Bacillamide D produced by Bacillus cereus from the mouse intestinal bacterial collection (miBC) is a potent cytotoxin in vitro.

The gut microbiota influences human health and the development of chronic diseases. However, our understanding of potentially protective or harmful microbe-host interactions at the molecular level is still in its infancy. To gain further insights into the hidden gut metabolome and its impact, we identified a cryptic non-ribosomal peptide BGC in the genome of Bacillus cereus DSM 28590 from the mouse intestine ( www.dsmz.de/miBC ), which was predicted to encode a thiazol(in)e substructure. Cloning and heterologous expression of this BGC revealed that it produces bacillamide D. In-depth functional evaluation showed potent cytotoxicity and inhibition of cell migration using the human cell lines HCT116 and HEK293, which was validated using primary mouse organoids. This work establishes the bacillamides as selective cytotoxins from a bacterial gut isolate that affect mammalian cells. Our targeted structure-function-predictive approach is demonstrated to be a streamlined method to discover deleterious gut microbial metabolites with potential effects on human health.

Enhanced biodegradation of phenol under Cr(VI) stress by microbial collaboration and potential application of machine learning for phenol biodegradation.

Cr(VI) and phenol commonly coexist in wastewater, posing a great threat to the environment and human health. However, it is still a challenge for microorganisms to degrade phenol under high Cr(VI) stress. In this study, the phenol-degrading strain Bacillus cereus ZWB3 was co-cultured with the Cr(VI)-reducing strain Bacillus licheniformis MZ-1 to enhance phenol biodegradation under Cr(Ⅵ) stress. Compared with phenol-degrading strain ZWB3, which has weak tolerance to Cr(Ⅵ), and Cr(Ⅵ)-reducing strain MZ-1, which has no phenol-degrading ability, the co-culture of two strains could significantly increase the degraded rate and capacity of phenol. In addition, the co-cultured strains exhibited phenol degradation ability over a wide pH range (7-10). The reduced content of intracellular proteins and polysaccharides produced by the co-cultured strains contributed to the enhancement of phenol degradation and Cr(Ⅵ) tolerance. The determination coefficients R2, RMSE, and MAPE showed that the BP-ANN model could predict the degradation of phenol under various conditions, which saved time and economic cost. The metabolic pathway of microbial degradation of phenol was deduced by metabolic analysis. This study provides a valuable strategy for wastewater treatment containing Cr(Ⅵ) and phenol.

A High-Homology Region Provides the Possibility of Detecting ß-Barrel Pore-Forming Toxins from Various Bacterial Species.

The pathogenicity of many bacteria, including Bacillus cereus and Staphylococcus aureus, depends on pore-forming toxins (PFTs), which cause the lysis of host cells by forming pores in the membranes of eukaryotic cells. Bioinformatic analysis revealed a region homologous to the Lys171-Gly250 sequence in hemolysin II (HlyII) from B. cereus in over 600 PFTs, which we designated as a "homologous peptide". Three ß-barrel PFTs were used for a detailed comparative analysis. Two of them-HlyII and cytotoxin K2 (CytK2)-are synthesized in Bacillus cereus sensu lato; the third, S. aureus α-toxin (Hla), is the most investigated representative of the family. Protein modeling showed certain amino acids of the homologous peptide to be located on the surface of the monomeric forms of these ß-barrel PFTs. We obtained monoclonal antibodies against both a cloned homologous peptide and a 14-membered synthetic peptide, DSFNTFYGNQLFMK, as part of the homologous peptide. The HlyII, CytK2, and Hla regions recognized by the obtained antibodies, as well as an antibody capable of suppressing the hemolytic activity of CytK2, were identified in the course of this work. Antibodies capable of recognizing PFTs of various origins can be useful tools for both identification and suppression of the cytolytic activity of PFTs.

Cereulide production capacities and genetic properties of 31 emetic Bacillus cereus group strains.

