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1.
Sci Rep ; 14(1): 7755, 2024 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-38565929

RESUMO

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.


Assuntos
Celulase , Paenibacillus , Bacillus cereus/metabolismo , Celulose/metabolismo , Reprodutibilidade dos Testes , Celulase/metabolismo , Paenibacillus/metabolismo , Fermentação
2.
Curr Microbiol ; 81(6): 153, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38652152

RESUMO

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.


Assuntos
Arsênio , Bacillus cereus , Microbiologia do Solo , Poluentes do Solo , Triticum , Triticum/crescimento & desenvolvimento , Triticum/microbiologia , Triticum/metabolismo , Bacillus cereus/metabolismo , Bacillus cereus/crescimento & desenvolvimento , Bacillus cereus/genética , Bacillus cereus/efeitos dos fármacos , Arsênio/metabolismo , Poluentes do Solo/metabolismo , RNA Ribossômico 16S/genética , Raízes de Plantas/microbiologia , Raízes de Plantas/crescimento & desenvolvimento , Biodegradação Ambiental , Sideróforos/metabolismo
3.
Sci Rep ; 14(1): 6354, 2024 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-38491100

RESUMO

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.


Assuntos
Bacillus cereus , Dióxido de Carbono , Bacillus cereus/metabolismo , Dióxido de Carbono/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier , Reatores Biológicos/microbiologia , Biomassa , Carbono
4.
Sci Total Environ ; 924: 171580, 2024 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-38462004

RESUMO

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.


Assuntos
Polietileno , Poliestirenos , Polietileno/metabolismo , Bacillus cereus/metabolismo , Índia , Biodegradação Ambiental , Plásticos/metabolismo
5.
Ecotoxicol Environ Saf ; 273: 116129, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38430580

RESUMO

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.


Assuntos
Chalconas , Glycyrrhiza uralensis , Glycyrrhiza uralensis/química , Glycyrrhiza uralensis/genética , Glycyrrhiza uralensis/metabolismo , Bacillus cereus/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Lignina/metabolismo , Estresse Salino , Flavonoides/farmacologia , Flavonoides/metabolismo
6.
Adv Food Nutr Res ; 108: 265-287, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38461001

RESUMO

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.


Assuntos
Bacillus cereus , Produtos da Carne , Humanos , Bacillus cereus/metabolismo , Manipulação de Alimentos , Esporos Bacterianos/metabolismo , Microbiologia de Alimentos
7.
Int J Biol Macromol ; 263(Pt 1): 130348, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38395274

RESUMO

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.


Assuntos
Acetiltransferases , Bacillus cereus , Acetiltransferases/química , Bacillus cereus/metabolismo , Sequência de Aminoácidos , Acetilcoenzima A/metabolismo , Proteínas Ribossômicas/metabolismo , Cristalografia por Raios X
8.
Nat Commun ; 15(1): 836, 2024 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-38282040

RESUMO

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.


Assuntos
Bacteriófagos , DNA , DNA/metabolismo , Bacteriófagos/genética , Bacteriófagos/metabolismo , Bacillus cereus/metabolismo , Endonucleases , Sistema Imunitário/metabolismo
9.
Int J Food Microbiol ; 413: 110600, 2024 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-38281435

RESUMO

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.


Assuntos
Bacillus cereus , Oryza , Bacillus cereus/genética , Bacillus cereus/metabolismo , Sequência de Bases , Odorantes/análise , Proteínas de Plantas/metabolismo , Pirróis/metabolismo
10.
Sci Total Environ ; 917: 169838, 2024 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-38232838

RESUMO

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.


Assuntos
Clorpirifos , Praguicidas , Tiazolidinas , Clorpirifos/metabolismo , Praguicidas/metabolismo , Compostos Organofosforados/metabolismo , Rizosfera , Microbiologia do Solo , Biodegradação Ambiental , Bacillus cereus/metabolismo
11.
Sci Total Environ ; 918: 170499, 2024 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-38296101

RESUMO

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.


Assuntos
Polipropilenos , Poluentes Químicos da Água , Humanos , Microplásticos , Plásticos , Espectroscopia de Infravermelho com Transformada de Fourier , Pandemias , Biodegradação Ambiental , Bacillus cereus/metabolismo , Poluentes Químicos da Água/análise
12.
Environ Res ; 246: 117929, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38157972

RESUMO

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.


Assuntos
Guanidinas , Inseticidas , Neonicotinoides , Tiazóis , Humanos , Tiametoxam , Inseticidas/toxicidade , Esgotos , Bacillus cereus/metabolismo , Ecossistema , Nitrocompostos/toxicidade , Nitrocompostos/metabolismo , Oxazinas/metabolismo , Oxazinas/toxicidade , Carbono , Ureia
13.
Int J Mol Sci ; 24(23)2023 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-38069290

RESUMO

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.


Assuntos
Bacillus , Bacteriocinas , Bacteriocinas/química , Bacillus/metabolismo , Antibacterianos/química , Bacillus cereus/metabolismo
14.
Sheng Wu Gong Cheng Xue Bao ; 39(12): 4939-4949, 2023 Dec 25.
Artigo em Chinês | MEDLINE | ID: mdl-38147993

RESUMO

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.


