• Title/Summary/Keyword: antagonistic strain

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Probiotic Properties of Lactobacillus brevis KU200019 and Synergistic Activity with Fructooligosaccharides in Antagonistic Activity against Foodborne Pathogens

  • Kariyawasam, Kariyawasam Majuwana Gamage Menaka Menike;Yang, Seo Jin;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.40 no.2
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    • pp.297-310
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    • 2020
  • This study aims to evaluate the probiotic properties of Lactobacillus brevis (L. brevis) KU200019 and the synergistic activity with prebiotics on antimicrobial activity, and the potential application as an adjunct culture in fermented dairy products. The commercial strain, L. brevis ATCC 14869 was used as reference strain. L. brevis KU200019 was showed higher viability in simulated gastric (99.38±0.21%) and bile (115.10±0.13%) conditions compared to reference strain. L. brevis KU200019 exhibited antimicrobial activity against various foodborne pathogens. The supplementation of fructooligosaccharides (FOS) enhanced viability of lactic acid bacteria (>8 Log CFU/mL) and antioxidant activity [2,2-diphenyl-2-picrylhydrazyl radical assay (DPPH) assay, 31.23±1.14%; 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assay, 38.82±1.46%] in fermented skim milk during refrigerated storage. L. brevis KU200019 was distinguished from the reference strain by its higher probiotic potential, antimicrobial activity, and higher antioxidant activity in fermented milk. Therefore, L. brevis KU200019 with FOS was demonstrated promising properties for further application in fermented dairy products with enhanced safety and quality.

Production, Purification and Antifungal Activity of Antibiotic Substances Produced by Pseudomonas aeruginosa Strain B5

  • Kim, Beom-Seok
    • Journal of Microbiology and Biotechnology
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    • v.3 no.1
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    • pp.12-18
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    • 1993
  • Pseudomonas aeruginosa strain B5 with antagonistic activity against Phytophthora capsici and Magnaporthe grisea, was isolated from pepper-growing soil. From the culture of P. aeruginosa strain B5 grown on King's medium B, antibiotic substances were purified using XAD-2 column chromatography. XAD-2 eluates inhibited not only the mycelial growth of P. capsid and M. grisea, but also the development of Phytophthora blight on pepper plants. The crude antibiotic substances were further purified by using silica gel column chromatography, Sephadex LH-20 column chromatography, thin layer chromatography on silica gel plates, and high performance liquid chromatography. Silica gel column chromatogrphy gave good separation of the four antibiotic substances. The pure antibiotics P1, P2, and P3 finally purified by preparative HPLC inhibited the mycelial growth of P. capsici, at concentrations from 7 to 10 $\mu g/ml$. Only P1 and P2 had antifungal activity against M. grisea at 8 $\mu g/ml$. P1 and P3 were highly inhibitory to the mycelial growth of Botryosphaeria dothidea and Botrytis cinerea at relatively low concentrations. However, the three antibiotics had no antifungal activity against Rhizoctonia solani. The chemical structures of these antibiotics are being identified.

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Characteristics of Bacteriocin Produced by a Lactobacillus plantarum Strain Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum 균주가 생산하는 박테리오신의 특성)

  • Chung, Jae Hyuk;Bae, Yun-Sook;Kim, Yeon-Joo;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.481-485
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    • 2010
  • Lactobacillus plantarum strain KK3 with tannase activity was isolated from Gochunipkimchi (red pepper leaf kimchi) and showed a high antagonistic activity against five kinds of food pathogens. Strain KK3 secreted antibacterial compound into culture medium and 24-h culture in MRS broth at $30^{\circ}C$ was enough for the antibacterial compound production. The crude antibacterial compound prepared from culture supernatant inhibited the growth of some Gram-negative bacteria and Bacillus cereus but not Listeria monocytogenes. The antibacterial activity was sensitive to proteinase K treatment, confirming its proteinaceous nature (bacteriocin). The crude bacteriocin was active in the pH range 3.5-8.5 and extremely stable after 15 min of heat treatment at $121^{\circ}C$. The strain KK3 produced equally active bacteriocin in Chinese cabbage juice as it produced in MRS broth.

Identification of the Oligotrophic Bacteria Strain 7F Biocontrolling Phytophthora Blight Disease of Red-pepper (고추 역병 방제를 위한 저영양 길항세균 7F 균주의 동정)

  • Kim, Dong-Gwan;Yeo, Yun-Soo;Kwon, Soon-Wo;Jang, Kil-Su;Lee, Chang-Muk;Lee, Mi-Hye;Kim, Soo-Jin;Koo, Bon-Sung;Yoon, Sang-Hong
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.41-47
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    • 2010
  • A total of 10,753 oligotrophic bacteria were isolated from the cultivated soils of red-pepper infected by Phytophthora blight disease in various regions of Korea (Chungju, Anmyon, Taean, Andong, Eumsung and Goesan). Seven bacteria isolates among these collected resources were selected by the first screening of in vitro antagonistic assay against major several plant pathogenic fungi including Phytophthora capsici. Finally, strain 7F was selected by pot assay for a possible biological control agent against Phytophthora blight disease of pepper seedling in the greenhouse. Strain 7F was identified as Bacillus subtilis on the basis of its 16S rDNA sequence analysis and as standardized biochemical characteristics assay kits such as API20 NE. In the experiment of P. capsici zoospore infected red-pepper on the pot test, infection rate of red-pepper with nonetreatment to Phytophthora blight disease was 87%, while the rate was only 6% in the pot treated with strain 7F. This result indicated that the Bacillus subtilis strain 7F will be useful as a potential biocontrol agent for Phytophthora blight disease of red-pepper.

