Search Results (13)

Brevibacillus brevis is one of the most common biocontrol strains with broad applications in the prevention and control of plant diseases and insect pests. In order to deepen our understanding of B. brevis genomes, describe their characteristics comprehensively, and mine secondary metabolites, we retrieved the genomic sequences of nine B. brevis strains that had been assembled into complete genomes from the NCBI database. These genomic sequences were analyzed using phylogenetic analysis software, pan-genome analysis software, and secondary metabolite mining software. Results revealed that the genome size of B. brevis strains ranged from 6.16 to 6.73 Mb, with GC content ranging from 47.0% to 54.0%. Phylogenetic analysis classified the nine B. brevis strains into three branches. The analyses of ANI and dDDH showed that B. brevis NEB573 had the potential to become a new species of Brevibacillus and needed further research in the future. The pan-genome analysis identified 10032 gene families, including 3257 core gene families, 3112 accessory gene families, and 3663 unique gene families. In addition, 123 secondary metabolite biosynthetic gene clusters of 20 classes were identified in the genomes of nine B. brevis strains. The major types of biosynthetic gene clusters were non-ribosomal peptide synthase (NRPS) and transAT polyketide synthase (transAT-PKS). Furthermore, a large number of untapped secondary metabolites were identified in B. brevis. In summary, this study elucidated the pan-genome characteristics of the biocontrol bacterium B. brevis and identified its secondary metabolites, providing valuable insights for its further development and utilization. Full article

To search for a novel thermostable esterase for optimized industrial applications, esterase from a thermophilic eubacterium species, Thermoanaerobacter tengcongensis MB4, was purified and characterized in this work. Sequence analysis of T. tengcongensis esterase with other homologous esterases of the same family revealed an apparent tail at the C-terminal that is not conserved across the esterase family. Hence, it was hypothesized that the tail is unlikely to have an essential structural or catalytic role. However, there is no documented report of any role for this tail region. We probed the role of the C-terminal domain on the catalytic activity and substrate preference of T. tengcongensis esterase EstA3 with a view to see how it could be engineered for enhanced properties. To achieve this, we cloned, expressed, and purified the wild-type and the truncated versions of the enzyme. In addition, a naturally occurring member of the family (from Brevibacillus brevis) that lacks the C-terminal tail was also made. In vitro characterization of the purified enzymes showed that the C-terminal domain contributes significantly to the catalytic activity and distinct substrate preference of T. tengcongensis esterase EstA3. All three recombinant enzymes showed the highest preference for paranitrophenyl butyrate (pNPC4), which suggests they are true esterases, not lipases. Kinetic data revealed that truncation had a slight effect on the substrate-binding affinity. Thus, the drop in preference towards long-chain substrates might not be a result of substrate binding affinity alone. The findings from this work could form the basis for future protein engineering allowing the modification of esterase catalytic properties through domain swapping or by attaching a modular protein domain. Full article

Rhizosphere microorganisms and the volatile organic compounds (VOCs) produced by them take part in the regulation of the chemotaxis of nematodes. A total of 150 strains of rhizosphere bacteria were screened via a chemotaxis experiment with Meloidogyne incognita. Some isolates affected the behavior of the nematodes, including attraction, randomness, and repulsion. Volatile metabolites produced via the selected bacteria were associated with the chemotaxis of nematodes. M. incognita was highly attracted to decanal. In addition, dimethyl disulfide, 2,5-dimethylpyrazine, pentadecanoic acid, and palmitic acid were found to attract weakly M. incognita. Furthermore, the chemotaxis of M. incognita was tested in a pot experiment. The bacteria Bacillus sp. 1-50, Brevibacillus brevis 2-35, B. cereus 5-14, Chryseobacterium indologens 6-4, and VOC decanal could regulate the movement of M. incognita in the pot with or without plants. The results provide insights into rhizosphere microorganisms and their VOCs and how they regulate the chemotaxis of the nematodes. Full article