Assuntos
Bacillus cereus , Proteínas de Bactérias , Bovinos , Animais , Camundongos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bacillus cereus/genética , Bacillus cereus/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Fatores de Virulência/metabolismo , Enterotoxinas/metabolismo
15.
Int J Mol Sci ; 24(21)2023 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-37958921

RESUMO

Leucyl-aminopeptidase (LAP), an important metallopeptidase, hydrolyses amino acid residues from the N-terminus of polypeptides and proteins, acting preferentially on the peptide bond formed by N-terminus leucine. A new leucyl-aminopeptidase was found in Bacillus cereus CZ. Its gene (bclap) contained a 1485 bp ORF encoding 494 amino acids with a molecular weight of 54 kDa. The bcLAP protein was successfully expressed in E. coli BL21(DE3). Optimal activity is obtained at pH 9.0 and 58 °C. The bcLAP displays a moderate thermostability and an alkaline pH adaptation range. Enzymatic activity is dramatically enhanced by Ni2+. EDTA significantly inhibits the enzymatic activity, and bestatin and SDS also show strong inhibition. The three-dimensional model of bcLAP monomer and homohexamer is simulated byPHYRE2 server and SWISS-MODEL server. The docking of bestatin, Leu-Trp, Asp-Trp and Ala-Ala-Gly to bcLAP is performed using AutoDock4.2.5, respectively. Molecular docking results show that the residues Lys260, Asp265, Lys272, Asp283, Asp342, Glu344, Arg346, Gly372 and His437 are involved in the hydrogen bonding with the ligands and zinc ions. There may be two nucleophilic catalytic mechanisms in bcLAP, one involving His 437 or Arg346 and the other involving His437 and Arg346. The bcLAP can hydrolyse the peptide bonds in Leu-Trp, Asp-Trp and Ala-Ala-Gly.


Assuntos
Bacillus cereus , Leucil Aminopeptidase , Sequência de Aminoácidos , Bacillus cereus/genética , Bacillus cereus/metabolismo , Simulação de Acoplamento Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Peptídeos
16.
Int J Mol Sci ; 24(22)2023 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-38003626

RESUMO

Hemolysin II (HlyII)-one of the pathogenic factors of Bacillus cereus, a pore-forming ß-barrel toxin-possesses a C-terminal extension of 94 amino acid residues, designated as the C-terminal domain of HlyII (HlyIICTD), which plays an important role in the functioning of the toxin. Our previous work described a monoclonal antibody (HlyIIC-20), capable of strain-specific inhibition of hemolysis caused by HlyII, and demonstrated the dependence of the efficiency of hemolysis on the presence of proline at position 324 in HlyII outside the conformational antigenic determinant. In this work, we studied 16 mutant forms of HlyIICTD. Each of the mutations, obtained via multiple site-directed mutagenesis leading to the replacement of amino acid residues lying on the surface of the 3D structure of HlyIICTD, led to a decrease in the interaction of HlyIIC-20 with the mutant form of the protein. Changes in epitope structure confirm the high conformational mobility of HlyIICTD required for the functioning of HlyII. Comparison of the effect of the introduced mutations on the effectiveness of interactions between HlyIICTD and HlyIIC-20 and a control antibody recognizing a non-overlapping epitope enabled the identification of the amino acid residues N339 and K340, included in the conformational antigenic determinant recognized by HlyIIC-20.


Assuntos
Bacillus cereus , Proteínas Hemolisinas , Humanos , Bacillus cereus/genética , Bacillus cereus/metabolismo , Proteínas Hemolisinas/metabolismo , Substituição de Aminoácidos , Epitopos/genética , Epitopos/metabolismo , Hemólise/genética , Aminoácidos/genética , Aminoácidos/metabolismo
17.
Vet Q ; 43(1): 1-10, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38010068

RESUMO

An outbreak of a disease with a high mortality rate occurred in a Chinese Softshell Turtle (Pelodiscus sinensis) farm in Hubei Province. This study isolated a highly pathogenic Bacillus cereus strain (Y271) from diseased P. sinensis. Y271 has ß hemolysis, containing both Hemolysin BL (hblA, hblC, and hblD), Non-hemolytic enterotoxin, NHE (nheA, nheB, and nheC), and Enterotoxin FM (entFM) genes. Y271 is highly pathogenic against P. sinensis with an LD50 = 6.80 × 103 CFU/g weight. B. cereus was detected in multiple tissues of the infected P. sinensis. Among them, spleen tissue showed the highest copy number density (1.54 ± 0.12 × 104 copies/mg). Multiple tissues and organs of diseased P. sinensis exhibited significant pathological damage, especially the spleen, liver, kidney, and intestine. It showed obvious tissue structure destruction, lesions, necrosis, red blood cells, and inflammatory cell infiltration. B. cereus proliferating in the spleen, liver, and other tissues was observed. The intestinal microbiota of the diseased P. sinensis was altered, with a greater abundance of Firmicutes, Fusobacterium, and Actinomyces than in the healthy group. Allobaculum, Rothia, Aeromonas, and Clostridium abundance were higher in the diseased group than in the healthy group. The number of unique microbial taxa (472) in the disease group was lower than that of the healthy group (705). Y271 was sensitive to multiple drugs, including florfenicol, enrofloxacin, neomycin, and doxycycline. B. cereus is the etiological agent responsible for the massive death of P. sinensis and reveals its potential risks during P. sinensis cultivation.


