Screening, isolation and purification of antibacterial agents from marine actinomycetes

Attimarad, Sunil L; Ediga, Gaviraj N; Karigar, A. A.; Karadi, R; Chandrashekhar, Nagesh; Shivanna, Chandrashekara

doi:10.3329/icpj.v1i12.12448

Cited by 29 publications

(17 citation statements)

References 13 publications

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“…This is contrary to studies carried elsewhere [20] of variation in the population composition of actinomycetes and therefore need for different nutrients from those isolated from other regions. [24] asserts that the soil nutrient composition leads to differences in nutrient requirements in actinomycetes.…”

Section: Discussioncontrasting

confidence: 55%

Antimicrobial Properties of Actinomycetes Isolated from Menengai Crater in Kenya

Waithaka¹,

Mwaura²,

Wagacha³

et al. 2017

CellBio

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A study was carried out to isolate and screen actinomycetes for antimicrobials from Menengai Crater in Kenya. The actinomycetes were isolated using starch casein agar, Luria Bertani agar and starch nitrate agar. Primary screening for antagonism was carried out using perpendicular method while secondary screening was done using agar disk technique. Extraction of the antimicrobials was carried out using ethyl acetate. Sensitivity testing of the crude extracts against Staphylococcus aureus, Bacillus subtilis, Escherichia faecalis, Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Xanthomonas campestris, Erwinia carotovora, Candida albicans, Alternaria alternate and Fusarium oxysporum was carried out using agar well technique. Biochemical tests and carbon source requirements were used in characterization of the selected antimicrobial producers. M1 was the best agar medium for isolation of actinomycetes. The number of actinomycetes from regions A, B, C, and D in the crater varied significantly (F = 27.50 P = 0.000). Out of the 156 actinomycetes isolates, 20 isolates were positive for both primary and secondary screening for antimicrobials. There was no significant difference in the zones of inhibition in primary screening of the actinomycetes for antagonistic properties against the test pathogens (F = 1.6957 P = 0.0838). The zones of inhibition after secondary screening varied significantly (F = 2.4473 P = 0.0089). Likewise, there was a significant difference (F = 6.6046 P = 0.001338) in the zones of inhibition after exposing the pathogens to ethyl extracts of the selected antagonistic actinomycetes. There is need to purify and characterize the antimicrobials obtained from the present study.

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Section: Discussioncontrasting

confidence: 55%

Antimicrobial Properties of Actinomycetes Isolated from Menengai Crater in Kenya

Waithaka¹,

Mwaura²,

Wagacha³

et al. 2017

CellBio

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“…The obtained results revealed that oatmeal culture medium was the best medium for the isolation of bioactive marine actinomycetes according to the obtained growth and bioactivity for 36 hour incubation period at 37°C and pH 7. Unlike the study of Attimarad et al [10] showed the Starch-casein agar supplemented with nystatin and nalidixic acid was found to be suitable for isolating actinomycetes from marine sediments, which were collected from the coastal areas of Gokharna and Muradeshwara of Karnataka state.…”

Section: Discussioncontrasting

confidence: 42%

“…Four recommended actinomycetes culture media; Oatmeal (shofan) Nitrate Medium, Starch Casein Nitrate Medium, Starch Nitrate Medium and Potato Dextrose Medium were tested in order to select the most suitable culture medium for antitumor production [10][11][12]. The supernatant of each actinomycetes isolate was obtained using a micro-centrifuge for 15 min at 12000 rpm then undergo antitumor activity test.…”

Section: Isolation Of Different Actinomycetesmentioning

confidence: 99%

Production of Antitumor Agents from Novel Marine Actinomycetes Strains Isolated from Alexandria, Egypt

