• Title/Summary/Keyword: antifungal metabolite

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Isolation and Characterization of Bacillus sp. Producing Broad-Spectrum Antibiotics Against Human and Plant Pathogenic Fungi

  • Chen, Na;Jin, Min;Qu, Hong-Mei;Chen, Zhi-Qiang;Chen, Zhao-Li;Qiu, Zhi-Gang;Wang, Xin-Wei;Li, Jun-Wen
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.256-263
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    • 2012
  • A strain of bacterium producing antifungal antibiotic was isolated and identification of the strain was attempted. We could identify the bacterium as being a Bacillus sp., based on morphological observation, physiological characteristics, and 16S rDNA sequence analysis, thus leading us to designate the strain as Bacillus sp. AH-E-1. The strain showed potent antibiotic activity against phytopathogenic and human pathogenic fungi by inducing mycelial distortion and swelling and inhibiting spore germination. The antibiotic metabolite produced by the strain demonstrated excellent thermal and pH (2-11) stability, but was labile to autoclaving. From these results, we could find a broader antifungal activity of Bacillus genus. Isolation and characterization of the active agent produced by the strain are under progress.

Production, Purification, and Characterization of Antifungal Metabolite from Pseudomonas aeruginosa SD12, a New Strain Obtained from Tannery Waste Polluted Soil

  • Dharni, Seema;Alam, Mansoor;Kalani, Komal;Abdul-Khaliq, Abdul-Khaliq;Samad, Abdul;Srivastava, Santosh Kumar;Patra, Dharani Dhar
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.674-683
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    • 2012
  • A new strain, SD12, was isolated from tannery waste polluted soil and identified as Pseudomonas aeruginosa on the basis of phenotypic traits and by comparison of 16S rRNA sequences. This bacterium exhibited broad-spectrum antagonistic activity against phytopathogenic fungi. The strain produced phosphatases, cellulases, proteases, pectinases, and HCN and also retained its ability to produce hydroxamate-type siderophore. A bioactive metabolite was isolated from P. aeruginosa SD12 and was characterized as 1-hydroxyphenazine ((1-OH-PHZ) by nuclear magnetic resonance (NMR) spectral analysis. The strain was used as a biocontrol agent against root rot and wilt disease of pyrethrum caused by Rhizoctonia solani. The stain is also reported to increase the growth and biomass of Plantago ovata. The purified compound, 1-hydroxyphenazine, also showed broad-spectrum antagonistic activity towards a range of phytopathogenic fungi, which is the first report of its kind.

Effect of Antibiotic Down-Regulatory Gene wblA Ortholog on Antifungal Polyene Production in Rare Actinomycetes Pseudonocardia autotrophica

  • Kim, Hye-Jin;Kim, Min-Kyung;Kim, Young-Woo;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1226-1231
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    • 2014
  • The rare actinomycete Pseudonocardia autotrophica was previously shown to produce a solubility-improved toxicity-reduced novel polyene compound named $\underline{N}ystatin$-like $\underline{P}seudonocardia$ $\underline{P}olyene$ (NPP). The low productivity of NPP in P. autotrophica implies that its biosynthetic pathway is tightly regulated. In this study, $wblA_{pau}$ was isolated and identified as a novel negative regulatory gene for NPP production in P. autotrophica, which showed approximately 49% amino acid identity with a global antibiotic down-regulatory gene, wblA, identified from various Streptomycetes species. Although no significant difference in NPP production was observed between P. autotrophica harboring empty vector and the S. coelicolor wblA under its native promoter, approximately 12% less NPP was produced in P. autotrophica expressing the wblA gene under the strong constitutive $ermE^*$ promoter. Furthermore, disruption of the $wblA_{pau}$ gene from P. autotrophica resulted in an approximately 80% increase in NPP productivity. These results strongly suggest that identification and inactivation of the global antibiotic down-regulatory gene wblA ortholog are a critical strategy for improving secondary metabolite overproduction in not only Streptomyces but also non-Streptomyces rare actinomycete species.

Investigation and utilization of unique natural products from endemic tree species in Taiwan

