scholarly journals Extraction, purification and characterization of antifungal and antibacterial substances produced from Bacillus velezensis CP 1604

2016 ◽  
Vol 14 (3) ◽  
pp. 563-571
Author(s):  
Trịnh Thành Trung ◽  
Đinh Thị Tuyết Vân ◽  
Nguyễn Phương Liên ◽  
Đào Thị Lương ◽  
Dương Văn Hợp
Keyword(s):  
Molecular Weight ◽  
Plant Pathogens ◽  
Hydrolytic Enzymes ◽  
Antimicrobial Activities ◽  
Low Ph ◽  
Proteinase K ◽  
Mass Spectrometry Analysis ◽  
Antibacterial Substance ◽  
Bacillus Velezensis ◽  
Antifungal Substance

Bacillus velezensis is one of the members of the Bacillus subtilis species complex which is generally considered as safety organism. The bacterium possesses several beneficial properties for crops, especially production of antifungal and antibacterial agents against plant pathogens. In this study, we aimed to investigate the biological properties of antimicrobial subtances produced from B. velezensis CP 1604. From liquid culture, substances with activity against Fusarium oxysporum and Xanthomonas oryzae were extracted by means of adsorption with Amberlite XAD-7, extraction from lyophilized powder using ethanol, precipitation at low pH and extraction with organic solvents of 1-butanol and 2-pentanol. The substances were subsequently purified using High Performance Liquid Chromatography (HPLC). Antifungal substance was eluted at 5.328 min while antibacterial substance was observed at 15.313 min. Mass spectrometry analysis showed that antifungal substance was iturin A with molecular weight of 1042 Da and antibacterial substance was macrolactin A with molecular weight of 402 Da. The antifungal substance was stable at high temperature but antibacterial activity was significantly reduced when treated at 100° C for 2 hours. Both substances reduced the activity at low pH but the activities still maintained at high pH. The antimicrobial activities against the fungal and bacterium were not affected when treated with hydrolytic enzymes such as trypsin, α-chymotrypsin, amylase, lipase and proteinase K. Further safety and efficiency investigations of the antifungal and antibacterial substances from B. velezensis CP 1604 on the plant disease control are required in order to seek for the potential application on sustainable agricultural production.

scholarly journals Bacillus velezensis PEA1 Inhibits Fusarium oxysporum Growth and Induces Systemic Resistance to Cucumber Mosaic Virus

Agronomy ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1312 ◽  
Author(s):  
Ahmed Abdelkhalek ◽  
Said I. Behiry ◽  
Abdulaziz A. Al-Askar
Keyword(s):  
Antiviral Activity ◽  
Fusarium Oxysporum ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Plant Pathogens ◽  
Plant Diseases ◽  
Mass Spectrometry Analysis ◽  
Systemic Resistance ◽  
Bacillus Velezensis ◽  
Antiviral Activities

Bacillus velezensis manifests robust biocontrol activity against fungal plant pathogens; however, its antiviral activity has rarely been investigated. Bacillus velezensis strain PEA1 was isolated, characterized, and evaluated for antifungal and antiviral activities against Fusarium oxysporum MT270445 and cucumber mosaic virus (CMV) MN594112. Our findings proved that strain PEA1 had intense antagonist activity against F.oxysporum. Under greenhouse conditions, the antiviral activities (protective, curative, and inactivation) of PEA1-culture filtrate (CF) on Datura stramonium plants were assayed, using a half-leaf method. The inactivation treatment exhibited the highest inhibition rate (97.56%) and the most considerable reduction of CMV-CP accumulation levels (2.1-fold) in PEA1-CF-treated plants when compared with untreated plants (26.9-fold). Furthermore, PEA1-CF induced systemic resistance with significantly elevated transcriptional levels of PAL, CHS, HQT, PR-1, and POD genes in D. stramonium leaves after all treatments. Gas chromatography‒mass spectrometry analysis showed that pyrrolo[1,2-a]pyrazine-1,4-dione is the main compound in the PEA1-CF ethyl acetate extract, which may act as an elicitor molecule that induces plant systemic resistance and inhibits both fungal growth and viral replication. Consequently, B. velezensis can be considered as a potential source for the production of bioactive compounds for the management of plant diseases. To our knowledge, this is the first report of the antiviral activity of B. velezensis against plant viral infection.

scholarly journals Simultaneous Production of Multiple Antimicrobial Compounds by Bacillus velezensis ML122-2 Isolated From Assam Tea Leaf [Camellia sinensis var. assamica (J.W.Mast.) Kitam.]

