Partial purification of Egg white Lysozyme from four different Bird species by Ethanol precipitation method and their Antibacterial Activity Assay: A Comparative study
Mainak Bhattacharjee*, Monojit Banerjee2, Dipankar Mahato3
Chowbaga Road, Ananadapur, P.O-East Kolkata Township, Kolkata-700107, West Bengal (India) E-mail-microman.mainak@gmail.com, Contact No.(M)- 09836492558, Former Lecturer, Department of Microbiology, Duragapur College of Commerce and Science (DCCS), G.T. Road, Rajbandh, Durgapur-713212, West Bardhhaman Distt., West Bengal (India) 2. Deptt. of Biochemistry, Triveni Devi Bhalotia (TDB) College, Raniganj, Dist-West Bardhhaman, West Bengal-713347 (India) 3. Department of Biotechnology, Duragapur College of Commerce and Science (DCCS), G.T. Road, Rajbandh, Durgapur-713212, West Bardhhaman Dist., West Bengal (India) *(Corresponding Author), Deptt.of Biotechnology, Heritage Institute of Technology,
ABSTRACT
In the present study, egg white lysozyme (EWL) was partially purified by ethanol precipitation method using varying concentrations of ethanol like 40%, 60% and 80% from the eggs of four different bird species including broiler-bred poultry hen, Aseel breed desi (country) hen, domesticated duck and pigeon in order to find out the most suitable concentration of ethanol for partial purification of EWL. In addition to this, a comparative anti-bacterial assay was also performed to determine the most effective EWL among the four different egg samples utilizing Bacillus subtilis as the test organism. It was found that 80% ethanol was most efficient for partial purification process of EWL when the incubation time is fixed for 6 hrs.after the ethanol treatment and prior to separation of EWL by centrifugation at acidic pH. It was also assessed that the EWL of broiler hen possessed the best lysozomal activity followed by Aseel hen, duck and pigeon. The egg white of common Indian domesticated pigeon was also found to exhibit lysozyme activity, although to a lesser extent. The lysozomal activities of the ethanolprecipitated crude EWLs were further confirmed by Lysoplate Assay using the cells of Micrococcus lysodeikiticus as the substrate. The study indicates that because of the considerably high lysozyme activity, the egg whites of poultry hen could be used as a potential source of pure lysozyme as the eggs are easily available and much cheaper. The eggs of Aseel breed of hens also contain potential lysozomal activity but they are not economically viable for mass scale production of lysozyme for their higher cost and lower availability.