  • Chu, Fang-Hua
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.23-23
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    • 2018
  • Taiwan, formerly known as Formosa, located on tropical and subtropical climate zones with abundant biological resources. According to the latest version of the Flora of Taiwan, there are 4339 species of vascular plants including 1054 endemic species. First, Taiwania (Taiwania cryptomerioides), named after its native island of Taiwan, have been isolated more than 500 secondary metabolites, including lignans, terpenoids, steroids, and flavonoids. Several of the metabolites are reported to have antibacterial, antifungal, antimite, antitermite and antitumor activities. In order to investigate plant secondary metabolic diversity toward industrial applications, we established deep transcriptome resources for non-model plants and fungi to produce terpenoid metabolites of economic importance. Second, many plants of Lauraceae have been utilized in folk medicine for their exciting bioactivities. The twigs and leaves from 27 tree species of Lauraceae grown in Taiwan were performed to evaluate potential bioactivity. The leaves of Cinnamomum osmophloeum are traditionally used in folk medicines, and many biological activities have been identified, such as antibacterial, antifungal, antitermite, antidiabetic, antihyperuricemia, antiinflammatory, and antioxidant activities. However, C. osmophloeum has nine chemotypes with various secondary metabolite profiles. In order to efficiently produce active compounds, we established the genetic markers to identify the chemotype plants. Finally, Cinnamomum kanehirae is the host of the medicinal mushroom Antrodia cinnamomea. Several in vivo and in vitro studies indicated that A. cinnamomea possesses a diverse range of biological activities. Because of the potential pharmacological application, we established the transformation system to enhance the triterpenoid contents production.

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Proteomic Reference Map and Comparative Analysis between Streptomyces griseus S4-7 and wbiE2 Transcription Factor-Mutant Strain

  • Kim, Jisu;Kwon, Young Sang;Bae, Dong-Won;Kwak, Youn-Sig
    • The Plant Pathology Journal
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    • v.36 no.2
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    • pp.185-191
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    • 2020
  • Streptomyces griseus S4-7, a well-characterized keystone taxon among strawberry microbial communities, shows exceptional disease-preventing ability. The whole-genome sequence, functional genes, and bioactive secondary metabolites of the strain have been described in previous studies. However, proteomics studies of not only the S4-7 strain, but also the Streptomyces genus as a whole, remain limited to date. Therefore, in the present study, we created a proteomics reference map for S. griseus S4-7. Additionally, analysis of differentially expressed proteins was performed against a wblE2 mutant, which was deficient in spore chain development and did not express an antifungal activity-regulatory transcription factor. We believe that our data provide a foundation for further in-depth studies of functional keystone taxa of the phytobiome and elucidation of the mechanisms underlying plant-microbe interactions, especially those involving the Streptomyces genus.

Genomics and LC-MS Reveal Diverse Active Secondary Metabolites in Bacillus amyloliquefaciens WS-8

  • Liu, Hongwei;Wang, Yana;Yang, Qingxia;Zhao, Wenya;Cui, Liting;Wang, Buqing;Zhang, Liping;Cheng, Huicai;Song, Shuishan;Zhang, Liping
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.417-426
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    • 2020
  • Bacillus amyloliquefaciens is an important plant disease-preventing and growth-promoting microorganism. B. amyloliquefaciens WS-8 can stimulate plant growth and has strong antifungal properties. In this study, we sequenced the complete genome of B. amyloliquefaciens WS-8 by Pacific Biosciences RSII (PacBio) Single Molecule Real-Time (SMRT) sequencing. The genome consists of one chromosome (3,929,787 bp) and no additional plasmids. The main bacteriostatic substances were determined by genome, transcriptome, and mass spectrometry data. We thereby laid a theoretical foundation for the utilization of the strain. By genomic analysis, we identified 19 putative biosynthetic gene clusters for secondary metabolites, most of which are potentially involved in the biosynthesis of numerous bioactive metabolites, including difficidin, fengycin, and surfactin. Furthermore, a potential class II lanthipeptide biosynthetic gene cluster and genes that are involved in auxin biosynthesis were found. Through the analysis of transcriptome data, we found that the key bacteriostatic genes, as predicted in the genome, exhibited different levels of mRNA expression. Through metabolite isolation, purification, and exposure experiments, we found that a variety of metabolites of WS-8 exert an inhibitory effect on the necrotrophic fungus Botrytis cinerea, which causes gray mold; by mass spectrometry, we found that the main substances are mainly iturins and fengycins. Therefore, this strain has the potential to be utilized as an antifungal agent in agriculture.

Diffusible and Volatile Antifungal Compounds Produced by an Antagonistic Bacillus velezensis G341 against Various Phytopathogenic Fungi

  • Lim, Seong Mi;Yoon, Mi-Young;Choi, Gyung Ja;Choi, Yong Ho;Jang, Kyoung Soo;Shin, Teak Soo;Park, Hae Woong;Yu, Nan Hee;Kim, Young Ho;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.33 no.5
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    • pp.488-498
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    • 2017
  • The aim of this study was to identify volatile and agardiffusible antifungal metabolites produced by Bacillus sp. G341 with strong antifungal activity against various phytopathogenic fungi. Strain G341 isolated from four-year-old roots of Korean ginseng with rot symptoms was identified as Bacillus velezensis based on 16S rDNA and gyrA sequences. Strain G341 inhibited mycelial growth of all phytopathogenic fungi tested. In vivo experiment results revealed that n-butanol extract of fermentation broth effectively controlled the development of rice sheath blight, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew, and red pepper anthracnose. Two antifungal compounds were isolated from strain G341 and identified as bacillomycin L and fengycin A by MS/MS analysis. Moreover, volatile compounds emitted from strain G341 were found to be able to inhibit mycelial growth of various phytopathogenic fungi. Based on volatile compound profiles of strain G341 obtained through headspace collection and analysis on GC-MS, dimethylsulfoxide, 1-butanol, and 3-hydroxy-2-butanone (acetoin) were identified. Taken together, these results suggest that B. valezensis G341 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

