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1.
Curr Microbiol ; 80(2): 81, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36662309

RESUMO

This study reports the draft genome sequence of Bacillus velezensis strain AAK_S6 as a valuable biocontrol agent with high genetic potential to harbor broad-spectrum secondary metabolite producing capacity. A genome data of 4,430,946 bp were generated with a GC content of 46.4% that comprised a total of 4861 genes including a total of 4757 coding sequences (CDS), 104 rRNAs, 85 tRNAs and 80 pseudo-genes. Based on the overall genome-based relatedness indices (OGRI), the strain AAK_S6 has been reassigned to its correct taxonomic position. The strain shared > 99% OrthoANI, > 98% ANIb, > 99% ANIm, > 0.9900 TETRA, > 93% dDDH and 0.08% GC content difference with model strains B. velezensis FZB42T and B. velezensis NRRL B-41580T thus delineating them as closely related species. The genome was mined for strain-specific secondary metabolites that revealed 20 gene clusters for the biosynthesis of several cyclic lipopeptides, saccharides, polyketides along with bacilysin. Thus, the comparative genome analysis of strain AAK_S6 with members of the genus Bacillus by phylogenomic approach revealed that the genomes were almost similar genetically and contained the core genome for B. velezensis. Genomic data strongly supported that the strain AAK_S6 represented an excellent potential candidate for the production of secondary metabolites that could serve as a basis for developing new biocontrol agents, plant growth promoters, and microbial fertilizers.


Assuntos
Bacillus , Genoma Bacteriano , Bacillus/metabolismo , Genômica , Desenvolvimento Vegetal
2.
Curr Microbiol ; 80(2): 80, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36662359

RESUMO

The application of mineral-solubilizing, plant growth-promoting bacteria as inoculants offers a promising alternative to chemical fertilizers. In the present study, lithic bacterial isolates were evaluated for mineral solubilization and plant growth-promoting potential. Among the 57 lithic bacterial isolates associated with different rock samples collected from various locations in Meghalaya, India, nine K-solubilizing isolates, six S-solubilizing isolates, five P- and Si-solubilizing isolates, and three Zn-solubilizing isolates with notable indole-3-acetic acid and siderophore production, and ACC deaminase activity were selected for further study. Based on 16S rRNA gene sequence analysis, isolates were affiliated to nine different genera (Arthrobacter, Acinetobacter, Pseudomonas, Halopseudomonas, Bacillus, Neobacillus, Peribacillus, Pantoea, and Priestia). On performing rice seed germination potentials, Pantoea agglomerans BL26, Priestia megaterium BL9, Bacillus subtilis GP2, Halopseudomonas xinjiangensis BL29, and Pseudomonas sp. BM1 were selected for in vitro pot experiments, being the most potent isolates. Following inoculation, all five isolates were found to significantly enhance growth of rice plants (P < 0.05). The maximum shoot length increased due to P. megaterium BL9, the maximum root length increased due to H. xinjiangensis BL29, and the maximum plant fresh weight increased due to P. megaterium BL9. The findings concluded that these five lithic bacterial isolates have potent plant growth-promoting potential with possible prospection through field trials. To the best of available literature, this is a first report on the characterization of lithic bacterial isolates as mineral solubilizers and plant growth promoters.


Assuntos
Bacillus , Bactérias , RNA Ribossômico 16S/genética , Desenvolvimento Vegetal , Bacillus/genética , Índia , Raízes de Plantas/microbiologia
3.
Sci Rep ; 13(1): 51, 2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36593253

