Isolation, Identification, and Evaluation of Antimicrobial Activity of Active Compound Produced by Marine Actinomycetes isolate A32 (original) (raw)
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2016
The continuation of new antibiotics exploration becomes an important research program in the world for pharmaceutical and agricultural applications. Marine filamentous bacteria such as actinomycetes have been widely used as an important biological tool to generate a variety of new secondary metabolites, such as antibiotic. The aim of this study was to obtain identified active compound and determine its antimicrobial activity. Isolation, identification, and antimicrobial activity assay of active compound produced by marine actinomycetes isolate A32 had been conducted. Production of active compound using isolate actinomycetes A32 was conducted involving glucose, yeast, peptone medium. The fermentation was carried out at 30 oC for 5 days. The broth of supernatant was extracted using ethyl acetate. Purification of active compound used chromatography column and eluted stepwise with the chloroform and methanol solvents. Antimicrobial activity was monitored using agar disc diffusion, and m...
Isolation of Marine Actinomycetes and Screening its Antibacterial Potential
International Journal for Research in Applied Science and Engineering Technology IJRASET, 2020
Antibacterial and pharmaceutical potential of marine isolates are considered as a significant objective in the present research. Marine actinomycetes were isolated and screened for its antibacterial activity. Five isolates showing chalky white and whitish grey colour colonies were isolated and designated as ACT 1 , ACT 2 , ACT 3 , ACT 4 and ACT 5. ACT 2 exhibited the inhibitory zones of 20mm, 19mm, 19mm and 22mm respectively against Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 25922), Bacillus subtilis (ATCC 6051) and Staphylococcus aureus (ATCC 9144). ACT 5 showed maximum inhibitory zones of 22mm, 18mm, 21mm and 24mm against the respective test organisms. Thus, the obtained results in the present research revealed the pharmaceutical applications of marine metabolites. The types of antibiotics and its structures synthesized by the marine isolates shall be studied as future study.
Journal of Pure and Applied Microbiology, 2019
The aim of the present study was to isolate antagonistic actinomycetes from marine sediment collected from the Red Sea coast at Hurghada city and the Suez Gulf. A total of 16 actinomycete isolates were obtained in October 2018 and their antagonistic activities were evaluated against Aeromonas hydrophila, Vibrio damsela, Staphylococcus aureus ATCC6538, Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa ATCC 9027, Salmonella typhimurium ATCC 14028, Escherichia coli ATCC 19404 and Candida albicans ATCC 10231 by using the agar well diffusion method. Among the 16 isolates, 14 (87.5%) isolates exhibited antimicrobial activity against most of the tested pathogens. The most potent isolate was identified by 16S rRNA sequence analysis as Streptomyces sp. and designated Streptomyces sp. MK388207. The experimental design of Plackett-Burman was implemented to optimize the culture conditions for antimicrobial agent(s) production by the most powerful isolate against C. albicans ATCC 10231 when grown in starch nitrate broth and the data revealed that negative (-) levels of KNO 3 , K 2 HPO 4 and pH and positive (+) levels of starch, MgSO 4 .7H 2 O, FeSO 4 and incubation period supported the production of the antimicrobial agent(s). Growth under the optimized culture conditions led to a 1.4-fold rise in antimicrobial activity. The ethyl acetate extract of Streptomyces sp.MK388207 was subjected to gasliquid chromatography mass spectrometry (GC MS) and revealed the presence of six fractions with the major component being phenol, 2,4-bis (1,1 dimethylethyl). The findings of this study suggested that the antagonistic marine Streptomyces, in particular Streptomyces sp MK388207, the antibacterial compounds produced by this isolate, could be used as antibiotics that could have future applications in the pharmaceutical industry.