Gray mold disease, caused by Botrytis cinerea, has reduced grape’s output and market quality globally. In this study, the antifungal activity of a mixed microbial combination of Brevibacillus brevis FJAT-0809-GLX and Bacillus sp. strains was investigated. The results showed that the inhibition rate against B. cinerea was 85.10% when B. brevis FJAT-0809-GLX was mixed with a mixture of B. brevis FJAT-10623 and Bacillus velezensis FJAT-55034 at a proportion of 80%:20%, which was significantly higher than those of other combination proportions. The inhibitory rates of the mixed microbial combination diluted 0 times, 10 times, and 100 times were 89.14%, 88.10%, and 86.33%, respectively, with no significant differences between each other. The mixed microbial combination appeared to be temperature-insensitive and significantly stable from pH 3 to pH 7. Furthermore, it was discovered that its antifungal activity was significantly stable with UV radiation for 30 min, 60 min, and 90 min, with values of 84.82%, 83.89%, and 82.22%, respectively. An amount of 0.025 mol mL⁻¹ of KCl, CuSO₄, and MgCl₂ had no effects on the antifungal activity of the mixed microbial combination, while 0.025 mol mL⁻¹ of NaCl, ZnSO₄, FeSO₄, CaCl₂, and MnSO₄ reduced the inhibition rate. The mixed microbial combination demonstrated antifungal activities against a variety of fungi, with inhibition rates ranging from 68.78% to 85.10%. The grape fruits and grape leaves treated with the mixed microbial combination decayed at 27.27% and 48.34%, respectively. Additionally, the mixed microbial combination improved grape fruit resistance by increasing the activity of defense enzymes polyphenol oxidase (PPO) and catalase (CAT). Therefore, the results indicated that the mixed microbial combination had great biocontrol potential against gray mold in grape fruits. Full article

The common bean (Phaseolus vulgaris L.) is a significant vegetable crop, grown because it is a rich source of protein, carbohydrates, and vitamin B complex. Fusarium solani and Rhizoctonia solani are the most widely known pathogens contributing to large yield losses for this crop. The use of cultural and chemical control practices has been ineffective. Therefore, a sustainable, affordable, and effective control method is urgently required. In this study, we aimed to isolate and characterize Bacillus velezensis from Lake Bogoria as a potential biocontrol agent for Fusarium solani. Bacteria were isolated from soil and sediments using the serial dilution technique. Molecular characterization was performed using the 16S rRNA gene. A total of 13 bacteria were isolated from soil and sediments. Based on the partial sequences, BLAST analysis showed two isolates, B20 (Bacillus velezensis strain QH03-23) and B30 (Bacillus velezensis strain JS39D), belonging to Bacillus velezensis. Other isolates were identified as Bacillus tequilensis, Brevibacillus brevis, Bacillus subtilis, Bacillus amyloliquefaciens, and Bacillus licheniformis. The effectiveness of their antifungal properties was determined via co-culturing, and we found mycelial inhibition rates of 28.17% (for B20) and 33.33% (for B30) for the Fusarium solani isolates. The characterization of the Bacillus velezensis strain revealed that they were Gram-positive and grew well at pH 7.0 and 8.5, although growth was recorded at pH 5.0 and 10.0. In terms of temperature, the optimal temperature conditions were 30−35 °C, with an optimum salinity of 0–0.5 M NaCl. When these isolates were tested for their ability to produce secondary metabolites, they were found to produce phosphate, pectinase, chitinase, protease, indole -3- acetic acid (IAA), and hydrogen cyanide (HCD), making them potential biocontrol agents. Full article