Assuntos
Bacillus cereus , Microbiologia de Alimentos , Animais , Bacillus cereus/genética , Bacillus cereus/metabolismo , Enterotoxinas/análise , Enterotoxinas/genética
18.
Mar Genomics ; 72: 101071, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38008533

RESUMO

Bacillus species have been considered as promising biological control agents due to their excellent antimicrobial ability. Bacillus cereus strain Z4 was isolated from 2000 m deep sea sediments of the Western Pacific Ocean, which possesses significant antifungal activity against Phytophthora nicotianae, the pathogenic fungus of tobacco black shank disease. To reveal the underlying antifungal genetic mechanisms, here, we report the complete genomic sequence of the strain Z4. The genome has one circular chromosome of 5,664,309 bp with a G + C content of 35.31%, 109 tRNAs, and 43 rRNAs. Genomic analysis identified 10 gene clusters related to the biosynthesis of biocontrol active compounds, including bacillibactin, petrobactin, fengycin, and molybdenum cofactor. Meanwhile, 6 gene clusters were responsible for the biosynthesis of metabolites with unknown functions. Strain Z4 also contains a large number of genes encoding carbohydrate-active enzymes and secreted proteins, respectively. The whole genomic analysis of Bacillus cereus Z4 may provide a valuable reference for elucidating its biocontrol mechanism against tobacco black shank.


Assuntos
Bacillus cereus , Bacillus , Bacillus cereus/genética , Bacillus cereus/metabolismo , Antifúngicos/metabolismo , /metabolismo , Oceano Pacífico , Bacillus/genética
19.
Int. microbiol ; 26(4): 961-972, Nov. 2023. graf
Artigo em Inglês | IBECS | ID: ibc-227484

RESUMO

Phytases are specialized enzymes meant for phytic acid degradation. They possess ability to prevent phytic acid indigestion, including its attendant environmental pollution. This study was aimed at investigating biochemical properties of purified phytase of B. cereus isolated from Achatina fulica. Phytase produced from Bacillus cereus that exhibited optimal phytate degrading-ability of all the bacteria isolated was purified in a three-step purification. The biochemical properties of the purified enzyme were also determined. The phytase homogeny of approximately 45 kDa exhibited 12.8-purification fold and 1.6% yield with optima phytate degrading efficiency and maximum stability at pH 7 and 50 °C. Remaining activity of 52 and 47% obtained between 60 and 70 °C after 2 h further established thermostability of the purified phytase. Mg2+ and Zn2+ enhanced phytate hydrolysis by the enzyme, while Na+ showed mild inhibition but Hg2+ severely inhibited the enzymatic activity. Km and Vmax were estimated to be 0.11 mM and 55.6 μmol/min/mL, displaying enzyme-high substrate affinity and catalytic efficiency, respectively. Phytase purified from Bacillus cereus, isolated from African giant snails, has shown excellent characteristics suitable for phytic acid hydrolysis and could be employed in industrial and biotechnological applications.(AU)


Assuntos
Humanos , Ácido Fítico/química , 6-Fitase/química , Trato Gastrointestinal , Bacillus cereus/metabolismo , Caramujos/metabolismo , Prótons , 6-Fitase/metabolismo , Microbiologia , Técnicas Microbiológicas , Ácido Fítico/metabolismo
20.
Microb Pathog ; 185: 106418, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37866551

RESUMO

"Fried rice syndrome" originated from the first exposure to a fried rice dish contaminated with Bacillus cereus. This review compiles available data on the prevalence of B. cereus outbreak cases that occurred between 1984 and 2019. The outcome of B. cereus illness varies dramatically depending on the pathogenic strain encounter and the host's immune system. B. cereus causes a self-limiting, diarrheal illness caused by heat-resistant enterotoxin proteins, and an emetic illness caused by the deadly toxin named cereulide. The toxins together with their extrinsic factors are discussed. The possibility of more contamination of B. cereus in protein-rich food has also been shown. Therefore, the aim of this review is to summarize the available data, focusing mainly on B. cereus physiology as the causative agent for "fried rice syndrome." This review emphasizes the prevalence of B. cereus in starchy food contamination and outbreak cases reported, the virulence of both enterotoxins and emetic toxins produced, and the possibility of contaminated in protein-rich food. The impact of emetic or enterotoxin-producing B. cereus on public health cannot be neglected. Thus, it is essential to constantly monitor for B. cereus contamination during food handling and hygiene practices for food product preparation.


Assuntos
Doenças Transmitidas por Alimentos , Oryza , Humanos , Bacillus cereus/metabolismo , Doenças Transmitidas por Alimentos/epidemiologia , Eméticos/metabolismo , Enterotoxinas/análise , Microbiologia de Alimentos , Contaminação de Alimentos/análise
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