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BIOLOG test results obtained by using Biolog Microbial Identification System developed by Biolog Inc., USA
Source publication
To isolate and characterize an antifungal protein from the culture broth of the bacterium Bacillus amyloliquefaciens.
The antifungal protein designated as baciamin was isolated and exhibited a molecular mass around 50 kDa. Baciamin manifested a broad spectrum of antifungal activity. Baciamin could induce membrane permeabilization of tested fungi. I...
Context in source publication
Context 1
... biochemical and physiological properties of the strain were tested by using Biolog Microbial Identification System. The Biolog results led to the identification of the strain as B. amyloliquefaciens (Table 2). When the results of the sequence of 16S rRNA and Biolog identification were viewed together, it can be corroborated that the strain used in this study was B. amyloliquefaciens. ...Similar publications
In the present study, six isolates of bacteria were isolated from the rhizosphere soil and assayed for its capability to show antagonism characteristics against the bacterial isolates Salmonella typhimurium, Pseudomonas aeruginosa and fungal isolates Aspergillus flavus and Aspergillus niger. Among the isolates, Bacillus amyloliquefaciens MS-3 was f...
Amylasen werden häufig in Waschmitteln eingesetzt. Diese Anwendung werden Amylasen benötigt, die eine hohen Stabilität und Aktivität bei alkalischem pH besitzen. Zudem ist eine hohe spezifische Aktivität wünschenswert, da so Enzymmenge und dadurch Kosten gespart werden können.
Ziel dieser Arbeit war die Etablierung und Anwendung von Methoden der ge...
A total of 118 bacterial strains were isolated from six different soil samples from different parts of Pakistan. 50 out of these strains exhibited antagonistic activities against at least two or more strain from a panel of pathogenic and non-pathogenic microorganisms. Tested strains included three sets; environmental, laboratory and multidrug resis...
The purpose of this study was to investigate the physicochemical properties and bioactivities of rice beans (Vigna umbellata) fermented by Bacillus amyloliquefaciens. The fermentation conditions were optimized on the basis of the fibrinolytic activity. Under the optimal fermentation conditions, the fibrinolytic activity reached a maximum of 78.0 FU...
Citations
... amyloliquefaciens, purified protein, and the SDS-PAGE (Coomassie blue-stained) gel showed a single band with a molecular weight of about 50 kDa for all samples (Original photographs in supplementary Fig. S3 A, B, and cropped photograph in Fig. 3). In another study on fungi, Wong et al. (2008) described an antifungal protein of B. amyloliquefaciens subsp. amyloliquefaciens which has a molecular mass of nearly 50 kDa and is designated as baciamin (Wong et al., 2008). ...
... In another study on fungi, Wong et al. (2008) described an antifungal protein of B. amyloliquefaciens subsp. amyloliquefaciens which has a molecular mass of nearly 50 kDa and is designated as baciamin (Wong et al., 2008). So, we confirmed that B. amyloliquefaciens subsp. ...
... amyloliquefaciens using ammonium precipitation. This protein is thermostable, salt-tolerant, trypsin, and pH stable (Wong et al., 2008). Very few antifungal proteins of bacterial origin have been reported up to this point, while antibacterial proteins have not. ...
Serratia rubidaea is an opportunistic Gram-negative pathogen that has developed antimicrobial resistance to a variety of commercial antibiotics. The spread of this multidrug-resistant pattern predicts that it will get harder and harder to treat S. rubidaea infections in the future. For this perception, antimicrobial proteins might represent a safe, effective, and biodegradable alternative because their site of action is on cyclic peptides. In this study, one candidate Bacillus amyloliquefaciens subsp. amyloliquefaciens was isolated from the soil of Sundarban mangrove forest, and its identification was confirmed both using the PCR (Polymerase chain reaction) method and the BIOLOG™ microbial identification system. The antibacterial protein, which has a molecular mass of about 50 kDa, was isolated from B. amyloliquefaciens subsp. amyloliquefaciens. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to confirm the extracted protein's purity. This potential protein was discovered to develop and exhibit antagonistic activity throughout a broad temperature, pH, and salinity range. At doses ranging from 300 to 400 μg/ml, this protein has antagonistic activity against multidrug resistant S. rubidaea and a wide range of other resistant pathogenic bacteria such as Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and so on. The research provides new insights to develop bio-control agents that can be applied for prevent, treat, and control infectious diseases caused by multidrug resistant S. rubidaea, as well as other pathogenic bacteria.
