أ.و.د. اخر هاشى كاظى و ب ذاء يهذي ػباس خايؼت كزبالء / قسى ػهىو انس اة / كه ت انؼهىو ӿ ا جؾش غز سعب خ بعغز ١ ش جبؽش ا ضب
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- Cecily Bell
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1 / ػه / 216 Optimization of condiction for Exopolysaccharid(EPS) production from Lactobacillus Plantarum localy isolated تسذ ذ انظزوف ان ثهى أل تاج يتؼذد انسكز اث انخارج خهىي Exopolysaccharid EPS) ( ي بكتز ا Lactobacillus Plantarum ان ؼزونت يسه ا ӿ 1 أ.و.د. اخر هاشى كاظى و ب ذاء يهذي ػباس 1 خايؼت كزبالء / قسى ػهىو انس اة / كه ت انؼهىو ӿ ا جؾش غز سعب خ بعغز ١ ش جبؽش ا ضب انخالطت دسعذ ا ظش ف ا ضسػ ١ خ ا ج ١ ئ ١ خ ا ض ال زبط زؼذد ا غىش ٠ بد ا خبسط خ ا ؼض خ ا ؾ ١ خ Lactobacillus Plantarum ا زخجخ ف دساعخ عبثمخ أ ػؾذ ا زبئظ ا أفؼ ز ا ظش ف وب ذ ثبعزؼ بي ا عؾ اإل زبع simplified synthetic medium ا ز ٠ ؾز ػ ػظ ١ ش ا ز ش ا ض ذ ثزشو ١ ض %5 )عىش ٠ بد خزض خ( ظذسا ىبسث غزخ ض ا خ ١ شح ثزشو ١ ض %.2 ظذسا ١ زش ع ١ دػ زا ا عؾ ث)وجش ٠ زبد ا غ ١ غ ١ ف عفبد ا ج ربع ١ ) ثزشو ١ ض و %.9 ظذسا أل الػ ا ؼذ ١ خ ػذي ا شل ا ١ ذس ع ١ ( ) ph االثزذائ ا 6.5 ؽغ مبػ %3 ؽغ / ؽغ ا عؾ ا ؾؼ ثظش ف ال ائ ١ خ ثذسعخ ؽشاسح 35 ذح 24 عبػخ ر اعزخالص EPS ثبعزؼ بي اال ٠ ضب ي ثزشو ١ ض %95 ا ى بد ا فزبؽ ١ خ : زؼذد ا غىش ٠ بد ا خبسط خ, الوز ثبع ظ ثال زش, ث ١ ش ؽ ١ Abstract The optimum cultural and ecological condition for Exopolysaccharid Production from Lactobacillus PLantarum localy isolated were studied Results revealed that, using production medium ( simplified synthetic medium)containing date juice (5%) as a carbon source and yeast extract (.2%) as a nitrogen source, the media was supplemented with a total concentration.9% of (magnesium sulphate and potassium phosphate ) as mineral salts. The primary ph was adjusted to 6.5 and the inoculum size was 3%(v/v), isolate were grown under anaerobic condition at 35 C o for 24 h and the EPS extracted with ethanol 95%. Key words: Exopolysaccharid, Lactobacillus Plantarum, biopolymer 1 -ان قذيت أد ا ط ت ا زضا ٠ ذ ف ا غ اد األخ ١ شح زؼذداد ا طج ١ ؼ ١ خ ف خز ف ا زطج ١ مبد ا ظ بػ ١ خ ا زم ١ بد ا ؾ ٠ ١ خ ا ص ٠ بدح اال ز ب ف ا زبط ا زؼذداد ا غىش ٠ خ ا خبسط خ ٠ خ (EPS) Exopolysaccharide األؽ ١ بء ا غ ش ٠ خ, رغزخذ ز ا زؼذداد و اد ا زظبص ؽ ١ غ فخ ػ ا إلصا خ اال ٠ بد ا ضم ١ خ فؼال ػ و ب ػ ا ؽب خ ؼالعبد) Cariar ( سار غبد زجبدي األ ٠ ) ( ion exchang resins (1). ٠ زظ EPS ا جبربد ا طؾب ت االؽ ١ بء ا غ ش ٠ خ ث ١ ذ أ اعزؼ بي االؽ ١ بء ا غ ش ٠ خ ال زبط ٠ ؼذ EPS أوضش الء خ ال ػ ائذ ا ؼ ١ خ اإل زبع ١ خ عش ٠ ؼخ ثؾظ ١ خ وج ١ شح ف ظش ف رخ ش غ ١ طش ػ ١ ب إر رغزغشق ػ ١ خ اإل زبط أ ٠ ب ب أ أعبث ١ غ ا ب ف ا جبربد فزغزغشق ػ ١ خ ا زبط ػذح أش ش, فؼال ػ ا زغ ١ شاد ا ع ١ خ ا غغشاف ١ خ ا ز لذ رؤصش ػ ا جبربد )2 ). و ب ا EPS ا زظ األؽ ١ بء ا غ ش ٠ خ خظبئض س ٠ ع ١ خ ع ١ ذح, وز ه مب خ ع ١ ذح زؾ ف دسعبد ا ؾشاسح ا ؼب ١ خ ا ضجبد ف ذ اعغ ا شل ا ١ ذس ع ١, ا أػزجبس ا ذوغزشا ا ضا ضب ا زظ ا جىزش ٠ ب آ غزائ ١ ب إداسح األغز ٠ خ ا ؼمبل ١ ش األ ش ٠ ى ١ خ ذ ا طش ٠ ك ذ اعغ ا زطج ١ مبد EPS ا ب ٠ ىش ث, ا ا غذ األلزظبد ٠ خ ا شرجطخ غ اال زبط ا ظ بػ ا ؼب ا شئ ١ ظ ا ز ٠ ؾذ اعزؼ بيEPS. إر ا غب جب ب ٠ ظبؽت رغ ٠ ك EPS ا ١ ىش ث ػ ١ خ إ زبع ١ خ ؽ ٠ خ ى فخ ؼ ب ر افم ب عال ز ب و ؼبفبد غزائ ١ خ ز ه فأ ث ١ األ اع ا جىز ١ ش ٠ خ ا خز فخ ا زغخ EPS أعزمطجذ ثىزش ٠ ب ؽب غ ا الوز ١ ه ا ز ب ب خبطب ر ه خظبئض زؼذد ا غىش ٠ بد ا خبسع ا ز ر زغ و ال ٠ ؾ أ خبؽش طؾ ١ خ وز ه فأ ثىزش ٠ ب ؽب غ ا الوز ١ ه رؼزجش آ خ غزائ ١ ب ( GRAS (. وز ه EPS ا زظ LAB رطج ١ مبد اعؼخ خ ف رؼض ٠ ض ا خظبئض ا ش ٠ ع ١ خ ا م ا ا زاق زغبد ا ؾ ١ ت ا خ شح ا ز رزؼ األ جب األعجب غ ١ ش ب (3). 31
2 / ػه / 216 فؼال ػ ب عجك ٠ غزؼ EPS ا ز ر زغ ثىزش ٠ ب ؽب غ ا الوز ١ ه ث طف ؾفضاد ؽ ٠ ١ خ prebiotic( ) رؼ ػ رؾف ١ ض ا ج ١ ذ ا طج ١ ؼ أل ؼبء ص ٠ بدح أػذاد ا جىزش ٠ ب ا ف ١ ذح ف األ ؼبء ض ثىزش ٠ ب Bifidobacterium ب ٠ شغغ ػ رضج ١ ؾ ا شػبد ا ١ ىش ث ١ خ ا زظبل ب (4 ; 5) و ب ٠ زبص ثمبث ١ ز ػ ا جمبء ف ا م بح ا ؼ ١ خ ذح ؽ ٠ خ ب ٠ ز ١ ؼ ا فشطخ زؾف ١ ض اعز ١ طب ثىزش ٠ ب ا ؼضصاد ا ؾ ٠ ١ خ) propbiotic ) )6(. ػ ا شغ ر ه فأ ا زؼذداد ا ؾ ٠ ١ خ ا طج ١ ؼ ١ خ الرضاي ؾذدح ثغضء طغ ١ ش أع اق ا زؼذداد ا ؾب ا غجت ٠ ؼ د ا ا زى فخ اإل زبع ١ خ ا ؼب ١ خ, ز ه ذفذ ز ا ذساعخ ا ا زبط EPS ثىزش ٠ ب ؾ ١ خ دساعخ ا ظش ف ا ض أل زبط أ ىب ١ خ إ زبع خالي اعزؼ بي اد أعبع ١ خ سخ ١ ظخ ا ض ز فشح عبءد ز ا ذساعخ ز بش ١ خ غ ا ذف. 2 -ان ىاد و طزائق انؼ م انؼزنت انبكت ز ت أعزخذ ذ ا ؼض خ ا جىز ١ ش ٠ خ.L Plantarum ا ؼض خ ا شخظخ ف دساعخ عبثمخ ال زبط EPS ب ا تاج EPS ي بكتز ا Lactobacillus Plantarum أعزؼ عؾ medium( )Simplified synthetic ؽغت ا طش ٠ مخ ا ط فخ لج ) 9 ( و عؾ ال زبط EPS ثىزش ٠ ب.L Plantarum إر ٠ زى زا ا عؾ ا اد ا زب ١ خ )غ / زش ) ف عفبد ا ظ د ٠ أؽبد ٠ خ ا ١ ذس ع ١ ) ) Na 2 HPO 4 2 غ, وجش ٠ زبد ا غ ١ غ ١ 6 غ, عزشاد األ ١ ا ضالص ١ خ 5 غ, ف عفبد ا ج ربع ١ ص بئ ١ خ ا ١ ذس ع ١ ( 4 ) KH 2 PO Trace element.إر ا 1 Trace element solution 1 غ, عىش ص 5 غ ؾ ي األ الػ ا ضسح ) MgSO 4 ( ٠ زى ا اد ا زب ١ خ زاثخ ف زش ا بء ا مطش ( وجش ٠ زبد ا ؾذ ٠ ذ ) 4 ) FeSO 5 غ,وجش ٠ زبد ا غ ١ ض solution ( 4 2)MnSOغ, و س ٠ ذا ض ه 1 غ, و س ٠ ذ ا ى ث ذ 1 غ. ص صع ا عؾ ف ػج اد صعبع ١ خ Vials( ) ؼم خ عؼخ 2. ثأػبفخ 2 ب ٠ ىش ١ زش ث الغ 9.98 ى ػج ح ثضالس ىشساد ى ػضي مؼ استخالص يتؼذد انسكز اث انخارخ أرجؼذ ا طش ٠ مخ ا ط فخ لج )1 ) ف اعزخالص زؼذد ا غىش ٠ بد ا خبسع غ ثؼغ ا زؾ ٠ شاد إر ثؼذ ا ز بء ذح ا ؾؼ ر ػغ عؾ ا زخ ش ف ؽ ب بئ ثذسعخ 9 ئ ٠ خ ذح 1 دلبئك ر فظ ا خال ٠ ب ػ عؾ ا زخ ش ثبعزخذا ع بص ا طشد ا شوض ثغشػخ 8 د سح / دل ١ مخ ذح 1 دلبئك, عففذ ا خال ٠ ب غشع ؽغبة ا ص ا غبف ىز خ ا ؾ ٠ ١ خ ف ؽ ١ أعزؼ ا شاشؼ إر أػ ١ ف ا ١ TCA( Trichloroacetic acid ) ثزشو ١ ض % 8 )ؽغ / ؽغ ) رشن ذح 3 عبػبد ثذسعخ 4 ص أعش ٠ ذ ػ ١ خ ؽشد شوض ثغشػخ 8 د سح / دل ١ مخ ذح 1 دلبئك ر ه زشع ١ ت ا جش ر ١ ا ع د ف ا عؾ ثؼذ ر ه ا ا شاعت أخز ا ششؼ أػ ١ ف ا ١ ؽغ ١ اإل ٠ ضب ي ا جشد ثزشو ١ ض %95 رشن ثذسعخ 4 ذح 24 عبػخ ص فظ EPS ث عبؽخ ا طشد ا شوض ثغشػخ 12 د سح / دل ١ مخ ذح 12 دل ١ مخ إر أ ا شاشؼ عفف ا شاعت ثذسعخ 4 ذح 24 عبػخ غشع ؽغبة ا ص ا غبف. إل زبط ر اعزؼ بي عؾ Simplified synthetic medium ىع ان ذ ب ان ستخذو ف استخالص وتزس ب :EPS EPS ثؼذ ا ز بء ذح ا ؾؼ ر اعزؼ بي ػذد ا ز ٠ جبد ( األ ٠ ضب ي ا ١ ضب ي األع ١ ز ا ى س ف س أع ١ ز ف س ب ذ ب ٠ ذ اال ٠ ض ثش ثب ي ) زشع ١ ت. EPS ص اعزؼ ذ رشاو ١ ض خز فخ اال ٠ ضب ي زشع ١ ت EPS ا شاشؼ ا غزخ ض إر ؽؼشد رشاو ١ ض خز فخ اإل ٠ ضب ي ( 35, (,95,8,65 5 % ر أػبفخ ؽغ ١ و رشو ١ ض ا ؽغ ا شاشؼ ا غزخ ض. تسذ ذ انظزوف ان ثهى إل تاج EPS ي انؼزنت ان تخبت أ عؾ : ر اخزجبس صالس أ عبؽ إ زبع ١ خ شبئؼخ االعزؼ بي إل زبط EPS ثىزش ٠ ب L.Plantarum ىع انىسظ اإل تاخ :.. (Skim Milk Medium) عؾ ) MRS عؾ) broth Simplified synthetic medium 35 فزشاد مؼ عؾ اإل زبط ثؾغ مبػ %2 ػغ ف ا ؾبػ خ ف ظش ف ال ائ ١ خ ثذسعخ ؽشاسح تأث ز يذة انسض : عبػخ. ص ١ خ خز فخ 18( ( 96, 72, 48, 24, ذح 24 عبػخ ثظش ف ال ائ ١ خ ف ا مؼ ثجىزش ٠ ب L.Plantarum ؽؼ عؾ ا زبط EPS تأث ز درخت ززارة انسض : دسعبد ؽشاسح ؽؼ خز فخ )3,25,45) 4, 37,35, زؾذ ٠ ذ دسعخ ا ؾؼ ا ض إل زبط. اعزؼ ذ ظبدس وبسث ١ خ خز فخ ( ػظ ١ ش ا ز ش ا ششػ ا غىش ص ا فشوز ص ا ب ز ص ىع ان ظذر انكاربى وتزك ز : ا ىبالوز ص ػذ ع د ظذس وبسث ) ج ١ ب رأص ١ ش ا ظذس ا ىبسث ف ا زبط.EPS ص اعزؼ ذ رشاو ١ ض خز فخ ػظ ١ ش ا ز شا ض ذ ا ز اعزؼ و ظذس ىبسث ف ا عؾ اال زبع ١ ؼط رشاو ١ ض زذسعخ ا غىش ٠ بد ا خزض خ ( 2.5,,7.5,5 % زؾذ ٠ ذ ا زشو ١ ض األ ض ػظ ١ ش ا ز ش إل زبط EPS ا ؼض خ ا زخجخ إر لذسد ا غىش ٠ بد ا خزض خ ف عؾ,12.5) 1 ػظ ١ ش ا ز ش ثطش ٠ مخ.Miller,(1959) ىع ان ظذر ان تزوخ وتزك ز : اعزؼ ذ وجش ٠ زبد األ ١ عزشاد األ ١ ا ضالص ١ خ غزخ ض ا خ ١ شح ا زشثز ا ١ س ٠ ب و ظبدس ١ زش ع ١ ١ خ ف ا عؾ اإل زبع,ص اعزؼ ذ رشاو ١ ض زذسعخ غزخ ض ا خ ١ شح ).,,,.3,.2.1 (.5.4, % زؾذ ٠ ذ ا زشو ١ ض األ ض ا ظذس ا ١ زش ع ١ إل زبط EPS ا ؼض خ ا جىز ١ ش ٠ خ ا زخجخ. ىع األيالذ ان ؼذ ت وتزك زها: دسط رأص ١ ش وجش ٠ زبد ا غ ١ غ ١ ا بئ ١ خ ) ) MgSO.4 7 H 2 ف عفبد ا ج ربع ١ ص بئ ١ خ ا ١ ذس ع ١ ( KH 2 PO 4 ) ؾ ي ا ؼ بطش ا ضسح solution) ) Trace element و ظبدس ال الػ ا ؼذ ١ خ زؾذ ٠ ذ ع 32
3 ملغم / EPSملغم / EPS لتر لتر / ػه / 216 اال الػ ا ؼذ ١ خ األفؼ إل زبط, EPS ص اعزؼ ذ وجش ٠ زبد ا غ ١ غ ١ ف عفبد ا ج ربع ١ ص بئ ١ خ ا ١ ذس ع ١ ؼب و ظذس أل الػ ا ؼذ ١ خ ث غت خز فخ انزقى انه ذروخ االبتذائ : صع ا عؾ اإل زبع ف د اسق صعبع ١ خ ػذي ا شل ا ١ ذس ع ١ ف ز ا ذ اسق ا )4.5 ),7.5,7,6.5 6, 5 5.,,5 ثؼذ ر م ١ ؾ ثب ؼض خ ا زخجخ ؽؼ ا عؾ ف ظش ف ال ائ ١ خ ثذسعخ ؽشاسح ؽؼ 35 ذح 24 عبػخ ص أعزخ ض EPS ا عؾ عفف ص ص زؾذ ٠ ذ ا شل ا ١ ذس ع ١ األ ض ال زبط. EPS زدى انهقاذ : مؼ ا عؾ اال زبع ثؾغ مبػ خز فخ 1( ) 5, 4, 3, 2, % ؽغ ا عؾ ص أعزخ ض EPS ا عؾ ر رغف ١ ف ص ج ١ ب رأص ١ ش ؽغ ا مبػ ف ا زبط EPS ا ؼض خ ا زخجخ. انتسه م األزظائ أعزؼ ا زظ ١ ا ؼش ائ ا ىب ( Design ) Completely Randomized ار أػز ذد ل ١ خ ( )L.S.D. ػ غز اؽز ب ١ خ.1 ف ع ١ غ ا زغبسة )11 (. ان تائح وان اقشت 1 - ىع ان ذ ب ان ستخذو وتزك ز : ٠ زؼؼ ا شى ) 1 ( ا و ١ خ EPS ا غزخ ض وب ذ )835 ) غ / زش ػ ذ اعزؼ بي اال ٠ ضب ي )38, ) 26 غ / زش ػ ذ اعزؼ بي) االع ١ ز ا ف س ب ذ ٠ ب ٠ ذ( ػ ا ز ا ف ؽ ١ ا ا ١ ضب ي ا ى س ف س اال ٠ ض ثش ثب ي ٠ ظ ش أ رأص ١ ش ف رشع ١ ت. EPS رزفك زبئظ ز ا ذساعخ غ ب عذ )14( ار أػط اال ٠ ضب ي أػ فؼب ١ خ زشع ١ ت EPS ا غزخ ض ثىزش ٠ ب Bacillus subtilis ثب مبس خ غ ا ز ٠ جبد األخش ا غزؼ خ ف ر ه ا ذساعخ. اعزؼ ذ رشاو ١ ض خز فخ اال ٠ ضب ي ( 35 ) 1, 95, 8, 65, 5, % العزخالص EPS ا عؾ ا شى )2( ٠ زؼؼ ا أفؼ رشع ١ ت EPS ػ ذ إػبفخ اال ٠ ضب ي ثزشو ١ ض( 1), 95 % أعزؼ ا زشو ١ ض %95 ف ا زغبسة ا الؽمخ ؼذ ع د فشق ؼ المذيبات L.S.D= انشكم )1( :تأث ز ىع ان ذ ب ان ستخذو ألستخالص EPS ان تح ي بكتز ا.L plantarum EPS mg/l 1 % تركيز االيثانول L.S.D = انشكم )2(: تأث ز تزك ز اال ثا ىل ان ستخذو ف استخالص EPS ان تح ي بكتز ا.L plantarum 33