The highly potent toxin cereulide is a frequent cause of foodborne intoxications. This extremely resistant toxin is produced by Bacillus cereus group strains carrying the plasmid encoded cesHPTABCD gene cluster. It is known that the capacities to produce cereulide vary greatly between different strains but the genetic background of these variations is not clear. In this study, cereulide production capacities were associated with genetic characteristics. For this, cereulide levels in cultures of 31 strains were determined after incubation in tryptic soy broth for 24 h at 24 °C, 30 °C and 37 °C. Whole genome sequencing based data were used for an in-depth characterization of gene sequences related to cereulide production. The taxonomy, population structure and phylogenetic relationships of the strains were evaluated based on average nucleotide identity, multi-locus sequence typing (MLST), core genome MLST and single nucleotide polymorphism analyses. Despite a limited strain number, the approach of a genome wide association study (GWAS) was tested to link genetic variation with cereulide quantities. Our study confirms strain-dependent differences in cereulide production. For most strains, these differences were not explainable by sequence variations in the cesHPTABCD gene cluster or the regulatory genes abrB, spo0A, codY and pagRBc. Likewise, the population structure and phylogeny of the tested strains did not comprehensively reflect the cereulide production capacities. GWAS yielded first hints for associated proteins, while their possible effect on cereulide synthesis remains to be further investigated.

Screening of cellulose-degrading bacteria and optimization of cellulase production from Bacillus cereus A49 through response surface methodology.

Cellulose-degrading microorganisms hold immense significance in utilizing cellulose resources efficiently. The screening of natural cellulase bacteria and the optimization of fermentation conditions are the hot spots of research. This study meticulously screened cellulose-degrading bacteria from mixed soil samples adopting a multi-step approach, encompassing preliminary culture medium screening, Congo red medium-based re-screening, and quantification of cellulase activity across various strains. Particularly, three robust cellulase-producing strains were identified: A24 (MT740356.1 Brevibacillus borstelensis), A49 (MT740358.1 Bacillus cereus), and A61 (MT740357.1 Paenibacillus sp.). For subsequent cultivation experiments, the growth curves of the three obtained isolates were monitored diligently. Additionally, optimal CMCase production conditions were determined, keeping CMCase activity as a key metric, through a series of single-factor experiments: agitation speed, cultivation temperature, unit medium concentration, and inoculum volume. Maximum CMCase production was observed at 150 rpm/37 °C, doubling the unit medium addition, and a 5 mL inoculation volume. Further optimization was conducted using the selected isolate A49 employing response surface methodology. The software model recommended a 2.21fold unit medium addition, 36.11 °C temperature, and 4.91 mL inoculant volume for optimal CMCase production. Consequently, three parallel experiments were conducted based on predicted conditions consistently yielding an average CMCase production activity of 15.63 U/mL, closely aligning with the predicted value of 16.41 U/mL. These findings validated the reliability of the model and demonstrated the effectiveness of optimized CMCase production conditions for isolate A49.

Impact of Bacillus cereus SPB-10 on Growth Promotion of Wheat (Triticum aestivum L.) Under Arsenic-Contaminated Soil.

This study investigates the impact of bacteria on arsenic reduction in wheat plants, highlighting the potential of microbe-based eco-friendly strategies for plant growth. In the present study, bacterial isolate SPB-10 was survived at high concentration against both form of arsenic (As3+ and As5+). SPB-10 produced 5.2 g/L and 11.3 g/L of exo-polysaccharide at 20 ppm of As3+ and As5+, respectively, whereas qualitative examination revealed the highest siderophores ability. Other PGP attributes such as IAA production were recorded 52.12 mg/L and 95.82 mg/L, phosphate solubilization was 90.23 mg/L and 129 mg/L at 20 ppm of As3+ and As5+, respectively. Significant amount of CAT, APX, and Proline was also observed at 20 ppm of As3+ and As5+ in SPB-10. Isolate SPB-10 was molecularly identified as Bacillus cereus through 16S rRNA sequencing. After 42 days, wheat plants inoculated with SPB-10 had a 25% increase in shoot length and dry weight, and 26% rise in chlorophyll-a pigment under As5+ supplemented T4 treatment than control. Reducing sugar content was increased by 24% in T6-treated plants compared to control. Additionally, SPB-10 enhanced the content of essential nutrients (NPK), CAT, and APX in plant's-leaf under both As3+ and As5+ stressed conditions after 42 days. The study found that arsenic uptake in plant roots and shoots decreased in SPB-10-inoculated plants, with the maximum reduction observed in As5+ treated plants. Bio-concentration factor-BCF was reduced by 90.89% in SPB-10-inoculated treatment T4 after 42 days. This suggests that Bacillus cereus-SPB-10 may be beneficial for plant growth in arsenic-contaminated soil.

