زﺎﺳ ﺺﻟﺎﺧ ي
ﺒﺴﻧ ﯽ ﺳرﺮﺑ و ﯽ
ﺻﻮﺼﺧ ﯿ
ﺮﺘﮐﺎﺑ تﺎ ﯾ
ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ هﺪﺷ ﺪ
ا ود ﻂﺳﻮﺗ ﯾ
ﻪﻟوﺰ ﺳﺎﺑﻮﺘﮐﻻ ﯿ
مورﺎﺘﻧﻼﭘ سﻮﻠ ﻣﻮﺑ
ﯽ ﺎﺑرذآ ﯾ
ﻗﺮﺷ نﺎﺠ ﯽ
ﺷﯿ هدازﺪﻟﺎﺧ اﻮ ﻣا ﺪﻤﺤﻣ ،
1ﯿ زﺎﺠﺣ ﻦ
2
ي ﻣﻮﺼﻌﻣ ﺎﺿر ،
*ﯽ ﺪﻧﺎﻬﺟ ﯾﺰ
2
ي
1 - ﺳﺎﻨﺷرﺎﮐ ﻪﺘﺧﻮﻣآ ﺶﻧاد ﯽ
ز ﺪﺷرا ﯾ روﺎﻨﻓ ﺖﺴ ي - ﺑ ﯿ ژﻮﻟﻮﻨﮑﺗﻮ ي ﻣ ﯿ ﺑوﺮﮑ ﯽ
، ﺎﭘ مﻮﻠﻋ هﺪﮑﺸﻧاد ﯾ
،ﻪ ﻪﻏاﺮﻣ هﺎﮕﺸﻧاد
2 - ﺸﻧاد ﯿ ﺑ هﺪﮑﺸﻫوﮋﭘ ،رﺎ ﯿ
ژﻮﻟﻮﻨﮑﺗﻮ ي زروﺎﺸﮐ ي لﺎﻤﺷ رﻮﺸﮐ بﺮﻏ و بﺮﻏ
3 - دﺎﺘﺳا ﯾ ز هوﺮﮔ رﺎ ﯾ روﺎﻨﻓ ﺖﺴ ي - ﺑ ﯿ ژﻮﻟﻮﻨﮑﺗﻮ ي ﻣ ﯿ ﺑوﺮﮑ ﯽ ﻪﻏاﺮﻣ هﺎﮕﺸﻧاد
) رﺎﺗ ﯾ ﺦ رد ﯾ ﺖﻓﺎ : 20 / 05 / 95 رﺎﺗ ﯾ ﺦ ﺬﭘ ﯾ شﺮ : 29 / 07 / 97 (
ﮑﭼ ﯿ هﺪ
ﺧﺮﺑ ،ﻖﯿﻘﺤﺗ ﻦﯾا رد ﯽ
ﺻﻮﺼﺧ ﯿ ﺮﺘﮐﺎﺑ تﺎ ﯾ ﺳﻮ ﯿ ﻦ ﺎﻫ ي ﻟﻮﺗ ﯿ ﻮﺳ ود ﻂﺳﻮﺗ هﺪﺷ ﺪ ﯾ
ﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﺳرﺮﺑ
ﯽ ﺪﻧﺪﺷ . ﻮـﺳود ،اﺪـﺘﺑا ﯾ
ـﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ سﻮﻠ
مورﺎﺘﻧﻼﭘ و BL1
ﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﻣﻮﺑEL3
ﯽ ﺎﺑرذآ ﯾ ﻗﺮﺷ نﺎﺠ ﯽ ﺴﮑﻠﮐ زا ﯿ ﻣ نﻮ ﯿ ﺑوﺮﮑ ﯽ زروﺎﺸﮐ هﺪﮑﺸﻫوﮋﭘ ي
لﺎﻤﺷ و ،بﺎﺨﺘﻧا رﻮﺸﮐ بﺮﻏ و بﺮﻏ
ﺻﺎﺧ ﯿ ﺮﺘﮐﺎﺑ ﺪﺿ ﺖ ﯾﺎﯾ
ﯽ ﻃ و ﯿ رﺎﻬﻣ ﻒ ي ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﻌﺗ،ﺎﻫ نآ ﻂﺳﻮﺗ هﺪﺷ ﺪ ﯿﯿ
ﺪﺷ ﻦ . ﺳﺎﺴﺣ ﺲﭙﺳ ﯿ
ﺮﺘﮐﺎﺑ ﺖ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﺰـﻧآ ود ﻪـﺑ هﺪﺷ ﺪ ﯾ
ﻢ
زﺎﺌﺗوﺮﭘ ي ) ﺌﺗوﺮﭘ ﯿ زﺎﻨ ﮐ ﺎﻔﻟآ و K ﯿ ﺮﺗﻮﻤ ﯾ ﺴﭙ ﯿ ﻦ ( ﻟﺎﻌﻓ و ﯿ ﺮﺘﮐﺎﺑ ﺖ ي ﺸﮐ ﯽ ﯾ ﺪﺷر رﺎﻬﻣ ﺎ ي
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﻧ و ،هﺪﺷ ﺪ ﯿ ﺎﭘ ﺰ ﯾ راﺪ ي ﺗراﺮﺣ ﯽ نآ ﺎﻫﺎﻣد رد ﺎﻫ ي
ﻌﺗ ﻒﻠﺘﺨﻣ ﯿﯿ ﺪﺷ ﻦ . ﻟﻮﮑﻟﻮﻣ نزو ﯽ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻨﮑﺗ ﺎﺑ ،ﺎﻫ ﻦ ﯿ
ﮏ SDS-PAGE ﻤﺨﺗ
ﯿ ﺪﺷ هدز ﻦ . ﺎﺘﻧ ﻖﺒﻃ ﯾ ﺮﺘﮐﺎﺑ ﻪﮐ ﺪﺷ ﺺﺨﺸﻣ هﺪﻣآ ﺖﺳد ﻪﺑ ﺞ ﯾ
ﺳﻮ ﯿ ﺎﻫ ﻦ ي
ﻟﻮﺗ ﯿﺪ ي ﻮﺳ ود ﺮﻫ ﯾ ﻠﻋ ﻪ ﯿ ﺎﻫ نژﻮﺗﺎﭘ مﺎﻤﺗ ﻪ ي
ﻃ و هدﻮﺑ لﺎﻌﻓ ﺶﻫوﮋﭘ رد ﻪﺘﻓر رﺎﮐ ﻪﺑ ﯿ
رﺎﻬﻣ ﻒ ي ﺳو ﯿ ﻨﭽﻤﻫ و ،ﺪﻨﺘﺷاد ﻊ ﯿ
ﺗ لﺎﻤﻋا زا ﺪﻌﺑ ﻦ ﯿ
ﺎﻫرﺎﻤ ي ﺎـﻣد ﯾ ﯽ
ﻟﺎﻌﻓ نﺎﻨﭽﻤﻫ ﻒﻠﺘﺨﻣ ﯿ
ﺮﺘﮐﺎﺑ ﺪﺿ ﺖ ﯾﺎﯾ
ﯽ ﺪﻧدﺮﮐ ﻆﻔﺣ ار دﻮﺧ .
ﻨﭽﻤﻫ ﯿ ﺪﺷ هﺪﻫﺎﺸﻣ ﻦ ﺮﺘﮐﺎﺑ ﻪﮐ
ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﺰﻧآ ﻪﺑ هﺪﺷ ﺪ ﯾ
ﺌﺗوﺮﭘ ﻢ ﯿ زﺎﻨ و هدﻮـﺑ سﺎﺴﺣ K
ﺮﺨﺗ ﻼﻣﺎﮐ ﯾ ﺪﻧﺪﺷ ﺐ . ﮐ ﺎﻔﻟآ ،ﻞﺑﺎﻘﻣ رد ﯿ
ﺮﺗﻮﻤ ﯾ ﺴﭙ ﯿ ﺮﺨﺗ ﻪﺑ ردﺎﻗ ﻦ ﯾ
ﺮﺘﮐﺎﺑ ﻞﻣﺎﮐ ﺐ ﯾ
ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ دﻮﺒﻧ هﺪﺷ ﺪ .