4 الوزن الجاف للEPS ملغم/لتر نتز يهغى/ / ػه / 216 ؾ EPS ثىزش ٠ ب.L plantarum ثبعزؼ بي صالصخ أ اع األ عبؽ اإل زبع ١ خ إر أظ شد ل س ا زبط 2 - ىع انىسظ اإل تاخ : أظ شد ا زبئظ ا ج ١ خ ف ا شى )3( ا أػ ا زبط زؼذد وب ثبعزؼ بي عؾ.Simplified synthetic medium زبئظ دساعبد أخش ا ؽظ ١ خ ا زبط EPS رؼز ذ ػ ع ا عؾ اإل زبع,فمذ ث ١ (12 ( ا ر ١ خ ف ع ا ششػ ا ذػ ث غزخ ض ا خ ١ شح ا غىش ص اػط ثىزش ٠ ب Lactobacillus plantarum,strain 3112S ا غزخ ض ثىزش ٠ ب.L plantarum ثبعزؼ بي عؾ ث ١ ب ث غذ و ١ خ EPS ؽظ ١ خ ا زبط EPS ث غذ 145 غ / زش MRS ا غبئ 659 غ / زش) 13 ( Simplified MRS synthetic medium الوسط االنتاجي skim milk L.S.D = انشكم )3( :تأث ز ىع انىسظ ف ا تاج EPS ي بكتز ا.L plantarum 3 -تأث ز درخت ززارة انسض : دسط رأص ١ ش دسعخ ؽشاسح ا ؾؼ ف ا زبط EPS ثىزش ٠ ب.L Plantarum أظ شد ا زبئظ ا ػؾخ ف ا شى )4( ا أػ ا زبط EPS ػ ذ دسعخ ؽشاسح 35 ار ث غذ و ١ خ EPS ا زظ 936 غ / زش, ب ٠ ش ١ ش ا ا دسعخ ا ؾشاسح ا ض ال زبط 35, ا شى )4( ٠ الؽع ا دسعخ ا ؾشاسح ا ض ا جىزش ٠ ب 37 ر ه ٠ زج ١ ثأ افؼ ا زبط ٠ ى EPS ف دسعبد ؽشاس ٠ خ أل ا ذسعخ ا ض ا ىبئ ا غ ش ا زظ EPS. ٠ ى رفغ ١ ش ر ه ثأ رجبؽئ ا خال ٠ ب ال خفبع دسعخ ا ؾشاسح ٠ ؤد ا رجبؽئ رظ ١ غ ث ١ شاد ا غذاس ا خ ثز ه فب ف عفبد األ ٠ ض ثش ٠ ٠ ذ isoprenoid phosphate عزى ز فشح زؼ و بل ف رظ ١ غ )EPS 15 ). 4 -تأث ز يذة انسض :ر ؽغبة ا ص ا غبف EPS ا غزخ ض ثىزش ٠ ب Lactibacillus plantarum خالي فزشاد ص ١ خ خز فخ و ب ج ١ ف ا شى )5(, ٠ الؽع ا ا ألظ ا زبط EPS وب ثؼذ 24 عبػخ ا ضسػخ ار ط ا 935 غ / زش ز ا ز ١ غخ رزفك غ ب عذ ) 16 ( ا ز روش ا ػ ذ ر ١ خ ثىزش ٠ ب Bifidobacterium longum subsp. longum CRC 2 ف عؾ ٠ ؾز ػ ا الوز ص و ظذس ىبسث فب أػ ا زبط EPS وب ثؼذ فزشح ؽؼ 24 عبػخ إر ث غ 18 غ / زش. ثؼذ ذح ا ؾؼ ا ضب ١ خ ثذأ ا زبط EPS ثبال خفبع رذس ٠ غ ١ ب ؽز ا خفغ ا 21 غ / زش ثؼذ 96 عبػخ ا ؾؼ.ا ا خفبع ا زبط EPS ف ذح ا ؾؼ ا ط ٠ خ ٠ ؼض ا ع د اال ض ٠ بد ا ؾ خ degradation( )enzymes ا ز رغجت رفىه ا زؼذد ا غىش )17 ) درخت انسزارة L.S.D = انشكم )4( :تأث ز درخت ززارة انسض ف ا تاج EPS ي بكتز ا.L Plantarum 34
5 لتر / ملغم لتر / ملغم / ػه / يذة انسض )ساػت ) L.S.D = انشكم )5( :تأث ز يذة انسض ف ا تاج EPS ي بكتز ا.L plantarum تأثير نوع المصدر الكاربوني وتركيزه : أظ شد ا زبئظ ا ج ١ خ ف ا شى )6( ا أػ ا زبط EPS وب ػ ذ اعزؼ بي ا غىش ص و ظذس ىبسث إر ث غذ و ١ خ اإل زبط 941 غ / زش ث ١ ب ث غذ و ١ خ EPS ا زغخ ػ ذ اعزؼ بي ػظ ١ ش ا ز ش و ظذس ىبسث 887 غ / زش, ٠ جذأ اإل زبط ثبال خفبع اعزؼ بي ا ظبدس ا ىبسث ١ خ األخش ر ه زبئظ ٠ زؼؼ ا ػظ ١ ش ا ز ش ا ض ذ ٠ ؼذ ا ظذس ا ضب ؽ ١ ش األ ١ خ ف ا زبط EPS كاالكتوز سكروز الشرش فركتوز مالتوز عصير التمر يظذر انكاربى بدون وجود مصدر كاربون L.S.D =1.334 انشكم )6( : تأث ز ىع ان ظذر انكاربى ف ا تاج EPS ي بكتز ا.L plantarum ٠ زج ١ ا زبئظ أػال ا ا زبط ا زؼذداد ا غىش ٠ خ ا خبسط خ ٠ خ ٠ خز ف ثأخزالف ع ا ظذس ا ىبسث لذ أش ١ ش ا ر ه ف دساعبد أخش فمذ روش )18( ا ا أػ ا زبط EPS أعزؾظ ػ ١ ػ ذ ر ١ خ ثىزش ٠ ب Streptococcus thermophiles LY3 ال