Biodegradation of polyethylene (PE), polypropylene (PP), and polystyrene (PS) microplastics by floc-forming bacteria, Bacillus cereus strain SHBF2, isolated from a commercial aquafarm.

The ubiquitous proximity of the commonly used microplastic (MP) particles particularly polyethylene (PE), polypropylene (PP), and polystyrene (PS) poses a serious threat to the environment and human health globally. Biological treatment as an environment-friendly approach to counter MP pollution has recent interest when the bio-agent has beneficial functions in their ecosystem. This study aimed to utilize beneficial floc-forming bacteria Bacillus cereus SHBF2 isolated from an aquaculture farm in reducing the MP particles (PE, PP, and PS) from their environment. The bacteria were inoculated for 60 days in a medium containing MP particle as a sole carbon source. On different days of incubation (DOI), the bacterial growth analysis was monitored and the MP particles were harvested to examine their weight loss, surface changes, and alterations in chemical properties. After 60 DOI, the highest weight loss was recorded for PE, 6.87 ± 0.92%, which was further evaluated to daily reduction rate (k), 0.00118 day-1, and half-life (t1/2), 605.08 ± 138.52 days. The OD value (1.74 ± 0.008 Abs.) indicated the higher efficiency of bacteria for PP utilization, and so for the colony formation per define volume (1.04 × 1011 CFU/mL). Biofilm formation, erosions, cracks, and fragments were evident during the observation of the tested MPs using the scanning electron microscope (SEM). The formation of carbonyl and alcohol group due to the oxidation and hydrolysis by SHBF2 strain were confirmed using the Fourier transform infrared spectroscopic (FTIR) analysis. Additionally, the alterations of pH and CO2 evolution from each of the MP type ensures the bacterial activity and mineralization of the MP particles. The findings of this study have confirmed and indicated a higher degree of biodegradation for all of the selected MP particles. B. cereus SHBF2, the floc-forming bacteria used in aquaculture, has demonstrated a great potential for use as an efficient MP-degrading bacterium in the biofloc farming system in the near future to guarantee a sustainable green aquaculture production.

Microbial degradation of low-density polyethylene (LDPE) and polystyrene using Bacillus cereus (OR268710) isolated from plastic-polluted tropical coastal environment.

The study focused on marine bacteria, specifically Bacillus cereus, sourced from heavily polluted coastal areas in Tamil Nadu, aiming to assess their efficacy in degrading low-density polyethylene (LDPE) and polystyrene over a 42-day period. When LDPE and polystyrene films were incubated with Bacillus cereus, they exhibited maximum weight losses of 4.13 ± 0.81 % and 14.13 ± 2.41 %, respectively. Notably, polystyrene exhibited a higher reduction rate (0.0036 day-1) and a shorter half-life (195.29 days). SEM images of the treated LDPE and polystyrene unveiled surface erosion with cracks. The energy dispersive X-ray (EDX) analysis revealed elevated carbon content and the presence of oxygen in the treated LDPE and polystyrene films. The ATR-FTIR spectra exhibited distinctive peaks corresponding to functional groups, with observable peak shifts in the treated films. Notable increases were detected in carbonyl, internal double bond, and vinyl indices across all treated groups. Additionally, both treated LDPE and polystyrene showed reduced crystallinity. This research sheds light on Bacillus cereus (OR268710) biodegradation capabilities, emphasizing its potential for eco-friendly waste management in coastal regions.

