• 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.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.1
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• pp.91-96
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• 2015

A lipopeptide extract of Bacillus amyloliquefaciens BACY1 (BLE) was found to induce cell death in human oral squamous cell carcinoma (OSCC) cell lines, SCC4 and SCC25, in this study. The results of MTT assay showed that BLE inhibited OSCC cell proliferation in a dose-dependent manner. BLE was also effective in increasing the sub-G1 phases. Furthermore, when membrane damage in SCC4 cells treated with BLE was monitored by LDH assay, release of LDH was significantly increased. The protein and mRNA levels of pro-apoptotic Bax, and caspase-3 were up-regulated by BLE. Taken together, these results suggest that BLE induces apoptosis and then inhibits the cell proliferation of human OSCC cells.

• Journal of Microbiology
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• v.43 no.3
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• pp.272-276
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• 2005

Bacillus subtilis ATCC 6633 was grown in BHIB medium supplemented with $Mn^{2+}$ for 96 h at $37^{\circ}C$ in a shaker incubator. After removing the microbial biomass, a lipopeptide biosurfactant was extracted from the supernatant. Its structure was established by chemical and spectroscopy methods. The structure was confirmed by physical properties, such as Hydrophile-Lipophile Balance (HLB), surface activity and erythrocyte hemolytic capacity. The critical micelle concentration (cmc) and erythrocyte hemolytic capacity of the biosurfactant were compared to those of surfactants such as SDS, BC (benzalkonium chloride), TTAB (tetradecyltrimethylammonium bromide) and HTAB (hexadecyltrimethylammonium bromide). The maximum hemolytic effect for all surfactants mentioned was observed at concentrations above cmc. The maximum hemolytic effect of synthetic surfactants was more than that of the biosurfactant produced by B. subtilis ATCC 6633. Therefore, biosurfactant would be considered a suitable surface-active agent due to low toxicity to the membrane.

• Journal of Microbiology and Biotechnology
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• v.24 no.1
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• pp.13-18
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• 2014

Bacillus subtilis JW-1 was isolated from rhizosphere soil as a potential biocontrol agent of bacterial wilt caused by Ralstonia solanacearum. Seed treatment followed by a soil drench application with this strain resulted in >80% reduction in bacterial wilt disease compared with that in the untreated control under greenhouse conditions. The antibacterial compound produced by strain JW-1 was purified by bioactivity-guided fractionation. Based on mass spectroscopy and nuclear magnetic resonance spectral data ($^1H$, $^{13}C$, $^1H-^1H$ correlation spectroscopies, rotating frame nuclear Overhauser effect spectroscopy, and heteronuclear multiple-bond correlation spectroscopy), the structure of this compound was elucidated as a cyclic lipopeptide composed of a heptapeptide (Gln-Leu-Leu-Val-Asp-Leu-Leu) bonded to a ${\beta}$-hydroxy-iso-hexadecanoic acid arranged in a lactone ring system.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.716-723
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• 2006

A biosurfactant-producing bacterial strain was selected from oil-contaminated soil because of its ability to degrade crude oil and tributyrin $(C_{4:0})$. The strain was identified as Bacillus subtilis A8-8 based on its morphological, biochemical, and physiological characteristics. When B. subtilis A8-8 was grown with crude oil as the sole carbon source, the biosurfactant from the strain emulsified crude oil, vegetable oil, and hydrocarbons. Soybean oil was the optimum substrate for the emulsifying activity and emulsion stability of the biosurfactant, both of which were superior to those of several commercially available surfactants. The biosurfactant was purified by a procedure including HCl precipitation, methanol treatment, and silica-gel chromatography. The partially purified biosurfactant was analyzed by TLC (thin-layer chromatography), SDS-PAGE, and HPLC and it reduced the surface tension of water from 72 mN/m to 26 mN/m at a concentration of 30 mg/l. Therefore, the purified lipopeptide biosurfactant has strong properties as an emulsifying agent and acts as an emulsion-stabilizing agent.

