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BACTERIOCIN PRODUCTION AND CHARACTERIZATION FROM LACTIC ACID BACTERIA FOR STRAIN DEVELOPMENT BY PHYSICO-CHEMICAL METHODS

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BACTERIOCIN PRODUCTION AND CHARACTERIZATION FROM LACTIC ACID BACTERIA FOR STRAIN DEVELOPMENT BY PHYSICO-CHEMICAL METHODS
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  BACTERIOCIN PRODUCTION AND CHARACTERIZATION FROMLACTIC ACID BACTERIA FOR STRAIN DEVELOPMENT BYPHYSICO-CHEMICAL METHODS Dedicated i  TOSomeone Who always encouraged me for the achievement of thisGoal of my life without whom it would be impossibleFor me to play the chess of life   TABLE OF CONTENT1INTRODUCTION 1 1.1) Inhibitory Characteristics Of acteria !pplied In Food FormationOr Fermentation....................................................................................................... "ii  1.#) !pplications Of acteriocin............................................................................. $ 1.#.1 Food conservation........................................................................................ $ 1.%) acterial Impedance..................................................................................... &  2. LITERATURE REVIEW ' (iterature eview********************* ' 3 MATERIALS AND METHODS ## %.1) Isolation Of acteriocin +roducing acterial ,train........................ ##%.1.1 ,election of strain.............................................................................. ##%.1.# +rocurement of samples............................................................. ##%.1.% ,terili-ation of glass ware......................................................... ##%.1." edia +reparation.................................................................. ##%.1./ ,ample preparation and dilutions............................................... #% %.1.$ Inoculation and Incubation..................................................... #%%.# acteriocin +roduction................................ #"%.#.1 +reparation of bacterial inoculum 0test organism)................................ #"%.%) !ntimicrobial !nalysis y Well +late ethod....... #"%.") Optimi-ation Of Growth Conditions.......................... #"0a) ,ensitivity of bacteriocin to p....................................................... #"0b) ,ensitivity of bacteriocin to heat.................................................. #/0c) ,ensitivity of bacteriocin to en-ymes........................................... #/%./) 2ffect of 3aCl on bacteriocin production................. #/%.$) +artial purification.............................................. #/ %.$.1 +artial purification by ammonium sulphate precipitation.................. #/%.&) +urification by gel filtration..................... #$a) ,welling of gel......................................................... #$ b) +ac4ing the column............................................ #$c) 25uilibration of the column.................................. #$iii  d) !pplication of sample.......................................... #$e) 2lution ......................................................... #$%.6) +urification by ion e7change chromatoghraphy ............. #&%.' !pplication of sample and elution........................ #&%.18 ,9,:+olyacrylamide Gel 2lectrophoraresis.............. #&%.11) ,train 9evelopment y ;sing +hysico:chemical utagenic !gents............................................................ #6 %.1#) ,tatistical !naysis............................................ #6 4. RESULTS .......................................................................... #6".1 +roduction Of acteriocin .................................... #6 ".# Characteri-ation Of acteriocin..................... #' a) 2ffect of temperature ................................................ #' b) 2ffect of p ...................................................... #'c) 2ffecet of en-ymes............................................ %8d) 2ffect of 3aCl....................................................... %8".%) +artial +urification.................................. %#a) +recipitation of protein by gel filtration pattern............ %% b) Ion e7change chromatography................................ %%"." ,train 9evelopment by ;sing +hysico:chemical utagenic !gents........................................................................................... %" 5. DISCUSSION .................................................................... %' REFERENCES ..................................................................... "# LIST OF TABLES <ables +age <able 0".#.a) 2ffect of temperature on the activity of bacteriocins................. #'<able ".# b. 2ffect of p on bacteriocin activity......................................... %8iv  <able ".# c. 2ffect of proteinase:=> chlroform and trypsin on bacteriocin activity against ,traphforiase............................................................................ %8<able ".# d. 2ffect of different concentrations of 3aCl on bacteriocin production by (actic acid  bacteria against ,trapforease.................................................. %1<able ".#. e. Grading of antimicrobial activity in symbolic and digitali-ed form along with their interpretation for the activity.................................................. %1FIG.".%.a. Gel filtration pattern of !mmonium ,ulphate precipitated residues of bacteriocin by lactic acid bacteria on ,ephade7 G:188.................................. %#<able ".% !ntimicrobial activity of different samples from different steps of purification samples of (actic acid bacterial species........................................................... %%<able ".". 9etails of dose and time of ;? e7posure for physical mutagenesis....... %/<able "./. 9 etails of dose of 3itrosoguanidine for chemical mutagenesis.............. %/<able ".$. Comparison of the amount of ?itamin 1# produced by parental and different physically mutant strains at %8@C........................................................ %$<able ".&. Comparison of the amount of ?itamin 1# produced by parental and different physically mutant strains at %#@C................................................. %$<able ".6. Comparison of the amount of ?itamin 1# produced by parental and chemically induced mutant strains at %8@C................................................. %&<able ".'. Comparison of the amount of ?itamin 1# produced by parental and chemically induced mutant strains at %#@C................................................ %& (I,< OF !2?I!<IO3, (! (actic acid bacteriaact acteriocinsv
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