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The Profile and Antimicrobial Activity of Bacillus Lipopeptide Extracts of Five Potential Biocontrol Strains.

ABSTRACT: In this study the efficacy of two different methods for extracting lipopeptides produced by five Bacillus strains-ethyl acetate extraction, and acid precipitation followed by methanol extraction-was investigated using mass spectrometry. High performance thin layer chromatography (HPTLC) was also used for the simultaneous separation of complex mixtures of lipopeptide extracts and for the determination of antimicrobial activity of their components. The mass spectra clearly showed well-resolved groups of peaks corresponding to different lipopeptide families (kurstakins, iturins, surfactins, and fengycins). The ethyl acetate extracts produced the most favorable results. The extracts of SS-12.6, SS-13.1, and SS-38.4 showed the highest inhibition zones. An iturin analog is responsible for the inhibition of Xanthomonas arboricola and Pseudomonas syringae phytopathogenic strains. HPTLC bioautography effectively identified the active compounds from a mixture of lipopeptide extracts, proving in situ its potential for use in direct detection and determination of antimicrobials. In the test of potential synergism among individual extracts used in different mixtures, stronger antimicrobial effects were not observed. Biochemical and phylogenetic analysis clustered isolates SS-12.6, SS-13.1, SS-27.2, and SS-38.4 together with Bacillus amyloliquefaciens, while SS-10.7 was more closely related to Bacillus pumilus.

SUBMITTER: Dimkic I

PROVIDER: S-EPMC5440568 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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The Profile and Antimicrobial Activity of <i>Bacillus</i> Lipopeptide Extracts of Five Potential Biocontrol Strains.

Dimkić Ivica I Stanković Slaviša S Nišavić Marija M Petković Marijana M Ristivojević Petar P Fira Djordje D Berić Tanja T

Frontiers in microbiology 20170523

In this study the efficacy of two different methods for extracting lipopeptides produced by five <i>Bacillus</i> strains-ethyl acetate extraction, and acid precipitation followed by methanol extraction-was investigated using mass spectrometry. High performance thin layer chromatography (HPTLC) was also used for the simultaneous separation of complex mixtures of lipopeptide extracts and for the determination of antimicrobial activity of their components. The mass spectra clearly showed well-resol ...[more]

PMID: 28588570

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Project description:BACKGROUND:The use of chemical fungicides against fungal pathogens adversely affects soil and plant health thereby resulting in overall environmental hazards. Therefore, biological source for obtaining antifungal agents is considered as an environment-friendly alternative for controlling fungal pathogens. RESULTS:In this study, seven endophytic bacteria were isolated from sugarcane leaves and screened for its antifungal activity against 10 fungal isolates belonging to the genera Alternaria, Cochliobolus, Curvularia, Fusarium, Neodeightonia, Phomopsis and Saccharicola isolated from diseased leaves of sugarcane. Among the seven bacterial isolates, SCB-1 showed potent antagonistic activity against the tested fungi. Based on the phenotypic data, Fatty Acid Methyl Esters (FAME) and 16S rRNA gene sequence analysis, the isolate SCB-1 was identified as Bacillus subtilis. The bacterial isolate was screened negative for chitinase production; however, chloroform and methanol extracts of the bacterial culture caused significant inhibition in the growth of the fungal isolates on semisolid media. Volatile component assay showed highest inhibitory activity against Saccharicola bicolor (SC1.4). A PCR based study detected the presence of the genes involved in biosynthesis of surfactin, bacillaene, difficidin, macrolactins and fengycin. Mass spectrometric analysis of the bacterial extract detected the presence of antifungal lipopeptide surfactin, but other metabolites were not detected. The biocontrol activity of the bacterial isolate was established when bacterial pretreated mung bean seeds were able to resist Fusarium infection, however, the untreated seeds failed to germinate. CONCLUSION:The antifungal potential of isolate Bacillus subtilis SCB-1 was established against taxonomically diverse fungal pathogens including the genera Saccharicola, Cochliobolus, Alternaria and Fusarium. The potent antifungal compound surfactin as well as volatiles produced by the bacterial isolate could be responsible for its bio-control activity against fungal infections.