زبط ا زؼذد ف عؾ ٠ ؾز ػ ا الوز ص و ظذس ؽ ١ ذ ىبسث. ٠ الؽع ب عجك روش اخزالف ا ظبدس ا ىبسث ١ خ ا بعجخ ال زبط EPS ثبخزالف ا ىبئ ا غ ش ػض ( 19) عجت ر ه ا اخزالف لبث ١ خ األؽ ١ بء ا غ ش ٠ خ ػ اعز الن ا بدح األعبط ا غبس ا ؾ ١ ا ز رغ ى, إػبفخ ا ر ه فب خط اد األ ٠ غ األػبف ١ خ ا ز رذخ ف رخ ١ ك EPS ا ظبدس ا ىبسث ١ خ ا خز فخ لذ رؤصش ف ؼذي ا زبط. EPS مذ وب بن ا ز ب ب وج ١ شا العزؼ بي ا ظبدس ا خب ف دساعبد ا زبط EPS األؽ ١ بء ا غ ش ٠ خ ظشا أل و فخ ا بدح األعبط ا غزخذ خ و ظذس ىبسث ف ا عؾ ا ز ر ف ١ األؽ ١ بء ا غ ش ٠ خ ٠ شى ػجئب ػ ا ى فخ ا ى ١ خ ف ا زبط EPS ز ه فمذ ر اعزؼ بي ػظ ١ ش ا ز ش و ظذس ىبسث ف ع ١ غ ا زغبسة ا الؽمخ. ا ب ثب غجخ زأص ١ ش رشو ١ ض ػظ ١ ش ا ز ش و ظذس ؽ ١ ذ ىبسث ف ا زبط EPS فمذ أظ شد ا زبئظ ا ج ١ خ ف ا شى )7( ا أػ و ١ خ ا زبط EPS ػ ذ ا زشاو ١ ض ( 5 ) 7.5 % إر ث غذ ( 884 ) 893 غ / زش ؼذ ع د فشق اؽظبئ ؼ ث ١ ب فمذ اعزؼ ا زشو ١ ض %5 وأفؼ رشو ١ ض ػظ ١ ش ا ز ش ر اعزخذا ف ع ١ غ شاؽ ا ذساعخ ا الؽمخ. لذ أ ػؼ (2) ا عجت ا مظب ف و ١ خ EPS ا زظ ا ىز خ ا ؾ ٠ ١ خ جىزش ٠ ب Rhodotorula glutinis ا بح ف عؾ ٠ ؾز ػ ا ى و ص و ظذس ىبسث ثأ ا ض ٠ بدح ف رشو ١ ض ا ى و ص ف ا عؾ ا ضسػ ا غبئ ٠ ؤد ا ص ٠ بدح ض عخ ا عؾ ثغجت ص ٠ بدح ا زؼذد ا غىش ا خبسع ا زغ غ ا ا ض ٠ بدح ف ض عخ ا عؾ رؤد ا ل خ األ وغغ ١ ا ز فش ر ص ٠ غ غ ١ ش طؾ ١ ؼ اد ف ا عؾ ( mixing )imperfect رضج ١ ؾ ا م ث ١ ا خال ٠ ب ا اد ا غزائ ١ خ ثب زب مظب ف ا زبط.EPS
6 لتر / ملغم نتز / يهغى / ػه / ا ص ا غبف خال ٠ ب ص EPS تركيز عصير التمر % L.S.D = انشكم )7(: تأث ز تزك ز ػظ ز انت ز ف ا تاج EPS ي بكتز اplantarum.L 6 -تأث ز ىع ان ظذر ان تزوخ وتزك ز :أظ شد ا زبئظ ا ػؾخ ف ا شى )8( ا غزخ ض ا خ ١ شح ا ظذس ا ١ زش ع ١ األوفأ ف ا زبط EPS مبس خ ثجبل ا ظبدس ا ١ زش ع ١ ١ خ ا غزخذ خ ار ث غذ و ١ خ ا زؼذد ا زظ 998 غ / زش ث ١ ب وب ال ا زبط زؼذد ػ ذ اعزؼ بي ا ١ س ٠ ب و ظذس ١ زش ع ١ ف ا عؾ إر ث غذ 29 غ / زش زا فمذ ر اػز بد ز ا ز ١ غخ ر اعزخذا ب ف ع ١ غ شاؽ ا ذساعخ ا الؽمخ بدون اضافة مصدر نيتروجين كبريتات األمونيو م سترات األمونيوم الثالثية تربتون يوريا مستخلص الخميرة يظذر ان تزوخ L.S.D = انشكم )8( : تأث ز ىع يظذر ان تزوخ ف ا تاج EPS ي بكتز ا.L plantarum لذ ٠ ؼض ر ه ا اؽز اء زا ا ظذس ا ١ زش ع ١ ا ؼؼ ػ األؽ بع األ ١ ١ خ ا ججز ١ ذاد األ الػ ا ىض ١ ش ػ ا ا ا ز رؤصش ف ا ىبئ ا غ ش إ زبطEPS )21). ؾبعخ ثىزش ٠ ب Lactobacilli ا ا ف ١ زب ١ بد ا خز فخ إ زبط )22( EPS فأ غزخ ض ا خ ١ شح ظذس بعت ١ زش ع ١ أل ب ر فش ا ى بد األعبع ١ خ ض ا ف ١ زب ١ بد األؽ بع ا ٠ خ )13(. ثؼذ ا ر رؾذ ٠ ذ ا ظذس ا ١ زش ع ١ األ ض أل زبط EPS دسط رأص ١ ش رشاو ١ ض خز فخ غزخ ض ا خ ١ شح زؾذ ٠ ذ ا زشو ١ ض األ ض أل زبط ا زؼذد, لذ ث ١ ذ ا زبئظ ا ػؾخ ف ا شى )9( ا ا زبط EPS ثذأ ٠ شرفغ ثض ٠ بدح رشو ١ ض غزخ ض ا خ ١ شح ف ا عؾ إر وب أػ ا زبط زؼذد ػ ذ اعزؼ بي غزخ ض ا خ ١ شح ثزشو ١ ض).3(.2 % ار ث غ ( ( غ / زش ػ ا ز ا اػز بدا ػ ا زبئظ ا غزؾظ ػ ١ ب فمذ ر اعزؼ بي غزخ ض ا خ ١ شح ثزشو ١ ض %.2 ف شاؽ ا ذساعخ ا الؽمخ ؼذ ع د فشق ؼ ف اإل زبط ػ ذ اعزؼ بي ا زشو ١ ض ٠ ( ).3.2 %. 36
