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Antibacterial activity of Pseudonocardia sp. JB05, a rare salty soil actinomycete against Staphylococcus aureus.

Jafari N, Behroozi R, Farajzadeh D, Farsi M, Akbari-Noghabi K - Biomed Res Int (2014)

Bottom Line: Finally, the partial structure of the resultant antibacterial compound was characterized by Fourier transform infrared spectroscopy.JB05, based on the results of biochemical and 16S rDNA gene sequence analysis.The results presented in this work, although at the initial stage in bioactive product characterization, will possibly contribute toward the Pseudonocardia scale-up for the production and identification of the antibacterial compounds.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran ; Department of Plant Biotechnology, Buali-Sina University of Hamedan, Iran.

ABSTRACT
Staphylococcus aureus is a Gram-positive bacterium that causes many harmful and life-threatening diseases. Some strains of this bacterium are resistant to available antibiotics. This study was designed to evaluate the ability of indigenous actinomycetes to produce antibacterial compounds against S. aureus and characterize the structure of the resultant antibacterial compounds. Therefore, a slightly modified agar well diffusion method was used to determine the antibacterial activity of actinomycete isolates against the test microorganisms. The bacterial extracts with antibacterial activity were fractionated by silica gel and G-25 sephadex column chromatography. Also, the active fractions were analyzed by thin layer chromatography. Finally, the partial structure of the resultant antibacterial compound was characterized by Fourier transform infrared spectroscopy. One of the isolates, which had a broad spectrum and high antibacterial activity, was designated as Pseudonocardia sp. JB05, based on the results of biochemical and 16S rDNA gene sequence analysis. Minimum inhibitory concentration for this bacterium was 40 AU mL(-1) against S. aureus. The antibacterial activity of this bacterium was stable after autoclaving, 10% SDS, boiling, and proteinase K. Thin layer chromatography, using anthrone reagent, showed the presence of carbohydrates in the purified antibacterial compound. Finally, FT-IR spectrum of the active compound illustrated hydroxyl groups, hydrocarbon skeleton, and double bond of polygenic compounds in its structure. To the best of our knowledge, this is the first report describing the efficient antibacterial activity by a local strain of Pseudonocardia. The results presented in this work, although at the initial stage in bioactive product characterization, will possibly contribute toward the Pseudonocardia scale-up for the production and identification of the antibacterial compounds.

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(a) Antibacterial activity of fractions numbers 1–15 of Pseudonocardia sp. JB05 extract. (b) Inhibition clear halo zone of S. aureus around the antibacterial fractions numbers 3, 4, 5, and 6 on agar plates.
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fig3: (a) Antibacterial activity of fractions numbers 1–15 of Pseudonocardia sp. JB05 extract. (b) Inhibition clear halo zone of S. aureus around the antibacterial fractions numbers 3, 4, 5, and 6 on agar plates.

Mentions: Antimicrobial purification was performed for Pseudonocardia sp. JB05 extract because of its highest inhibition zone against S. aureus. The antibacterial supernatant was vacuum-evaporated to dryness and then extracted with dH2O and/or organic solvents (n-hexane and/or ethyl acetate). Our results showed that the dH2O extract was the most active extract against S. aureus, compared to those extracted using organic solvents. As an appropriate polar solvent, dH2O was used to extract the compound before being compared with the one from the organic solvent [18]. The dH2O extract was fractionated according to the following procedure: a total of 15 fractions were separated through a silica gel column (Figure 3(a)). When evaluated by an inhibition test, fractions numbers 3, 4, 5, and 6 indicated a clear inhibition zone (Figure 3(b)). They were pooled and loaded on G25 column chromatography and, after fractionation, fraction number 5 showed the largest clear zone against S. aureus.


Antibacterial activity of Pseudonocardia sp. JB05, a rare salty soil actinomycete against Staphylococcus aureus.

Jafari N, Behroozi R, Farajzadeh D, Farsi M, Akbari-Noghabi K - Biomed Res Int (2014)

(a) Antibacterial activity of fractions numbers 1–15 of Pseudonocardia sp. JB05 extract. (b) Inhibition clear halo zone of S. aureus around the antibacterial fractions numbers 3, 4, 5, and 6 on agar plates.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150441&req=5

fig3: (a) Antibacterial activity of fractions numbers 1–15 of Pseudonocardia sp. JB05 extract. (b) Inhibition clear halo zone of S. aureus around the antibacterial fractions numbers 3, 4, 5, and 6 on agar plates.
Mentions: Antimicrobial purification was performed for Pseudonocardia sp. JB05 extract because of its highest inhibition zone against S. aureus. The antibacterial supernatant was vacuum-evaporated to dryness and then extracted with dH2O and/or organic solvents (n-hexane and/or ethyl acetate). Our results showed that the dH2O extract was the most active extract against S. aureus, compared to those extracted using organic solvents. As an appropriate polar solvent, dH2O was used to extract the compound before being compared with the one from the organic solvent [18]. The dH2O extract was fractionated according to the following procedure: a total of 15 fractions were separated through a silica gel column (Figure 3(a)). When evaluated by an inhibition test, fractions numbers 3, 4, 5, and 6 indicated a clear inhibition zone (Figure 3(b)). They were pooled and loaded on G25 column chromatography and, after fractionation, fraction number 5 showed the largest clear zone against S. aureus.

Bottom Line: Finally, the partial structure of the resultant antibacterial compound was characterized by Fourier transform infrared spectroscopy.JB05, based on the results of biochemical and 16S rDNA gene sequence analysis.The results presented in this work, although at the initial stage in bioactive product characterization, will possibly contribute toward the Pseudonocardia scale-up for the production and identification of the antibacterial compounds.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran ; Department of Plant Biotechnology, Buali-Sina University of Hamedan, Iran.

ABSTRACT
Staphylococcus aureus is a Gram-positive bacterium that causes many harmful and life-threatening diseases. Some strains of this bacterium are resistant to available antibiotics. This study was designed to evaluate the ability of indigenous actinomycetes to produce antibacterial compounds against S. aureus and characterize the structure of the resultant antibacterial compounds. Therefore, a slightly modified agar well diffusion method was used to determine the antibacterial activity of actinomycete isolates against the test microorganisms. The bacterial extracts with antibacterial activity were fractionated by silica gel and G-25 sephadex column chromatography. Also, the active fractions were analyzed by thin layer chromatography. Finally, the partial structure of the resultant antibacterial compound was characterized by Fourier transform infrared spectroscopy. One of the isolates, which had a broad spectrum and high antibacterial activity, was designated as Pseudonocardia sp. JB05, based on the results of biochemical and 16S rDNA gene sequence analysis. Minimum inhibitory concentration for this bacterium was 40 AU mL(-1) against S. aureus. The antibacterial activity of this bacterium was stable after autoclaving, 10% SDS, boiling, and proteinase K. Thin layer chromatography, using anthrone reagent, showed the presence of carbohydrates in the purified antibacterial compound. Finally, FT-IR spectrum of the active compound illustrated hydroxyl groups, hydrocarbon skeleton, and double bond of polygenic compounds in its structure. To the best of our knowledge, this is the first report describing the efficient antibacterial activity by a local strain of Pseudonocardia. The results presented in this work, although at the initial stage in bioactive product characterization, will possibly contribute toward the Pseudonocardia scale-up for the production and identification of the antibacterial compounds.

Show MeSH
Related in: MedlinePlus