... In addition to tiny chemical molecules with antifungal action, antifungal proteins have received attention. [2] Antifungal proteins are now known to be produced by a wide range of animals, including humans, other vertebrates, invertebrates, plants, fungi, and bacteria. [3] There is a diverse range of antifungal proteins with various architectures. ...
Fungal infections have dramatically increased over the past few years, as evidenced by an increase in the number of deaths from fungal infections including AIDS, cancer, etc. New antifungal treatments must be developed to meet the global challenge of preventing fungal infection and mycotoxin contamination of food and feed. Due to their characteristics like diversity and function, antifungal spectrum, mechanism of action, high stability, and the availability of biotechnological production methods, antimicrobial peptides and proteins (AMPs) with antifungal activity are attracting significant attention as natural antifungal agents. This study focused on the isolation of antifungal protein-producing microorganisms by using a crowded plate technique from the soil along with the purification of antifungal protein by methods like ammonium sulfate precipitation, solvent extraction, etc. along with this characterization of antimicrobial protein was carried out by polyacrylamide gel electrophoresis. Antifungal potential of purified protein was tested using the agar well diffusion method against respective pathogenic fungi. Bacillus species having good antifungal activity have been isolated from the soil. Efficient extraction of antifungal compound produced by isolated Bacillus species has been done using ammonium sulfate precipitation as well as a solvent extraction method. The purified antifungal protein exhibits good antifungal activity against food-borne pathogens like Aspergillus niger and Candida albicance. Therefore, the isolated Bacillus strain and its potent antifungal protein could be used in various food and biomedical applications.
... Besides antibacterial activity of bacteriocin, it has been found to have antifungal activity. For example, baciamin, an antifungal protein produced by B. amyloliquefaciens was reported to be active against various fungi like Botrytis cinerea, Helminthosporium t u r c i c u m , H a r p o p h o r a m a y d i s , V a l s a m a l i , Mycosphaerella arachidicola, Pythium aphanidermatum, Rhizoctonia solani, and Fusarium oxysporum (14,48). Bacteriocin producing bacterial strains can be used as probiotic supplement for human and animals as they can inhibit the intestinal pathogens such as Clostridium perfringens, Clostridium difficile and others (49,50). ...
Antimicrobial peptides (AMPs) are inherently occurring proteins that are produced by microorganisms as secondary metabolites. Members of genus Bacillus produce many types of AMPs by ribosomal (bacteriocins) and non-ribosomal (polymyxins and iturins) mechanisms. Bacteriocins are ribosomally synthesized peptides that inhibit the growth of closely related bacterial strains. Moreover, bacteriocins produced by Bacillus species have been widely used in pharmaceutical, food industry, fishery, livestock as well as in agriculture sector. The objective of this review is to assess the characterization of the Bacillus-derived bacteriocins, their potential use in different sectors and structure-activity relationships.
... Bacteriocins with antimicrobial activity, greater than 10 kDa of class III, were already detected in different Bacillus species [49]. For example, baciamin, bacisubin, CAMT2, and Bac14B showed high antifungal and antimicrobial activity [52][53][54][55]. Further biochemical and molecular characterization after complete purification will be undertaken. ...
In the present study, we identified the Bacillus subtilis strain annotated Fa17.2 isolated from Bromelia flower inflorescences collected from the subtropical humid mesothermal region, Santo Domingo de Los Tsachilas Province, Ecuador. The probiotic capacity and antimicrobial potential against four foodborne pathogens were assessed. The cell culture of Fa17.2 is highly resistant to synthetic gastric acid (pH 2.5, 3.0, and 3.5), bile salts (0.3%), tolerating different sodium chloride concentrations (1, 3, and 5%), and growth conditions (15 °C and 45 °C), suggesting its potential probiotic features. The isolate showed no antibiotic resistance and was considered safe as no hemolysis was detected on sheep blood agar. The optimum medium for bacterial growth and the release of antimicrobial compounds was MRS with 10% glucose. The active components released in the neutralized crude extract (NCE) were insensitive to organic solvents, surfactants, and nonproteolytic enzymes and sensitive to proteolytic enzymes suggesting their proteinaceous nature. The antimicrobial activity was enhanced by heat and maintained active over a wide range of pH (2.0–8.0). Moreover, the crude extract (CE) showed inhibitory activity against several Gram-negative and Gram-positive bacteria. The molecular weight of partially purified precipitated bacteriocin-like substances (BLISs) was about 14 kDa in 20% Tricine-SDS-PAGE. The CE obtained from Fa17.2 inhibits the growth of four foodborne pathogens, Staphylococcus aureus, Escherichia coli, Kosaconia cowanii, and Shigella dysenteriae, which implies its potential as an antimicrobial producer strain.
