrane, leading to the leakage of intracellular molecules and death of the cell. This study provides a feasible approach to afford modified lignin with enhanced antimicrobial performance, which would facilitate the high-value valorization of lignin as biological materials.

Nutrient condition modulates the antibiotic tolerance of Pseudomonas aeruginosa.

The variation in nutrient content across diverse environments has a significant impact on the survival and metabolism of microorganisms. In this study, we examined the influence of nutrients on the antibiotic tolerance of the PAO1 strain of Pseudomonas aeruginosa. Our findings indicate that under nutrient-rich conditions, this strain exhibited relatively high tolerance to ceftazidime, chloramphenicol, and tetracycline, but not aminoglycosides and fluoroquinolones. Transcriptome analysis revealed that genes associated with antibiotic tolerance were expressed more efficiently in nutrient-rich media, including ribosomal protein genes and multidrug efflux pump genes, which conferred higher tetracycline tolerance to the strain. Furthermore, the genes responsible for translation, biosynthesis, and oxidative phosphorylation were suppressed when nutrients were limited, resulting in decreased metabolic activity and lower sensitivity to ciprofloxacin. Artificial interference with ATP synthesis utilizing arsenate confirmed that the curtailment of energy provision bolstered the observed tolerance to ciprofloxacin. In general, our results indicate that this strain of P. aeruginosa tends to activate its intrinsic resistance mechanisms in nutrient-rich environments, thereby enhancing resistance to certain antibiotics. Conversely, in nutrient-limited environments, the strain is more likely to enter a dormant state, which enables it to tolerate antibiotics to which it would otherwise be sensitive. These findings further suggest that antibiotics released in environments with varying eutrophication levels may have divergent effects on the development of bacterial antibiotic resistance.

Fundicoccus culcitae sp. nov., a novel potential bacteriocin producing bacterium isolated from a spoiled eye mask.

A Gram-positive, facultatively anaerobic, oval beaded-shape, oxidase-negative, and non-motile bacterium designated DM20194951T was isolated from a spoiled eye mask obtained from Guangdong, China. Based on the 16S rRNA gene sequence, phylogenetic analysis indicated that strain DM20194951T showed the highest sequence similarity (95.8%) to Fundicoccus ignavus WS4937T. Meanwhile, strain DM20194951T could be distinguished from the type strains in the genus Fundicoccus by distinct phenotypic and genotypic traits. Strain DM20194951T grew variably with 1-2% (w/v) NaCl and tolerated pH 6.0-10.0. Growth was observed from 28 to 37 °C. The diagnostic diamino acids in the cell-wall peptidoglycan consisted of aspartic and glutamic acids as well as alanine. The predominant fatty acids were C18:1 ω9c, C16:0, and C16:1 ω9c. In the polar lipid profile, two glycolipids, three phospholipids, one phosphatidylglycerol, and one diphosphatidylglycerol were found. No respiratory quinones were detected. The DM20194951T genome is 3.2 Mb in size and contains a G + C content of 38.1%. A gene cluster for lactococcin 972 family bacteriocin production was found in the DM20194951T genome. Based on morphological, genotypic, and phylogenetic data, strain DM20194951T should be considered to represent a novel species in the genus Fundicoccus, for which the name Fundicoccus culcitae sp. nov. is proposed with the type strain DM20194951T (= KCTC 43472T = GDMCC 1.3614T).

Tannic Acid Interfacial Modification of Prochloraz Ethyl Cellulose Nanoparticles for Enhancing the Antimicrobial Effect and Biosafety of Fungicides.