El-Naggar¹,

El-Assar²,

Shata³

2017

J Antimicrob Agents

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Production of antitumor agent(s) from alternative natural marine microbial resources may decrease or avoid the bad side effects of the used anti-tumour chemotherapeutic agents and may increase the specificity of the antitumor agents to be safer for human application. So, this study aimed to search for new alternative antitumor agent producers using two recommended bioassays (Red Potato Disc Bioassay and Sulforhodamine B Bioassay). The obtained results indicated the isolation of three new marine actinomycetes strains from the western harbor of Alexandria, they identified using the 16S rRNA gene sequence as Streptomyces sp. strain AMS11, Nocardiopsis halotolerans strain AMS10b and Nocardiopsis halotolerans strain AMS10a. The bio-toxicity test was carried out using the biomarker Artemia salina, they showed a moderate toxicity level (LC 50 <1000 ppm). The antitumor activities of their supernatants against the colon cancer cell line (HCT116) and the liver cancer cell line (HEPG2) showed promising results in comparison to the recommended chemotherapeutic drug 5-flourouracil used for the treatment of both liver and colon cancer cells. The optimization for antitumor activity was performed using the Plackett -Burman experimental design. [6,7]. The bio-toxicity of the potent supernatants was performed using the biomarker Artemia salina [8]. Finally, the optimization to reach the ideal conditions for maximum bioactivity was carried out using a Plackett-Burman experimental design [9]. Production of Antitumor Agents from Novel Marine Actinomycetes Strains Materials and Methods Isolation of different actinomycetesIsolation of different marine actinomycetes was carried out from 10 stations of the western harbor of Alexandria, Egypt. Four recommended actinomycetes culture media; Oatmeal (shofan) Nitrate Medium, Starch Casein Nitrate Medium, Starch Nitrate Medium and Potato Dextrose Medium were tested in order to select the most suitable culture medium for antitumor production [10][11][12]. The supernatant of each actinomycetes isolate was obtained using a micro-centrifuge for 15 min at 12000 rpm then undergo antitumor activity test. Screening for the antitumor activity Red Potato Disc bioassayThroughout this assay the potato discs has been infected by a tumor inducer bacterium "Agrobacterium tumefaciens". The examined potato discs were inoculated with 50 µl/disc of; B: sterile seawater, C 1 : A. tumefaciens suspension with OD=1 (the bacterial infection effect), C 2 : A. tumefaciens+1% Ampicillin (the T iplasmid effect only) and T: C 2 +the supernatant (1:2) of the tested actinomycetes The positive antitumor activity of each supernatant was detected by the results of five examined potato discs as replicates and confirmed through two successive experiments. All discs were incubated at 28°C for two weeks. The antitumor activity % was calculated according to Coker et al. [13] and El-Masry et al. [14]. The anti-tumor activity percentage was calculated after flooding the discs with the Gram iodine solution for one min. The ...

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“…Aluminum plates pre-coated with silica gel (20×20 cm, 0.25 mm Alugram® SIL G/UV 254, Macherey and Nagel, Duren) and two mobile phases [ethyl acetate : methanol (6:4) and Petroleum ether : Chloroform (1:1)] (Attimarad et al, 2012).…”

Section: Thin Layer Chromatography (Tlc)mentioning

confidence: 99%

Streptomyces: isolation, optimization of culture conditions and extraction of secondary metabolites

Khattab

Babiker²,

Saeed

2016

Int Curr Pharm J

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The objectives of this study were to isolate and identify Streptomyces from soil sediments as well as to optimize cultural growth conditions for maximum antibacterial productivity. A total of fifty soil sediments were collected from Red Sea, Sudan. The soil sediments were pretreated and cultivated on agar medium. Promising Streptomyces spp. were isolated by agar overlay method using indicator organisms. Optimization of chemical and physical culture conditions was carried out. The later was judged by assessment of antibacterial activity. Ethyl acetate was used to extract the secondary metabolite compounds. The separation of the active ingredients was performed using both thin layer chromatography (TLC) and gas chromatography-mass spectrometer (GC-MS). The results revealed nine strains of Streptomyces. Of them two (PS1 and PS28) isolates exhibited high activity against pathogenic bacteria. The optimum growth conditions were pH 7.5, temperature at 30°C, soyabean concentration 2.5 g/l, incubation period in 7 days, MgSO4.7H2O conc. 1g/l and K2HPO4 conc. 2.5g/l. TLC test showed three and two fragments from metabolites of PS1 and PS28 respectively, while the GC-MS analysis revealed eight and eleven compounds with antibacterial activity of PS1 and PS28 respectively. It is concluded that marine is promising source of secondary metabolites.Khattab et al., International Current Pharmaceutical Journal, February 2016, 5(3): 27-32

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Screening, isolation and purification of antibacterial agents from marine actinomycetes

Cited by 29 publications

References 13 publications

Antimicrobial Properties of Actinomycetes Isolated from Menengai Crater in Kenya

Antimicrobial Properties of Actinomycetes Isolated from Menengai Crater in Kenya

Production of Antitumor Agents from Novel Marine Actinomycetes Strains Isolated from Alexandria, Egypt

Streptomyces: isolation, optimization of culture conditions and extraction of secondary metabolites

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