2021 ◽  
Vol 12 ◽  
Author(s):  
Patthanasak Rungsirivanich ◽  
Elvina Parlindungan ◽  
Paula M. O’Connor ◽  
Des Field ◽  
Jennifer Mahony ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Camellia Sinensis ◽  
Gene Clusters ◽  
Antimicrobial Activities ◽  
Transcriptional Analysis ◽  
Mass Spectrometry Analysis ◽  
Bacillus Velezensis ◽  
Spectrometry Analysis ◽  
Wide Range ◽  
Assam Tea

Bacillus velezensis ML122-2 is an antimicrobial-producing strain isolated from the leaf of Assam tea or Miang [Camellia sinensis var. assamica (J.W.Mast.) Kitam.]. The cell-free supernatant (CFS) of strain ML122-2 exhibits a broad-spectrum antimicrobial activity against various Gram-positive and Gram-negative bacteria as well as the mold Penicillium expansum. The genome of B. velezensis ML122-2 was sequenced and in silico analysis identified three potential bacteriocin-associated gene clusters, that is, those involved in the production of mersacidin, amylocyclicin, and LCI. Furthermore, six gene clusters exhibiting homology (75–100% DNA sequence identity) to those associated with the secondary metabolites bacilysin, bacillibactin, surfactin, macrolactin H, bacillaene, and plipastatin were identified. Individual antimicrobial activities produced by B. velezensis ML122-2 were purified and characterized by Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry analysis, revealing three antimicrobial peptides with molecular masses corresponding to surfactin, plipastatin, and amylocyclicin. Transcriptional analysis of specific genes associated with mersacidin (mrsA), amylocyclicin (acnA), plipastatin (ppsA), and surfactin (srfAA) production by B. velezensis ML122-2 showed that the first was not transcribed under the conditions tested, while the latter three were consistent with the presence of the associated peptides as determined by mass spectrometry analysis. These findings demonstrate that B. velezensis ML122-2 has the genetic capacity to produce a wide range of antimicrobial activities that may support a specific community structure and highlight the biotechnological properties of Assam tea.

scholarly journals Antimicrobial activities of Bacillus velezensis strains isolated from stingless bee products against methicillin-resistant Staphylococcus aureus

PLoS ONE ◽  
2021 ◽  
Vol 16 (5) ◽  
pp. e0251514
Author(s):  
Mohamad Malik Al-adil Baharudin ◽  
Mohamad Syazwan Ngalimat ◽  
Fairolniza Mohd Shariff ◽  
Zetty Norhana Balia Yusof ◽  
Murni Karim ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Peptide ◽  
High Performance ◽  
Pathogenic Bacteria ◽  
Antimicrobial Activities ◽  
Proteinase K ◽  
Tween 20 ◽  
Bacillus Velezensis ◽  
Promising Alternative ◽  
Methicillin Resistant

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have reached epidemic proportions globally. Therefore, there is an urgent need for a continuous supply of antibiotics to combat the problem. In this study, bacteria initially identified as species belonging to the Bacillus amyloliquefaciens operational group were re-identified based on the housekeeping gene, gyrB. Cell-free supernatants (CFS) from the strains were used for antimicrobial tests using the agar well diffusion assay against MRSA and various types of pathogenic bacteria. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and physicochemical characteristics of the CFS were determined. Based on gyrB sequence analysis, five strains (PD9, B7, PU1, BP1 and L9) were identified as Bacillus velezensis. The CFS of all B. velezensis strains showed broad inhibitory activities against Gram-negative and -positive as well as MRSA strains. Strain PD9 against MRSA ATCC 33742 was chosen for further analysis as it showed the biggest zone of inhibition (21.0 ± 0.4 mm). The MIC and MBC values obtained were 125 μl/ml. The crude antimicrobial extract showed bactericidal activity and was stable at various temperatures (40–80°C), pH (4–12), surfactants (Tween 20, Tween 80, SDS and Triton X-100) and metal ions (MgCI2, NaCI2, ZnNO3 and CuSO4) when tested. However, the crude extract was not stable when treated with proteinase K. All these properties resembled the characteristics of peptides. The antimicrobial compound from the selected strain was purified by using solvent extraction method and silica gel column chromatography. The purified compound was subjected to High Performance Liquid Chromatography which resulted in a single peak of the anti-MRSA compound being detected. The molecular weight of the anti-MRSA compound was determined by using SDS-PAGE and zymogram. The size of the purified antimicrobial peptide was approximately ~ 5 kDa. The antimicrobial peptide produced from B. velezensis strain PD9 is a promising alternative to combat the spread of MRSA infections in the future.