Contamination by ochratoxigenic fungi and its prevention during the pile-fermentation of post-fermented tea have always been a concern. The present study aimed to elucidate the anti-fungal effect and mechanism of polypeptides produced by B. brevis DTM05 (isolated from post-fermented tea) on ochratoxigenic fungi, and to to evaluate their use in the pile-fermentation process of post-fermented tea. The results showed that polypeptides (produced by B. brevis DTM05) with a strong antifungal effect against A. carbonarius H9 mainly had a molecular weight between 3 and 5 kDa. The Fourier-transform infrared spectra of this polypeptide extract showed that it was a mixture consisting mainly of polypeptides and small amounts of lipids and other carbohydrates. The polypeptide extracts significantly inhibited the growth of A. carbonarius H9, and its minimum inhibitory concentration (MIC) was 1.6 mg/L, which significantly reduced the survival rate of spores. The polypeptides also effectively controlled the occurrence and ochratoxin A (OTA) production of A. carbonarius H9 on the tea matrix. The lowest concentration of polypeptides that significantly inhibited the growth of A. carbonarius H9 on the tea matrix was 3.2 mg/L. The enhancement of the fluorescence staining signal in the mycelium and conidiospore showed that the polypeptides with a concentration of more than 1.6 mg/L increased the permeability of the mycelium membrane and conidial membrane of A. carbonarius H9. The significant increase in the extracellular conductivity of mycelia suggested the outward leakage of intracellular active substances, and also further indicated an increase in cell membrane permeability. Polypeptides with a concentration of 6.4 mg/L significantly down-regulated the expression level of the polyketide synthase gene related to OTA production (acpks) in A. carbonarius H9, which may be the fundamental reason why polypeptides affect OTA production. In conclusion, reasonable use of the polypeptides produced by B. brevis can destroy the structural integrity of the cell membrane, make the intracellular active substances leak outward, accelerate the death of fungal cells and down-regulate the expression level of the polyketide synthase gene in A. carbonarius; thus, they can effectively control the contamination of ochratoxigenic fungi and OTA production during the pile-fermentation of the post-fermented tea. Full article

Brevibacillus laterosporus (Bl) is a Gram-positive and spore-forming bacterium belonging to the Brevibacillus brevis phylogenetic cluster. Globally, insect pathogenic strains of the bacterium have been isolated, characterised, and some activities have been patented. Two isolates, Bl 1821L and Bl 1951, exhibiting pathogenicity against the diamondback moth and mosquitoes, are under development as a biopesticide in New Zealand. However, due to the suspected activity of putative antibacterial proteins (ABPs), the endemic isolates often grow erratically. Various purification methods, including size exclusion chromatography, sucrose density gradient centrifugation, polyethylene glycol precipitation, and ammonium sulphate precipitation employed in this study, enabled the isolation of two putative antibacterial proteins of ~30 and ~48 kD from Bl 1821L and one putative antibacterial protein of ~30 kD from Bl 1951. Purification of the uninduced cultures of Bl 1821L and Bl 1951 also yielded protein bands of ~30 and ~48 kD on SDS-PAGE, which indicated their spontaneous induction. A disc diffusion assay was used to determine the antagonistic activities of the putative ABPs. Subsequent transmission electron microscope (TEM) examination of a purified putative antibacterial protein-containing solution showed the presence of encapsulin (~30 kD) and polysheath (~48 kD)-like structures. Although only the ~30 kD protein was purified from Bl 1951, both structures were seen in this strain under TEM. Furthermore, while assessing the antibacterial activity of some fractions of Bl 1951 against Bl 1821L in the size exclusion chromatography method, the population of Bl 1821L persister cells was noted. Overall, this work added a wealth of knowledge about the purification of the high-molecular-weight (HMW) proteins (bacteriocins) of Gram-positive bacteria including Bl. Full article

