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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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Phylogenetic tree of the 16S rDNA nucleotide sequences. Numbers above branches represent bootstrap values (1000 replicates) using neighbor joining.
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fig2: Phylogenetic tree of the 16S rDNA nucleotide sequences. Numbers above branches represent bootstrap values (1000 replicates) using neighbor joining.

Mentions: Actinomycetes specific primers were used to amplify 16S rDNA gene (500–625 bp). These primers that were earlier used by Stach [15], these specific primers, can easily identify actinomycetes from other bacteria (Figure 1(b)). The 16S rDNA sequences of the four antibacterial-producing isolates (number 012-1, number 012-2, number 010-31, and number 025-26) were blasted using megablast tool of GenBank (http://www.ncbi.nlm.nih.gov/) and were deposited in the NCBI database as Streptomyces sp. JB07 (HQ896734), Pseudonocardia sp. JB06 (HQ896733), Pseudonocardia sp. JB05 (HQ896675), and Pseudonocardia sp. JB02 (HQ398191). Analysis of phylogenetic tree by using 16S rDNA sequences had led to the formation of three main clades, including Streptomyces, Pseudonocardia, and Nocardiopsis (Figure 2). These isolated bacteria belong to the two main families of actinomycetes; Streptomycetaceae, with a frequency of 95.3%, and Pseudonocardiaceae, with a frequency of 0.06% [24].


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)

Phylogenetic tree of the 16S rDNA nucleotide sequences. Numbers above branches represent bootstrap values (1000 replicates) using neighbor joining.
© Copyright Policy
Related In: Results  -  Collection

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

fig2: Phylogenetic tree of the 16S rDNA nucleotide sequences. Numbers above branches represent bootstrap values (1000 replicates) using neighbor joining.
Mentions: Actinomycetes specific primers were used to amplify 16S rDNA gene (500–625 bp). These primers that were earlier used by Stach [15], these specific primers, can easily identify actinomycetes from other bacteria (Figure 1(b)). The 16S rDNA sequences of the four antibacterial-producing isolates (number 012-1, number 012-2, number 010-31, and number 025-26) were blasted using megablast tool of GenBank (http://www.ncbi.nlm.nih.gov/) and were deposited in the NCBI database as Streptomyces sp. JB07 (HQ896734), Pseudonocardia sp. JB06 (HQ896733), Pseudonocardia sp. JB05 (HQ896675), and Pseudonocardia sp. JB02 (HQ398191). Analysis of phylogenetic tree by using 16S rDNA sequences had led to the formation of three main clades, including Streptomyces, Pseudonocardia, and Nocardiopsis (Figure 2). These isolated bacteria belong to the two main families of actinomycetes; Streptomycetaceae, with a frequency of 95.3%, and Pseudonocardiaceae, with a frequency of 0.06% [24].

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