Project description:Oxidative stress is among the major triggers for many important human functional disorders, which often lead to various metabolic or tissue diseases. The aim of the study is to obtain five standardized vegetal extracts (Cynarae extractum-CE, Rosmarini extractum-RE, Taraxaci extractum-TE, Cichorii extractum-CHE, and Agrimoniae extractum-AE) that contain active principles with an essential role in protecting liver cells against free radicals and quantify their antioxidant actions. The compounds of therapeutic interest from the analyzed extracts were identified and quantified using the UHPLC-HRMS/MS technique. Thus, the resulting identified compounds were 28 compounds in CE, 48 compounds in RE, 39 compounds in TE, 43 compounds in CHE, and 31 compounds in AE. These compounds belong to the class of flavonoids, isoflavones, phenolic acids and dicarboxylic acids, depsides, diterpenes, triterpenes, sesquiterpenes, proanthocyanidins, or coumarin derivatives. From the major polyphenolic compounds quantified in all the extracts analyzed by UHPLC-HRMS/MS, considerable amounts have been found for chlorogenic acid (619.8 µg/g extract for TE-2032.4 µg/g extract for AE), rutoside (105.1 µg/g extract for RE-1724.7 µg/g extract for AE), kaempferol (243 µg/g extract for CHE-2028.4 µg/g extract for CE), and for naringenin (383 µg/g extract for CHE-1375.8 µg/g extract for AE). The quantitative chemical analysis showed the highest content of total phenolic acids for AE (24.1528 ± 1.1936 g chlorogenic acid/100 g dry extract), the highest concentration of flavones for RE (6.0847 ± 0.3025 g rutoside/100 g dry extract), and the richest extract in total polyphenols with 31.7017 ± 1.2211 g tannic acid equivalent/100 g dry extract for AE. Several methods (DPPH, ABTS, and FRAP) have been used to determine the in vitro total antioxidant activity of the extracts to evaluate their free radical scavenging ability, influenced by the identified compounds. As a result, the correlation between the content of the polyphenolic compounds and the antioxidant effect of the extracts has been demonstrated. Statistically significant differences were found when comparing the antiradical capacity within the study groups. Although all the analyzed extracts showed good IC50 values, which may explain their antihepatotoxic effects, the highest antioxidant activity was obtained for Agrimoniae extractum (IC50ABTS = 0.0147 mg/mL) and the lowest antioxidant activity was obtained for Cynarae extractum (IC50ABTS = 0.1588 mg/mL). Furthermore, the hepatoprotective potential was evaluated in silico by predicting the interactions between the determined phytochemicals and key molecular targets relevant to liver disease pathophysiology. Finally, the evaluation of the pharmacognostic and phytochemical properties of the studied extracts validates their use as adjuvants in phytotherapy, as they reduce oxidative stress and toxin accumulation and thus exert a hepatoprotective effect at the cellular level.

Project description:Lipopeptide biosurfactant producing Bacillus strains have many useful applications in biotechnology and agriculture, based on their emulsifying, surface activity and antimicrobial properties. In the current study, lipopeptide production kinetics, and biocontrol potentials of two new B. velezensis strains, ES1-02 and EFSO2-04 were analyzed and compared with those of commercial strains QST713 and FZB42. ES1-02 and EFSO2-04 showed higher specific growth rates than FZB42, but lower growth rates than QST713. All strains produced surfactin lipopetides, while fengycin production was not observed in ES1-02 and EFSO2-04. Production of fengycin A, B, X and Y were however confirmed in strains QST713 and FZB42. Significant differences were observed in the production of lipopeptides of the iturin family. While ES1-02 and EFSO2-04 produced bacillomycin L, QST713 produced iturin A, and FZB42 produced bacillomycin D. This was in line with the PCR analysis of corresponding genes encoding the identified lipopeptides. Highest surfactin titer of 97.4 mg/L was observed in ES1-02, while QST713 produced highest amount of iturin/bacillomycin (8.5 mg/L). Surfactin isoforms C12 to C17, and iturin/bacillomycin isoforms C11 to C17 were identified by mass spectrometry. ES1-02 and EFSO2-04 showed biocontrol potentials comparable with that of QST713 against Diaporthe spp., while FZB42 showed superior antifungal potentials. Up to 41%, 43%, 47 % and 68.9 % inhibition of D. caulivora were achieved by ES1-02, EFSO2-04, QST713 and FZB42 respectively. Upon exposure to B. velezensis strains, morphological changes to Diaporthe hyphae in form of swellings, distortion, and complete disruption occurred. Interaction of D. longicolla DPC_HOH20 with ES1-02 and EFSO2-04 induced 10-fold and 5-fold increase in surfactin synthesis, respectively. Antagonist interaction with D. longicolla induced significant changes in the proteome of ES1-02 including an increased abundance of several proteins associated with biosynthesis of antimicrobial compounds and fatty acids, while proteins associated with phosphate uptake were decreased in abundance.

Dataset Information

The Profile and Antimicrobial Activity of Bacillus Lipopeptide Extracts of Five Potential Biocontrol Strains.

Publications

The Profile and Antimicrobial Activity of <i>Bacillus</i> Lipopeptide Extracts of Five Potential Biocontrol Strains.

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