Isolation of actinomycetes with antibacterial activity against multi-drug resistant bacteria
Malaysian Journal of Microbiology
Aims: To isolate and characterise actinomycetes from various sources of soil samples (fruit orchard, dipterocarp forest and oil palm plantation) and to screen these isolates for antibacterial activity against multi-drug resistant bacteria. Methodology and results: A total of 158 fast-growing actinomycete isolates with different colony morphology were subjected to primary cross-streak and secondary well diffusion screening. Six isolates (OP1E, OP7A, OP2A-C, MG1A, UT9C1 and UT7E) were found to inhibit at least one of the seven multi-drug resistant (MDR) bacteria. MG1A exhibited the strongest and broadest spectrum of antibacterial activity against 6 MDR bacteria tested. These isolates were identified as Streptomyces species based on 16S rRNA gene sequence analysis. Further morphological and biochemical analysis revealed that MG1A was highly similar to S. griseocarneus (98.36%) whereas OP1E and OP2A-C were similar to S. parvulus (99.93% and 99.51% respectively). Preliminary identification using LCMS/MS and database search revealed that the major compound in the extract of OP2A-C could be dactinomycin (1255.4170 g/mol). Other antibacterial compounds in the extracts remain to be identified. Conclusion, significance and impact of study: Soil actinomycetes with antibacterial activity against MDR bacteria were isolated not only from undisturbed natural soils but cultivated soils. These isolates were characterised, identified and the antibacterial compounds were extracted for further study. The isolates could serve as a potential source for the development of new and sustainable compounds against MDR bacteria.
Journal of Medical Sciences(Faisalabad), 2001
Two pure bioactive compounds, MB-1a and MB-1b were isolated from a bacterial isolate and the antibacterial activities and brine shrimp lethality bioassay were evaluated in the present investigation. The isolate was collected from soils of an agricultural field located in Rajshahi University campus and characterized as a species of Streptomyces genus by polyphasic approach. Two pure compounds were isolated and purified from fermentation broth of this bacterium by solvent extraction followed by chromatographic separation. The MIC and MBC of these compounds were studied against six pathogenic bacteria. The MIC and MBC of both MB1a and MB1b were in the range of 4-8 µg/ml. Same values of MIC and MBC indicates the bactericidal characteristics of both of the compounds. Brine shrimp lethality was evaluated and LC 50 values of MB1a, MB1b and crude extract was obtained as 9.55 µg/ml, 12.30 µg/ml, and 44.67 µg/ml, respectively.
Caerulomycin A—An Antifungal Compound Isolated from Marine Actinomycetes
Advances in Microbiology, 2014
Actinomycetes have been prolific sources of novel secondary metabolites with a range of biological activities that may ultimately find application as therapeutic compounds. Hence several drug discovery companies are engaged in isolation of novel bioactive metabolites from these microbial sources. Antibiotics form the major class of such bioactive metabolites and have been widely used for treating infectious diseases. One of the most critical problems in clinical practice is the increase of prevalence of drug resistant strains, especially azole resistance among fungi. Due to this, there is a constant need for development of new antifungal antibiotics having novel scaffolds and/or mechanism of action. In our in-house screening program in the quest of novel and superior antifungal compounds, an actinomycetes strain PM0525875 was isolated from a marine invertebrate. The extracts of this microbe showed potent in-vitro antifungal activity against drug resistant fungal strains. The antifungal active peak from the extract obtained by shake flask fermentation was identified by chromatographic and other analytical techniques during bioactivity guided isolation. Later the fermentation conditions were optimized in 30 L fermentor for the production of sufficient amount antifungal compound for complete structural characterization. Consequently the fermented broth extract was subjected to bioactivity-guided fractionation, to isolate the active principle using different preparative chromatographic techniques followed by its characterization. The active principle was characterized to be Caerulomycin A. Minimum inhibitory concentration (MIC) of the compound was found in the range of 0.39 -1.56 µg/ml against pathogenic fungal test strains. The phylogenetic analysis of producer strain using 16S rRNA sequence showed closest match with Actinoalloateichus cyanogriseus. Herewith we report the isolation of Caerulomycin A from marine invertebrate-associated Actinoalloteichus sp. using optimized medium and fermentation conditions. * Corresponding author.