7 زش / غ لتر / ملغم / ػه / تركيز مستخلص الخميرة% L.S.D = انشكم )9(: تأث ز تزك ز يستخهض انخ زة ف ا تاج EPS ي بكتز ا.L plantarum ا ا ز ١ غخ ا غزؾظ خ ز ا ذساعخ رزفك غ ب سد ف ا ؼذ ٠ ذ ا ذساعبد فمذ أظ شد زبئظ دساعخ )24( ا ػ ذ ر ١ خ ثىزش ٠ ب Rhodotorula glutinins ف عؾ ٠ ؾز ػ غزخ ض ا خ ١ شحثزشو ١ ض %3 و ظذس ١ زش ع ١ ر ا ؾظ ي ػ أػ ا زبط زؼذد (EPS) إر ث غذ و ١ ز 2.6 غ / زش. 7 - ىع األيالذ ان ؼذ ت وتزك زها دسط رأص ١ ش ع األ الػ ا ؼذ ١ خ ف ا زبط EPS ثىزش ٠ ب.L plantarum إر أظ شد ا زبئظ ا ج ١ خ ف ا شى )1( ا أفؼ ا زبط زؼذد 997 غ / زش ػ ذ اؽز اء ا عؾ ػ وجش ٠ زبد ا غ ١ غ ١ ف عفبد ا ج ربع ١ ص بئ ١ خ ا ١ ذس ع ١ ؾ ي ا ؼ بطش ا ضسح ٠ ى بن فشق وج ١ ش ف اإل زبط ػ ذ اؽز اء ا عؾ ػ وجش ٠ زبد ا غ ١ غ ١ ف عفبد ا ج ربع ١ ص بئ ١ خ ا ١ ذس ع ١ فمؾ إر ث غذ و ١ خ اإل زبط 989 غ / زش, ف ؽ ١ ا خفغ ا زبط ا زؼذد ػ ذ اعزؼ بي وجش ٠ زبد ا غ ١ غ ١ ف عفبد ا ج ربع ١ ؾ ي ا ؼ بطش ا ضسح ( وال ػ ؽذ (. ا ر فش األ الػ ا ؼذ ١ خ ف عؾ ا زخ ش ٠ ؼذ أؽذ ا ؼ ا ا ؼش س ٠ خ أل زبط EPS األؽ ١ بء ا غ ش ٠ خ إر روش ) 25 (ا األ الػ ا ؼذ ١ خ ا ؼبفخ رشى ظذسا أل ٠ بد ا ز رؤصش ػ ا فؼب ١ خ ا زؾف ١ ض ٠ خ إل ض ٠ بد ا الص خ ا خال ٠ ب إ زبط. EPS و ب ٠ ؼذ أ ٠ ا غ ١ غ ١ أؽذ ا ى بد ا شئ ١ غ ١ خ ف عؾ ا زخ ش ا ز رذػ ا خال ٠ ب ػ ١ بد األ ٠ غ اإل زبع ١ خ ا خز فخ, أ ٠ ؼب د س وج ١ ش ف رفؼ ١ ظب ا زظبص ا غىش) 26 ( نوع األمالح المعدنية L.S.D = انشكم )1( : تأث ز ىع األيالذ ان ؼذ ت ف ا تاج EPS ي بكتز ا.L plantarum 37
8 لتر / ملغم لتر / ملغم / ػه / 216 و ب ر دساعخ رأص ١ ش ا زشاو ١ ض ا زذسعخ أل الػ ا ؼذ ١ خ ف ا زبط EPS إر أظ شد ا زبئظ ا ػؾخ ف ا شى )11) ا ا زبط زا ا زؼذد ٠ ضداد ثض ٠ بدح رشاو ١ ض اا ؾ ١ ا ى ١ خ ف ا عؾ,إر ا ا زبط EPS ث غ ألظب ػ ذ ا زشو ١ ض ٠ ) ( %,ا ب ػ ذ ص ٠ بدح ا زشو ١ ض ف ١ الؽع ا خفبع و ١ خ EPS ا زظ تركيز االمالح% L.S.D = انشكم )11(: تأث ز تزك ز األيالذ ان ؼذ ت ( كبز تاث ان غ س ىو +فىسفاث انبىتاس ىو ) ف ا تاج EPS ي بكتز ا.L plantarum 8 -تأث ز انزقى انه ذروخ االبتذائ ا رأص ١ ش ا شل ا ١ ذس ع ١ األثزذائ عؾ ا ضسػ ف ا زبط EPS ثىزش ٠ ب.L plantarum ػؼ ف ا شى )12( إر ٠ الؽع ا ا زبط ا زؼذد ثشى أفؼ ف ذ سل ١ ذس عPH( ١ ) ٠ زشا ػ ث ١ 7-6 اال ا أػ ا زبط زؼذد رؾمك ػ ذ ا شل ا ١ ذس ع 6.5 ١ إر ث غذ و ١ خ EPS ا زظ 1278 غ / زش.ا ب ػ ذ ا شل ا ١ ذس ع ١ 5 ف ١ الؽع ا خفبع ف ا زبط ا زؼذد, لذ ٠ ؼض ر ه ا اسرفبع ؽ ػخ ا عؾ ب ٠ غجت ا ؼغؾ ا ؾب ؼ stress) ) acid ػ ا خال ٠ ب )27(, ا ا جىزش ٠ ب ا زغخ زؼذد ا غىش لذ رؾ فغ ب ارغب ا ؼغؾ ا ج ١ ئ stress) ) environmental خالي ر ظ ١ ؽشػ EPS ا ز ر زغ ( 28( الرقم الهيدروجيني االبتدائي L.S.D = انشكم )12( : تأث ز انزقى انه ذروخ االبتذائ ف ا تاج EPS ي بكتز اplantarum.L 9 -تأث ز زدى انهقاذ : ج ١ ب رأص ١ ش ؽغ ا مبػ ف ا زبط EPS ثىزش ٠ ب.L plantarum مؼ عؾ اإل زبط ثبعزؼ بي ؽغ مبػ زذسعخ )1 (% ث ١ ذ ا زبئظ ا ػؾخ ف ا شى ) 13 ( ا ا زبط ٠ ضداد EPS ثض ٠ بدح ؽغ ا مبػ ؽز ٠ ظ ا أػ ا زبط زؼذد 129 غ / زش ػ ذ اعزؼ بي ؽغ ا مبػ %3. ا زبئظ ز ا ذساعخ رزفك غ دساعبد أخش ار اعزؼ ؽغ ا مبػ %3 ال زبط EPS ثىزش ٠ ب )29( Lactococccus lactis NCDC. ث ١ ب اعزؼ ؽغ ا مبػ %2 ال زبط EPS ثىزش ٠ ب )3( Lactobacillus bulgaricus. 38