RESUMO

The bacterial nanocellulose has been used in a wide range of biomedical applications including carriers for drug delivery, blood vessels, artificial skin and wound dressing. The total of ten morphologically different bacterial strains were screened for their potential to produce bacterial nanocellulose (BNC). Among these isolates, Bacillus sp. strain SEE-3 exhibited potent ability to produce the bacterial nanocellulose. The crystallinity, particle size and morphology of the purified biosynthesized nanocellulose were characterized. The cellulose nanofibers possess a negatively charged surface of - 14.7 mV. The SEM images of the bacterial nanocellulose confirms the formation of fiber-shaped particles with diameters of 20.12‒47.36 nm. The TEM images show needle-shaped particles with diameters of 30‒40 nm and lengths of 560‒1400 nm. X-ray diffraction show that the obtained bacterial nanocellulose has crystallinity degree value of 79.58%. FTIR spectra revealed the characteristic bands of the cellulose crystalline structure. The thermogravimetric analysis revealed high thermal stability. Optimization of the bacterial nanocellulose production was achieved using Plackett-Burman and face centered central composite designs. Using the desirability function, the optimum conditions for maximum bacterial nanocellulose production was determined theoretically and verified experimentally. Maximum BNC production (20.31 g/L) by Bacillus sp. strain SEE-3 was obtained using medium volume; 100 mL/250 mL conical flask, inoculum size; 5%, v/v, citric acid; 1.5 g/L, yeast extract; 5 g/L, temperature; 37 °C, Na2HPO4; 3 g/L, an initial pH level of 5, Cantaloupe juice concentration of 81.27 percent and peptone 11.22 g/L.


Assuntos
Bacillus , Cucumis melo , Nanofibras , Bactérias/química , Celulose/química , Meios de Cultura/química
4.
PeerJ ; 11: e14621, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36643649

RESUMO

Wheat (Triticum aestivum L.) is a major source of calorific intake in its various forms and is considered one of the most important staple foods. Improved wheat productivity can contribute substantially to addressing food security in the coming decades. Soil salinity is the most serious limiting factor in crop production and fertilizer use efficiency. In this study, 11 bacteria were isolated from wheat rhizosphere and examined for salt tolerance ability. WGT1, WGT2, WGT3, WGT6, WGT8, and WGT11 were able to tolerate NaCl salinity up to 4%. Bacterial isolates were characterized in vitro for plant growth-promoting properties including indole-3-acetic acid (IAA) production, phosphate solubilization, nitrogen fixation, zinc solubilization, biofilm formation, and cellulase-pectinase production. Six isolates, WGT1, WGT3, WGT4, WGT6, WGT8, and WGT9 showed IAA production ability ranging from 0.7-6 µg m/L. WGT8 displayed the highest IAA production. Five isolates, WGT1, WGT2, WGT5, WGT10, and WGT11, demonstrated phosphate solubilization ranging from 1.4-12.3 µg m/L. WGT2 showed the highest phosphate solubilization. Nitrogen fixation was shown by only two isolates, WGT1 and WGT8. Zinc solubilization was shown by WGT1 and WGT11 on minimal media. All isolates showed biofilm formation ability, where WGT4 exhibited maximum potential. Cellulase production ability was noticed in WGT1, WGT2, WGT4, and WGT5, while pectinase production was observed in WGT2 and WGT3. Phylogenetic identification of potential bacteria isolates confirmed their close relationship with various species of the genus Bacillus. WGT1, WGT2, and WGT3 showed the highest similarity with B. cereus, WGT6 with B. tianshenii, WGT8 with B. subtilis, and WGT11 with B. thuringiensis. Biofertilizer characteristics of salt-tolerant potential rhizospheric bacteria were evaluated by inoculating wheat plants under controlled conditions and field experiments. B. cereus WGT1 and B. thuringiensis WGT11 displayed the maximum potential to increase plant growth parameters and enhance grain yield by 37% and 31%, respectively. Potential bacteria of this study can tolerate salt stress, have the ability to produce plant growth promoting substances under salt stress and contribute significantly to enhance wheat grain yield. These bacterial isolates have the potential to be used as biofertilizers for improved wheat production under salinity conditions and contribute to the sustainable agriculture.


Assuntos
Bacillus , Rizosfera , Estresse Salino , Triticum , Celulases , Fosfatos , Filogenia , Poligalacturonase , Triticum/crescimento & desenvolvimento , Triticum/microbiologia , Zinco
5.
Curr Microbiol ; 80(2): 79, 2023 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-36656344