2000
In the course of identification and screening for antimicrobial activities of 100 actinomycete isolates from 35 soil and sand samples collected from the shore of Samed Island, Rayong province, 79 isolates were identified as Streptomyces, 20 were Micromonospora, and one was Nocardia based on their morphological, cultural, physiological and biochemical characteristics. On primary screening for antimicrobial activities, 54 isolates of Streptomyces, 14 of Micromonospora, and one of Nocardia showed the activities against Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 6633, and Candida albicans ATCC 10231, while the activities against Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853 could be observed in few isolates. On secondary screening of 30 isolates, the strain PC4 -3 with good antimicrobial activity was selected for secondary metabolite fermentation. This strain contained LL -diaminopemelic acid in cell wall and was similar to Streptomyces sporoclivatus LMG 20312 T with 99.1% 16S rDNA sequence similarity. In addition, the representative strains of genera, MS7 -3 and SC36 -1 were closed to M. matsumotoense JCM 9104 T and Nocardia araoensis DSM 44729 T based on 98.6% and 99.3% of 16S rDNA sequence similarity, respectively. The ethyl acetate extract from fermentation broth of PC4 -3 showed antimicrobial activities against S. aureus ATCC 25923, B. subtilis ATCC 6633 and C. albicans ATCC 10231. Antimicrobial assay -guided fractionation of the ethyl acetate extract yielded 2 known ansamycins, including geldanamycin and 17 -O -demethylgeldanamycin. The chemical structures of the isolated compounds were elucidated through extensive analyses of UV, IR, MS and NMR spectroscopic data and comparison with the literatures. Geldanamycin exhibited antimicrobial activity against S. aureus ATCC 25923 and C. albicans ATCC 10231.
Indonesian Journal of Chemistry
Purification and structure elucidation of antibiotic produced by marine actinomycetes (Streptomyces sp A11) was conducted. Production of antibiotic was carried out by liquid fermentation using yeast and peptone medium for 5 days fermentation. Purification of antibiotic was carried out by silica gel 60 (Merck, 0.063-0.200 mm) column chromatography and preparative HPLC. Structure elucidation was carried out using ESI-MS, 1H NMR, 13C NMR, DEPT 13C NMR, and FTIR. This antibiotic was identified as cyclo (tyrosyl-prolyl) / (C14H16N2O3). This antibiotic had biological activity to Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Bacillus subtilis ATCC 66923, Pseudomonas aeruginosa ATCC27853, and produced by extracellular secretion. Keywords: antibiotic, actinomycetes, purification, structure elucidation
Indonesian Journal of Chemistry, 2010
Purification and structure elucidation of antibiotic produced by marine actinomycetes (Streptomyces sp A11) was conducted. Production of antibiotic was carried out by liquid fermentation using yeast and peptone medium for 5 days fermentation. Purification of antibiotic was carried out by silica gel 60 (Merck, 0.063-0.200 mm) column chromatography and preparative HPLC. Structure elucidation was carried out using ESI-MS, 1H NMR, 13C NMR, DEPT 13C NMR, and FTIR. This antibiotic was identified as cyclo (tyrosyl-prolyl) / (C14H16N2O3). This antibiotic had biological activity to Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Bacillus subtilis ATCC 66923, Pseudomonas aeruginosa ATCC27853, and produced by extracellular secretion. Keywords: antibiotic, actinomycetes, purification, structure elucidation
The Journal of Antibiotics, 2011
Two new antimycin antibiotics, that is antimycins A 19 (1) and A 20 (2), were isolated from a cultured broth of marine actinomycete Streptomyces antibioticus H74-18 together with antimycins A 1a (3a) and A 1b (3b), A 2a (4), A 3a (5a) and A 3b (5b). Their structures were determined by spectroscopic methods in combination with X-ray diffraction. Antimycin A 19 possessed a chiral acyl chain and an alkyl branch. The absolute configuration of chiral acyl chain in 1 was determined by X-ray diffraction analysis. Antimycin A 20 (2) has the shortest and simplest acetoxy acyl chain in the antimycins family. All the antimycins (1-5) showed potential antifungal activities against Candida albicans with MIC of about 5-10 lg ml -1 .