د فﺮﻃ زا ﯾ ﺮﺘﮐﺎﺑ ﻪـﮐ ﺪـﺷ ﺺﺨﺸﻣﺮﮕ ﯾ
ـﺳﻮ ﯿ ﻦ
ﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ور BL1
ي ﺮﮕﻧﺎﺸـﻧ نژﻮﺗﺎـﭘ ﺮﺷا
ـﯾ ﺸ ﯿ ﻠﮐﺎ ﯽ ) ATCC 1399 (
ا ﮔﺪﻨﺸـﮐ ﺮـﺛ ﯽ
ﻟﺎـﺣ رد ﻪﺘـﺷاد ﯽ
ﺪـﺷر رﺎـﻬﻣ ﺮـﺛا ﻪـﮐ ي
ﻠﻋ ﺮـﺑ ـﯿ ﻪ
ﯾ ﺳﺮ ﯿﻨﯿ ﻟﻮﮐوﺮﺘﻧاﺎ ﯿﺘﯿ ﺎﮑ ) ATCC 35669 (
داد نﺎﺸﻧ . ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻮﺳ ﻦ ﯾ ﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﻧ ﯿEL3
ور ﺮﺑ ﺰ ي رﺎـﻬﻣ ﺮـﺛا رﻮﮐﺬـﻣ ﺮﮕﻧﺎﺸﻧ نژﻮﺗﺎﭘ ود ﺮﻫ
ﺪﺷر ي داد نﺎﺸﻧ . ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﻮـﺳ ود ﺮـﻫ ﻂـﺳﻮﺗ هﺪـﺷ ﺪ ـﺳﺎﺑﻮﺘﮐﻻ ﯾﻪ
ﯿ مورﺎـﺘﻧﻼﭘ سﻮﻠ ﻟﻮـﮑﻟﻮﻣ نزو ،ﻪـﻌﻟﺎﻄﻣ درﻮـﻣ
ﯽ زا ﺮـﺘﻤﮐ 10 ﮐ ﯿ نﻮـﺘﻟادﻮﻠ
) 10 ﮐﯿ نﻮﺘﻟادﻮﻠ ﺮﺘﮐﺎﺑ <
ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ هﺪﺷ ﺪ 5<
ﮐ ﯿ نﻮﺘﻟاد ﻮﻠ ( ﻌﺗ هدوﺪﺤﻣ رد و ﻪﺘﺷاد ﯿﯿ
ﺮﺘﮐﺎﺑ هﺪﺷ ﻦ ﯾ
ﺳﻮ ﯿ ﺎﻫ ﻦ ي هوﺮﮔ ﺪﻨﺘﻓﺮﮔ راﺮﻗ IIb .
ﻠﮐ ﯿ ﺪ نﺎﮔژاو : ﺮﺘﮐﺎﺑ ي ﺎﻫ ي ﺳا ﯿ ﺘﮐﻻ ﺪ ﯿ ﺳﺎﺑﻮﺘﮐﻻ ،ﮏ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﺘﮐﺎﺑ ،
ﺮﯾ ﺳﻮ ﯿ ﺺﻟﺎﺧ،ﻦ زﺎﺳ ي ﺒﺴﻧ ﯽ ﻟﺎﻌﻓ،
ﯿ ﺮﺘﮐﺎﺑﺪﺿ ﺖ ﯾﺎﯾ
ﯽ
لﻮﺌﺴﻣ*
تﺎﺒﺗﺎﮑﻣ :
1 - ﻪﻣﺪﻘﻣ
ﺮﺘﮐﺎﺑ ي ﺎﻫ ي ﺳا ﯿ ﺘﮐﻻ ﺪ ﯿ ﻪـﺑ اﺬـﻏ ﺖﻌﻨـﺻ رد هدﺮﺘﺴـﮔ رﻮﻃ ﻪﺑ ﮏ
ﺎﻫ ﺖﺸﮐ ناﻮﻨﻋ ي
اﺮﺑ ﺮﮔزﺎﻏآ ي
آﺮﻓ ﯾ ﺎﻫ ﺪﻨ ي ﻤﺨﺗ ﯿﺮ ي ﻣ رﺎﮐ ﻪﺑ ، ﯽ
ﺪﻧور . ﺳﺎﺑﻮﺘﮐﻻ ﯿ ﻠﻋ ﺮﺑ ﻪﮐ ﺖﺳﺎﻫ ﻪﻫد ﺎﻫ سﻮﻠ ـﯿ
ﺑ ﻪ ﯿ رﺎـﻤ ي ﺎـﻫ ي
ﻧﻮﻔﻋ ﯽ ﻣ هدﺎﻔﺘﺳا ﯽ ﺪﻧﻮﺷ 1] ﺎﻧاﻮﺗ و[ ﯾ ﯽ اﺮـﺑ ﺎـﻫ نآ ي
ﺖـﻇﺎﻔﺣ
ﻠﻋ ﺮﺑ ﻒﻠﺘﺨﻣ داﻮﻣ ﯿ
ﺳو رﻮﻃ ﻪﺑ ،ﺎﻫ نژﻮﺗﺎﭘ ﻪ ﯿ
راﺮﻗ ﻪﻌﻟﺎﻄﻣ درﻮﻣ ﻊ
ﺖﺳا ﻪﺘﻓﺮﮔ .
ﺴﺑ ﻪﮐ هﺪﺷ ﺺﺨﺸﻣ ﯿ
رﺎ ي ا زا ﯾ ﺮﺘﮐﺎﺑ ﻦ ي ﺎﻫ ي ﺳا ﯿ ﺪ
ﺘﮐﻻ ﯿ ﻣ ﺪﺿ داﻮﻣ ﮏ ﯿ
ﺑوﺮﮑ ﯽ ﺮﺘﮐﺎﺑ ﻪﻠﻤﺟ زا ﯾ
ﺳﻮ ﯿ ﻟﻮﺗ ﺎـﻫ ﻦ ـﯿ ـﻣ ﺪ ﯽ
ﺮﺘﮐﺎﺑ ﺪﺷر رﺎﻬﻣ ﻪﺑ ردﺎﻗ ،ﻪﮐ ﺪﻨﻨﮐ ي
ﺎـﻫ ي ﺑ ﯿ رﺎـﻤ ي از ي ـﻔﻠﺘﺨﻣ ﯽ
ﻣ ﯽ ﺪﻨﺷﺎﺑ . ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺮﺘﮐﺎﺑ زا هﺪﺷ اﺪﺟ ﻦ ي
ﺎـﻫ ي ـﺳا ﯿ ﺘﮐﻻﺪ ﯿ ،ﮏ
ﺘﭙﭘ ﯿ ﺎﻫ ﺪ ﯾ ﺌﺗوﺮﭘ ﺎ ﯿ ﺎﻫ ﻦ ي ﻣ ﺪﺿ ﮏﭼﻮﮐ ﯿ
ﺑوﺮﮑ ﯽ ﺒﻃ ﯿﻌ ﯽ ﻪﮐ ﺪﻨﺘﺴﻫ
ﻟﺎﻌﻓ ﯿ ﺮﺘﮐﺎﺑ ﺖ ي ﺸﮐ ﯽ ﯾ ﺮﺘﮐﺎﺑ ﺪﺷر رﺎﻬﻣ ﺎ ﯾﺎﯾ
ﯽ ﻠﻋ ﺮﺑ ﯿ ﺎـﻫ ﻪﻧﻮﮔ ﻪ ي
ﺘﻧژ ﻂﺒﺗﺮﻣ ﯿ ﮑ ﯽ ﻣ نﺎﺸﻧ دﻮﺧ زا ﯽ
ﺪﻨﻫد 2] .[ ﻣ زا ﯿ ﺮﺘﮐﺎﺑ نﺎ ﯾ ـﺳﻮ ﯿ ﻦ
ﺎﻫ ﯾ ﯽ ﻧﺎﮔرا ﻂﺳﻮﺗ ﻪﮐ ﯿ
ﺎـﻫ ﻢﺴ ي ـﻣ ﺰﺘﻨـﺳ ﺖـﺒﺜﻣ مﺮـﮔ ﯽ
،ﺪﻧﻮـﺷ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﺳﺎﺑﻮﺘﮐﻻ ﯿ رﺎـﺠﺗ شزرا ﺎـﻫ سﻮﻠ ي
ﺪـﻧراد 3] .[
ﻔﻠﺘﺨﻣ عاﻮﻧا ﯽ
ﺮﺘﮐﺎﺑ زا ﯾ ﺳﻮ ﯿ ﺮﺘﮐﺎﺑ زا ﺎﻫ ﻦ ي
ﺎـﻫ ي ـﺳا ﯿ ﺘﮐﻻ ﺪ ـﯿ ﮏ
ﺎﺳﺎﻨﺷ ،اﺬﻏ هاﺮﻤﻫ ﯾ
ﯽ ﻌﺗ و ﯿﯿ ﺻﻮﺼﺧ ﻦ ﯿ
ﻪﻠﻤﺟ زا ﻪﮐ ﺪﻧا هﺪﺷ ﺖ
ﺮﺘﻤﻬﻣ ﯾ ﺎﻧ ﺎﻫ نآ ﻦ ﯾ
ﺴ ﯿ د ،ﻦ ﯾ ﺴﮐﻮﮐﻮﻠﭙ ﯿ ﻓوﺪﺳا ،ﻦ ﯿﻠ ﯿ رﺎﮕﻟﻮﺑ ،ﻦ ﯾ ،نﺎﮑ
ﺗﻮﻠﻫ ــﯿ ﺴ ﯿ ــﺳﺎﺘﮐﻻ ،ﻦ ﯿ
ــﺳارﺎﺘﻧﻼﭘ ﻒــﻠﺘﺨﻣ عاﻮــﻧا و ﻦ ﯿ
ــﻣ ﺎــﻫ ﻦ ﯽ