scholarly journals Biocontrol of tomato bacterial wilt by a new strain Bacillus velezensis FJAT-46737 and its lipopeptides

2019 ◽  
Author(s):  
Meichun Chen ◽  
Jieping Wang ◽  
Bo Liu ◽  
Yujing Zhu ◽  
Rongfeng Xiao ◽  
...  
Keyword(s):  
Bacterial Wilt ◽  
Plant Pathogens ◽  
Antimicrobial Agents ◽  
Gene Clusters ◽  
Antimicrobial Activities ◽  
Whole Genome Sequence ◽  
Dependent Manner ◽  
Bacillus Velezensis ◽  
Tomato Bacterial Wilt

Abstract Background There is an urgent need to discover alternative antimicrobial agents to control bacterial wilt. The objectives of this study were to report a new lipopeptide-producding biocontrol strain FJAT-46737 and its lipopeptides, and to investigate their antagonistic effects.Results Based on the whole genome sequence analysis, the new strain FJAT-46737 was finally identified as Bacillus velezensis and seven gene clusters that responsible for synthesis of bioactive secondary metabolite in the FJAT-46737 were predicted. Antimicrobial results demonstrated that FJAT-46737 exhibited broad-spectrum antimicrobial activities in vitro against bacteria and fungi. The pot experiments showed that the tomato bacterial wilt control efficiencies of the whole cultures, the 2-fold diluted supernatants and the crude lipopeptide of FJAT-46737 were 66.2%, 82.0%, and 96.2%, respectively. The above results suggested that one of the antagonistic mechanisms of FJAT-46737 was secretion of lipopeptides that consisted of iturins, fengycins and surfactins. The crude lipopeptides had significant antagonistic activities on several pathogen in a dosage-dependent manner, and the fengycins confirmed to play a major role in antibacterial abilities in vitro of the lipopeptides against Ralstonia solanacearum . Furthermore, it was found that the rich organic nitrogen sources (expecially yeast extracts) in the media were benificial for FJAT-46737 to produce fengycin and surfactin. The secretion of these two lipopeptides were also adjusted by the culture temperature: the content of the fengycins decreased by 96.6% and that of the surfactins ascended by 59.9% from 20 o C to 40 o C. And 20 o C~25 o C is the suitable temperature for FJAT-46737 to produce lipopeptide.Conclusions The B. velezensis strain FJAT-46737 and its lipopeptides would used as new sources of potential biocontrol agents against plant pathogens, especially the bacterial wilt pathogen R. solanacearum .

scholarly journals Biocontrol of tomato bacterial wilt by a new strain Bacillus velezensis FJAT-46737 and its lipopeptides

2019 ◽  
Author(s):  
Meichun Chen ◽  
Jieping Wang ◽  
Bo Liu ◽  
Yujing Zhu ◽  
Rongfeng Xiao ◽  
...  
Keyword(s):  
Bacterial Wilt ◽  
Plant Pathogens ◽  
Antimicrobial Agents ◽  
Gene Clusters ◽  
Antimicrobial Activities ◽  
Whole Genome Sequence ◽  
Dependent Manner ◽  
Bacillus Velezensis ◽  
Tomato Bacterial Wilt

Abstract Background: There is an urgent need to discover alternative antimicrobial agents to control bacterial wilt. The objectives of this study were to report a new lipopeptide-producing biocontrol strain FJAT-46737 and its lipopeptides, and to investigate their antagonistic effects. Results: Based on the whole genome sequence analysis, the new strain FJAT-46737 was finally identified as Bacillus velezensis and seven gene clusters that responsible for synthesis of bioactive secondary metabolite in the FJAT-46737 were predicted. Antimicrobial results demonstrated that FJAT-46737 exhibited broad-spectrum antimicrobial activities in vitro against bacteria and fungi . The pot experiments showed that the tomato bacterial wilt control efficiencies of the whole cultures, the 2-fold diluted supernatants and the crude lipopeptide of FJAT-46737 were 66.2%, 82.0%, and 96.2%, respectively. The above results suggested that one of the antagonistic mechanisms of FJAT-46737 was secretion of lipopeptides that consisted of iturins, fengycins and surfactins. The crude lipopeptides had significant antagonistic activities on several pathogen in a dosage-dependent manner, and the fengycins confirmed to play a major role in antibacterial abilities in vitro of the lipopeptides against Ralstonia solanacearum . Furthermore, it was found that the rich organic nitrogen sources (especially yeast extracts) in the media were beneficial for FJAT-46737 to produce fengycin and surfactin. The secretion of these two lipopeptides were also adjusted by the culture temperature: the content of the fengycins decreased by 96.6% and that of the surfactins ascended by 59.9% from 20 o C to 40 o C. The temperature of 20 o C~25 o C is the suitable temperature for FJAT-46737 to produce lipopeptide. Conclusions: The B. velezensis strain FJAT-46737 and its lipopeptides would used as new sources of potential biocontrol agents against plant pathogens, especially the bacterial wilt pathogen R. solanacearum .