... Bacillus amyloliquefaciens could produce several extracellular enzymes to augment the digestibility and absorption of nutrients in addition to the overall intestinal immune function (54). Additionally, B. amyloliquefaciens exerted antagonistic activities against pathogens by producing diverse bioactive metabolites including lipopeptides, fengycin, and iturin (55). One study reported that the fengycin secreted by B. amyloliquefaciens could competitively combine the receptor protein accessory gene regulator (AgrC) of bacterial quorum-sensing systems to inhibit the pathogens colonized in animal intestines (8). ...
This study aimed to investigate the protective effects of Bacillus amyloliquefaciens (BA40) against Clostridium perfringens (C. perfringens) infection in mice. Bacillus subtilis PB6 was utilized as a positive control to compare the protective effects of BA40. In general, a total of 24 5-week-old male C57BL/6 mice were randomly divided into four groups, with six mice each. The BA40 and PB6 groups were orally dosed with resuspension bacteria (1 × 10⁹ CFU/ml) once a day, from day 1 to 13, respectively. In the control and infected groups, the mice were orally pre-treated with phosphate-buffered saline (PBS) (200 μl/day). The mice in the infected groups, PB6 + infected group and BA40 + infected group, were orally challenged with C. perfringens type A (1 × 10⁹ CFU/ml) on day 11, whereas the control group was orally dosed with PBS (200 μl/day). The results showed that the BA40 group ameliorated intestinal structure damage caused by the C. perfringens infection. Furthermore, the inflammatory responses detected in the infected groups which include the concentrations of IL-1β, TNF-α, IL-6, and immunoglobulin G (IgG) in the serum and secretory immunoglobulin (SigA) in the colon, and nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) activity in the jejunum, were also alleviated (P < 0.05) by BA40 treatment. Similarly, cytokines were also detected by quantitative PCR (qPCR) in the messenger RNA (mRNA) levels, and the results were consistent with the enzyme-linked immunosorbent assay (ELISA) kits. Additionally, in the infected group, the mRNA expression of Bax and p53 was increasing and the Bcl-2 expression was decreasing, which was reversed by BA40 and PB6 treatment (P < 0.05). Moreover, the intestinal microbiota imbalance induced by the C. perfringens infection was restored by the BA40 pre-treatment, especially by improving the relative abundance of Verrucomicrobiota (P < 0.05) and decreasing the relative abundance of Bacteroidetes (P < 0.05) in the phyla level, and the infected group increased the relative abundance of some pathogens, such as Bacteroides and Staphylococcus (P < 0.05) in the genus level. The gut microbiota alterations in the BA40 group also influenced the metabolic pathways, and the results were also compared. The purine metabolism, 2-oxocarboxylic acid metabolism, and starch and sucrose metabolism were significantly changed (P < 0.05). In conclusion, our results demonstrated that BA40 can effectively protect mice from C. perfringens infection.
... Bacillus spp., in particular B. amyloliquefaciens, have been reported to produce a wide variety of antimicrobial compounds including hydrogen cyanide (HCN), phenazines, pyrrolnitrin, 2,4-diacetylphloroglucinol (2,4-DAPG), lipopeptides (bacillomycin, iturins, and fengycins), and cell wall degrading enzymes (Lin et al., 2014;Mounia et al., 2014). B. amyloliquefaciens has been reported as one of the most effective biocontrol agent of plant pathogens, due to its ability to reduce plant infections and survive in "harsh" environmental conditions (Stein et al., 2004;Wong et al., 2008). Overall, our findings agree with the reports by other investigators. ...
