Page 55 - 2019
P. 55

Ph.D.
                                                                                     (Science)
          ISOLATION AND CHARACTERIZATION OF HALOPHILIC BACTERIA
          AND THEIR ENZYME PROFILING

          Ph.D. Scholar : Patel Gayatriben Bharatbhai
          Research Supervisor : Dr. Nikunjana A. Patel



                                                                                Regi. No.: 13146061002
          Abstract :
          The Narara Island is supersaturated with NaCl making ever more extreme environment
          for microbial life. Total twenty five isolates of Halophilic and Halotolerant bacteria were
          isolated from the salted area, mangroves area, and near creek area of Narara, Jamnagar,
          India. All the isolates showed different cultural characteristics like size, shape, elevation
          and pigmentation etc. The isolates were characterized by biochemical, physiological and
          genetic  characterization.  The  isolates  were  also  checked  for  extracellular  enzyme
          production in presence of 15% and 20% of sodium chloride concentration on solid agar
          medium  supplemented  with  specific  substrates.  Isolate  GB24  was  identified  as  a
          Halobacterium  sp.  and  GB18  were  shows  prominent  Amylase  activity  in  presence  of
          NaCl.  Isolate  GB6  was  identified  as  a  Virgobacillus  sp.  was  produced protease  activity
          while isolate GB12 shows lipolytic activity. The crude enzymes from each isolates were
          partially purified by Ammonium sulphate precipitation and Gel filtration chromatography
          using sephadex-G25 column. The organic solvent stability and detergent compatibility of
          purified enzymes were checked in presence of sodium chloride. The molecular mass of
          purified enzymes from different fractions was analyzed by SDSPAGE analysis. Based on
          16s rRNA sequencing isolates GB24 and GB6 showed 99% similarity with Halobacterium
          sp. and Virgobacillus sp. respectively. Isolate GB18 is similar to Halomonas sp with 63%
          similarity  and  isolate  GB12  shows  82%  relatedness  to  Marinobacter  sp.  so  it  may  be
          extended for further confirmation using detail genetic analysis. This work may be further
          explored to extend for the biotechnological applications in different fields such as color
          industries, paper industries, pulp industries, detergent industries etc

          Keywords:  Halophiles,  Enzyme  profiling,  16s  rRNA  sequencing,  Gel  filtration
          chromatography, SDS –PAGE analysis













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