Characterization of Bacillus luciferensis Strain KJ2C12 from Pepper Root, a Biocontrol Agent of Phytophthora Blight of Pepper

  • Kim, Hye-Sook;Sang, Mee-Kyung;Myung, Inn-Shik;Chun, Se-Chul;Kim, Ki-Deok
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.62-69
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    • 2009
  • In this study, we characterized the bacterial strain KJ2C12 in relation with its biocontrol activity against Phytophthora capsici on pepper, and identified this strain using morphological, physiological, biochemical, fatty acid methyl ester, and 16S rRNA gene sequence analyses. Strain KJ2C12 significantly (P=0.05) reduced both final disease severity and areas under the disease progress curves of 5-week-old pepper plants inoculated with P. capsici compared to buffer-treated controls. As for the production of antibiotics, biofilms, biosurfactant, extracellular enzyme, HCN, and swarming activity, strain KJ2C12 produced an extracellular enzyme with protease activity, but no other productions or swarming activity. However, Escherichia coli produced weak biofilm only. Strain KJ2C12 could colonize pepper roots more effectively in a gnotobiotic system using sterile quartz sand compared to E. coli over 4 weeks after treatments. However, no bacterial populations were detected in 10 mM $MgSO_4$ buffer-treated controls. Strain KJ2C12 produced significantly higher microbial activity than the $MgSO_4$-treated control or E. coli over 4 weeks after treatments. Bacterial strain KJ2C12 was identified as Bacillus luciferensis based on morphological, physiological, and biochemical characteristics as well as FAME and 16S rRNA gene sequence analyses. In addition, these results suggested that B. luciferensis strain KJ2C12 could reduce Phytophthora blight of pepper by protecting infection courts through enhanced effective root colonization with protease production and an increase of soil microbial activity.

The Biocontrol Activity of Chromobacterium sp. Strain C-61 against Rhizoctonia solani Depends on the Productive Ability of Chitinase

  • Park, Seur-Kee;Lee, Myung-Chul;Harman, Gary E.
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.275-282
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    • 2005
  • A chitinolytic bacterium, Chromobacterium sp. strain C-61, was found strongly antagonistic to Rhizoctonia solani, a causal agent of damping-off of eggplant. In this study, the biocontrol activity and enzymatic characteristics of strain C-61 were compared with its four Tn5 insertion mutants (C61-A, -B, -C, and -D) that had lower chitinolytic ability. The chitinase activity of a 2-day old culture was about $76\%,\;49\%\;and\;6\%$ level in C61-A, C61-B and in C61-C, respectively, compared with that of strain C-61. The $\beta-N-acetylhexosaminidase$(Nahase) activity was little detected in strain C-61 but increased largely in C-61A, C61-B and C61-C. Activities of chitinase and Nahase appeared to be negatively correlated in these strains. Another mutant, C-61D, produced no detectable extracellular chitinase and Nahase. The in vitro and in vivo biocontrol activities of strain C-61 and its mutants were closely related to their ability to produce chitinase but not Nahase. No significant differences in population densities between strain C-61 and its mutants were observed in soil around eggplant roots. The results of SDS-PAGE and isoelectrofocusing showed that a major chitinase of strain C-61 is 54-kDa with pI of approximately 8.5. This study provides evidence that the biocontrol activity of Chromobacterium sp. strain C-61 against Rhizoctonia solani depends on the ability to produce chitinase with molecular weight of 54-kDa and pI of 8.5.

Purification and Structural Analysis of Surfactin Produced by Endophytic Bacillus subtilis EBS05 and its Antagonistic Activity Against Rhizoctonia cerealis

  • Wen, Cai-Yi;Yin, Zhi-Gang;Wang, Kai-Xuan;Chen, Jian-Guang;Shen, Shun-Shan
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.342-348
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    • 2011
  • Bacillus subtilis EBS05, an endophytic bacteria strain isolated from a medicinal plant Cinnamomum camphor, can produce antagonistic compounds that effectively inhibit plant pathogenic fungi. The greenhouse experiments showed that wheat sharp eyespot disease (WSED) was reduced by 91.2%, 88.2% and 43.0% after the treatment with fermentation broth, bacteria-free filter and a fungicide fludioxonil, respectively. The culture broth of strain EBS05 can more effectively control WSED than can fludioxonil. The fermentation broth and bacteria-free filter ability to suppress WSED was not significantly different, suggesting that an active secreted substance played a major role in controlling WSED. Separation and purification of the active compounds was carried out by serial processes, including hydrochloric acid (pH 2.0) treatment, methanol extraction and Sephadex LH-20 column chromatography, silica gel column chromatography and reverse-phase high-pressure liquid chromatography (HPLC), respectively. The purified compounds, one of active peaks in the HPLC spectrum, were obtained from the collection. Analysis of the chemical structures by time-of-flight mass spectrometry (TOF-MS) and electrospray ionization mass spectrometry/mass spectrometry (ESI-MS/MS) showed that the active substances produced by the endophytic bacteria EBS05 are mixture of the ${\beta}$-hydroxy-C12~C15-$Leu^7$ surfactin A isomers with 1035.65 Da, 1021.64 Da, 1007.63 Da and 993.65 Da molecular weights, respectively.