9 / لتر ملغم / ػه / حجم اللقاح % L.S.D = انشكم )13( : تأث ز زدى انهقاذ ف ا تاج EPS ي بكتز ا Lactibacillus plantarum ان ظادر: 1- Donot, F.; Fontana, A.; Baccou, J.C.; Schorr-Galindo, S. (212) Microbial exopolysaccharides: main examples of synthesis, excretion, genetics and extraction. Carbohyd Polym, 87: Garai-Ibabe G, Aeizaga J, Aznar R, Elizaquivel P, Prieto A, Irastorza A & Dueñas MT (21) Screening and selection of 2-branched (1,3)-β-D-glucan producing lactic acid bacteria and exopolysaccharide characterization. Journal of Agricultural and Food Chemistry,58: Callaway T. R., Edrington T. S., Anderson R. C., Harvey R. B.,Genovese K. J., Kennedy C. N.,Venn D.W. and Nisbet D. J.(28). Probiotics, prebiotics and competitive exclusion for prophylaxis against bacterial disease, Animal Health Research Reviews. 9(2): Macfarlane S., Macfarlane G. T. and Cummings J. H. (26). Review article- Prebiotics in the gastrointestinal tract,alimentary pharmacology and Therapeutics, 24: Yadav,V.; Prappulla,S.G.; Jha,A. & Poonia,A.(211). A novel exopolysaccharide from probiotic Lactobacillus fermentum CFR 2195: Production, purification and characterization. Research Article, Biotechnol. Bioinf. Bioeng. 1(4): انطائ, عؼبد سػب زؼت.) 211(. ا زبط ر م ١ خ ؽب غ ا الوز ١ ه خال ٠ ب م ١ ذح جىزش ٠ ب Lactobacillus ا ؼض خ ؾ ١ ب". سعب خ بعغز ١ ش. و ١ خ ا ؼ / عب ؼخ وشثالء. 8- Holt, J.H.; Krieg, N.R.; Sneath, P.H.A.; Staley, J.T. & Williams, S.T. (1984). Group 17: grampositive cocci. In W. R. Hensyl (Ed.) Bergey s Manual of Determinative Bacteriology (9th ed.). Baltimore, MD, USA: Williams & Wilkins., Vijayendra, S.V.N.; Palanivel, G. Mahadevamma, S.& Tharanathan, R.N.(28). Physicochemical characterization of 4 an exopolysacchairde produced by a non-ropy strain of Leuconostoc sp. CFR : Kanmani P., Satishkumar R., Yuvaraj N., Paari K. A., Pattukumar V., Arul V., (211). Production and purification of a novel exopolysaccharide from lactic acid bacterium functional characteristics activity in vitro, Bioresource Streptococcus phocae PI8 and its Technology,12( 7): Levesque, R. (27). Spss Programing and Data Management : Aguid for SPSS and SAD Users, Fourth Edition, SPSS Inc., Chicago. 12- Tsuda,H.&Miymoto,t.( 29).Production of Exopolysaccharide by Lactobacillus plantarum and the Prebiotic Activity of the Exopolysaccharide. Food Sci. Technol. Res., 16 (1), Haroun,B.M.; El- Menoufy, H.A.; Amin, H.A.& A. El-Waseif,A.(213). Biosynthesis and Morphology of an Exopolysaccharide from a Probiotic Lactobacillus plantarum under different growth conditions. Journal of Applied Sciences Research, 9(2): , ISSN X. 14-Razack, S.A.; Velauyutham,V.; Thangavelu,v.(213). Medium optimization for the production ofexopolysaccharide by Bacillus subtilis usingsynthetic sources and agro wastes. Turk J Biol, 37:
10 / ػه / Sutherland, I.W. (21). Microbial polysaccharides from Gram-negative bacteria. International Dairy Journal, 11: Audy, J.; Labrie, S.; Roy, D.& LaPointe, G.(21). Sugar source modulates exopolysaccharide biosynthesis in Bifidobacterium longum subsp. longum CRC 2. Microbiology.156: Zisu,B.& Shah,N.P.