Structures and activation mechanism of the Gabija anti-phage system.

Prokaryotes have evolved intricate innate immune systems against phage infection1-7. Gabija is a highly widespread prokaryotic defence system that consists of two components, GajA and GajB8. GajA functions as a DNA endonuclease that is inactive in the presence of ATP9. Here, to explore how the Gabija system is activated for anti-phage defence, we report its cryo-electron microscopy structures in five states, including apo GajA, GajA in complex with DNA, GajA bound by ATP, apo GajA-GajB, and GajA-GajB in complex with ATP and Mg2+. GajA is a rhombus-shaped tetramer with its ATPase domain clustered at the centre and the topoisomerase-primase (Toprim) domain located peripherally. ATP binding at the ATPase domain stabilizes the insertion region within the ATPase domain, keeping the Toprim domain in a closed state. Upon ATP depletion by phages, the Toprim domain opens to bind and cleave the DNA substrate. GajB, which docks on GajA, is activated by the cleaved DNA, ultimately leading to prokaryotic cell death. Our study presents a mechanistic landscape of Gabija activation.

Reforming CO2 bio-mitigation utilizing Bacillus cereus from hypersaline realms in pilot-scale bubble column bioreactor.

The bubble column reactor of 10 and 20 L capacity was designed to bio-mitigate 10% CO2 (g) with 90% air utilizing thermophilic bacteria (Bacillus cereus SSLMC2). The maximum biomass yield during the growth phase was obtained as 9.14 and 10.78 g L-1 for 10 and 20 L capacity, respectively. The maximum removal efficiency for CO2 (g) was obtained as 56% and 85% for the 10 and 20 L reactors, respectively. The FT-IR and GC-MS examination of the extracellular and intracellular samples identified value-added products such as carboxylic acid, fatty alcohols, and hydrocarbons produced during the process. The total carbon balance for CO2 utilization in different forms confirmed that B. cereus SSLMC2 utilized 1646.54 g C in 10 L and 1587 g of C in 20 L reactor out of 1696.13 g of total carbon feed. The techno-economic assessment established that the capital investment required was $286.21 and $289.08 per reactor run of 11 days and $0.167 and $0.187 per gram of carbon treated for 10 and 20 L reactors, respectively. The possible mechanism pathways for bio-mitigating CO2 (g) by B. cereus SSLMC2 were also presented utilizing the energy reactions. Hence, the work presents the novelty of utilizing thermophilic bacteria and a bubble column bioreactor for CO2 (g) bio-mitigation.

Bacillus cereus G2 alleviate salt stress in Glycyrrhiza uralensis Fisch. by balancing the downstream branches of phenylpropanoids and activating flavonoid biosynthesis.

The salinity environment is one of the biggest threats to Glycyrrhiza uralensis Fisch. (G. uralensis) growth, resulting from the oxidative stress caused by excess reactive oxygen species (ROS). Flavonoids are the main pharmacodynamic composition and help maintain ROS homeostasis and mitigate oxidative damage in G. uralensis in the salinity environment. To investigate whether endophytic Bacillus cereus G2 can improve the salt-tolerance of G. uralensis through controlling flavonoid biosynthesis, the transcriptomic and physiological analysis of G. uralensis treated by G2 in the saline environment was conducted, focused on flavonoid biosynthesis-related pathways. Results uncovered that salinity inhibited flavonoids synthesis by decreasing the activities of phenylalanine ammonialyase (PAL) and 4-coumarate-CoA ligase (4CL) (42% and 39%, respectively) due to down-regulated gene Glyur000910s00020578 at substrate level, and then decreasing the activities of chalcone isomerase (CHI) and chalcone synthase (CHS) activities (50% and 42%, respectively) due to down-regulated genes Glyur006062s00044203 and Glyur000051s00003431, further decreasing isoliquiritigenin content by 53%. However, salt stress increased liquiritin content by 43%, which might be a protective mechanism of salt-treated G. uralensis seedlings. Interestingly, G2 enhanced PAL activity by 27% whereas reduced trans-cinnamate 4-monooxygenase (C4H) activity by 43% which could inhibit lignin biosynthesis but promote flavonoid biosynthesis of salt-treated G. uralensis at the substrate level. G2 decreased shikimate O-hydroxycinnamoyltransferase (HCT) activity by 35%, increased CHS activity by 54% through up-regulating the gene Glyur000051s00003431 encoding CHS, and increased CHI activity by 72%, thereby decreasing lignin (34%) and liquiritin (24%) content, but increasing isoliquiritigenin content (35%), which could mitigate oxidative damage and changed salt-tolerance mechanism of G. uralensis.