Application of Rhizobacteria for Plant Growth Promotion Effect and Biocontrol of Anthracnose Caused by Colletotrichum acutatum on Pepper

  • Lamsal, Kabir;Kim, Sang Woo;Kim, Yun Seok;Lee, Youn Su
    • Mycobiology
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    • v.40 no.4
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    • pp.244-251
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    • 2012
  • In vitro and greenhouse screening of seven rhizobacterial isolates, AB05, AB10, AB11, AB12, AB14, AB15 and AB17, was conducted to investigate the plant growth promoting activities and inhibition against anthracnose caused by Colletotrichum acutatum in pepper. According to identification based on 16S rDNA sequencing, the majority of the isolates are members of Bacillus and a single isolate belongs to the genus Paenibacillus. All seven bacterial isolates were capable of inhibiting C. acutatum to various degrees. The results primarily showed that antibiotic substances produced by the selected bacteria were effective and resulted in strong antifungal activity against the fungi. However, isolate AB15 was the most effective bacterial strain, with the potential to suppress more than 50% mycelial growth of C. acutatum in vitro. Moreover, antibiotics from Paenibacillus polymyxa (AB15) and volatile compounds from Bacillus subtilis (AB14) exerted efficient antagonistic activity against the pathogens in a dual culture assay. In vivo suppression activity of selected bacteria was also analyzed in a greenhouse with the reference to their prominent in vitro antagonism efficacy. Induced systemic resistance in pepper against C. acutatum was also observed under greenhouse conditions. Where, isolate AB15 was found to be the most effective bacterial strain at suppressing pepper anthracnose under greenhouse conditions. Moreover, four isolates, AB10, AB12, AB15, and AB17, were identified as the most effective growth promoting bacteria under greenhouse conditions, with AB17 inducing the greatest enhancement of pepper growth.

Increased Antifungal Activity with Genetic Development of Antagonistic Pseudomonas stutzeri YPL-1 against Fusariym solani (식물근부균 Fusarium Solani에 길항하는 생물방제균 Pseudomonas stutzeri YPL-1의 유전공학적 개발)

  • 임호성;김상달
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.437-441
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    • 1990
  • For the genetic development of more powerful antagonistic Pseudomom - YPL-1 as a biocontxol agent against soilborne plant pathogenic Fuaarium solani causing root rot of many important crops, mutants improving the productivity of chitinase were obtained by mutation with UV radiation or NTG treatment, P. stutzeri YPL-M26 (UV mutant) and P. stutzeri YPL-MI78 (NTG mutant) could improve the productivity of chitinase by 2.5 and 2.0 times, and its antifungal activity by 1.7 and 1.5 times, respectively. The antifungal mechanism of P. stutzeri YPL-M26 was caused by lysis of the fungal cell wall by hydrolytic enzymes such as chitinase. The antifungal activity of crude chitinase of P. stutzeri YPLM26 on the mycelial growth of F. solani was observed to be much higher than that of the original strain. The enzymes produced by P. stutzeri YPL-M26 were the same as the original strain in enzymatic properties such as optimal pH and temperature.

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Culture Optimization for Bacillus lentimorbus G-74 by Using a Miscanthus purpurascens Juice Medium (억새즙액 배지를 이용한 Bacillus lentimorbus G-74 균주의 배양 최적화)

  • Kang, Sun-Chul;Seo, Hae-Jeong
    • Korean Journal of Environmental Agriculture
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    • v.23 no.1
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    • pp.68-74
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    • 2004
  • Miscanthus purpurascens Juice containing potassium (37,952 mg/L), nitrogen (14,000 mg/L), phosphorus (6,800 mg/L), magnesium (5,969 mg/L), calcium (5,910 mg/L), etc., was investigated to develop a novel meidum far the mass cultivation of useful microorganisms. For this research, we first isolated an antagonistic bacterium G-74 from soil, which showed strong growth inhibition against two phytopathogenic fungi, Rhizoctonia solani and Botrytis cinerea, and identified as Bacillus lentimorbus G-74 based on the morphological characteristics and MIDI analysis. Culture conditions for G-74 strain in the M. purpurascens juice medium were optimized. Dilution rate of the medium, temperature and initial pH for the optimum growth of G-74 strain were 30% (V/V), $35^{\circ}C$ and 5.0, respectively. It was found that additions of 2.0% (W/V) corn starch as a carbon source and 1.0% (W/V) yeast extract as a nitrogen source in this medium increased B. lentimorbus G-74 growth to 66% more efficient than Luria Bertani medium.