RESUMO

Two aerobic, Gram-staining-positive, rod-shaped, endospore-forming, thermophilic bacterial strains, designated FJAT-47801T and FJAT-47835, were isolated from the sediment collected from Zhangjiang Estuary Mangrove National Nature Reserve in Fujian Province, China. Growth was observed at 25-55 °C (optimum, 50 °C) and pH 7.0-9.0 (optimum, pH 7.0), with up to 4.0% (w/v) NaCl (optimum, without NaCl). Strains FJAT-47801T and FJAT-47835 showed the highest 16S rRNA gene sequence similarity to Bacillus oleivorans (98.5%). The 16S rRNA gene sequence similarity between FJAT-47801T and FJAT-47835 was 99.9% indicating they were the same species. Phylogenetic (based on 16S rRNA gene sequences) and phylogenomic (based on 120 conserved bacterial single-copy genes) trees showed that strains FJAT-47801T and FJAT-47835 should be affiliated to the genus Bacillus. The of menaquinone of strain FJAT-47801T was MK-7. The major fatty acids of strain FJAT-47801T were iso-C15:0, anteiso-C15:0, iso-C17:0, and C16:0. The major polar lipids strain FJAT-47801T were phosphatidylethanolamine (PE), diphosphatidylglycerol (DPG), and phosphatidylglycerol (PG). The genomic DNA G+C content of strain FJAT-47801T was 39.3%. The average nucleotide identity (84.3%) and the digital DNA-DNA hybridization value (28.1%) between strain FJAT-47801T and B. oleivorans CCTCC AB 2013353T were below the cut-off level for species delineation. Based on the above results, strain FJAT-47801T represents a novel species of the genus Bacillus, for which the name Bacillus litorisediminis sp. nov., is proposed. The type strain is FJAT-47801T (=GDMCC 1.2712T = JCM 34875T).


Assuntos
Bacillus , Fosfolipídeos , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Cloreto de Sódio/análise , DNA Bacteriano/genética , DNA Bacteriano/análise , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Parede Celular/química , Ácido Diaminopimélico/análise , Ácido Diaminopimélico/química , Peptidoglicano/análise , Análise de Sequência de DNA , Ácidos Graxos/química
6.
Sci Rep ; 13(1): 831, 2023 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-36646757

RESUMO

Monitoring the presence of pathogenic Bacillus spores is important for industrial applications, as well as necessary for ensuring human health. Bacillus thuringiensis is used as a biopesticide against several insect pests. Bacillus cereus spores are a significant cause of food poisoning, and Bacillus anthracis is a recognized biosecurity threat. Laboratory-based methods, such as polymerase chain reaction, enzyme-linked immunosorbent assay, or matrix-assisted laser desorption ionization spectroscopy provide sensitive detection of bacteria and spores, but the application of those methods for quasi-continuous environmental monitoring presents a significant challenge requiring frequent human intervention. To address this challenge, we developed a workstation for quasi-autonomous monitoring of water reservoirs for the presence of bacteria and spores, and designed and validated the functionality of a microprocessor-controlled module capable of repetitive collection and pre-concentration of spores in liquid samples tested with fiberglass (FG), polyether sulfone and polyvinylidene fluoride filters. The best results were obtained with FG filters delivering a 20× concentration of B. thuringiensis and B. cereus spores from saline suspensions. The successful 20× pre-concentration of Bacillus spores demonstrated with FG filters could be repeated up to 3 times when bleach decontamination is applied between filtrations. Taken together, our results demonstrate an attractive instrument suitable for semi-automated, quasi-continuous sampling and pre-processing of water samples for biosensing of bacterial spores originating from a complex environment.


Assuntos
Bacillus anthracis , Bacillus thuringiensis , Bacillus , Humanos , Bacillus cereus , Esporos Bacterianos/química
7.
Food Res Int ; 163: 112291, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36596196

RESUMO

Thermosonication (TS) is a novel and viable technique employed to replace conventional thermal processing. TS treatment combined with pasteurization was used to kill the residual heat-resistant Bacillus in pasteurized milk and extend the shelf life of pasteurized milk and compared with High Temperture Shoort Time (HTST) pasteurization to study its decontamination effect on Bacillus subtilis and the quality of treated milk. The results showed that after 40 kHz, 240 W, 25 min ultrasonic treatment and 50 °C heating decontamination treatment, the number of B. subtilis in the medium and milk medium decreased by 4.17 log CFU/mL and 4.09 log CFU/mL respectively. The results of cell membrane permeability showed that the leakage of DNA and protein in the HTST-TS group increased by 52.3 % and 34 %, respectively, when compared to that in the HTST group. In addition, transmission electron microscopy (TEM) analysis showed that the bacterial cell membrane of the HTST-TS group swelled up, the cell wall was ruptured, and the cell content was accumulated in the cells. The results showed that HTST-TS treatment significantly inhibited the activities of ATPase (47 %), succinate dehydrogenase (SDH) (68.6 %), and malate dehydrogenase (MDH) (54.4 %). The physical and chemical sensory evaluation of milk treated with HTST-TS showed that HTST-TS treatment could improve the L* value (2.24 %), zeta potential (64.19 %), and colloidal particle size (14.49 %) of milk but had no significant effect on oral sensitivity. In conclusion, this study provides new insights, which may be helpful in implementing this new combined decontamination method in the dairy industry to improve the quality of pasteurized milk and extend the its shelf life.