Bacterial spot, caused by Xanthomonas cucurbitae, is an important disease of cucurbits worldwide. Considering the absence of resistant cultivars, along with the limited effects of chemical treatments, we evaluated the use of potential biocontrol agents for management of the disease. We collected 1,134 bacterial isolates associated with bacterial spot symptomatic leaves and fruits and screened in vitro for their antagonistic activity against X. cucurbitae. A total of 271 isolates inhibited colony formation of X. cucurbitae in a disc diffusion assay. Species of these antagonistic isolates were identified based on 16s rRNA primers and phylogenetic analysis using four housekeeping genes. Based on results from the in vitro assays, the strong inhibitors of X. cucurbitae were selected for greenhouse and field evaluations. We found that Bacillus amyloliquefaciens, Bacillus velezensis, Pantoea agglomerans, and Pseudomonas putida were able to prevent or limit development of bacterial spot on pumpkin plants in the greenhouse and field studies. Additionally, Bacillus amyloliquefaciens and Pseudomonas putida increased growth of ‘Howden’ pumpkin (Cucurbita pepo). Our results suggested that these biocontrol agents can be used for management of the bacterial spot disease and enhance plant growth of pumpkins.
... Other useful activities shown by Baciamin were antitumor and immuno-enhancing activities. Its antifungal activity was retained after exposure to the temperature range 0-100°C for 20 min, and to the pH range of 6-11 for 30 min, however, the activity was nullified at pH values below 5 [22]. ...
Bacillus amyloliquefaciens is one of the many Bacillus species which is used as a probiotic, exerting health benefits to the host. B. amyloliquefaciens is a gram-positive, rod shaped bacteria with peritrichous flagella. The species also produces enzymes viz. cellulases, amylases, and proteases, which aid in digestion. The species is principally utilized as a feed additive substance for domestic animals and birds. It is gaining importance as plant growth promoter rhizobacteria (PGPR). Various strains of B. amyloliquefaciens are used as a biocontrol agent against Peach gummosis, Fusarium oxysporum growth, algal blooms, and anthracnose rot caused by Colletotrichumacutatum. Recently, it is being used to induce defense mechanism in plants against various plant pathogens. B. amyloliquefaciens is capable of producing a broad-spectrum antifungal protein, Baciamin, along with a lipopeptide compound known as Bacillomycin D, which also shows an antifungal property. The contribution of B. amyloliquefaciens in Gene expression regulation in rice and inducing tolerance against various abiotic stresses is crucial in agricultural field.
... The antifungal activities of the rF2 culture filtrates treated as detailed above were determined according to the method described by Zhao et al. [37]. The formula reported by Wong et al. was used to calculate the relative activity of the treated rF2 protein [38]. Three replicates for each treatment were assessed in three repeated experiments. ...
Background
Wheat sheath blight, a soil borne fungal disease caused by Rhizoctonia cerealis, is considered as one of the most serious threats to wheat worldwide. Bacillus subtilis Z-14 was isolated from soil sampled from a wheat rhizosphere and was confirmed to have strong antifungal activity against R. cerealis.
Results
An antifungal protein, termed F2, was isolated from the culture supernatant of Z-14 strain using precipitation with ammonium sulfate, anion exchange chromatography, and reverse phase chromatography. Purified F2 had a molecular mass of approximately 8 kDa, as assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Edman degradation was used to determine the amino acid sequence of the N-terminus, which was NH2ASGGTVGIYGANMRS. This sequence is identical to a hypothetical protein RBAM_004680 (YP_001420098.1) synthesized by B. amyloliquefaciens FZB42. The recombinant F2 protein (rF2) was heterologously expressed in the yeast host Pichia pastoris, purified using a Niaffinity column, and demonstrated significant antifungal activity against R. cerealis. The purified rF2 demonstrated broad spectrum antifungal activity against different varieties of fungi such as Fusarium oxysporum, Verticillium dahliae, Bipolaris papendorfii, and Fusarium proliferatum. rF2 was thermostable, retaining 91.5% of its activity when incubated for 30 min at 100 °C. Meanwhile, rF2 maintained its activity under treatment by proteinase K and trypsin and over a wide pH range from 5 to 10.