The Gram-positive and spore-forming bacterium Brevibacillus laterosporus (Bl) belongs to the Brevibacillus brevis phylogenetic cluster. Isolates of the species have demonstrated pesticidal potency against a wide range of invertebrate pests and plant diseases. Two New Zealand isolates, Bl 1821L and Bl 1951, are under development as biopesticides for control of diamondback moth and other pests. However, due to the often-restricted growth of these endemic isolates, production can be an issue. Based on the previous work, it was hypothesised that the putative phages might be involved. During investigations of the cause of the disrupted growth, electron micrographs of crude lysate of Bl 1821L showed the presence of phages’ tail-like structures. A soft agar overlay method with PEG 8000 precipitation was used to differentiate between the antagonistic activity of the putative phage and phage tail-like structures (bacteriocins). Assay tests authenticated the absence of putative phage activity. Using the same method, broad-spectrum antibacterial activity of Bl 1821L lysate against several Gram-positive bacteria was found. SDS-PAGE of sucrose density gradient purified and 10 kD MWCO concentrated lysate showed a prominent protein band of ~48 kD, and transmission electron microscopy revealed the presence of polysheath-like structures. N-terminal sequencing of the ~48 kD protein mapped to a gene with weak predicted amino acid homology to a Bacillus PBSX phage-like element xkdK, the translated product of which shared >90% amino acid similarity to the phage tail-sheath protein of another Bl published genome, LMG15441. Bioinformatic analysis also identified an xkdK homolog in the Bl 1951 genome. However, genome comparison of the region around the xkdK gene between Bl 1821L and Bl 1951 found differences including two glycine rich protein encoding genes which contain imperfect repeats (1700 bp) in Bl 1951, while a putative phage region resides in the analogous Bl 1821L region. Although comparative analysis of the genomic organisation of Bl 1821L and Bl 1951 PBSX-like region with the defective phages PBSX, PBSZ, and PBP 180 of Bacillus subtilis isolates 168 and W23, and Bacillus phage PBP180 revealed low amino acids similarity, the genes encode similar functional proteins in similar arrangements, including phage tail-sheath (XkdK), tail (XkdO), holin (XhlB), and N-acetylmuramoyl-l-alanine (XlyA). AMPA analysis identified a bactericidal stretch of 13 amino acids in the ~48 kD sequenced protein of Bl 1821L. Antagonistic activity of the purified ~48 kD phage tail-like protein in the assays differed remarkably from the crude lysate by causing a decrease of 34.2% in the number of viable cells of Bl 1951, 18 h after treatment as compared to the control. Overall, the identified inducible phage tail-like particle is likely to have implications for the in vitro growth of the insect pathogenic isolate Bl 1821L. Full article

Pharmaceutical products that mix natural raw materials are subject to unavoidable contamination with microorganisms from the environment and animals. This study focused on the effect of radiation on the quality of primary packaged pills, which are crude drug products. The pills, which were sealed in a sack for primary packaging laminated with polyethylene terephthalate, polyethylene, and aluminum foil, were irradiated by gamma rays or electron beam (EB). The survival counts of bacteria were reduced to 10³ CFU/g or less by 6 kGy of irradiation. The counts of the spore-forming bacteria Bacillus megaterium, B. cereus, and Brevibacillus brevis in the pills were reduced to not over 100 CFU/g after 10 kGy irradiation. Although some of the cinnamaldehyde in the pills was oxidized to cinnamic acid, the decomposition of swertiamarin, berberine, glycyrrhizin, and cinnamaldehyde in the pills after 10 kGy irradiation were within the analytical accuracy by high-performance liquid chromatography. Gamma-ray or EB treatment at the final production of crude drug preparations was within the permissible standard value for the non-aqueous preparations for oral administration, with no statistically significant change in the indicator ingredients of crude drugs. Full article

Breast milk is the combination of bioactive compounds and microflora that promote newborn’s proper growth, gut flora, and immunity. Thus, it is always considered the perfect food for newborns. Amongst their bioactives, probiotic communities—especially lactic acid bacteria (LAB)—are characterized from breast milk over the first month of parturition. In this study, seven LAB were characterized phenotypically and genotypically as Levilactobacillus brevis BDUMBT08 (MT673657), L. gastricus BDUMBT09 (MT774596), L. paracasei BDUMBT10 (MT775430), L. brevis BDUMBT11 (MW785062), L. casei BDUMBT12 (MW785063), L. casei BDUMBT13 (MW785178), and Brevibacillus brevis M2403 (MK371781) from human breast milk. Their tolerance to lysozyme, acid, bile, gastric juice, pancreatic juice, and NaCl and potential for mucoadhesion, auto-aggregation, and co-aggregation with pathogens are of great prominence in forecasting their gut colonizing ability. They proved their safety aspects as they were negative for virulence determinants such as hemolysis and biofilm production. Antibiogram of LAB showed their sensitivity to more than 90% of the antibiotics tested. Amongst seven LAB, three isolates (L. brevis BDUMBT08 and BDUMBT11, and L. gatricus BDUMBT09) proved their bacteriocin producing propensity. Although the seven LAB isolates differed in their behavior, their substantial probiotic properties with safety could be taken as promising probiotics for further studies to prove their in vivo effects, such as health benefits, in humans. Full article