Assuntos
Bacillus , Animais , Leite/química , Subtilisina/análise , Descontaminação , Pasteurização/métodos
8.
BMC Microbiol ; 23(1): 8, 2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36627563

RESUMO

BACKGROUND: Verticillium wilt, caused by the fungus Verticillium dahliae, leads to significant losses in cotton yield worldwide. Biocontrol management is a promising means of suppressing verticillium wilt. The purpose of the study was to obtain and analyze endophytic bacteria with Verticillium wilt-resistant activities from the roots of Gossypium barbadense 'Xinhai15' and to explore the interactions between the soil and plants. RESULTS: An endophytic bacterium Bacillus sp. T6 was obtained from the Verticillium wilt-resistant cotton G. barbadense 'Xinhai15', which showed significant antagonistic abilities against cotton Verticillium wilt. The bioassay results indicated that the strain possessed strong antagonistic abilities that inhibited V. dahliae spore germination and mycelial growth without contact, and thus it was speculated that the active factor of the bacteria might be volatile compounds. A total of 46 volatile substances were detected via headspace solid-phase microextraction and gas chromatography-mass spectrometry analysis. The pure product verification experiment confirmed that the styrene produced by the T6 strain was the main virulence factor. Transcriptome analysis showed that following styrene induction, 247 genes in V. dahliae, including four hydrolase genes, eight dehydrogenase genes, 11 reductase genes, 17 genes related to transport and transfer were upregulated. Additionally, 72 genes, including two chitinase genes, two protease genes, five transport-related genes, and 33 hypothetical protein genes, were downregulated. The quantitative real-time PCR results confirmed that the expression of the four genes VDAG_02838, VDAG_09554, VDAG_045572, and VDAG_08251 was increased by 3.18, 78.83, 2.71, and 2.92 times, respectively, compared with the uninduced control group. CONCLUSIONS: The research provides a new reference for the development and application of the volatile compounds of endophytic bacteria as new biocontrol agents for the control of Verticillium wilt and as biological preservatives for agricultural products.


Assuntos
Bacillus , Verticillium , Verticillium/metabolismo , Gossypium/microbiologia , Bacillus/genética , Bactérias , Estirenos/metabolismo , Doenças das Plantas/microbiologia
9.
Trop Anim Health Prod ; 55(1): 38, 2023 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-36640209

RESUMO

Probiotics are non-pathogenic microorganisms that are potentially important non-antibiotic alternatives. This study aimed to compare novel multi-strain and single-strain Bacillus probiotics and their respective influences on broiler chickens' performance, gut health, litter quality, immune response, and NBN and TLR gene expression. A total of 1200 Arbor-Acres 1-day-old broiler chicks were randomly allocated into three treatments (T1 was a control, T2 was supplemented with a combined Bacillus coagulans (2 × 109 cfu/g) and Bacillus licheniformis (8 × 109 cfu/g) probiotic strains (0.2 kg/ton of feed), and T3 was supplemented with Bacillus licheniformis (3.2 × 109 cfu/g) probiotic (0.5 kg/ton of feed) with eight replicas of each. Supplementing the broiler diet with either the single-strain (T3) or the multi-strain (T2) Bacillus-based probiotic raised the overall birds' body weight, body weight gain, feed conversion ratio, and European production efficiency factor compared to the control (T1), with a significant enhancement achieved by the multi-strain Bacillus product (P = 0.005). T2 and T3 exhibited significantly improved cholesterol, Alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and alkaline phosphatase levels than the control (P ≤ 0.05). The transcript levels of both NBN and TLR genes were upregulated in the liver in the T2 and T3 groups. The T2 group experienced significant reductions in gut bacterial counts, especially for Clostridia, and recorded the lowest litter moisture and nitrogen. In conclusion, supplementing broiler diets with probiotics of multiple Bacillus strains increased production profitability by promoting bird growth, improving feed intake, enhancing gut mucosa and immune organs, and upregulating genes responsible for immunity. All these inhibit the overgrowth of enteric pathogens and sustain litter quality.


