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      Anti-Methicillin Resistant Staphylococcus aureus Activity and Optimal Culture Condition of Streptomyces sp. SUK 25

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          Abstract

          Background:

          The potential of secondary metabolites extracted from Streptomyces sp. to treat bacterial infections including infections with Staphylococcus aureus is previously documented. The current study showed significant antimicrobial activities associated with endophytic Streptomyces sp. isolated from medicinal plants in Peninsular Malaysia.

          Objectives:

          The current study aimed to determine anti-methicillin-resistant- Staphylococcus aureus (MRSA) activities of Streptomyces sp. isolates.

          Materials and Methods:

          Disc diffusion and Minimum Inhibitory Concentration (MIC) assay were used to determine the antibacterial activity of Streptomyces sp. isolates. Optimization of fermentation parameters for the most potent anti-MRSA extract in terms of medium type, pH, aeration rate, and culture period was also carried out. Lastly, toxicity of the extract against Chang liver cells was determined employing the MTT, 2- (3, 5- diphenyltetrazol-2-ium-2-yl) -4, 5-dimethyl -1, 3 - thiazole; bromide assay.

          Results:

          The results indicated Streptomyces sp. SUK 25 isolates showed the most potent anti-MRSA activity. Disc diffusion assay revealed that spread plate technique was more efficient in screening anti-MRSA activity compared to pour plate (P < 0.05). To determine anti–MRSA MIC of Streptomyces sp. SUK 25, Thronton media was used. Therefore, MIC was determined as 2.44 ± 0.01 µg/mL, and accordingly, the lowest MIC was 1.95 µg/mL based on a seven-day culture, pH7, and aeration rate of 140 rpm. The crude extract was not toxic against Chang liver cells (IC 50 = 43.31 ± 1.24 µg/mL).

          Conclusions:

          The Streptomyces sp. SUK 25 culturing was optimized using Thronton media, at pH 7 and aeration of 140 rpm. Further isolation and identification of bioactive compounds will develop anti-MRSA therapeutics.

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          Most cited references36

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          Genome-scale analysis of Streptomyces coelicolor A3(2) metabolism.

          Streptomyces are filamentous soil bacteria that produce more than half of the known microbial antibiotics. We present the first genome-scale metabolic model of a representative of this group--Streptomyces coelicolor A3(2). The metabolism reconstruction was based on annotated genes, physiological and biochemical information. The stoichiometric model includes 819 biochemical conversions and 152 transport reactions, accounting for a total of 971 reactions. Of the reactions in the network, 700 are unique, while the rest are iso-reactions. The network comprises 500 metabolites. A total of 711 open reading frames (ORFs) were included in the model, which corresponds to 13% of the ORFs with assigned function in the S. coelicolor A3(2) genome. In a comparative analysis with the Streptomyces avermitilis genome, we showed that the metabolic genes are highly conserved between these species and therefore the model is suitable for use with other Streptomycetes. Flux balance analysis was applied for studies of the reconstructed metabolic network and to assess its metabolic capabilities for growth and polyketides production. The model predictions of wild-type and mutants' growth on different carbon and nitrogen sources agreed with the experimental data in most cases. We estimated the impact of each reaction knockout on the growth of the in silico strain on 62 carbon sources and two nitrogen sources, thereby identifying the "core" of the essential reactions. We also illustrated how reconstruction of a metabolic network at the genome level can be used to fill gaps in genome annotation.
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            Cultivation-independent population analysis of bacterial endophytes in three potato varieties based on eubacterial and Actinomycetes-specific PCR of 16S rRNA genes.