ﺪﻨﺷﺎﺑ 4] .[ ﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﺮـﺗ هﺪﺷ ﻪﺘﺧﺎﻨﺷ ناﻮﻨﻋ ﻪﺑ
ﯾ ﻦ
ﺮﺘﮐﺎﺑ ﻮﻀﻋ ي
ﺎﻫ ي ﺳا ﯿ ﺘﮐﻻﺪ ﯿ ﺎﻫ مﻮﺑ زا ،ﮏ ي
ﻋﻮﻨﺘﻣ ﯽ هﺪـﺷ اﺪـﺟ
ﺳارﺎﺘﻧﻼﭘ و ﺪﻧا ﯿ
ﺎﻫ ﻦ ي دﺪﻌﺘﻣ ي ﻮﺳ زا ﯾ ﺎـﻫ ﻪ ي ـﺷ رد دﻮـﺟﻮﻣ ﯿ
ﺮ
6] - 5 ﻨﭘ ،[ ﯿﺮ 7] ﻮﺠﺑآ ،[ 8] ﺧ و [ ﯿ ﻤﺨﺗ رﺎ ـﯿ هﺪـﺷ ﺮ 9] ﺎـﺳﺎﻨﺷ[ ﯾ ﯽ
ﺪﻧا هﺪﺷ ﺳﺎﺑﻮﺘﮐﻻ . ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﻞﻗاﺪﺣ
6 ﺮﺘﮐﺎﺑ ﯾ ـﺳﻮ ﯿ اﺰـﺠﻣ ﻦ
ﻟﻮﺗ ﯿ ﻣ ﺪ ﯽ ﺪﻨﻨﮐ . ﺎﺷ ﻪﻠﻤﺟ زا ﯾ
ﺮﺗ ﻊ ﯾ ﺳارﺎﺘﻧﻼﭘ ﻦ ﯿ
ﺎﻫ ﻦ ﯾ ﯽ ﻂﺳﻮﺗ ﻪﮐ
ﺳﺎﺑﻮﺘﮐﻻ ﯿ مرﺎﺘﻧﻼﭙﺳﻮﻠ ﻟﻮﺗ
ﯿ ـﻣ ﺪﻧا هﺪﺷ ﺪ ﯽ
ـﺳارﺎﺘﻧﻼﭘ ﻪـﺑ ناﻮـﺗ ﯿ
ﻦ
ﺳارﺎﺘﻧﻼﭘ ،EF ﯿ ﻦ ﺳارﺎﺘﻧﻼﭘ ،W ﯿ ﻦ ـﺳارﺎﺘﻧﻼﭘ و JK ﯿ
ﻦ هرﺎـﺷا S
ﮕﻤﻫ ﻪﮐ دﺮﮐ ﯽ
ﺘﮐﺎﺑ وﺰﺟ ﺮﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﺘﭙﭘ ود ﯿ ﺪ ي ﻣ ﯽ ﺪﻨﺷﺎﺑ 10] .[
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﺳﺎﺑﻮﺘﮐﻻ ﯿ ﺎـﻫ هوﺮـﮔ رد سﻮﻠ ﯾ
ﯽ ناﻮـﻨﻋ ﺖـﺤﺗ
ﻪﺘﺳد ﺮﺘﮐﺎﺑ I ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ) ﺘﻧﻻ ﯽ ﺑ ﯿ ﺗﻮ ﯿ ﺎﻫ ﮏ ( ﻪﺘﺳد ، ﺮﺘﮐﺎﺑ II ﯾ ﺳﻮ ﯿ ﻦ
ﺎﻫ ) ﺮﺘﮐﺎﺑ ﻞﻣﺎﺷ ﯾ
ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻏ ﯿ ﺘﻧﻻ ﺮ ﯽ ﺑ ﯿ ﺗﻮ ﯿ تراﺮﺣ ﻪﺑ موﺎﻘﻣ ،ﮏ
ﻟﻮﮑﻟﻮﻣ نزو ﺎﺑ ﯽ
10kDa ﺮﺘﮐﺎﺑ<
ﯾ ﺳﻮ ﯿ ﻦ
<
5kDa ـﻣ ﯽ ﺪﻨـﺷﺎﺑ ( ،
ﻪﺘﺳد ) III ﺌﺗوﺮﭘ ﯿ ﺎﻫ ﻦ ي ﻟﻮﮑﻟﻮﻣ نزو ﺎﺑ تراﺮﺣ ﻪﺑ سﺎﺴﺣ ﯽ
ﻻﺎﺑ
ﻣ ﻞﻣﺎﺷ ار ﯽ ﺪﻧﻮﺷ ( ﻪﺘﺳد و ) IV
ﺌﺗوﺮﭘ ﯿ ﺎﻫ ﻦ ﯾ ﯽ ﺎـﺑ هاﺮـﻤﻫ ﻪـﮐ
ﯾ ﻟ ﺶﺨﺑ ﮏ ﯿﭙﯿ
ﺪ ي ﯾ ﻫﻮﺑﺮﮐ ﺎ ﯿ ﺗارﺪ ﯽ ﻣ ﯽ ﻟﻮـﮑﻟﻮﻣ نزو و ﺪﻨـﺷﺎﺑ ﯽ
ﻻﺎﺑ ﯾ ﯽ ﺪﻧراد ( ﺪﻨﺑ ﻪﺘﺳد ، ي ﻣ ﯽ ﺪﻧﻮﺷ 11] .[ ا زا فﺪﻫ
ﯾ ﺑﺮﺠﺗ ﺶﻫوﮋﭘ ﻦ ﯽ
زﺎـﺳ ﺺﻟﺎﺧ ، ي
ﺒﺴـﻧ ﯽ ـﺳرﺮﺑ و ﯽ
ﺧﺮﺑ ﯽ ﺻﻮﺼﺧ ﯿ ﮐ تﺎ ﯿﻔ ﯽ ﻤﮐ و ﯽ ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ـﯿ هﺪـﺷ ﺪ
ا ود ﻂـﺳﻮﺗ ﯾ
ﻪـﻟوﺰ ـﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎـﺘﻧﻼﭘ سﻮﻠ ﻣﻮـﺑ
ﯽ ﺎﺑرذآ ﯾ نﺎــﺠ
ﻗﺮﺷ ﯽ دﻮﺑ .
2 - ﺎﻫ شور و داﻮﻣ
2 - 1 - ﻮﺳ ﯾ ﺎﻫ ﻪ ي ﺮﺘﮐﺎﺑ ﯾﺎ ﯾ ﯽ
ﻮــﺳ ود ﯾ ــﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎــﺘﻧﻼﭘ سﻮﻠ و BL1
ــﺳﺎﺑﻮﺘﮐﻻ ﯿ سﻮﻠ
مورﺎﺘﻧﻼﭘ ﻮﺳ ناﻮﻨﻋ ﻪﺑEL3
ﯾ ﺎﻫ ﻪ ي ﻟﻮﺗ ﯿ ﺮﺘﮐﺎﺑ هﺪﻨﻨﮐ ﺪ ﯾ
ـﺳﻮ ﯿ زا ،ﻦ
ﺴﮑﻠﮐ ﯿ ﻣ نﻮ ﯿ ﺑوﺮﮑ ﯽ ﺑ هﺪﮑﺸﻫوﮋﭘ ﯿ
ژﻮﻟﻮﻨﮑﺗﻮ ي زروﺎﺸﮐ ي لﺎﻤﺷ -
ﺳرﺮﺑ ﺖﻬﺟ رﻮﺸﮐ بﺮﻏ و بﺮﻏ ﯽ
ﺻﻮﺼﺧ ﯿ ﺮﺘﮐﺎﺑ تﺎ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ
ﺪﻧﺪﺷ بﺎﺨﺘﻧا .
ﺞﻨـﭘ ﻣﻮﻤﺴـﻣ ﻞـﻣﺎﻋ نژﻮﺗﺎـﭘ ﯿ
اﺬـﻏ ﺖ ﯾ ﯽ ﻞﻣﺎـﺷ
ﺮﺷا ﯾ ﺸ ﯿ ﻠﮐﺎ
3ﯽ ) ATCC 1399 (
ﺳﺎﺑ ، ﯿ سﻮﺋﺮﺳ سﻮﻠ )4
ATCC
(1431 ﻓﺎﺘﺳا ، ﯿ سﻮﺋرآ سﻮﮐﻮﮐﻮﻠ )5
ATCC 29213 (
ﯾ ، ﺳﺮ ﯿﻨﯿ ﺎ
ﻟﻮﮐوﺮﺘﻧا ﯿﺘﯿ ﺎـــﮑ )6
ATCC 35669 (
ـــﺷ و ﯿ ﺮﻨﺴـــﮑﻠﻓ ﻼﮕ
7ي
) PTCC 1234 (
ﺮﺘﮐﺎﺑ ناﻮﻨﻋ ﻪﺑ ي
ﺎﻫ ي فﺪﻫ ) ﺮﮕﻧﺎﺸﻧ ( ﻧﯿ زا ﺰ
ﺴﮑﻠﮐ ﯿ ﻣ نﻮ ﯿ ﺑوﺮﮑ ﯽ ﺪﻧﺪﺷ بﺎﺨﺘﻧا .