Assuntos
Bacillus coagulans , Bacillus licheniformis , Bacillus , Probióticos , Animais , Galinhas , Bacillus licheniformis/fisiologia , Dieta/veterinária , Probióticos/farmacologia , Peso Corporal , Expressão Gênica , Ração Animal/análise
10.
PLoS One ; 18(1): e0277983, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36701319

RESUMO

Fusarium graminearum is the causal agent of Fusarium Head Blight, a serious disease affecting grain crops worldwide. Biological control involves the use of microorganisms to combat plant pathogens such as F. graminearum. Strains of Bacillus velezensis are common biological control candidates for use against F. graminearum and other plant pathogens, as they can secrete antifungal secondary metabolites. Here we study the interaction between B. velezensis E68 and F. graminearum DAOMC 180378 by employing a dual RNA-seq approach to assess the transcriptional changes in both organisms. In dual culture, B. velezensis up-regulated genes related to sporulation and phosphate stress and down-regulated genes related to secondary metabolism, biofilm formation and the tricarboxylic acid cycle. F. graminearum up-regulated genes encoding for killer protein 4-like proteins and genes relating to heavy metal tolerance, and down-regulated genes relating to trichothecene biosynthesis and phenol metabolism. This study provides insight into the molecular mechanisms involved in the interaction between a biocontrol bacterium and a phytopathogenic fungus.


Assuntos
Bacillus , Fusarium , Fusarium/genética , Fusarium/metabolismo , Bacillus/genética , Perfilação da Expressão Gênica , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologia
11.
Curr Microbiol ; 80(2): 77, 2023 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-36652029

RESUMO

Rhizobacteria that are helpful to plants can lessen the impacts of salt stress, and they may hold promise for the development of sustainable agriculture in the future. The present study was intended to explicate consortia of salt-tolerant plant-beneficial rhizobacteria for the amelioration of salinity stress in Arabidopsis plants. Inoculation with both the consortia positively influenced the growth of plants as indicated by total chlorophyll content, MDA content, and antioxidant enzyme activities under stressful conditions. Both the multi-trait consortia altered the expression profiles of stress-related genes including CSD1, CAT1, Wrky, Ein, Etr, and ACO. Furthermore, the metabolomic analysis indicated that inoculated plants modulated the metabolic profiles to stimulate physiological and biochemical responses in Arabidopsis plants to mitigate salt stress. Our study affirms that the consortia of salt-tolerant bacterial strains modulate the transcriptional as well as metabolic machinery of plants to protect them from salinity stress. Nevertheless, the findings of this study revealed that consortia are composed of salt-tolerant bacterial strains viz. Bacillus safensis NBRI 12M, B. subtilis NBRI 28B, and B. subtilis NBRI 33N demonstrated significant improvement in Arabidopsis plants under saline stress conditions.


Assuntos
Arabidopsis , Bacillus , Tolerância ao Sal/genética , Arabidopsis/genética , Bacillus/metabolismo , Estresse Salino , Bactérias/metabolismo , Antioxidantes/metabolismo , Estresse Fisiológico
12.
Food Microbiol ; 111: 104211, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36681393