Conclusions
A novel antifungal protein, F2, was purified from biocontrol Bacillus subtilis Z-14 strain fermentation supernatant and heterologously expressed in Pichia pastoris to verify its antifungal activity against R. cerealis and the validity of the gene encoding F2. Considering its significant antifungal activity and stable characteristics, protein F2 presents an alternative compound to resist fungal infections caused by R. cerealis.
... These results are similar to the ones found by Fernandes et al. [67] where surfactants produced by Bacillus were more active against Gram-positive bacteria than Gram-negative. The genus Bacillus has the capability to produce lipopeptide types of biosurfactants with potent antimicrobial activity [68,69]. Lipopeptides with strong antibacterial activity have the ability to destroy the bacterial cell membranes by detergent-like action on cell membranes. ...
Biofouling causes major problems and economic losses to marine and shipping industries. In the search for new antifouling agents, marine bacteria with biosurfactants production capability can be an excellent option, due to the amphipathic surface-active characteristic that confers antimicrobial and antibiofilm activities. The aim of this study was to evaluate the antifouling activity of biosurfactants producing marine bacteria from the Gulf of California. The cell free culture supernatant (CFCS) of Bacillus niabensis (S-69), Ralstonia sp. (S-74) (isolated from marine sediment) and of B. niabensis (My-30) (bacteria associated to the sponge Mycale ramulosa) were screened for production of biosurfactants (using hemolysis and drop collapse test, oil displacement and emulsifying activity). The toxicity and antifouling activity were evaluated against biofoulers (bacteria forming biofilm and macrofoulers) both in laboratory and field assays. The results indicate that all bacteria were biosurfactant producers, but the higher capability was shown by B. niabensis (My-30) with high emulsifying properties (E24) of 71%. The CFCS showed moderate toxicity but were considered non-toxic against Artemia franciscana at low concentrations. In the antifouling assay, the CFCS of both strains of B. niabensis showed the best results for the reduction of the biofilm formation (up 50%) against all Gram-positive bacteria and most Gram-negative bacteria with low concentrations. In the field assay, the CFCS of B. niabensis (My-30) led to the reduction of 30% of biofouling compared to the control. The results indicate that the biosurfactant produced by B. niabensis (My-30) has promising antifouling activity.
... The antifungal activities of the rF2 culture ltrates treated as detailed above were determined according to the method described by Zhao et al. [37]. The formula reported by Wong et al. was used to calculate the relative activity of the treated rF2 protein [38]. Three replicates for each treatment were assessed in three repeated experiments. ...
Background: Wheat sheath blight, a soil borne fungal disease caused by Rhizoctonia cerealis, is considered as one of the most serious threats to wheat worldwide. Bacillus subtilis Z-14 was isolated from soil sampled from a wheat rhizosphere and was confirmed to have strong antifungal activity against R. cerealis.
Results: An antifungal protein, termed F2, was isolated from the culture supernatant of Z-14 strain using precipitation with ammonium sulfate, anion exchange chromatography, and reverse phase chromatography. Purified F2 had a molecular mass of approximately 8 kDa, as assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Edman degradation was used to determine the amino acid sequence of the N-terminus, which was NH2‑ASGGTVGIYGANMRS. This sequence is identical to a hypothetical protein RBAM_004680 (YP_001420098.1) synthesized by B. amyloliquefaciens FZB42. The recombinant F2 protein (rF2) was heterologously expressed in the yeast host Pichia pastoris, purified using a Ni‑affinity column, and demonstrated significant antifungal activity against R. cerealis. The purified rF2 demonstrated broad spectrum antifungal activity against different varieties of fungi such as Fusarium oxysporum, Verticillium dahliae, Bipolaris papendorfii, and Fusarium proliferatum. rF2 was thermostable, retaining 91.5% of its activity when incubated for 30 min at 100 °C. Meanwhile, rF2 maintained its activity under treatment by proteinase K and trypsin and over a wide pH range from 5 to 10.
Conclusions: A novel antifungal protein, F2, was purified from biocontrol Bacillus subtilis Z-14 strain fermentation supernatant and heterologously expressed in Pichia pastoris to verify its antifungal activity against R. cerealis and the validity of the gene encoding F2. Considering its significant antifungal activity and stable characteristics, protein F2 presents an alternative compound to resist fungal infections caused by R. cerealis.