With the poorly soluble and intrinsically unstable feature, prochloraz (Pro) was confronted with lower bioavailability in the crop defense against fungal erosion. Therefore, it was a challenging project to explore the innovative antifungal compound delivery system for improving bioavailability. The superior adhesive fungicide formulation was supposed to be an efficient pathway to enhance transmembrane permeability and biological activity. According to abundant phenolic hydroxyl groups, tannic acid (TA) was an ideal modified adhesive biomaterial to improve interfacial interactions. The fundamental purpose of this research was focused on the synergistic mechanism of TA-interfacial-modified Pro-ethyl cellulose (EC) nanoparticles for improving bioavailability and biosafety. In the stability test, TA-modified Pro-EC nanoparticles had the capacity to reduce Pro initial release burst, extending a persistent validity and improving anti-photodegradation property. The toxicity index of Pro-EC and Pro-EC-TA was approximately 2.93-fold and 4.96-fold that of Pro technical against Fusarium graminearum (F. graminearum), respectively. Compared with nonmodified EC nanoparticles, TA-modified EC nanoparticles obtained eminent transmembrane permeability and superior adherence ability to F. graminearum, for hydroxyl and carboxyl groups of TA to enhance interaction with target cell membranes. The contents of cellular reactive oxygen species induced by Pro-EC and Pro-EC-TA nanoparticles were about 2.31 times and 3.00 times that of the control check (CK), respectively. Compared to the CK group, the membrane potential and ergosterol values of F. graminearum treated with Pro-EC-TA nanoparticles were drastically reduced by 74.91 and 56.20%, respectively. In the biosafety assay, the maximum half-lethal concentration value of the TA-modified Pro-EC nanoparticles indicated that the acute toxicity of the Pro-EC-TA nanoparticles to adult zebrafish was approximately 8.34-fold reduced compared to that of the Pro technical. These findings demonstrated that the successful interfacial modification of Pro-EC nanoparticles with TA was a highly efficient, environmentally safe, and promising alternative for sustainable agricultural application, thus making the fungicide formulation process more simplified, easier fabrication, and lower cost.

Outer Membrane Porins Contribute to Antimicrobial Resistance in Gram-Negative Bacteria.

Gram-negative bacteria depend on their cell membranes for survival and environmental adaptation. They contain two membranes, one of which is the outer membrane (OM), which is home to several different outer membrane proteins (Omps). One class of important Omps is porins, which mediate the inflow of nutrients and several antimicrobial drugs. The microorganism's sensitivity to antibiotics, which are predominantly targeted at internal sites, is greatly influenced by the permeability characteristics of porins. In this review, the properties and interactions of five common porins, OmpA, OmpC, OmpF, OmpW, and OmpX, in connection to porin-mediated permeability are outlined. Meanwhile, this review also highlighted the discovered regulatory characteristics and identified molecular mechanisms in antibiotic penetration through porins. Taken together, uncovering porins' functional properties will pave the way to investigate effective agents or approaches that use porins as targets to get rid of resistant gram-negative bacteria.

Geraniol attenuates virulence factors by inhibiting quorum sensing of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a ubiquitous opportunistic pathogen that can cause severe respiratory tract infections. Geraniol, a chemical component of essential oils, has antimicrobial and anti-inflammatory activities, along with low toxicity. However, the effect and mechanism of geraniol against P. aeruginosa virulence factors are rarely studied. In this study, we investigated the quorum sensing (QS) inhibitory effects and mechanisms of geraniol against P. aeruginosa PAO1, using physiological and biochemical techniques, quantitative reverse transcription polymerase chain reaction, and transcriptomics. Geraniol slightly affected P. aeruginosa PAO1 growth, prolonged the lag phase, and delayed growth periods in a concentration-dependent manner. Geraniol inhibited three QS systems of P. aeruginosa, las, rhl, and pqs by suppressing the expression level of their key genes, including the three signal synthetase encoding genes of lasI, rhlI, and pqsABCDEH, and the corresponding signal receptor encoding genes of lasR, rhlR, and pqsR. Geraniol also suppressed certain virulence genes regulated by these three QS systems, including rhlABC, lasAB, lecAB, phzABMS, and pelABG, resulting in the attenuation of the related virulence factors, rhamnolipids, exoprotease LasA, elastase, lectin, pyocyanin, and biofilm. In conclusion, geraniol can suppress the virulence factors of P. aeruginosa PAO1 by inhibiting the three QS systems of las, rhl, and pqs. This study is significant for improving the treatment of bacterial infections caused by P. aeruginosa.

Effect of γ-polyglutamic acid on the physicochemical properties of soybean protein isolate-stabilized O/W emulsion.

An increased interest has been observed in the application of soybean protein isolate (SPI) into O/W emulsion because of the amphipathic characteristics of SPI. However, at pH around 4.5, SPI was almost lost its hydrophilic characteristic, thus greatly limiting its application in emulsion under an acidic environment. Therefore, this drawback of SPI needs to be urgently solved. This study aims to investigate the effect of γ-polyglutamic acid (γ-PGA) on physicochemical properties of SPI-stabilized O/W emulsion. The results suggested that the interaction between γ-PGA and SPI improved the SPI solubility in solution, and increased emulsifying properties of SPI in the pH range of 4.0-5.0 via electrostatic interaction. Meanwhile, the charge neutralisation between SPI emulsions with γ-PGA was confirmed via ζ-potentiometry. With the presence of γ-PGA in emulsion at pH 4.0 and 5.0, the electrostatic complexation between SPI and anionic γ-PGA exhibited decreased the viscosity of SPI emulsion, which might be related to the phenomenon as indicated by the confocal laser scanning microscope measurements. Therefore, the electrostatic complexation between SPI and γ-PGA suggested that the promising potential of γ-PGA to be used in SPI-stabilized O/W emulsion under an acidic environment.