RESUMO

Daqu is a solid-state fermentation and saccharification starter for the Chinese liquor baijou. During the daqu stage, amylolytic and proteolytic enzymes are produced by Bacillus and fungi. Bacillus spp. also produce lipopeptides with a broad spectrum of antimicrobial activities but direct evidence for their impact on community assembly in daqu is lacking. This study aimed to study the interaction between Bacillus spp. and fungi in daqu models. The antifungal activity of surfactin, fengycin, and iturin A was initially assessed in vitro. Iturin A displayed the strongest antifungal activity (MIC = 10-50 mg/L). In situ antifungal activity of B. amyloliquefaciens and B. velezensis against molds was observed in a simple daqu model inoculated with single strains of Bacillus species. Formation of lipopeptides in situ was supported by quantification of mRNA encoding for enzymes for surfactin, fengycin, and iturin A biosynthesis. In situ antifungal activity of Bacillus species was also observed in a complex daqu model that was inoculated with 8 bacterial or fungal strains plus one of the three strains of Bacillus. A relationship of lipopeptides to in situ antifungal activity was further supported by detection of the lipopeptides by liquid chromatography coupled to mass spectrometry. Both results indicated that B velezensis FUA2155 had higher antifungal activity in the daqu model, and was the only strain that produced multiple iturin A congeners in situ. Taken together, this study provides evidence that production of lipopeptides by Bacillus species in daqu may impact community assembly and hence product quality.


Assuntos
Bacillus , Bacillus/química , Antifúngicos/farmacologia , Antifúngicos/química , Fermentação , Bactérias/metabolismo , Fungos/metabolismo , Lipopeptídeos/farmacologia , Lipopeptídeos/análise , Lipopeptídeos/química
13.
World J Microbiol Biotechnol ; 39(3): 84, 2023 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-36693977

RESUMO

The coexistence of heavy metals (HMs) and petroleum hydrocarbons (PHs) exacerbates ecotoxicity and impair the drivers of eco-functionalities that stimulate essential nutrients for the productivity of the impacted environment. Profiling the bacteria that stem the ecological impact via HMs sequestration and PHs catabolism with nitrogen fixation is imperative to bioremediation of the polluted sites. The sediment of site that was consistently contaminated with industrial wastewaters was analysed for ecological toxicants and the bacterial strains that combined HMs resistance with PHs catabolism in a nitrogen-limiting system were isolated from the sediment and characterized. The geochemistry of the samples revealed the co-occurrence of the above-benchmark concentrations of HMs with the derivatives of hydrocarbons. Notwithstanding, nickel and mercury (with 5% each of the total metal concentrations in the polluted site) exhibited probable effect concentrations on the biota and thus hazardous to the ecosystem. Approx. 31% of the bacterial community, comprising unclassified Planococcaceae, unclassified Bradyrhizobiaceae, Rhodococcus, and Bacillus species, resisted 160 µmol Hg2+ in the nitrogen-limiting system within 24 h post-inoculation. The bacterial strains adopt volatilization, and sometimes in combination with adsorption/bioaccumulation strategies to sequester Hg2+ toxicity while utilizing PHs as sources of carbon and energy. Efficient metabolism of petroleum biomarkers (> 87%) and Hg2+ sequestration (≥ 75% of 40 µmol Hg2+) displayed by the selected bacterial strains portend the potential applicability of the bacilli for biotechnological restoration of the polluted site.


Assuntos
Bacillus , Mercúrio , Metais Pesados , Petróleo , Petróleo/metabolismo , Ecossistema , Metais Pesados/metabolismo , Bactérias/genética , Bactérias/metabolismo , Biodegradação Ambiental , Hidrocarbonetos/metabolismo , Bacillus/metabolismo
14.
Lett Appl Microbiol ; 76(1)2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36688758

RESUMO

D-amino acids, the important components of the bacterial cell walls, are valuable molecular and genetic markers of bacterial-derived organic material in the environment. D-serine, a racemization product of L-serine is one such amino acid present in various prokaryotes and eukaryotes. It is a well-recognized regulator of various activities in the human nervous system. In plants, it has a role in the nitrogen cycle regulation and pollen tube growth. Serine enantiomers are present in different concentrations and few bacterial strains are reported to contribute to D-serine in the environment. During the present study, soil samples from different places in North India were collected and processed to isolate and screen the bacteria on M9 minimal media (Himedia) for D-serine synthesis. Thin-layer chromatography (TLC Silica gel 60 F 254 (20 × 20 cm, Merck, Darmstadt, Germany) and Mass spectroscopic analysis (Bruker MICROTOF II spectrometer) studies, etc were performed. D-serine-producing isolates were characterized as per standard procedures. Bacterial isolate A1C1 with maximum D-serine (0.919 ± 0.02 nM) synthesis under optimal growth conditions (37°C ± 0.5, 150 ± 0.5 RPM, and 7 ± 0.5 pH) was identified as Bacillus tequilensis based on 16sRNA sequencing. The isolate could be a valuable serine racemization tool for various industrial and environmental applications.