            Abstract Endophytic bacteria are ubiquitous in most plants and colonise plants without exhibiting pathogenicity. Studies on the diversity of bacterial endophytes have been mainly approached by characterisation of isolates obtained from internal tissues. Despite the broad application of culture-independent techniques for the analysis of microbial communities in a wide range of natural habitats, little information is available on the species diversity of endophytes. In this study, microbial communities inhabiting stems, roots and tubers of three potato varieties were analysed by 16S rRNA-based techniques such as terminal restriction fragment length polymorphism analysis, denaturing gradient gel electrophoresis as well as 16S rDNA cloning and sequencing. Two individual plant experiments were conducted. In the first experiment plants suffered from light deficiency, whereas healthy and robust plants were obtained in the second experiment. Plants obtained from both experiments showed comparable endophytic populations, but healthy potato plants possessed a significantly higher diversity of endophytes than stressed plants. In addition, plant tissue and variety specific endophytes were detected. Sequence analysis of 16S rRNA genes indicated that a broad phylogenetic spectrum of bacteria is able to colonise plants internally including alpha-, beta-, and gamma-Proteobacteria, high-GC Gram-positives, microbes belonging to the Flexibacter/Cytophaga/Bacteroides group and Planctomycetales. Group-specific analysis of Actinomycetes indicated a higher abundance and diversity of Streptomyces scabiei-related species in the variety Mehlige Mühlviertler, which is known for its resistance against potato common scab caused by S. scabiei.
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              Antimicrobial potential of Actinomycetes species isolated from marine environment.

              To evaluate the antimicrobial activity of Actinomycetes species isolated from marine environment. Twenty one strains of Actinomycetes were isolated from samples of Royapuram, Muttukadu, Mahabalipuram sea shores and Adyar estuary. Preliminary screening was done using cross-streak method against two gram-positive and eight gram-negative bacteria. The most potent strains C11 and C12 were selected from which antibacterial substances were extracted. The antibacterial activities of the extracts were performed using Kirby-Bauer disc diffusion method. Molecular identification of those isolates was done. All those twenty one isolates were active against at least one of the test organisms. Morphological characters were recorded. C11 showed activity against Staphylococcus species (13.0±0.5 mm), Vibrio harveyi (11.0±0.2 mm), Pseudomonas species (12.0±0.3 mm). C12 showed activity against Staphylococcus species (16.0±0.4 mm), Bacillus subtilis (11.0±0.2 mm), Vibrio harveyi (9.0±0.1 mm), Pseudomonas species (10.0±0.2 mm). 16S rRNA pattern strongly suggested that C11 and C12 strains were Streptomyces species. The results of the present investigation reveal that the marine Actinomycetes from coastal environment are the potent source of novel antibiotics. Isolation, characterization and study of Actinomycetes can be useful in discovery of novel species of Actinomycetes.
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                Author and article information

                Journal
                Jundishapur J Microbiol
                Jundishapur J Microbiol
                10.5812/jjm
                Kowsar
                Jundishapur Journal of Microbiology
                Kowsar
                2008-3645
                2008-4161
                31 May 2015
                May 2015
                : 8
                : 5
                : e16784
                Affiliations
                [1 ]School of Diagnostic and Biomedical Sciences, Faculty of Medicine and Health Sciences, University Sultan Zainal Abidin, Kuala Terengganu, Terengganu, Malaysia
                [2 ]School of Bioscience and Biotechnology Study, Faculty of Science and Technology, University Kebangsaan Malaysia, Selangor, Malaysia
                [3 ]School of Diagnostic and Applied Health Sciences, Faculty of Health Sciences, University Kebangsaan Malaysia, Kuala Lumpur, Malaysia
                Author notes
                [* ]Corresponding author: Mohamad Zin Noraziah, School of Diagnostic and Applied Health Sciences, Faculty of Health Sciences, University Kebangsaan Malaysia, Kuala Lumpur, Malaysia. Tel: +60-392897373, E-mail: noraziah.zin@ 123456ukm.edu.my
                Article
                10.5812/jjm.16784
                4458357
                26060562
                c3816119-648f-4f08-bc25-72a6c1f53fe8
                Copyright © 2015, Ahvaz Jundishapur University of Medical Sciences.

                This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License ( http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.

                History
                : 21 December 2013
                : 25 March 2014
                : 06 May 2014
                Categories
                Research Article

                streptomyces sp.,mrsa,culture
                streptomyces sp., mrsa, culture

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