Phylogenic Analysis of Alternaria brassicicola Producing Bioactive Metabolites

  • Jung, Dong-Sun;Na, Yeo-Jung;Ryu, Ki-Hyun
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.289-294
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    • 2002
  • The fungal strain SW-3 having antimicrobial activity was isolated from soil of crucified plants in Pocheon, Kyungki-Do, Korea. Strain SW-3 was identified as Alternaria brassicicola by its morphological characteristics, and confirmed by the analysis of the 18S gene and ITS regions of rDNA. The fungus showed a similarity of 99% with Alternaria brassicicola in the 18S rDNA sequence analysis. A. brassicicola has been reported to produce an antitumor compound, called depudecin. We found that strain SW-3 produced antimicrobial metabolites, in addition to depudecin, during sporulation under different growth conditions. The metabolite of the isolated fungus was found to have strong antifungal activity against Microsporium canis and Trichophyton rubrum, and antibacterial activity against Staphylococcus aureus and Pseudomonas aerogenes. The amount and kind of metabolites produced by the isolate were affected by growth conditions such as nutrients and growth periods.

Isolation and Characterization of Actinomycete Strain BK185 Possessing Antifungal Activity against Ginseng Root Rot Pathogens (인삼 뿌리썩음병균에 항균활성이 있는 방선균 BK185의 분리 및 특성)

  • Kim, Byung-Yong;Bae, Mun-Hyung;Ahn, Jae-Hyung;Weon, Hang-Yeon;Kim, Sung-Il;Kim, Wan-Kyu;Oh, Dong-Chan;Song, Jaekyeong
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.396-403
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    • 2014
  • Ginseng (Panax ginseng C. A. Meyer) is an economically valuable pharmaceutical crop in Korea. In order to find promising biocontrol agents for soil-borne fungal pathogens which infect ginseng roots, we have isolated actinomycete, BK185 from soil. The isolate was investigated for the antifungal activity against to ginseng rot pathogens prior to testing genetic and chemical properties. The strain was identified as Streptomyces sp. using phylogenetic analysis based on 16S rRNA gene sequence. The most closely related species was S. sporoclivatus and S. geldanamycininus with high similarities (>99%). The isolate, BK185 showed positive reaction for PCR detection targeting biosynthetic gene clusters of PKS (Type-I polyketide synthase) and NRPS (Non-ribosomal polypeptide synthetase) genes. Major metabolite from the BK185 was analyzed by The LC/MS and identified to geldamycin, which was known to contained broad antibacterial, antifungal or anticancer activities. The results provide evidences that the strain, BK185 can be promising biocontrol agent for ginseng organic farming.

In vitro Evaluation of the Mechanism of Antagonism and Phosphate Solubilization by the Insect Gut Bacteria Pseudomonas sp. PRGB06 that Exhibits Plant Growth Promotion and Bio-Fertilizing Traits (배추좀나방 내장에서 분리한 식물생장촉진미생물 Pseudomonas sp. PRGB06의 길항기작과 인산가용화의 기내 평가)

  • Kim, Kyoung-A;Indiragandhi, P.;Anandham, R.;Palaniappan, P.;Trivedi, P.;Madhaiyan, M.;Han, Gwang-Hyun;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.1
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    • pp.18-25
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    • 2008
  • Pseudomonas sp. PRGB06, a bacterial strain isolated from diamondback moth (Plutella xylostella) gut, was examined for its plant growth promotion and biofertilizing traits. The bacteria growth was observed under various conditions of carbon sources, temperature, pH and salt concentrations. In addition, the mechanisms of antagonism and phosphate solubilization were investigated. The bacterial strain PRGB06, grew well using most of the tested carbon sources. The best growth was observed at $30^{\circ}C$ and pH 7. The inhibition of the pathogenic fungi was likely due to the volatile antifungal metabolite and ammonia gas produced by the bacteria. A significant positive relationship was found between the phosphate solubilization and acid production. When inoculated with PRGB06 in vitro and in gnotobiotic condition, red pepper and maize showed increase in root length, seedling vigor and dry bio-mass.