• Title/Summary/Keyword: Antifungal lipopeptide

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Biocontrol of Peach Gummosis by Bacillus velezensis KTA01 and Its Antifungal Mechanism

  • Tae-An Kang;GyuDae Lee;Kihwan Kim;Dongyup Hahn;Jae-Ho Shin;Won-Chan Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.296-305
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    • 2024
  • Peach tree gummosis is a botanical anomaly distinguished by the secretion of dark-brown gum from the shoots of peach trees, and Botryosphaeria dothidea has been identified as one of the fungal species responsible for its occurrence. In South Korea, approximately 80% of gummosis cases are linked to infections caused by B. dothidea. In this study, we isolated microbes from the soil surrounding peach trees exhibiting antifungal activity against B. dothidea. Subsequently, we identified several bacterial strains as potential candidates for a biocontrol agent. Among them, Bacillus velezensis KTA01 displayed the most robust antifungal activity and was therefore selected for further analysis. To investigate the antifungal mechanism of B. velezensis KTA01, we performed tests to assess cell wall degradation and siderophore production. Additionally, we conducted reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis based on whole-genome sequencing to confirm the presence of genes responsible for the biosynthesis of lipopeptide compounds, a well-known characteristic of Bacillus spp., and to compare gene expression levels. Moreover, we extracted lipopeptide compounds using methanol and subjected them to both antifungal activity testing and high-performance liquid chromatography (HPLC) analysis. The experimental findings presented in this study unequivocally demonstrate the promising potential of B. velezensis KTA01 as a biocontrol agent against B. dothidea KACC45481, the pathogen responsible for causing peach tree gummosis.

Assessment of the Contribution of Antagonistic Secondary Metabolites to the Antifungal and Biocontrol Activities of Pseudomonas fluorescens NBC275

  • Dutta, Swarnalee;Yu, Sang-Mi;Lee, Yong Hoon
    • The Plant Pathology Journal
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    • v.36 no.5
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    • pp.491-496
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    • 2020
  • An understanding of the contribution of secondary metabolites (SMs) to the antagonistic and biocontrol activities of bacterial biocontrol agents serves to improve biocontrol potential of the strain. In this study, to evaluate the contribution of each SM produced by Pseudomonas fluorescens NBC275 (Pf275) to its antifungal and biocontrol activity, we combined in silico analysis of the genome with our previous study of transposon (Tn) mutants. Thirteen Tn mutants, which belonged to 6 biosynthetic gene clusters (BGCs) of a total 14 BGCs predicted by the antiSMASH tool were identified by the reduction of antifungal activity. The biocontrol performance of Pf275 was significantly dependent on 2,4-diacetylphloroglucinol and pyoverdine. The clusters that encode for arylpolyene and an unidentified small linear lipopeptide influenced antifungal and biocontrol activities. To our knowledge, our study identified the contribution of SMs, such as a small linear lipopeptide and arylpolyene, to biocontrol efficacy for the first time.

Biosurfactant as a microbial pesticide

  • Lee, Baek-Seok;Choi, Sung-Won;Choi, Ki-Hyun;Lee, Jae-Ho;Kim, Eun-Ki
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.40-44
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    • 2003
  • Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

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Isolation and Characterization of Three Kinds of Lipopeptides Produced by Bacillus subtilis JKK238 from Jeot-Kal of Korean Traditional Fermented Fishes (한국 전통젓갈에서 분리한 Bacillus subtilis JKK238 균주 유래 세 종류 Lipopeptide의 분리 및 특성)

  • Yoon Sang-Hong;Kim Jung-Bong;Lim Yoong-Ho;Hong Seong-Ryeul;Song Jae-Kyeung;Kim Sam-Sun;Kwon Soon-Wo;Park In-Cheol;Kim Soo-Jin;Yeo Yun-Soo;Koo Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.295-301
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    • 2005
  • About seven hundred bacterial strains were collected from Jeot-Kal, a Korean traditional fermented fishes, in various Korean districts. One of the strains designated JKK238 has its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The JKK238 strain was isolated from Oh-Jeot, a kind of fermented shrimps, of Kangkyeung in Korea, and was identified as Bacillus subtilis based on its physiological characteristics, fatty acids compositions of cellular wall, and 16S rDNA sequence analysis. We isolated simply antimicrobial lipopeptides (AMLP) by $25\%$ ammonium sulfate precipitation of 3 days-old tryptic soy broth cultures of the JKK238 strain. Further analysis of AMLP revealed that B. subtilis JKK238 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families simultaneously. Above results indicate that the JKK238 strain can be added to the limited number B. subtilis strains reported to co-produce the three kinds of lipopeptide families.