Effect of novel and conventional food processing technologies on Bacillus cereus spores.

This chapter provides a summary of the effect of thermal and non-thermal processing technologies on Bacillus cereus spores, a well-known pathogenic bacterium associated with foodborne illnesses. B. cereus has been frequently detected in rice, milk products, infant food, liquid eggs products and meat products all over the world. This Gram positive, rod-shaped, facultative anaerobe can produce endospores that can withstand pasteurization, UV radiation, and chemical reagents commonly used for sanitization. B. cereus spores can germinate into vegetative cells that can produce toxins. The conventional regime for eliminating spores from food is retorting which uses the application of high temperature (121 °C). However, at this temperature, there could be a significant amount of loss in the organoleptic and functional qualities of the food components, especially proteins. This leads to the research on the preventive measures against germination and if possible, to reduce the resistance before using a non-thermal technology (temperatures less than retorting-121 °C) for inactivation. This chapter reviews the development and success of several food processing technologies in their ability to inactivate B. cereus spores in food.

Functional and structural characterisation of RimL from Bacillus cereus, a new Nα-acetyltransferase of ribosomal proteins that was wrongly assigned as an aminoglycosyltransferase.

Enzymes of the GNAT (GCN5-relate N-acetyltransferases) superfamily are important regulators of cell growth and development. They are functionally diverse and share low amino acid sequence identity, making functional annotation difficult. In this study, we report the function and structure of a new ribosomal enzyme, Nα-acetyl transferase from Bacillus cereus (RimLBC), a protein that was previously wrongly annotated as an aminoglycosyltransferase. Firstly, extensive comparative amino acid sequence analyses suggested RimLBC belongs to a cluster of proteins mediating acetylation of the ribosomal protein L7/L12. To assess if this was the case, several well established substrates of aminoglycosyltransferases were screened. The results of these studies did not support an aminoglycoside acetylating function for RimLBC. To gain further insight into RimLBC biological role, a series of studies that included MALDI-TOF, isothermal titration calorimetry, NMR, X-ray protein crystallography, and site-directed mutagenesis confirmed RimLBC affinity for Acetyl-CoA and that the ribosomal protein L7/L12 is a substrate of RimLBC. Last, we advance a mechanistic model of RimLBC mode of recognition of its protein substrates. Taken together, our studies confirmed RimLBC as a new ribosomal Nα-acetyltransferase and provide structural and functional insights into substrate recognition by Nα-acetyltransferases and protein acetylation in bacteria.

Genome sequencing and protein modeling unraveled the 2AP biosynthesis in Bacillus cereus DB25.

2-Acetyl-1-pyrroline (2AP) is an important and major flavor aroma compound responsible for the fragrance of basmati rice, cheese, wine, and several other food products. Biosynthesis of 2AP in aromatic rice and a few other plant species is associated with a recessive Betaine aldehyde dehydrogenase 2 (BADH2) gene. However, the literature is scant on the relationship between the functional BADH2 gene and 2AP biosynthesis in prokaryotic systems. Therefore, in the present study, we aimed to explore the functionality of the BADH2 gene for 2AP biosynthesis in 2AP synthesizing rice rhizobacterial isolate Bacillus cereus DB25 isolated from the rhizosphere of basmati rice (Oryza sativa L.). Full-length BcBADH2 sequence was obtained through whole genome sequencing (WGS) and further confirmed through traditional PCR and Sanger sequencing. Then the functionality of the BcBADH2 gene was evaluated in-silico through bioinformatics analysis and protein docking studies and further experimentally validated through enzyme assay. The sequencing and bioinformatics analysis results revealed a full-length 1485 bp BcBADH2 coding sequence without any deletion or premature stop codons. Full-length BcBADH2 was found to encode a fully functional protein of 54.08 kDa with pI of 5.22 and showed the presence of the conserved amino acids responsible for enzyme activity. The docking studies confirmed a good affinity between the protein and its substrate whereas the presence of BcBADH2 enzyme activity confirmed the functionality of BADH2 enzyme in B. cereus DB25. In conclusion, the findings of the present study suggest that B. cereus DB25 is able to synthesize 2AP despite a functional BADH2 gene and there may be a different molecular mechanism responsible for 2AP biosynthesis in bacterial systems, unlike that found in aromatic rice and other eukaryotic plant species.