High value valorization of lignin as environmental benign antimicrobial.

Lignin is a natural aromatic polymer of p-hydroxyphenylpropanoids with various biological activities. Noticeably, plants have made use of lignin as biocides to defend themselves from pathogen microbial invasions. Thus, the use of isolated lignin as environmentally benign antimicrobial is believed to be a promising high value approach for lignin valorization. On the other hand, as green and sustainable product of plant photosynthesis, lignin should be beneficial to reduce the carbon footprint of antimicrobial industry. There have been many reports that make use of lignin to prepare antimicrobials for different applications. However, lignin is highly heterogeneous polymers different in their monomers, linkages, molecular weight, and functional groups. The structure and property relationship, and the mechanism of action of lignin as antimicrobial remains ambiguous. To show light on these issues, we reviewed the publications on lignin chemistry, antimicrobial activity of lignin models and isolated lignin and associated mechanism of actions, approaches in synthesis of lignin with improved antimicrobial activity, and the applications of lignin as antimicrobial in different fields. Hopefully, this review will help and inspire researchers in the preparation of lignin antimicrobial for their applications.

N,N-dimethyl chitosan oligosaccharide (DMCOS) promotes antifungal activity by causing mitochondrial damage.

By modifying chitosan oligosaccharide (COS) with the Eschweiler-Clarke reaction, the chitosan oligosaccharide derivative DMCOS was synthesized. FT-IR, 1D and 2D NMR spectra, MALDI-ToF MS, and elemental analysis were applied to analyze the structure of DMCOS, which revealed that the primary amines were converted into tertiary amines after methylation. DMCOS displayed less thermal stability than COS. In comparison to COS, it was discovered that DMCOS possessed more potent antimicrobial activity against four bacterial strains (Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa) and three yeast strains (Candida albicans, Candida tropicalis, and Candida parapsilosis). The antioxidant studies indicated that DMCOS had less antioxidant activity than COS. Consequently, ROS level elevated in C. albicans cells following treatment with DMCOS, which decreased mitochondrial membrane potential. It was recalled that DMCOS may kill C. albicans by causing mitochondrial damage. In addition, DMCOS was demonstrated to be non-cytotoxic.

Proteomic signatures of synergistic interactions in antimicrobials.

Mounting evidence has shown that antimicrobial agents can interfere synergistically with bacterial viability and proliferation when acting together at both the planktonic and biofilm levels without clear underlying molecular mechanisms. Here, multiplexed proteomics by iTRAQ was used to study the interplay between two biocides, the isothiazolone 1,2-benzisothiazolin-3-one (BIT) and the chelating agent disodium ethylenediaminetetraacetic acid (EDTA-2Na), employing the Citrobacter werkmanii as a model system. We first confirmed that these two biocides act synergistically on this bacterial species and then extracted the proteomic profiles of C. werkmanii cells in the presence of BIT, EDTA-2Na, and their combinations. In particular, we identified 43 core proteins that are differentially expressed in all three conditions simultaneously. Meanwhile, we found that these core proteins are consistently up-regulated when these two biocides are present, but not for single biocides, where we found a balanced mix of up- and down-regulation. Meanwhile, most of the deletion mutants of the core DEPs exhibited biofilm growth inhibition under joint biocide action, while their response was very heterogenous, with respect to the wild-type strain. Together, our results show that while BIT and EDTA-2Na act on multiple protein targets, they interact synergistically at the protein level in a very consistent manner. SIGNIFICANCE: Our preliminary experiments have demonstrated that a combination of 1,2-benzisothiazolin-3-one (BIT) and EDTA-2Na shows higher inhibitory effects on planktonic growth and biofilm formation in both C. werkmanii and Staphylococcus aureus than when these two biocides act alone. However, the mechanistic basis of such synergistic interaction is still unknown. Therefore, the key proteins involved in the above-mentioned enhanced antimicrobial synergy were elucidated using multiplexed proteomics analysis by isobaric tags for relative and absolute quantification (iTRAQ). Our results reveal that the joint action of BIT and EDTA-2Na induces consistent protein expression alteration in a set of core proteins of C. werkmanii, which underlies a strong synergistic antimicrobial effect, which increase our understanding of the action modes of BIT and EDTA-2Na as well as their combinations.

ompX contribute to biofilm formation, osmotic response and swimming motility in Citrobacter werkmanii.