(23). Effects of ph, Temperature, Supplementation with Whey Protein Concentrate, and Adjunct Cultures on the Production of Exopolysaccharides by Streptococcus thermophilus J. Dairy Sci. 86: Degeest, B., L. De Vuyst. (2). Correlation of activities of the enzymes α- phosphoglucomutase, UDP-galactose 4-epimerase, and UDP-glucose pyrophosphorylase with exopolysaccharide biosynthesis by Streptococcus thermophilus LY3. Applied and Environmental Microbiology 66(8): Lindberg, B. (199). Components of bacterial polysaccharides. Adv. Carbohyd. Chem.Biochem.48: Cho, D.H.; Chae,H.J.& Kim, E.Y. ( 21). Synthesis and characterization of a novel Extracelluular polysaccharide by Rhodotorula glutinis. Applied Biochemistry and Biotechnology. 95: Polak-berecka,M.; Wasko,A.;Szwajgier,D.& Choma,A.(213). Bifidogenic and Antioxidant Activity of Exopolysaccharides Produced by Lactobacillus rhamnosus E/N Cultivated on Different Carbon Sources. Polish Journal of Microbiology Vol. 62, No 2, Grobben G.J.; Chin-Joe,I.; Kitzen, V.A. ; Boels, I.C. ; Boer, F. ; Sikkema, J. ; Smith, M.R. & De Bont, J.A.M. ( 1998). Enhancement of exopolysaccharide production by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772 with a simplified defined medium. Appl. Environ. Microbiol. 64: Amrane A. & Pringent,Y.( 1998). Influence of yeast extract concentration on batch cultures of Lactobacillus helveticus: Growth and production coupling. World J. Microbiol. Biotechnol. 14: Ibrahim,G.S.; Mahmoud,M.G, ; Asker,M.M.S. & Ghazy,E.A.(212). Production and Biological Evaluation of Exopolysaccharide From Isolated Rhodotorula glutinins. Australian Journal of Basic and Applied Sciences, 6(3): 41-48, ISSN Kaur, V.; Bera, M.B. ; Panesara, P.S. & Chopra, H.K.(213). Production and Characterization of Exopolysaccharide Produced by Alcaligenes Faecalis B14 Isolated from Indigenous Soil. International Journal of Biotechnology and Bioengineering Research.ISSN , Vol: 4( 4), pp El Enshasy, H.; Then, C.; Othman, N. Z. ;Al Homosany, H; Sabry, M.; Sarmidi, M. R.& Aziz, R. A. (211). Enhanced xanthan production process in shake flasksand pilot scale bioreactors using industrial semi-defined medium. Afr. J.Biotech. 1(6): p Vaningelgem, F.; Van der Meulen, R.; Zamfir, M.; Adriany, T.;, Laws, A.P.; & De Vuyst, L. (24a). Streptococcus thermophilus ST 111 producesa stable high-molecular-mass exopolysaccharide in milk-based medium.international Dairy Journal, 14, Broadbent, D.J. ; McMahon, D.L.; Welker, C.J.; Oberg & S. Moineau. )23(. Biochemistry, genetics, and applications of exopolysaccharide production in Streptococcus thermophilus. Journal of Dairy Science.86: Prathima1, P.C.; Lule1,V.K.; Tomar,S.K. & Singh,A.K.(214). Optimization of Exopolysaccharide production by Lactococccus lactis NCDC 91 by Response Surface Methodology. 3- EL-Gizawy, S.A.; Olfat; Barakat,S.; Sharaf,M.; EL-Shafei,K.; Fathy,F.A. & EL- Sayed,S.H.(213). Effect of growth conditions on the production of exopolysaccharides by microencapsulated Lactobacillus bulgaricus and use it to improve quality of Kareish cheese. Journal of Applied Sciences Research, 9(2):
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