scholarly journals Biocontrol of tomato bacterial wilt by the new strain Bacillus velezensis FJAT-46737 and its lipopeptides

2020 ◽  
Author(s):  
Meichun Chen ◽  
Jieping Wang(New Corresponding Author) ◽  
Bo Liu(Former Corresponding Author) ◽  
Yujing Zhu ◽  
Rongfeng Xiao ◽  
...  
Keyword(s):  
Bacterial Wilt ◽  
Plant Pathogens ◽  
Antimicrobial Agents ◽  
Nitrogen Sources ◽  
Gene Clusters ◽  
Antimicrobial Activities ◽  
Whole Genome Sequence ◽  
Bacillus Velezensis ◽  
Tomato Bacterial Wilt

Abstract Background: There is an urgent need to discover alternative antimicrobial agents to control bacterial wilt. This study reports on a new lipopeptide-producing biocontrol strain FJAT-46737 and explores its lipopeptidic compounds, and this study investigates the antagonistic effects of these compounds. Results: Based on a whole genome sequence analysis, the new strain FJAT-46737 was identified as Bacillus velezensis, and seven gene clusters responsible for the synthesis of bioactive secondary metabolites in FJAT-46737 were predicted. The antimicrobial results demonstrated that FJAT-46737 exhibited broad-spectrum antimicrobial activities in vitro against three bacteria and three fungi. Pot experiments showed that the control efficiencies for tomato bacterial wilt of the whole cultures, the 2-fold diluted supernatants and the crude lipopeptide of FJAT-46737 were 66.2%, 82.0%, and 96.2%, respectively. The above results suggested that one of the antagonistic mechanisms of FJAT-46737 was the secretion of lipopeptides consisting of iturins, fengycins and surfactins. The crude lipopeptides had significant antagonistic activities against several pathogens (including Ralstonia solanacearum, Escherichia coli and Fusarium oxysporum) and fengycins were the major antibacterial components of the lipopeptides against R. solanacearum in vitro. Furthermore, the rich organic nitrogen sources (especially yeast extracts) in the media promoted the production of fengycin and surfactin by FJAT-46737. The secretion of these two lipopeptides was related to temperature fluctuations, with the fengycin content decreasing by 96.6% and the surfactins content increasing by 59.9% from 20 oC to 40 oC. The optimal temperature for lipopeptide production by FJAT-46737 varied between 20 oC and 25oC. Conclusions: The B. velezensis strain FJAT-46737 and its lipopeptides represent new sources of potential biocontrol agents against plant pathogens, especially the bacterial wilt pathogen R. solanacearum.

scholarly journals Weissellicin 110, a Newly Discovered Bacteriocin from Weissella cibaria 110, Isolated from Plaa-Som, a Fermented Fish Product from Thailand

2007 ◽  
Vol 73 (7) ◽  
pp. 2247-2250 ◽  
Author(s):  
Sirinat Srionnual ◽  
Fujitoshi Yanagida ◽  
Li-Hsiu Lin ◽  
Kuang-Nan Hsiao ◽  
Yi-sheng Chen
Keyword(s):  
Proteolytic Enzymes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Band ◽  
Proteinase K ◽  
Mass Spectrometry Analysis ◽  
Fermented Fish ◽  
Spectrometry Analysis ◽  
Weissella Cibaria ◽  
Fish Product

ABSTRACT Weissella cibaria 110, isolated from the Thai fermented fish product plaa-som, was found to produce a bacteriocin active against some gram-positive bacteria. Bacteriocin activity was not eliminated by exposure to high temperatures or catalase but was destroyed by exposure to the proteolytic enzymes proteinase K and trypsin. The bacteriocin from W. cibaria 110 was purified, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the purified bacteriocin contained one protein band that was approximately 2.5 kDa in size. Mass spectrometry analysis showed the mass of the peptide to be approximately 3,487.8 Da. N-terminal amino acid sequence analysis was performed, and 27 amino acids were identified. Because it has no similarity to other known bacteriocins, this bacteriocin was defined as a new bacteriocin and termed weissellicin 110.