Citrobacter werkmanii, an aerobe and mesophilic Proteobacterium, is universal in industrial putrefaction, coastal water, and human blood. Our previous studies have discovered that outer membrane protein X (OmpX) of C. werkmanii is involved in calcium response, but the underlying mechanisms and its molecular characteristics remain elusive. To that end, the ompX gene was deleted from the genome of C. werkmanii and its phenotypic variations were thoroughly investigated in conjunction with the wild type (WT) and complementary strains using biochemical and molecular techniques such as RNA-Seq, respectively. The results demonstrated that deleting ompX reduces biofilm formation on polystyrene and glass surfaces. Meanwhile, ΔompX's swimming ability but not for its twitching or swarming abilities, was also reduced on semi-solid plates compared with WT, which was caused by inhibition of flagellar assembly genes, such as flgC, flhB, and fliE, etc. Furthermore, ompX inactivation altered susceptibility to various bactericide classes, as well as responses to Ca2+ and Mg2+ stress. In addition, when compared to WT, ΔompX captures a total of 1,357 deferentially expressed genes (DEGs), of which 465 were up-regulated and 892 were down-regulated, which can be enriched into various GO ontology and KEGG pathway terms. Furthermore, ompX, as well as ompD and ompW, can be modulated at the transcriptional levels by rbsR and tdcA. Overall, the ompX gene contributed to a variety of biological functions in C. werkmanii and could be served as a targeted site for controlling biofilm formation and developing new bactericides.

The interplays between epigallocatechin-3-gallate (EGCG) and Aspergillus niger RAF106 based on metabolism.

Epigallocatechin-3-gallate (EGCG) is a vital kind of catechin with high bioactive activities, however, limited research has been conducted to elucidate the molecular basis of EGCG biotransformation by Aspergillus niger and the underlying regulatory mechanisms. In this study, A. niger RAF106, isolated from Pu-erh tea, was applied to conduct the EGCG fermentation process, and the samples were collected at different fermentation times to determine the intermediary metabolites of EGCG and the metabolome as well as physiological activity changes of A. niger RAF106. The results demonstrated that EGCG enhances the growth of A. niger RAF106 by promoting conidial germination and hyphae branching. Meanwhile, metabolomic analyses indicated that EGCG significantly regulates the amino acid metabolism of A. niger RAF106. Furthermore, metabolomic analyses also revealed that the levels of original secondary metabolites in the supernatant of the cultures changed significantly from the fermentation stage M2 to M3, in which the main differentially changed metabolites (DCMs) were flavonoids. Most of these flavonoids exhibited antioxidant properties and thus increased the radical scavenging activity of the supernatant of the cultures. In addition, we also found several intermediary metabolites of EGCG, GA, and EGC, including oolonghomobisflavan A, (-)-Epigallocatechin 3, 5-di-gallate, (-)-Epigallocatechin 3-(3-methyl-gallate) (-)-Catechin 3-O-gallate, 4'-Methyl-(-)-epigallocatechin 3-(4-methyl-gallate), myricetin, prodelphinidin B, 7-galloylcatechin, and 3-hydroxyphenylacetic acid. These findings contribute to improving the bioavailability of EGCG and help mine highly active metabolites, which can be used as raw materials for the development of pharmaceutical intermediates or functional foods. In addition, the results also provide a theoretical basis for better control of the risk of A. niger origin and the regulatory mechanisms of the biotransformation process mediated by A. niger.

Microbiologically Influenced Corrosion of Q235 Carbon Steel by Ectothiorhodospira sp.

The biological sulfur cycle is closely related to iron corrosion in the natural environment. The effect of the sulfur-oxidising bacterium Ectothiorhodospira sp., named PHS-Q, on the metal corrosion behaviour rarely has been investigated. In this study, the corrosion mechanism of Q235 carbon steel in a PHS-Q-inoculated medium is discussed via the characterization of the morphology and the composition of the corrosion products, the measurement of local corrosion and the investigation of its electrochemical behaviour. The results suggested that, initially, PHS-Q assimilates sulfate to produce H2S directly or indirectly in the medium without sulfide. H2S reacts with Fe2+ to form an inert film on the coupon surface. Then, in localised areas, bacteria adhere to the reaction product and use the oxidation of FeS as a hydrogen donor. This process leads to a large cathode and a small anode, which incurs pitting corrosion. Consequently, the effect of PHS-Q on carbon steel corrosion behaviour is crucial in an anaerobic environment.

