National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
Marine water samples were collected from a coastal region of Pushpavanam, Nagapattinam District, Tamil Nadu, India. Marine Pseudomonas and Proteus were isolated and identified according to morphology, biochemical test and selective complex marine growth medium. The identified marine organisms were maintained with LB broth. The cultures were harvested by cooling centrifugation at 9,000x g for 20 min. This suspension was incubated at 28ºC for 24 hours after incubation the cells removed by centrifugation method. The supernatant was filter sterilized and antibacterial activity was zone of inhibition performed by measuring overlay assay technique and drop test assay method. Samples were suspended in sucrose – dye – solution and identification of antibacterial protein band resolved by polyacrylamide gel electrophoresis. The gel was washed with distilled water. The protein fraction was identified by the drop test. All the proteins were assayed with a bicinchoninic acid protein kit. The protein fraction elution was done with phosphate buffer solution. The protein fraction was stored at 20ºC after freeze – drying. The antibacterial protein was detected within SDS-PAGE treatment with Coomassie brilliant blue. The concentration level of antibacterial protein is necessary to result in cell death and to inhibit the growth of marine microorganisms in suitable medium. Bacterial cells were inoculated at time zero and the bacterial growth rates and cell viability were measured by colony forming unit.
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