In this study, 15 bacterial endophytes linked with the leaves of the native medicinal plant Pulicaria incisa were isolated and identified as Agrobacterium fabrum, Acinetobacter radioresistant, Brevibacillus brevis, Bacillus cereus, Bacillus subtilis, Paenibacillus barengoltzii, and Burkholderia cepacia. These isolates exhibited variant tolerances to salt stress and showed high efficacy in indole-3-acetic acid (IAA) production in the absence/presence of tryptophan. The maximum productivity of IAA was recorded for B. cereus BI-8 and B. subtilis BI-10 with values of 117 ± 6 and 108 ± 4.6 μg mL⁻¹, respectively, in the presence of 5 mg mL⁻¹ tryptophan after 10 days. These two isolates had a high potential in phosphate solubilization and ammonia production, and they showed enzymatic activities for amylase, protease, xylanase, cellulase, chitinase, and catalase. In vitro antagonistic investigation showed their high efficacy against the three phytopathogens Fusarium oxysporum, Alternaria alternata, and Pythium ultimum, with inhibition percentages ranging from 20% ± 0.2% to 52.6% ± 0.2% (p ≤ 0.05). Therefore, these two endophytic bacteria were used as bio-inoculants for maize seeds, and the results showed that bacterial inoculations significantly increased the root length as well as the fresh and dry weights of the roots compared to the control plants. The Zea mays plant inoculated with the two endophytic strains BI-8 and BI-10 significantly improved (p ≤ 0.05) the growth performance as well as the nutrient uptake compared with an un-inoculated plant. Full article

Intercropping has been widely used to control disease and improve yield in agriculture. In this study, maize and peanut were used for non-separation intercropping (NS), semi-separation intercropping (SS) using a nylon net, and complete separation intercropping (CS) using a plastic sheet. In field experiments, two-year land equivalent ratios (LERs) showed yield advantages due to belowground interactions when using NS and SS patterns as compared to monoculture. In contrast, intercropping without belowground interactions (CS) showed a yield disadvantage. Meanwhile, in pot experiments, belowground interactions (found in NS and SS) improved levels of soil-available nutrients (nitrogen (N) and phosphorus (P)) and enzymes (urease and acid phosphomonoesterase) as compared to intercropping without belowground interactions (CS). Soil bacterial community assay showed that soil bacterial communities in the NS and SS crops clustered together and were considerably different from the CS crops. The diversity of bacterial communities was significantly improved in soils with NS and SS. The abundance of beneficial bacteria, which have the functions of P-solubilization, pathogen suppression, and N-cycling, was improved in maize and peanut soils due to belowground interactions through intercropping. Among these bacteria, numbers of Bacillus, Brevibacillus brevis, and Paenibacillus were mainly increased in the maize rhizosphere. Burkholderia, Pseudomonas, and Rhizobium were mainly increased in the peanut rhizosphere. In conclusion, using maize and peanut intercropping, belowground interactions increased the numbers of beneficial bacteria in the soil and improved the diversity of the bacterial community, which was conducive to improving soil nutrient (N and P) supply capacity and soil microecosystem stability. Full article

A Gram-positive, facultatively anaerobic, endospore-forming and rod-shaped bacterium was isolated from soil samples and designated strain LQQ. This organism strongly quenches the acylhomoserine lactone quorum-sensing signal. The LQQ strain exhibits phenotypic characteristics consistent with its classification in the genus Bacillus. It is positive in catalase and no special growth factor is needed. It uses glucose as sole carbon source. The DNA G + C content is 39.8 mol %. The closest relatives based on the 16S rRNA gene sequence are Bacillus anthracis, Bacillus thuringiensis, and Brevibacillus brevis (syn. Bacillus brevis) with the similarity of 96.5%. The DNA–DNA hybridization data indicates a low level of genomic relatedness with the relative type strains of Bacillus thuringiensis (6.1%), Bacillus anthracis (10.5%) and Brevibacillus brevis (8.7%). On the basis of the phenotypic and phylogenetic data together with the genomic distinctiveness, the LQQ strain represents a novel species of the genus Bacillus, for which the name Bacillus marcorestinctum sp. nov. is proposed. The type strain is LQQ^T. Full article