Exogenous Indole Regulates Lipopeptide Biosynthesis in Antarctic Bacillus amyloliquefaciens Pc3

  • Ding, Lianshuai;Zhang, Song;Guo, Wenbin;Chen, Xinhua
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.784-795
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    • 2018
  • Bacillus amyloliquefaciens Pc3 was isolated from Antarctic seawater with antifungal activity. In order to investigate the metabolic regulation mechanism in the biosynthesis of lipopeptides in B. amyloliquefaciens Pc3, GC/MS-based metabolomics was used when exogenous indole was added. The intracellular metabolite profiles showed decreased asparagine, aspartic acid, glutamine, glutamic acid, threonine, valine, isoleucine, hexadecanoic acid, and octadecanoic acid in the indole-treated groups, which were involved in the biosynthesis of lipopeptides. B. amyloliquefaciens Pc3 exhibited a growth promotion, bacterial total protein increase, and lipopeptide biosynthesis inhibition upon the addition of indole. Besides this, real-time PCR analysis further revealed that the transcription of lipopeptide biosynthesis genes ituD, fenA, and srfA-A were downregulated by indole with 22.4-, 21.98-, and 26.0-fold, respectively. It therefore was speculated that as the metabolic flux of most of the amino acids and fatty acids were transferred to the synthesis of proteins and biomass, lipopeptide biosynthesis was weakened owing to the lack of precursor amino acids and fatty acids.

Antifungal Compound Produced by Bacillus sp. TBM912 (Bacillus sp. TBM912가 생산하는 항균물질)

  • 주우홍;한수지;최용락;정영기
    • Journal of Life Science
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    • v.14 no.1
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    • pp.193-197
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    • 2004
  • A continuous enrichment culture procedure was used to isolate bacteria from various soil sources capable of suppressing large patch disease of turfgrass. Six isolates consistently suppressed large patch in turfgrass, and ranged in the spectrum of extracellular enzymes that they expressed. The best disease- suppressing isolate, TBM912, expressed protease, CMCase, and pectinase activity and inhibited the growth of Rhizectonin solani and Betrytis cinerea in vitro. Here we show that this strain also produces an antibiotic that was identified by TLC, SDS-PACE and HPLC analysis as lipopeptide.

Postharvest Biological Control of Colletotrichum acutatum on Apple by Bacillus subtilis HM1 and the Structural Identification of Antagonists

  • Kim, Hae-Min;Lee, Kui-Jae;Chae, Jong-Chan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1954-1959
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    • 2015
  • Bacillus subtilis HM1 was isolated from the rhizosphere region of halophytes for its antifungal activity against Colletotrichum acutatum, the causative agent of anthracnose. Treatment of postharvest apples with the cell culture or with a cell-free culture supernatant reduced disease severity 80.7% and 69.4%, respectively. Both treatments also exhibited antifungal activity against various phytopathogenic fungi in vitro. The antifungal substances were purified and analyzed by acid precipitation, gel filtration, high-performance liquid chromatography, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Three compounds were identified as fengycin, iturin, and surfactin. The MALDI-TOF/TOF mass spectrum revealed the presence of cyclized fengycin homologs A and B, which were distinguishable on the basis of the presence of either alanine or valine, respectively, at position 6 of the peptide sequence. In addition, the cyclized structure of fengycin was shown to play a critical role in antifungal activity.

Production of Antifungal Lipopeptide Iturin by Bacillus subtilis (Bacillus subtilis로부터 항진균 리포펩타이드 물질 Iturin의 생산)

  • 손광현;이항우
    • KSBB Journal
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    • v.9 no.2
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    • pp.224-229
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    • 1994
  • Iturin, an antifungal lipopeptide, fermentation by Bacillus subtilis was investigated focusing on the effeats of nutrients aeration and specific cell growth rate on iturin production. Cell growth and product formation were not affected by different kinds of carbon sources such as sucrose, glucose and fructose. Soytone concentration above 20g/$\ell$ did not influence iturin production. Diauxic growth pattern appeared when only soytone was used as a sole nitrogen source probably due to the shortage of amino acids and/or peptides in soytone which could be favorably assimilated by the cells. The composition of three major components in iturin was not changed significantly by the variation of dissolved oxygen concentration of the culture broth but changed substantially by the change of specific growth rate of the cells.

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Taxonomy, Purification and Physicochemical Properties of Novel Antifungal Antibiotics AF-011A (신규 항진균 물질 AF-011A의 생산균주 동정, 정제 및 물리 화학적 특성)

  • 김성호;현봉철;서정우;김창완;연창석;이덕근;김광표;정재경;임융호
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.556-563
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    • 1993
  • AF-011A is a novel lipopeptide with potent antifungal activity isolated from Pseudomonas cepacia AF6008 deposited as KFCC 10759. The compound was isolated from the fermentation broth by extraction with 50% isopropyl alcohol. Purification was effected by chromatography on Diaion HP-20, Alumina and C18 followed by HPTLC on silica gel. These techniques affored two closelt related compounds. AF-011A1 ans AF-011A2. The molecular weights of AF-011A1/A2 were determined by fast atom bombardment mass spectrometry(A1 m/z 1,215 : A2 m/z 1,199).

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