ﻮﺳ ﯾ ﺎﻫ ﻪ ي بﺎـﺨﺘﻧا نژﻮﺗﺎـﭘ
وﺎﺣ ﻢﻫ هﺪﺷ ي
ﺮﺘﮐﺎﺑ ي ﺎﻫ ي ﺮﺘﮐﺎﺑ ﻢﻫ و ﺖﺒﺜﻣ مﺮﮔ ي
ﺎﻫ ي مﺮﮔ
ﻔﻨﻣ ﯽ ﺪﻧدﻮﺑ .
2 - 2 - ﻟﻮﺗ ﯿ ﺮﺘﮐﺎﺑ ﺪ ﯾ ﺳﻮ ﯿ ﻦ
ﻬﺗ ﺖﻬﺟ ﯿ ﺎﻣ ﻪ ﯾ ﻊ ور ﯾ ﯽ وﺎﺣ ي ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻮﺳ زا ﻦ ﯾﻪ ﺎـﻫ ي ﻟﻮﺗ ـﯿ ﺪ
ﺮﺘﮐﺎﺑ هﺪﻨﻨﮐ ﯾ
ﺳﻮ ﯿ ﺮﺘﮐﺎﺑ ،ﻦ ي ﺎـﻫ ـﺳﺎﺑﻮﺘﮐﻻي ﯿ مورﺎـﺘﻧﻼﭘ سﻮﻠ BL1
ﺳﺎﺑﻮﺘﮐﻻو ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ اﺮﺷ ﺖﺤﺗ EL3
ﯾ ﺮﺘﺳا ﻂ ﯾ ﻣ ﻪـﺑ ﻞ ـﯿ ناﺰ
1 % ﺤﻣ رد ﯿ ﺖﺸﮐ ﻂ MRS Broth ﻘﻠﺗ
ﯿ تﺪﻣ ﻪﺑ و ﺪﻧﺪﺷ ﺢ 48
ﺎﻣد رد ﺖﻋﺎﺳ ي
C 37° ﺪﻧﺪﺷ ﻪﺑﻮﮑﻧا .
راﺬـﮔ ﻪﻧﺎﺨﻣﺮﮔ زا ﺪﻌﺑ ي
،
ﺴﻧﺎﭙﺳﻮﺳ ﯿ نﻮ ﺎﻫ ي ﺮﺘﮐﺎﺑ ـﯾ ﺎﯾ ﯽ تﺪـﻣ ﻪـﺑ ﺮـﺿﺎﺣ 20
ﻗد ﯿ رد ﻪـﻘ g
6000 ﺎﻣد رد ي 4°C ﺮﺘﻧﺎﺳ ﯾﻔ ﯿ ﻓ زا رﻮﺒﻋ ﺎﺑ و هﺪﺷ ژﻮ ﯿ
ﺮـﺳ ﺮـﺘﻠ
ﮕﻧﺮﺳ ﯽ m 22µ /0 ) ﺑﯿ ﻓﻮ ﯿ ﻞ ( ﺮﺘﺳا ﯾ ﺪﻧﺪﺷ ﻞ . ﺎﻣ ﯾ ﻊ ﺎﻫ ﯾ وﺮ ﯾ ﯽ رﺎﻋ ي
ﺮﺘﺳا لﻮﻠﺳ زا ﯾ
ﻮﺳ زا ماﺪﮐ ﺮﻫ ﻞ ﯾﻪ
ﻟﻮﺗ ﺖﻠﻋ ﺎﻫ ـﯿ
ـﺳا ﺪ ﯿ ﻂـﺳﻮﺗ ﺪ
ﻮـﺳ ﯾﻪ ﺎـﻫ ـﺳﺎﺑﻮﺘﮐﻻي ﯿ مورﺎــﺘﻧﻼﭘ سﻮﻠ و BL1
ارادEL3 ي pH
ﺳا ﯿ ﺪ ي زا هدﺎﻔﺘﺳا ﺎﺑ ﻪﮐ ،ﺖﺷاد NaOH 4 M
ـﺜﻨﺧ ﯽ و ،هﺪـﺷ
3. Eschrichia coli 4. Bacillus cereus 5. Staphylococcus areus 6.Yerssiniaenterocolitica 5. Shigellaflexneri 6. Disk diffussion
ﺎــﻬﻧ هرﺎﺼــﻋ ﯾ
ﯽ ﺮﺘﮐﺎﺑ ناﻮــﻨﻋ ﻪــﺑ ﯾ
ــﺳﻮ ﯿ ــﺳرﺮﺑ ﺖــﻬﺟ مﺎــﺧ ﻦ ﯽ
ﺻﻮﺼﺧ ﯿ ﻘﺑ رد تﺎ ﯿ ﺎﻣزآ ﻪ ﯾ ﺖﻓﺮﮔ راﺮﻗ هدﺎﻔﺘﺳا درﻮﻣ ﺎﻫ ﺶ 12]
.[
2 - 3 - ﺳرﺮﺑ ﯽ ﻟﺎﻌﻓ ﯿ ﺮﺘﮐﺎﺑ ﺪﺿ ﺖ ـﯾ
ﺎﯾ ﯽ ﻌﺗ و ـﯿ ﯿ ﻦ
ﻃﯿ رﺎﻬﻣ ﻒ ي
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻦ ﺎﻫ
ﻟﺎﻌﻓ ﯿ ﺮﺘﮐﺎﺑ ﺪﺿ ﺖ ﯾﺎﯾ
ﯽ ﻃ و ﯿ رﺎـﻬﻣ ﻒ ي ﺮﺘﮐﺎﺑ ﯾ ـﺳﻮ ﯿ ﻦ ﺎـﻫ ي ﻟﻮﺗ ـﯿ ﺪ
ﻮﺳ ود ﻂﺳﻮﺗ هﺪﺷ ﯾ
ﺖﺴـﺗ زا هدﺎﻔﺘـﺳا ﺎـﺑ ،ﻪـﻌﻟﺎﻄﻣ درﻮﻣ ﺪﻟﻮﻣ ﻪ
د زا رﺎﺸﺘﻧا ﯾ
8ﮏﺴ ﻌﺗ ﯿﯿ ﺪﺷ ﻦ 13] اﺮﺑ .[
ي د ﻪـﻠﺣﺮﻣ مﺎـﺠﻧا ـﯾ
ﮏﺴ
راﺬﮔ ي ﺤﻣ اﺪﺘﺑا ، ﯿ ﻫ ﺮﻟﻮﻣ ﺖﺸﮐ ﻂ ﯿ
مﺮـﻧ رﺎـﮔآ ﻞﮑـﺷ ﻪـﺒﻧﻮﺘﻨ ﺎـﺑ 9
ندوﺰﻓا 7/
0 % ﻬﺗ رﺎﮔآ ﯿ ﺮﺘﺳا ،ﻪ ﯾ دﺮﮔ ﮏﻨﺧ و ﻞ ﯾﺪ
. ﺲﭙﺳ 20 ﻣ ﯿﻠ ﯽ
ﻟﯿ ا زا ﺮﺘ ﯾ ﺤﻣ ﻦ ﯿ ﻣ ﻪﺑ ،ﺖﺸﮐ ﻂ ـﯿ
ناﺰ 1 % ﺎـﻫ نژﻮﺗﺎـﭘ ﺎـﺑ ي
درﻮـﻣ
تروﺪــﮐ ﺎــﺑﺮﻈﻧ 3/
0 ) OD600nm=0/3 (
ــﻘﻠﺗ ﯿ نورد و هﺪــﺷ ﺢ
ﻠﭘﯿ ﺖ ر ﺎﻫ ﯾ ﺪﺷ ﻪﺘﺨ . ﻠﭘﯿ ﺖ ﺎﻫ ـﯾ نورد ﺖﻋﺎـﺳ ﮏ ﯾ
ﻗﺎـﺑ لﺎـﭽﺨ ﯽ
دﻮﺷ ﻪﺘﺴﺑ ًﻼﻣﺎﮐ رﺎﮔآ ﺎﺗ ﺪﻧﺪﻧﺎﻣ .
ا مﺮـﻧ رﺎﮔآ زا هدﺎﻔﺘﺳا ﺖﻠﻋ ـﯾ
ﻦ
ﺮﺘﮐﺎﺑ ﻪﮐ ﺖﺳا ﯾ
ﺳﻮ ﯿ ﺘﺣار ﻪﺑ ﺪﻧاﻮﺘﺑ ﻦ ﯽ
ﻧا نآ رد رﺎﺸـﺘ ﯾ ﻪـﺑ و ﻪـﺘﻓﺎ
د فاﺮﻃا ﯾ ﺪﻨﮐ ذﻮﻔﻧ ﮏﺴ .