Assuntos
Bacillus , Serina , Humanos , Serina/análise , Serina/química , Serina/metabolismo , Aminoácidos/metabolismo , Bacillus/metabolismo , Cromatografia em Camada Delgada
15.
Int J Mol Sci ; 24(2)2023 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-36674766

RESUMO

Polyhydroxyalkanoates (PHAs) are widely used in medical and potentially in other applications due to their biocompatibility and biodegradability. Understanding PHA biosynthetic pathways may lead to the detection of appropriate conditions (substrates) for producing a particular PHA type by a specific microbial strain. The aim of this study was to establish a method enabling potentially interesting PHA bacterial producers to be found. In the study, all four classes of PHA synthases and other genes involved in PHA formation (fabG, phaA, phaB, phaG, and phaJ) were detected by PCR in 64 bacterial collection strains and food isolates. Acinetobacter, Bacillus, Cupriavidus, Escherichia, Klebsiella, Lelliottia, Lysinibacillus, Mammaliicoccus, Oceanobacillus, Pantoea, Peribacillus, Priestia, Pseudomonas, Rahnella, Staphylococcus, and Stenotrophomonas genera were found among these strains. Fructose, glucose, sunflower oil, and propionic acid were utilized as carbon sources and PHA production was detected by Sudan black staining, Nile blue staining, and FTIR methods. The class I synthase and phaA genes were the most frequently found, indicating the strains' ability to synthesize PHA from carbohydrates. Among the tested bacterial strains, the Pseudomonas genus was identified as able to utilize all tested carbon sources. The Pseudomonas extremorientalis strain was determined as a prospect for biotechnology applications.


Assuntos
Bacillus , Poli-Hidroxialcanoatos , Bactérias/metabolismo , Bacillus/metabolismo , Aciltransferases/genética , Aciltransferases/metabolismo , Carbono/metabolismo
16.
Int J Mol Sci ; 24(2)2023 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-36675293

RESUMO

S-layers are involved in the adaptation of bacteria to the outside environment and in pathogenesis, often representing special virulence factors. Vegetative cells of the entomopathogenic bacterium Brevibacillus laterosporus are characterized by an overproduction of extracellular surface layers that are released in the medium during growth. The purpose of this study was to characterize cell wall proteins of this bacterium and to investigate their involvement in pathogenesis. Electron microscopy observations documented the presence of multiple S-layers, including an outermost (OW) and a middle (MW) layer, in addition to the peptidoglycan layer covering the plasma membrane. After identifying these proteins (OWP and MWP) by mass spectrometry analyses, and determining their gene sequences, the cell wall multilayer-released fraction was successfully isolated and used in insect bioassays alone and in combination with bacterial spores. This study confirmed a central role of spores in bacterial pathogenicity to insects but also detected a significant virulence associated with fractions containing released cell wall multilayer proteins. Taken together, S-layer proteins appear to be part of the toxins and virulence factors complex of this microbial control agent of invertebrate pests.


Assuntos
Bacillus , Brevibacillus , Animais , Fatores de Virulência/metabolismo , Insetos/microbiologia
17.
Arch Microbiol ; 205(1): 52, 2023 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-36600085

RESUMO

This study aimed to investigate the biomass production of Bacillus subtilis in flask and bioreactor conditions. It is necessary to carry the culture from the shake flask to the pH, air, temperature and stirring controlled bioreactor in order to reduce the working time and increase the production efficiency and product quality. In this study, Bacillus was isolated from soil and grown under flask and bioreactor conditions as biocontrol agent against Botrytis cinerea and Fusarium oxysporum. In this process, a pH value of 7.5, 100% O2 saturation, 30% dissolved O2, at the temperature of 37 °C, total flow of 0.1 Lmin-1 and mixing speed of 150 min-1 were preferred for optimal concerning high production yield of B. subtilis in bioreactor. To test whether B. subtilis has antifungal activity on the growth of B. cinerea and F. oxysporum, a dual culture assay in a PDA medium was carried out. Ultimately, high biomass production in a short incubation period by reaching 2.2 µg/mL after 9 h in the bioreactor. It was observed that the bacteria produced in the bioreactor cultivation grew stronger and showed high antifungal activity which resulted 33.33% inhibition percentage against B. cinerea. It was concluded that B. subtilis can be used as a green-fungicide against B. cinerea and F. oxysporum, and bacterial metabolites from B. subtilis could pave the way for the development of next generation green/biopesticides.