• Journal of Microbiology and Biotechnology
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• v.26 no.4
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• pp.748-756
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• 2016

In the present study, a lipopeptide (named AXLP14) antagonistic to Xanthomonas oryzae pv. oryzae (Xoo) was obtained from the culture supernatant of Bacillus amyloliquefaciens B014. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis demonstrated that AXLP14 consisted of surfactin homologs. The minimum inhibition concentration and minimum bactericidal concentration of AXLP14 against Xoo were determined to be 1.25 and 2.50 mg/ml, respectively. At a concentration of 0.613 mg/ml, AXLP14 strongly inhibited the formation of Xoo biofilm. AXLP14 also inhibited the motility of Xoo in a concentration-dependent manner. Applying AXLP14 to rice seedlings significantly reduced the incidence and severity of disease caused by Xoo. In Xoo-infected rice seedlings, AXLP14 strongly and continuously up-regulated the expression of both OsNPR1 and OsWRKY45. In addition, AXLP14 effectively inhibited the Xoo-induced up-regulation of the expression of the abscisic acid biosynthesis gene OsNECD3 and the abscisic acid signalingresponsive gene OsLip9, indicating that AXLP14 may protect rice against Xoo-induced disease by enhancing salicylic acid defense and interfering with the abscisic acid response to virulence.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.52-59
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• 2013

The nutritional requirements for the maximum production of lipopeptides by Bacillus subtilis N7 (B. subtilis N7) were investigated and optimized using response surface methodology (RSM) under shake flask fermentation. A one-factor-at-a-time experimental setup was used to screen carbon and nitrogen sources. A Plackett-Burman design (PBD) was employed to screen the most critical variables for lipopeptides production amongst ten nutritional elements. The central composite experimental design (CCD) was finally adopted to elucidate the composition of the fermentation medium. Statistical analyses (analysis of variance, ANOVA) of the results showed that KCl, $MnSO_4$ and $FeSO_4{\cdot}6H_2O$ were important components and that their interactions were strong. Lipopeptide production was predicted to reach 709.87 mg/L after a 60 h incubation using an optimum fermentation medium composed of glucose 7.5 g/L, peanut oil 1.25 g/L, $MgSO_4$ 0.37 g/L, $KH_2PO_4$ 0.75 g/L, monosodium glutamate 6.75 g/L, yeast extract and $NH_4Cl$ (5:3 w/w) 10 g/L, KCl 0.16 g/L, $FeSO_4{\cdot}6H_2O$ 0.24 mg/L, $MnSO_4$ 0.76 mg/L, and an initial pH of 7.0. Lipopeptide production ($706.57{\pm}3.70$ mg/L) in the optimized medium confirmed the validity of the predicted model.

• KSBB Journal
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• v.14 no.6
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• pp.737-741
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• 1999

Phenanthrene degrading bacteria were isolated from the petroleum contaminated soil near an oil tank. Four of 15 strains decreased surface tension of culture broth of phenanthrene-containing minimal media. H6, one of the isolated bacteria decreased surface tension of culture broth below 33 dyne/cm during growth on glucose. H6 was identified as Bacillus subtilis and biosurfactant produced by H6 was lipopeptide. The biosurfactant was produced at 0.13 g/L in the mineral medium containing 2% glucose. Critical micelle concentration(CMC) of the biosurfactant was 52 mg/L. Foaming power was similar to Tween 80 and dispersing power was superior to Tween 80m SDS and Brij30. High thermal stability and emulsion index were also observed.

• 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.

• Journal of Mushroom
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• v.22 no.2
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• pp.37-47
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• 2024

Fluorescent bacteria were isolated from sporocarps that browned into various mushrooms during survey at places of the production in Korea. We examined the pathogenicity, biodiversity, and genetic characteristics of the 19 strains identified as Pseudomonas tolaasii by sequence analysis of 16S rRNA and White Line Assay. The results emphasize the importance of rpoB gene system, fatty acid profiles, specific and sensitive PCR assays, and lipopeptide detection for the identification of P. tolaasii. As a result of these various analyses, 17 strains (CHM03~CHM19) were identified as P. tolaasii. The phylogenetic analysis based on the 16S rRNA gene showed that all strains were clustered closest to P. tolaasii lineage, two strains (CHM01, CHM02) were not identified as P. tolaasii and have completely different genetic characteristics as a result of fatty acids profile, specific and sensitive PCR, lipopetide detection, rpoB sequence and REP-PCR analysis. Pathogenicity tests showed 17 strains produce severe brown discolouration symptoms to button mushrooms and watersoaking of sporophore tissue within three days after inoculation. But two strains did not produce discolouration symptoms. Therefore, these two strains will be further investigated for correct species identification by different biological and molecular characteristics.