،رﺮـﻘﻣ نﺎـﻣز تﺪـﻣ ﺖـﺷﺬﮔ زا ﺲـﭘ
دﯾ ﺎﻫ ﮏﺴ ي ﺮﺘﺳا ﮏﻧﻼﺑ ﯾ
ﺎﺑ ﻞ l 50µ ﺮﺘﮐﺎﺑ زا ﯾ ـﺳﻮ ﯿ ﻬﺗ مﺎـﺧ ﻦ ـﯿ
ﻪ
اﺮﺷ ﺖﺤﺗ و هﺪﺷ ﻪﺘﺸﻏآ ﻪﻧﺎﮔاﺪﺟ ترﻮﺻ ﻪﺑ هﺪﺷ ﯾ
ﺮﺘﺳا ﻂ ﯾ ﺮﺑ ﻞ
ور ي ﻠﭘ ﯿ ﺎﻫ ﺖ ي وﺎﺣ ي ﺤﻣ ﯿ تﺪﻣ ﻪﺑ و ﺪﻧﺪﺷ هداد راﺮﻗ ،ﺖﺸﮐ ﻂ
12 ﺎﻣد ﺎﺑ رﻮﺗﺎﺑﻮﮑﻧا رد ﺖﻋﺎﺳ ي
0C 37 ﺪﻧﺪﺷ ﻪﺑﻮﮑﻧا .
ﺎﺘﻧ ﯾ ﺎـﺑ ﺞ
ﮔ هزاﺪﻧا ﯿﺮ ي ﻣ ﺐﺴﺣ ﺮﺑ ﺪﺷر مﺪﻋ ﻪﻟﺎﻫ ﺮﻄﻗ ﯿﻠ
ﯽ ﺮﺘﻣ ) ( mm ﺎﺑ
ﻟﻮــﮐ زا هدﺎﻔﺘــﺳا ﯿ
ﺖــﻗد ﺎــﺑ ﺲ 1/
0 ﻣ ﯿ ــﻠ ﯽ ﮔ هزاﺪــﻧا ﺮــﺘﻣ ــﯿ
ﺮ ي
ﺪﻧﺪﺷ 13] .[
2 - 4 - ﺳرﺮﺑ ﯽ ﺎﭘ ﯾ راﺪ ي ﺗراﺮﺣ ﯽ ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ
اﺮﺑ ي ﺳرﺮﺑ ﯽ ﺎﭘ ﯾ راﺪ ي ـﺗراﺮﺣ ﯽ ﺮﺘﮐﺎﺑ ﯾ ـﺳﻮ ﯿ ﺎـﻫ ﻦ ي ﻟﻮﺗ ـﯿ هﺪـﺷ ﺪ
ﻮﺳ ود ﻂﺳﻮﺗ ﯾ
ﺮﺘﮐﺎﺑ ،ﻪﻌﻟﺎﻄﻣ درﻮﻣ ﻪ ﯾ
ﺳﻮ ﯿ زا هﺪـﻣآ ﺖﺳﺪﺑ مﺎﺧ ﻦ
ﻮﺳ ﯾ ﻬﺗ ﺪﺷ ﻪﺘﻔﮔ ﻪﭽﻧآ ﻖﺒﻃ ﺎﻫ ﻪ ﯿ
ﻪ 12] ﺎﭘ و ،[ ﯾ راﺪ ي ﺗراﺮﺣ ﯽ نآ
ﺎﻫﺎﻣد رد ﺎﻫ ي
ﻒﻠﺘﺨﻣ )
0C 121 ،
0C 100
،
0C 70 ( تﺪﻣ ﻪﺑ 15
ﻗد ﯿ ﺠﻨﺳ ،ﻪﻘ ﯿ ﺪﺷ هﺪ . ﺳرﺮﺑ ﺖﻬﺟ ﯽ
ﻟﺎﻌﻓ ﻆﻔﺣ ـﯿ
ﺮﺘﮐﺎﺑﺪـﺿ ﺖ ﯾﺎﯾ
ﯽ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺗ زا ﺪﻌﺑ ﺎﻫ ﻦ ﯿ
ﺎـﻫ رﺎـﻤ ي ﺎـﻣد ﯾ ﯽ زا رﺎﺸـﺘﻧا ﺖﺴـﺗ زا ،
دﯾ ﺪﺷ هدﺎﻔﺘﺳا ﮏﺴ 13]
.[
2 - 5 - ﺳرﺮﺑ ﯽ ﺳﺎﺴﺣ ﯿ ﺰﻧآ ﺖ ﯾﻤ ﯽ ﻌﺗ و ﯿﯿ ﻟﺎﻌﻓ ﻦ ﯿ ﺖ
ﺮﺘﮐﺎــﺑ ي ﺸــﮐ ﯽ ــﯾ ﺮﺘﮐﺎﺑ ﺪــﺷر رﺎــﻬﻣ ﺎ ــﯾ
ﺎﯾ ﯽ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ
6. Disk diffussion 7. Soft agar
ﻟﺎﻌﻓ ﯿ ﺮﺘﮐﺎﺑ ﺖ ي ﺸﮐ ﯽ ﯾ ﺮﺘﮐﺎﺑ ﺪﺷر رﺎﻬﻣ ﺎ ﯾﺎﯾ
ﯽ ﺮﺘﮐﺎﺑ ﯾ ـﺳﻮ ﯿ ﺎـﻫ ﻦ ي
ﻟﻮﺗ ﯿ ﻮﺳ ﻂﺳﻮﺗ هﺪﺷ ﺪ ﯾ
ﺎﻫ ﻪ ي ﺒﻃ ،ﻪـﻌﻟﺎﻄﻣ درﻮـﻣ ﯿ
ﺌﺗوﺮﭘ ﺖـﻌ ﯿ
ـﻨ ﯽ
ﻢــﻫ داﻮــﻣ ﺘــﺳ
ﯿ ﻟﻮﺗ ﺰ ــﯿ ــﺳﺎﺑﻮﺘﮐﻻ ﻂــﺳﻮﺗ هﺪــﺷ ﺪ ﯿ
و ﺎــﻫ سﻮﻠ
ﺖﺳا هﺪﺷ هداد نﺎﺸﻧ ﻼﺒﻗ ﺎﻫ سﻮﮐﻮﮐوﺮﺘﻧا 14 ]
.[ ا رد ﯾ ﻘﺤﺗ ﻦ ﯿ ﻖ
ﺳﺎﺴﺣ ﯿ ﺮﺘﮐﺎﺑ ﺖ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﺰﻧآ ﻪﺑ هﺪﺷ ﺪ ﯾ
ﺎﻫ ﻢ ي زﺎـﺌﺗوﺮﭘ ي
ﺳرﺮﺑ ﯽ ﻧ و ﺪﺷ ﯿ ﻟﺎﻌﻓ ﺰ ﯿ ﺮﺘﮐﺎﺑ ﺖ ي ﺸﮐ ﯽ ﯾ ﺮﺘﮐﺎﺑ ﺪـﺷر رﺎﻬﻣ ﺎ ـﯾ
ﺎﯾ ﯽ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﺗرﺎـﻣد ﻂـﺳﻮﺗ ﻪﭽﻧآ ﻖﺒﻃ هﺪﺷ ﺪ ﯿ
ﺰﻨ 15] [
ﺿﻮﺗ ﯿ ﻌﺗ ،هﺪـﺷ هداد ﺢ ـﯿ
ﯿ ﺪـﺷ ﻦ . ﻟﺎﻌﻓ ـﯿ ﺮﺘﮐﺎﺑ ﺪـﺿ ﺖ ـﯾ
ﺎﯾ ﯽ ﺎـﻣ ﯾ ﻊ
ور ﯾ ﯽ ﺎﻫ ي رﺎﻋ ي ﺮﺘﺳا لﻮﻠﺳ زا ﯾ
ﻞ ) ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ مﺎﺧ ﻦ ( ﻪـﺑ طﻮﺑﺮﻣ
ﻮﺳ ود زا ماﺪﮐ ﺮﻫ ﯾ
رد ،ﻪ ـﺜﻨﺧ pH
ﯽ ـﺳرﺮﺑ ﯽ زا ﺪـﻌﺑ و ﺪﻧﺪـﺷ
نﺎﻣز تﺪﻣ ﺖﺷﺬﮔ 12
ـﺳﺎﺑﻮﮑﻧا ﺖﻋﺎـﺳ ﯿ
ﺎـﻣد رد نﻮ ي
0C 37 و
ﻪﻟﺎﻫ هﺪﻫﺎﺸﻣ ﺎﻫ
ي ﻠﭘ ،ﺪﺷر مﺪﻋ ﯿ
ﺖ و هﺪﺷ جرﺎﺧ رﻮﺗﺎﺑﻮﮑﻧا زا ﺎﻫ
اﺮﺷ ﺖﺤﺗ ﯾ ﺮﺘﺳا ﻂ ﯾ ﺗ ﻞ ﯿ ﺰﻧآ رﺎﻤ ﯾﻤ ﯽ ﺰﻧآ ﺎﺑ ﯾ ﺎﻫ ﻢ ي زﺎـﺌﺗوﺮﭘ ي ﺮـﺑ
ور ي ﻠﭘ ﯿ ﺖ ﺎﻫ ي اراد ي ﺪﺷ مﺎﺠﻧا ،ﺪﺷر مﺪﻋ ﻪﻟﺎﻫ .