Bacterial screening in Indian coastal regions for efficient polypropylene microplastics biodegradation.

Polypropylene based medical devices significantly increased production and usage in COVID-19 pandemic states, and this material is very resilient in the environment. Thus, more than ever, rapid action is needed to reduce this pollution. This study focuses on the degradation of polypropylene microplastics (PP MPs) by unique marine bacterial strains obtained from the Thoundi (Bacillus tropicus, Bacillus cereus, Stenotrophomonas acidaminiphila, and Brucella pseudintermedia) and Rameshwaram coasts (Bacillus cereus). Those above five bacterial strains were chosen after preliminary screening of their hydrophobicity, biofilm-forming capabilities, and responsiveness to the zone of clearance technique. During the biodegradation process (28 days), the growth, metabolic activity, and viability of these five isolates were all raised. After the post-biodegradation process, the weight loss percentages of the mentioned bacterial strains treated with PP MPs gradually decreased, with values of 51.5 ± 0.5 %, 47.5 ± 0.5 %, 33 ± 1 %, 28.5 ± 0.5 and 35.5 ± 0.5 %, respectively. UV-Vis DRS and SEM analysis confirmed that bacterial strains adhering to MPs cause cracks and cavities on their surface. The degradation of PP MPs can be inferred from alterations in the FT-IR spectrum, specifically in the carbonyl group range of 1100-1700 cm-1, as well as changes in the 1H NMR spectrum, including chemical shift and proton peak pattern alterations. Bacterial strains facilitated the degradation of PP MPs through the secretion of hydrolase-categorized enzymes of protease, lipase, and esterase. The findings of this study indicate that marine bacteria may possess distinctive characteristics that facilitate the degradation of plastic waste and contribute to environmental conservation.

Structural and biochemical insights into the mechanism of the Gabija bacterial immunity system.

The Gabija system is a newly discovered bacterial immune system that consists of GajA and GajB. Here we report the cryo-EM structure of the Gabija complex from Bacillus cereus VD045 at 3.6 Å, which provides the direct evidence of interactions between GajA and GajB. The Gabija complex is an octameric ring structure with four GajA and four GajB. GajA is an OLD nucleases family protein, while GajB belongs to the SF1 helicases. The Gabija complex has sequence-specific DNA nuclease activity and prefers circular rather than linear DNA as substrate, its activity is more sensitive to concentrations change of nucleotides compared to GajA alone. Our data suggest a mechanism of Gabija immunity: the nuclease activity of Gabija complex is inhibited under physiological conditions, while it is activated by depletion of NTP and dNTP upon the replication and transcription of invading phages and cleave the circular DNA to prevent phage DNA replication.

Rhizosphere bacteria G-H27 significantly promoted the degradation of chlorpyrifos and fosthiazate.