Assuntos
Bacillus subtilis , Bacillus , Bacillus subtilis/metabolismo , Antifúngicos/farmacologia , Antifúngicos/metabolismo , Reatores Biológicos , Doenças das Plantas/microbiologia
18.
Arch Microbiol ; 205(1): 53, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36602580

RESUMO

The bacterial communities that colonize the cadaver environment of insects infected and killed by parasitic fungi can be selected by the sympatric fungi and provide novel impacts. In this study, we found that Bacillus cereus/thuringiensis predominate the bacterial community in Dolichoderus thoracicus ant cadavers colonized by O. pseudolloydii. The most predominant bacterial strains in these ant cadavers were hemolytic and able to produce hydrolytic enzymes for digesting the ant tissue. A relatively intense lethal effect on the co-cultured nematode was displayed by a hemolytic strain. Moreover, the antagonistic effect against pathogenic fungi detected in the bacteria sympatric with O. pseudolloydii was reported here. Naphthoquinones have been shown to confer antibacterial activities and produced by the ant-pathogenic Ophiocordyceps fungi. However, our results did not show the naphthoquinone tolerance we expected to be detected in the bacteria from the ant infected by O. pseudolloydii. The bacterial diversity in the samples associated with O. pseudolloydii infected ants as revealed in this study will be a step forward to the understanding of the roles playing by the microbial community in the native habitats of O. pseudolloydii.


Assuntos
Formigas , Bacillus thuringiensis , Bacillus , Animais , Formigas/microbiologia , Insetos
19.
Spectrochim Acta A Mol Biomol Spectrosc ; 289: 122216, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36527970

RESUMO

Accurately, rapidly, and noninvasively identifying Bacillus spores can greatly contribute to controlling a plenty of infectious diseases. Laser tweezers Raman spectroscopy (LTRS) has confirmed to be a powerful tool for studying Bacillus spores at a single cell level. In this study, we constructed a single-cell Raman spectra dataset of living Bacillus spores and utilized deep learning approach to accurately, nondestructively identify Bacillus spores. The trained convolutional neural network (CNN) could efficiently extract tiny Raman spectra features of five spore species, and provide a prediction accuracy of specie identification as high as 100 %. Moreover, the spectral feature differences in three Raman bands at 660, 826, and 1017 cm-1 were confirmed to mostly contribute to producing such high prediction accuracy. In addition, optimal CNN model was employed to monitor and identify sporulation process at different metabolic phases in one growth cycle. The obtained average prediction accuracy of metabolic phase identification was approximately 88 %. It can be foreseen that, LTRS combined with CNN approach have great potential for accurately identifying spore species and metabolic phases at a single cell level, and can be gradually extended to perform identification for many unculturable bacteria growing in soil, water, and food.


Assuntos
Bacillus , Aprendizado Profundo , Pinças Ópticas , Análise Espectral Raman/métodos , Esporos Bacterianos/química
20.
Microbiol Res ; 268: 127276, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36525789

RESUMO

There is currently a constant search for ecofriendly bioproducts, which could contribute to various biomedical applications. Among bioproducts, exopolysaccharides are prominent contemporary extracellular biopolymers that are produced by a great variety of bacterial species. These homo- or heteropolymers are composed of monomeric sugar units linked by glycosidic bonds, which are secreted to the external medium. Bacillus spp. are reported to be present in different ecosystems and produce exopolysaccharides with different biological properties such as antioxidant, antibacterial, antiviral anti-inflammatory, among others. Since a great diversity of bacterial strains are able to produce exopolysaccharides, a great variation in the molecular composition is observed, which is indeed present in some of the chemical structures predicted until date. These molecular characteristics and their relations with different biological functions are discussed in order to visualize future applications in biomedical section.


Assuntos
Bacillus , Polissacarídeos Bacterianos/química , Ecossistema , Bactérias , Antivirais
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