ا ﻪﺑ ﯾ ترﻮﺻ ﻦ
ﻪﮐ l 5µ ﺰﻧآ زا ﯾ ﺌﺗوﺮﭘ ﻢ ﯿ زﺎﻨ K10 ) mg.ml-1 20 ( و l 5µ ﺰﻧآ زا ﯾ ﻢ
ﮐ ﺎﻔﻟآ ﯿ ﺮﺗﻮﻤ ﯾ ﺴﭙ ﯿ
11ﻦ ) mg.ml-1 20 ( اﺮـﺷ ﺖـﺤﺗ ﯾ
ﺮﺘـﺳا ﻂ ﯾ ﺎـﺑ ﻞ
ﺎﻫ ﻪﻟﺎﻫ فاﺮﻃا رد اﺰﺠﻣ رﻮﻃ ﻪﺑ ﺮﻠﭙﻤﺳ ي
ﻪـﺑ طﻮـﺑﺮﻣ ﺪﺷر مﺪﻋ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻦ ﺎﻫ ي ﻟﻮﺗ ﯿ ﺪ ي ﻮﺳ زا ماﺪﮐ ﺮﻫ ﯾ
ر ،ﺎﻫ ﻪ ﯾ ﺪـﺷ ﻪـﺘﺨ )
رد
ﻠﭘ ﺮﻫ ﯿ اﺮﺑ ﺖ ي ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ مﺎﺧ ﻦ ﯾ ﻮﺳ ﮏ ﯾ هداد راﺮﻗ راﺮﮑﺗ ود ﻪ
ﺰﻧآ زا ماﺪﮐ ﺮﻫ و ﺪﺷ ﯾ
ﻢ ﻪﻟﺎﻫ فاﺮﻃا رد اﺰﺠﻣ رﻮﻃ ﻪﺑ ﺎﻫ ر ﺎﻫ
ﯾ ﻪﺘﺨ
ﺪﻧﺪﺷ .(
ﻠﭘ ﺲﭙﺳ ﯿ ﺖ تﺪـﻣ ﻪـﺑ هرﺎـﺑود ﺎﻫ 12
رد ﺖﻋﺎـﺳ
0C 37
ﺪﻧﺪﺷ ﻪﺑﻮﮑﻧا .
ـﺳرﺮﺑ ﻪـﺑ ،رﺮـﻘﻣ نﺎـﻣز تﺪـﻣ ﺖـﺷﺬﮔ زا ﺪﻌﺑ ﯽ
ﻟﺎﻌﻓ ﯿ ﺮﺘﮐﺎﺑ ﺖ ي ﺸﮐ ﯽ ﯾ ﺪـﺷر رﺎـﻬﻣ ﺎ ي
ﺮﺘﮐﺎـﺑ ي ﺎـﻫ ي رد نژﻮﺗﺎـﭘ
ﺪﺷ ﻪﺘﺧادﺮﭘ ﺎﻫ ﻪﻟﺎﻫ فاﺮﻃا .
ﻣﺎﻤﺗ ﯽ ﺎﻣزآ ﻞﺣاﺮﻣ ﯾ
ﻪﺑ و رﺎﺑ ﻪﺳ ﺶ
مﺎﺠﻧا ﻢﻫ زا ﻞﻘﺘﺴﻣ رﻮﻃ ﺪﻧﺪﺷ
15] .[
2 - 6 - ﻤﺨﺗ ﯿ ﻟﻮﮑﻟﻮﻣ نزو ﻦ ﯽ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻦ ﺎﻫ
اﺮﺑ ي ﻤﺨﺗ ﯿ ﻟﻮﮑﻟﻮﻣ نزو ﻦ ﯽ
ﺎﻣ ﯾ ور ﻊ ﯾ ﯽ ﺎﻫ ي ﺜﻨﺧ ﯽ رﺎـﻋ هﺪﺷ ي
ﺮﺘﺳا لﻮﻠﺳ زا ﯾ
ﻞ ) ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ مﺎﺧ ﻦ ( ود زا ماﺪـﮐ ﺮـﻫ ﻪـﺑ طﻮﺑﺮﻣ
ﻮﺳ ﯾ ﻣ ﻪﺑ ،ﻪ ﯿ ناﺰ 60 % ﻧﻮﻣآ تﺎﻔﻟﻮﺳ ردﻮﭘ ﺎﺑ ـﯿ
ﺖـﻬﺟ و ،عﺎﺒـﺷا مﻮ
ﺮﺘﮐﺎﺑ بﻮﺳر ﯾ
ﺳﻮ ﯿ تﺪـﻣ ﻪـﺑ ﻦ 24
رد ﺖﻋﺎـﺳ
0C 4 راﺪـﻬﮕﻧ ي
ﺪﻧﺪﺷ . ﺎﻫ نﻮﮑﻟﺎﻓ رﺮﻘﻣ نﺎﻣز تﺪﻣ ﺖﺷﺬﮔ زا ﺪﻌﺑ ي
ﻪـﺑ رﻮﮐﺬـﻣ
تﺪﻣ 30 ﻗد ﯿ رد ﻪﻘ 5500g ﺎﻣد رد ي 4°C ﺮﺘﻧﺎﺳ ﯾﻔ ﯿ و هﺪـﺷ ژﻮ
ﺎﻫ بﻮﺳر ﯾ ﯽ ﮑﺸﺗ ﺎﻫ نﻮﮑﻟﺎﻓ ﻪﺗ رد ﯿ
ﺪﻧﺪﺷ ﻞ . ﺎﻣ ﺲﭙﺳ ﯾ ور ﻊ ﯾ ﯽ
ر رود ﯾ ﻣ ﻞﻗاﺪـﺣ رد ﺎﻫ بﻮﺳر و هﺪﺷ ﻪﺘﺨ ـﯿ
تﺎﻔﺴـﻓ ﺮﻓﺎـﺑ ناﺰ
ﻟﺎــﺳ ﯿ ﻦ ) 4/
7 (pH:
تﺪــﻣ ﻪــﺑ 24
رد ﺖﻋﺎــﺳ 4°C
ﻞــﺣ
10. Proteinase K
ﺪﻧﺪﺷ 10] .[ ﻟﻮﮑﻟﻮﻣ نزو ﯽ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ﯾ ﯽ ﺒﺴﻧ ترﻮﺻ ﻪﺑ ﻪﮐ ﯽ
زﺎﺳ ﺺﻟﺎﺧ ي
ﺪﺷ ﺪﻧ ـﻠﭘ لژ زرﻮﻓوﺮﺘﮑﻟا زا هدﺎﻔﺘﺳا ﺎﺑ ، ﯽ
ﺮﮐآ ـﯾ ﻞ
ﻣآ ﯿ ﺪﺳ ﺪ ﯾ ـﺳد ود ﻢ ﯿ تﺎﻔﻟﻮـﺳ ﻞ )
SDS-PAGE (
شور ﻖـﺒﻃ
ــﻠﻣﻻ ﯽ ﻌﺗ ، ــﯿ ﯿ ﺪﻧﺪــﺷ ﻦ 16]
.[ ﮓــﻧر ﺎــﺑ لژ ،زرﻮــﻓوﺮﺘﮑﻟا زا ﺪــﻌﺑ
ﺳﺎﻣﻮﮐ ﯽ ﻮﻠﺑ R-250 ﻣآ ﮓﻧر ﯿﺰ ي ﺮﻃ زا و ﺪﺷ ـﯾ
ﺎـﺑ ﻮﺸـﺘﺴﺷ ﻖ
ﺘﺳا طﻮﻠﺨﻣ ﯿ
ﺳا ﮏ ﯿ ﺪ - ﺘﻣ ﯿ ﻞﮑﻟا ﻞ بآ – ) 5:5:1 ( تﺪﻣ ﻪﺑ 24
ﺮﺑ ﮓﻧر ،ﺖﻋﺎﺳ ي
ﺪﺷ . ﺮﮐرﺎﻣ زا ي ﻟﻮﮑﻟﻮﻣ نزو هدوﺪﺤﻣ ﺎﺑ ﯽ
ﺎﭘﯾ ﯿ ﺪﺷ هدﺎﻔﺘﺳا دراﺪﻧﺎﺘﺳا ﺮﮐرﺎﻣ ناﻮﻨﻋ ﻪﺑ ﻦ )
ﺳ ﯿ ا نژ ﺎﻨ ﯾ ناﺮ .(
3 - ﺎﺘﻧ ﯾ ﺞ
3 - 1 - ﺳرﺮﺑ ﯽ ﻟﺎﻌﻓ ﯿ ﺮﺘﮐﺎﺑ ﺪﺿ ﺖ ـﯾ
ﺎﯾ ﯽ ﻌﺗ و ـﯿ ﯿ ﻦ
ﻃﯿ رﺎﻬﻣ ﻒ ي
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻦ ﺎﻫ
ﺎﺘﻧ ﯾ ﻌﺗ زا ﻞﺻﺎﺣ ﺞ ـﯿ
ﯿ ﻃ ﻦ ـﯿ رﺎـﻬﻣ ﻒ ي لوﺪـﺟ رد ﻪـﮐ )
1 ( ﻧ ـﯿ ﺰ
ﻣ هﺪﻫﺎﺸﻣ ﯽ ﺮﺘﮐﺎﺑ ﻪﮐ داد نﺎﺸﻧ ،دﻮﺷ ﯾ
ـﺳﻮ ﯿ ﺎـﻫ ﻦ ي ﻟﻮﺗ ـﯿ هﺪـﺷ ﺪ
ﻮﺳ ود ﺮﻫ ﻂﺳﻮﺗ ﯾ
ﻪﻌﻟﺎﻄﻣدرﻮﻣ ﻪ ﺳﺎﺑﻮﺘﮐﻻ
ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ و BL1
ﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﻠﻋ ﺮﺑ ،EL3
ﯿ ﻣﺎﻤﺗ ﻪ ﯽ نژﻮﺗﺎﭘ ﺎﻫ ي مﺮـﮔ
ﻔﻨﻣ مﺮﮔ و ﺖﺒﺜﻣ ﯽ
ﺪﻧدﻮﺑ لﺎﻌﻓ ﺶﻫوﮋﭘ رد ﻪﺘﻓر رﺎﮐ ﻪﺑ .