Degradation of zearalenone in swine feed and feed ingredients by Bacillus subtilis ANSB01G

2014 ◽  
Vol 7 (2) ◽  
pp. 143-151 ◽  
Author(s):  
Y.P. Lei ◽  
L.H. Zhao ◽  
Q.G. Ma ◽  
J.Y. Zhang ◽  
T. Zhou ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Culture Supernatant ◽  
Antimicrobial Activities ◽  
Proteinase K ◽  
Rrna Gene ◽  
Distillers Grains With Solubles ◽  
Dried Distillers Grains ◽  
Cell Extracts ◽  
Gene Sequence Analysis ◽  
And Staphylococcus Aureus

Zearalenone (ZEA) and its derivatives are mycotoxins that can cause oestrogenic effects and impair the reproductive physiology of animals, especially in female swine. Strategies to reduce or eliminate ZEA contamination in foods and feeds are very much needed. Among 36 bacterial isolates obtained from a variety of animal intestinal chyme, mouldy foods and feeds, soils, etc., five isolates demonstrated the ability to reduce more than 50% of ZEA in a liquid medium; ANSB01G isolate taken from normal broiler intestinal chyme reduced ZEA the most, by 88.65%. Using physiological, biochemical, and 16S rRNA gene sequence analysis methods, the ANSB01G isolate was identified as Bacillus subtilis. Under simulated intestinal tract conditions, the ANSB01G B. subtilis isolate degraded 84.58, 66.34 and 83.04% of ZEA in naturally contaminated maize, dried distillers’ grains with solubles, and swine complete feed, respectively. The highest degradation of ZEA occurred when the mycotoxin was co-incubated with the whole bacterial culture, resulting in a reduction of 88.65%, followed by 75.60% using culture supernatant, 26.11% using cell extracts, and 15.06% using viable cells. Treatments consisting of both heating and addition of proteinase K significantly reduced the rate of ZEA degradation in the culture supernatant, indicating that the ZEA degradation might be enzymatic. B. subtilis ANSB01G displayed resistance to simulated gastrointestinal tract environments and antimicrobial activities against several common bacterial pathogens, including Escherichia coli, Salmonella typhimurium and Staphylococcus aureus. These properties of B. subtilis ANSB01G suggest the possibility of its potential to effectively degrade ZEA in feed and to develop functional feed products for livestock industries.

scholarly journals Pathogenicity and Volatile Nematicidal Metabolites from Duddingtonia flagrans against Meloidogyne incognita

2021 ◽  
Vol 9 (11) ◽  
pp. 2268
Author(s):  
Xiaoyu Mei ◽  
Xin Wang ◽  
Guohong Li
Keyword(s):  
Meloidogyne Incognita ◽  
Plant Pathogens ◽  
Plant Root ◽  
Nematicidal Activity ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Egg Mass ◽  
Duddingtonia Flagrans ◽  
Egg Hatching ◽  
Root Knot Nematodes

Plant parasitic nematodes, especially parasitic root-knot nematodes, are one of the most destructive plant pathogens worldwide. The control of plant root-knot nematodes is extremely challenging. Duddingtonia flagrans is a type of nematode-trapping fungi (NTF), which produces three-dimensional adhesive networks to trap nematodes. In this study, the pathogenicity and volatile organic compounds (VOCs) of the NTF D. flagrans against the plant root-knot nematode, Meloidogyne incognita, were investigated. The predatory process of D. flagrans trapping M. incognita was observed using scanning electron microscopy. Gas chromatography-mass spectrometry analysis of the VOCs from D. flagrans led to the identification of 52 metabolites, of which 11 main compounds were tested individually for their activity against M. incognita. Three compounds, cyclohexanamine, cyclohexanone, and cyclohexanol, were toxic to M. incognita. Furthermore, these three VOCs inhibited egg hatching of M. incognita. Cyclohexanamine showed the highest nematicidal activity, which can cause 97.93% mortality of M. incognita at 8.71 µM within 12 h. The number of hatched juveniles per egg mass after 3 days was just 8.44 when treated with 26.14 µM cyclohexanamine. This study is the first to demonstrate the nematicidal activity of VOCs produced by D. flagrans against M. incognita, which indicates that D. flagrans has the potential to biocontrol plant root-knot nematodes.

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