Microbial remediation of polluted environments is the most promising and significant research direction in the field of bioremediation. In this study, chlorpyrifos and fosthiazate were selected as representative organophosphorus pesticides, wheat was the tested plant, and fluorescently labeled degrading Bacillus cereus G-H27 were the film-forming bacteria. Exogenous strengthening technology was used to establish degrading bacterial biofilms on the root surface of wheat. The influence of root surface-degrading bacterial biofilms on the enrichment of chlorpyrifos and fosthiazate in wheat was comprehensively evaluated. First, the fluorescently-labeled degrading bacteria G-H27 was constructed, and its film-forming ability was investigated. Second, the growth- promoting characteristics and degradation ability of the bacteria G-H27 were investigated. Finally, the degradation effect of the root surface-degrading bacterial biofilm on chlorpyrifos and fosthiazate was determined. The above research provides an important material basis and method for the bioremediation of pesticide-contaminated soil.

Microbial degradation mechanism and pathway of the insecticide thiamethoxam by isolated Bacillus Cereus from activated sludge.

The high water solubility and ecotoxicity of thiamethoxam (TMX) is a potential hazard to ecosystems and human health. Here, a strain of Bacillus cereus with high TMX degradation activity was isolated from the sediment of the A2O process in the wastewater treatment plant and was able to utilize TMX as its sole carbon source. Under different environmental conditions, the degradation efficiency of TMX by Bacillus cereus-S1 (strain S1) ranged from 41.0% to 68.9% after 216 h. The optimum degradation conditions were DO = 3.5 mg/L and pH 9.0. The addition of an appropriate carbon-to-nitrogen ratio could accelerate the degradation of TMX. A plausible biodegradation pathway has been proposed based on the identified metabolites and their corresponding degradation pathways. TMX can be directly converted into Clothianidin (CLO), TMX-dm-hydroxyl and TMX-Urea by a series of reactions such as demethylation, oxadiazine ring cleavage and C=N substitution by hydroxy group. The main products were TMX-dm-hydroxyl and TMX-Urea, the amount of CLO production is relatively small. This study aims to provide a new approach for efficient degradation of TMX; furthermore, strain S1 is a promising biological source for in situ remediation of TMX contamination.

Characterization of the Novel Leaderless Bacteriocin, Bawcin, from Bacillus wiedmannii.

The rise of drug-resistant bacteria is a major threat to public health, highlighting the urgent need for new antimicrobial compounds and treatments. Bacteriocins, which are ribosomally synthesized antimicrobial peptides produced by bacteria, hold promise as alternatives to conventional antibiotics. In this study, we identified and characterized a novel leaderless bacteriocin, bawcin, the first bacteriocin to be characterized from a Bacillus wiedmannii species. Chemically synthesized and purified bawcin was shown to be active against a broad range of Gram-positive bacteria, including foodborne pathogens Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes. Stability screening revealed that bawcin is stable over a wide range of pH (2.0-10.0), temperature conditions (25-100 °C), and against the proteases, papain and pepsin. Lastly, three-dimensional structure homology modeling suggests that bawcin contains a saposin-fold with amphipathic helices and a highly cationic surface that may be critical for membrane interaction and the subsequent cell death of its targets. This study provides the foundational understanding of the activity and properties of bawcin, offering valuable insights into its applications across different antimicrobial uses, including as a natural preservative in food and livestock industries.

[Prokaryotic expression and biological activities of the hemolysin BL subunit of a pathogenic Bacillus cereus of cattle origin].

Bacillus cereus belongs to Gram-positive bacteria, which is widely distributed in nature and shows certain pathogenicity. Different B. cereus strains carry different subsets of virulence factors, which directly determine the difference in their pathogenicity. It is therefore important to study the distribution of virulence factors and the biological activity of specific toxins for precise prevention and control of B. cereus infection. In this study, the hemolysin BL triayl was expressed, purified, and characterized. The results showed that the bovine pathogenic B. cereus hemolysin BL could be expressed and purified in the prokaryotic expression system, and the bovine pathogenic B. cereus hemolysin BL showed hemolysis, cytotoxicity, good immunogenicity and certain immune protection in mice. In this study, the recombinant expression of hemolysin BL triayl was achieved, and the biological activity of hemolysin BL of bovine pathogenic ceroid spore was investigated. This study may facilitate further investigating the pathogenic mechanism of B. cereus hemolysin BL and developing a detection method for bovine pathogenic B. cereus disease.