Table 1 Inhibitory specterum of the bacteriocin produced by two studied L.plantarum species L.plantarum BL1 L.plantarum EL3
Neutralized Acidic
Neutralized Acidic
Selected species
14mm 12mm
14mm 11mm
E.coli (ATCC1399)
13mm 15mm 13mm
11mm S.flexneri (PTCC1234)
12mm 11mm 14mm 12mm
9mm 11mm 13mm
10mm 15mm 13mm
8mm 12mm Y.entrocolitica(ATCC35669)
B.cereus (ATCC1431)
S.aureus(ATCC29213)
Zone of inhibition> 2mm /(+)Zone of inhibition< mm ٢(-) ٢
ﺎﺘﻧ ﻖﺒﻃ ﯾ ﺖﺳﺪﺑ ﺞ ﺑ هﺪﻣآ
ﯿ ﺮﺘﺸ ﯾ ﻣ ﻦ ﯿ ﮔﺪﻧرادزﺎﺑ ناﺰ ﯽ
ﺮﺘﮐﺎﺑ ﯾ ـﺳﻮ ﯿ ﻦ
ﻟﻮﺗ ﯿ ﻮـﺳ ﻂـﺳﻮﺗ هﺪﺷ ﺪ ﯾ
ـﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎـﺘﻧﻼﭘ سﻮﻠ رد BL1
ﺜﻨﺧpH ﯽ ﻪﺑ طﻮﺑﺮﻣ ، ﺷﯿ
ﺮﻨﺴﮑﻠﻓ ﻼﮕ ي
) 3/mm 1 15± ( ﺮﺘﻤﮐ و ﯾ ﻦ
ﮔﺪــﻧرادزﺎﺑ ﯽ
ﻠﻋﺮﺑ ــﯿ ب ﻪ . سﻮﺋﺮــﺳ ) 1mm 11± ( درﻮــﻣ رد و،
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻮﺳ ﻦ ﯾ ﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﺑ ﯿEL3
ﺮﺘﺸ ﯾ ﻣ ﻦ ﯿ ناﺰ
ﮔﺪﻧرادزﺎﺑ ﯽ ﻠﻋﺮﺑ ﯿ ﻓﺎﺘـﺳا ﻪ ﯿ سﻮـﺋرآ سﻮﮐﻮﮐﻮﻠ )
1mm 15± ( و
ﺮﺘﻤﮐ ﯾ ﻣ ﻦ ـﯿ ﻪـﺑ طﻮـﺑﺮﻣ ناﺰ ـﺳﺎﺑ
ﯿ سﻮﺋﺮـﺳ سﻮﻠ )
1 mm 10± (
ﺪﺷ هﺪﻫﺎﺸﻣ .
3 - 2 - ﺳرﺮﺑ ﯽ ﺎﭘ ﯾ راﺪ ي ﺗراﺮﺣ ﯽ ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻦ ﺎﻫ
ﺎﺘﻧ ﯾ ﺳرﺮﺑ زا ﻞﺻﺎﺣ ﺞ ﯽ
ﺎﭘ ﯾ راﺪ ي ﺎﻣد ﯾ ﯽ ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎـﻫ ﻦ ي ﻟﻮﺗ ـﯿ ﺪ
ﺗ لﺎﻤﻋا زا ﺪﻌﺑ هﺪﺷ ﯿ
ﺎﻫرﺎﻤ ي ﺎﻣد ﯾ ﯽ ﻟﺎﻌﻓ ﺶﺠﻨﺳ و ﻒﻠﺘﺨﻣ ـﯿ
ﺖ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺮﺘﮐﺎﺑ ﻪﮐ داد نﺎﺸﻧ ،ﺎﻫ ﻦ ﯾ
ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﻂﺳﻮﺗ هﺪﺷ ﺪ
ﻮﺳ ود ﺮﻫ ﯾ ـﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎـﺘﻧﻼﭘ سﻮﻠ و EL3
ـﺳﺎﺑﻮﺘﮐﻻ ﯿ سﻮﻠ
مورﺎﺘﻧﻼﭘ ﺪـﻌﺑ BL1
ﺗﯿ ﺎـﻫرﺎﻤ ي ﺎـﻣد ﯾ ﯽ ﻟﺎﻌﻓ نﺎـﻨﭽﻤﻫ ـﯿ
ﺪـﺿ ﺖ
ﺮﺘﮐﺎﺑ ﯾﺎﯾ ﯽ ﺎﻫ نژﻮﺗﺎﭘ ﺪﺷر رﺎﻬﻣ ﻪﺑ ردﺎﻗ و ﺪﻧدﺮﮐ ﻆﻔﺣ ار دﻮﺧ ي
ﺋﺰﺟ ﺶﻫﺎﮐ ﻪﮐ ﺪﻨﭼﺮﻫ ،ﺪﻧدﻮﺑ ﺮﮕﻧﺎﺸﻧ ﯽ
ﻟﺎﻌﻓ ﯿ اﺰﻓا ﺎﺑ ﺖ ﯾ مواﺪﻣ ﺶ
ﺪﺷ هﺪﻫﺎﺸﻣ ﺎﻣد )
ﻞﮑﺷ 1 )(
لوﺪﺟ 2 .(
Fig 1 Effect of thermal treatment in stability of produced bacteriocins by two studied L.plantarum
species, (A) 120°C, (B) 100°C, (C) 70°C
Table 2 Effect of thermal treatment in stability of produced bacteriocins by two studied L.plantarum species
L.plantarumEL3 L.plantarum BL1
120°C 100°C
70°C 120°C
100°C 70°C
7 mm 9 mm
12 mm 8mm
10 mm 13 mm
E.coli
8 mm 10 mm
13 mm 8.5mm
8.5 mm 11 mm
Sh.flexneri
6.5 mm 9 mm
10 mm 9 mm
10 mm 13 mm
Y.entrocolitica
6.5 mm 8 mm
10.5 mm 6 mm
8 mm 8 mm
B.cereus
9 mm 11 mm
13 mm 8 mm
11 mm 14 mm
S.aureus
3 - 3 - ﺳرﺮﺑ ﯽ ﺳﺎﺴﺣ ﯿ ﺰﻧآ ﺖ ﯾﻤ ﯽ ﻌﺗ و ﯿﯿ ﻟﺎﻌﻓ ﻦ ﯿ ﺖ
ﺮﺘﮐﺎــﺑ ي ﺸــﮐ ﯽ ــﯾ ﺮﺘﮐﺎﺑ ﺪــﺷر رﺎــﻬﻣ ﺎ ــﯾ
ﺎﯾ ﯽ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ
ﺳرﺮﺑ رد ﯽ ﺳﺎﺴﺣ ﯿ ﺰﻧآ ﺖ ﯾﻤ ﯽ ﺮﺘﮐﺎﺑ ﯾ ـﺳﻮ ﯿ ﺎـﻫ ﻦ ي ﻟﻮﺗ ـﯿ و هﺪـﺷ ﺪ
ﻟﺎﻌﻓ ﯿ ﺮﺘﮐﺎﺑ ﺖ ي ﺸﮐ ﯽ ﯾ ﺮﺘﮐﺎﺑ ﺪـﺷر رﺎﻬﻣ ﺎ ـﯾ
ﺎﯾ ﯽ ﺖـﺷﺬﮔ زا ﺲـﭘ
ﺮﺘﮐﺎﺑ ﻪﮐ ﺪﺷ هﺪﻫﺎﺸﻣ ،رﺮﻘﻣ نﺎﻣز تﺪﻣ ﯾ
ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ـﯿ هﺪـﺷ ﺪ
ﻮﺳ ود ﺮﻫ ﻂﺳﻮﺗ ﯾ
ﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﻪﺑ ﻪﻌﻟﺎﻄﻣ درﻮﻣ
ﺰﻧآ ﯾ زﺎﺌﺗوﺮﭘ ﻢ ي ﺌﺗوﺮﭘ ﯿ زﺎﻨ ﺳﺎﺴﺣ ﻼﻣﺎﮐ K ﯿ
ﺰـﻧآ ﺎـﻣا ﺪﻨﺘﺷاد ﺖ ﯾ
ﻢ
ﮐ ﺎﻔﻟآ ﯿ ﺮﺗﻮﻤ ﯾ ﺴﭙ ﯿ ﺮﺨﺗ ﻪﺑ ردﺎﻗ ﻦ ﯾ
ﺮﺘﮐﺎﺑ ﺐ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ هﺪـﺷ ﺪ
ﺖﺳا هدﻮﺒﻧ ﻞﻣﺎﮐ ترﻮﺻ ﻪﺑ .