Outer membrane protein of OmpF contributes to swimming motility, biofilm formation, osmotic response as well as the transcription of maltose metabolic genes in Citrobacter werkmanii.

Bacterial outer membrane proteins (Omps) are essential for environmental sensing, stress responses, and substance transport. Our previous study discovered that OmpA contributes to planktonic growth, biocide resistance, biofilm formation, and swimming motility in Citrobacter werkmanii, whereas the molecular functions of OmpF in this strain are largely unknown. Thus, in this study, the ompF gene was firstly knocked out from the genome of C. werkmanii using a homologous recombination method, and its phenotypical alternations of ∆ompF were then thoroughly characterized using biochemical and molecular approaches with the parental wild type (WT) and complementary (∆ompF-com) strains. The results demonstrated that the swimming ability of ∆ompF on semi-solid plates was reduced compared to WT due to the down-regulation of flgC, flgH, fliK, and fliF. Meanwhile, ompF deletion reduces biofilm formation on both glass and polystyrene surfaces due to decreased cell aggregation. Furthermore, ompF inactivation induced different osmotic stress (carbon sources and metal ions) responses in its biofilms when compared to WT and ∆ompF-com. Finally, a total of 6 maltose metabolic genes of lamB, malE, malK, malG, malM, and malF were all up-regulated in ∆ompF. The gene knockout and HPLC results revealed that the MalEFGK2 cluster was primarily responsible for maltose transport in C. werkmanii. Furthermore, we discovered for the first time that the upstream promoter of OmpF and its transcription can be combined with and negatively regulated by MalT. Overall, OmpF plays a role in a variety of biochemical processes and molecular functions in C. werkmanii, and it may even act as a targeted site to inhibit biofilm formation.

Visible light-regulated cationic polymer coupled with photodynamic inactivation as an effective tool for pathogen and biofilm elimination.

BACKGROUND: Pathogenic microorganism pollution has been a challenging public safety issue, attracting considerable scientific interest. A more problematic aspect of this phenomenon is that planktonic bacteria exacerbate biofilm formation. There is an overwhelming demand for developing ultra-efficient, anti-drug resistance, and biocompatibility alternatives to eliminate stubborn pathogenic strains and biofilms. RESULTS: The present work aims to construct a visible light-induced anti-pathogen agents to ablate biofilms using the complementary merits of ROS and cationic polymers. The photosensitizer chlorin e6-loaded polyethyleneimine-based micelle (Ce6-TPP-PEI) was constructed by an amphiphilic dendritic polymer (TPP-PEI) and physically loaded with photosensitizer chlorin e6. Cationic polymers can promote the interaction between photosensitizer and Gram-negative bacteria, resulting in enhanced targeting of PS and lethality of photodynamic therapy, and remain active for a longer duration to prevent bacterial re-growth when the light is turned off. As expected, an eminent antibacterial effect was observed on the Gram-negative Escherichia coli, which is usually insensitive to photosensitizers. Surprisingly, the cationic polymer and photodynamic combination also exert significant inhibitory and ablative effects on fungi and biofilms. Subsequently, cell hemolysis assessments suggested its good biocompatibility. CONCLUSIONS: Given the above results, the platform developed in this work is an efficient and safe tool for public healthcare and environmental remediation.

The role of structural evolution in the complexation and flocculation of heavy metals by the microbial product poly-γ-glutamic acid.

The process and mechanism of heavy metal flocculation with extracellular polymeric substances (EPS) secreted by microorganisms, are crucial to their fate in natural environment, wastewater treatment and soil bioremediation applications. However, the structural features of EPS and the relationship between these features and the flocculation process and mechanism remain unclear. In the present study, structural features of the microbial product poly-γ-glutamic acid (γ-PGA) complexed with the heavy metal ions Pb2+ and Cu2+ were characterized and the evolution of these features was identified as having a key role in the flocculation process and mechanism. The secondary structure of the γ-PGA-Pb complex changed significantly, while that of the γ-PGA-Cu complex was only slightly altered. The significant structural change in γ-PGA-Pb was found to be responsible for the combination of residual COOH and Pb2+, promoting the bridging of inter-colloids and faster growth of hydrodynamic diameter. If the conformation changed sufficiently, such as with the γ-PGA-Pb complex in the pH range 4.6-6.2, pH had no impact on the conversion ratio. The unchanged structure of γ-PGA-Cu prevented the flocculation process, although the coordination mode of γ-PGA-Cu resulted in a higher biosorption capacity. This in-depth molecular-level study provides insight into the γ-PGA flocculation mechanism, promoting the use of γ-PGA and γ-PGA producing microorganisms for application in various remediation strategies.