ﺰـﻧآ ﯾ ﺌﺗوﺮﭘ ﻢ ﯿ زﺎـﻨ ـﺳو K ﯿ ﻃ ﻊ ـﯿ ﻒ
ﺮﺨﺗ ﻪﺑ ردﺎﻗ و هدﻮﺑ ﯾ
ﺮﺘﮐﺎﺑ ﺐ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ،ﺖـﺳا هدﻮﺑ هﺪﺷ ﺪ
ﮐ ﺎـﻔﻟآ زﺎـﺌﺗوﺮﭘ ﺎﻣا ﯿ
ﺮﺗﻮﻤ ﯾ ﺴـﭙ ﯿ ﻟد ﻪـﺑ ﻦ ـﯿ ﺻﺎﺼـﺘﺧا ﻞ ﯿ
رد ﻻﺎـﺑ ﺖ
ﺮﺨﺗ ﻪﺑ ردﺎﻗ ،دﺮﮑﻠﻤﻋ ﯾ
ﺮﺘﮐﺎﺑ ﻞﻣﺎﮐ ﺐ ﯾ
ﺳﻮ ﯿ و ﺖـﺳا هدﻮـﺒﻧ ﺎـﻫ ﻦ
ﻣ ﯽ ﺘﻧ ناﻮﺗ ﯿ ﺎﺑ رﺎﺘﺧﺎﺳ ﻪﮐ ﺖﻓﺮﮔ ﻪﺠ ﺮﺘﮐ
ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ،هﺪﺷ ﺪ
اﺮﺘﺴــــﺑﻮﺳ ي
ا ﺐــــﺳﺎﻨﺘﻣ ــــﯾ
ﺰــــﻧآ ﻦ ﯾ ﺪــــﻧا هدﻮــــﺒﻧ ﻢ .
) (A
) (B
Fig 2 Determination of bactericidal or bacteriostatic activity of produced bacterions, (A) Zone of inhibition before enzymatic treatment, (B) After enzymatic treatment, Bactericidal activity of plantaracinBL1 on E.coli
andbacteriostatic activity of plantaracinBL1 on Y.entrocolitica, and bacteriostatic activity of plantaracin EL3against E.coli and Y.entrocolitica are shown
ﻨﭽﻤﻫ ﯿ ﺎـﺘﻧ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ﻦ ﯾ
ﺪـﻌﺑ هﺪـﻣآ ﺖـﺳد ﻪـﺑ ﺞ ﺗ زا
ﯿ ﺎـﻫرﺎﻤ ي
ﺰﻧآ ﯾ ــﻤ ﯽ ﺮﺘﮐﺎﺑ ﻪــﮐ ﺪــﺷ ﺺﺨﺸــﻣ ، ﯾ
ــﺳﻮ ﯿ ﻟﻮﺗ ﻦ ــﯿ ﻂــﺳﻮﺗ هﺪــﺷ ﺪ
ﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ور ،BL1
ي نژﻮﺗﺎﭘ E.coli (ATCC
1399) ﮔﺪﻨﺸﮐ ﺮﺛا ﯽ
ﺗ زا ﺪﻌﺑ و ﻪﺘﺷاد ﯿ
ﺮﺘﮐﺎﺑ رﺎﻤ ﯾ ﺳﻮ ﯿ ﺰـﻧآ ﺎﺑ ﻦ ﯾ ﻢ
ﺎﻫ ي زﺎﺌﺗوﺮﭘ ي ﻫﺎﻣ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ، ﯿ
ﺌﺗوﺮﭘ ﺖ ﯿﻨ ﯽ ا ﯾ ﺘﭙﭘ ﻦ ـﯿ ﺎﻫﺪ ي ﺪـﺿ
ﺮﺘﮐﺎﺑ ﯾﺎﯾ ﯽ ﺰﺠﺗ و ﯾ نژﻮﺗﺎﭘ ،ﺎﻫ نآ ﻪ (ATCC 1399)E.coli
رد
ﺜﮑﺗ ﺪـﺷر مﺪﻋ ﻪﻟﺎﻫ ـﯿ
ﻧ ﺮ ﯿ ﺖـﻓﺎ . ـﻤﻫ ،ﻞـﺑﺎﻘﻣ رد ﯿ
ﺮﺘﮐﺎﺑ ﻦ ﯾ ـﺳﻮ ﯿ ﻦ
ــــﺻﺎﺧ ﯿ ﮔﺪــــﻨﻨﮐرﺎﻬﻣ ﺖ ﯽ
ور ﺮــــﺑ ﺪــــﺷر ي
ATCC
35669) ( Y.entrocolitica ﺗ زا ﺪــﻌﺑ و ﺖــﺷاد
ﯿ ﺎــﻫرﺎﻤ ي
ﺰﻧآ ﯾﻤ ﯽ ﺑ زا و ﯿ ﺮﺘﮐﺎﺑ ﻦﺘﻓر ﻦ ﯾ
ﺳﻮ ﯿ ا ،ﻦ ﯾ ﺮﺘﮐﺎـﺑ ﻦ ي مﺪـﻋ ﻪـﻟﺎﻫ رد
ﺜﮑﺗ ﺪــﺷر ــﯿ ﭘ ﺮ ــﯿ دﺮــﮐ اﺪ . ﺮﺘﮐﺎﺑ ﯾ ــﺳﻮ ﯿ ﻟﻮﺗ ﻦ ــﯿ ﻂــﺳﻮﺗ هﺪــﺷ ﺪ
ﻻ ﺳﺎﺑﻮﺘﮐ ﯿ مورﺎـﺘﻧﻼﭘ سﻮﻠ ور ﺮـﺑ EL3
ي ﻮـﺳ ود ﺮـﻫ ﯾ
نژﻮﺗﺎـﭘ ﻪ
ﺮﮕﻧﺎﺸﻧ ) E.coli و Y.entrocolitica (
ﺻﺎﺧ ﯿ ﮔﺪﻨﻨﮐ رﺎﻬﻣ ﺖ ﯽ
ﺗ زا ﺪﻌﺑ و داد نﺎﺸﻧ ﺪﺷر ﯿ
ﺰﻧآ ﺎﺑ رﺎﻤ ﯾ ﺎﻫ ﻢ ي زﺎﺌﺗوﺮﭘ ي ا ﺪﺷر ، ﯾ ﻦ
ﺎﻫ ﻪﻟﺎﻫ رد ﺎﻫ نژﻮﺗﺎﭘ ي
ﺪﺷ هﺪﻫﺎﺸﻣ ﺪﺷر مﺪﻋ .
3 - 4 - ﻤﺨﺗ ﯿ ﻟﻮﮑﻟﻮﻣ نزو ﻦ ﯽ
ﻟﻮﮑﻟﻮﻣ نزو ﯽ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﻮـﺳ ﻂـﺳﻮﺗ هﺪﺷ ﺪ ﯾ
ﺎـﻫ ﻪ ي
ﺳﺎﺑﻮﺘﮐﻻ ﯿ لژ زرﻮﻓوﺮﺘﮑﻟا ﺎﺑ ،ﻪﻌﻟﺎﻄﻣ درﻮﻣ مورﺎﺘﻧﻼﭘ سﻮﻠ SDS-
ﻌﺗ PAGE ﯿﯿ ﺪﺷ ﻦ ) ﻞﮑﺷ 3 .(
زا ﺪﻌﺑ ﮓﻧر ﻣآ ﯿﺰ ي ﺳﺎﻣﻮﮐ ﺎﺑ لژ ﯽ
ﺮﺘﮐﺎﺑ ،ﻮﻠﺑ ﯾ ﺳﻮ ﯿ ﺎـﻫ ﻦ ي ﻟﻮﺗ ـﯿ ﺪ
ﻮــﺳ ود ﺮــﻫ ﻂــﺳﻮﺗ هﺪــﺷ ﯾ
ﺌﺗوﺮﭘ ﺪــﻧﺎﺑ ﮏــﺗ ﻪ ﯿ
ــﻨ ﯽ هدوﺪــﺤﻣ رد
10kDa - 5 ﮑﺸﺗ ﯿ ﻌﺗ هدوﺪﺤﻣ ﻪﮐ ،ﺪﻧداد ﻞ ﯿﯿ
اﺮﺑ هﺪﺷ ﻦ ي
ﻪﺘـﺳد
ﺮﺘﮐﺎﺑ II ﯾ ﺳﻮ ﯿ ﻣ ﺎﻫ ﻦ ﯽ ﺪﺷﺎﺑ .
Fig 3.The SDS-PAGE analysis of partially purified bacteriocins produced by two studied
L.plantarum species
4 - ﺘﻧ و ﺚﺤﺑ ﯿ
ﮔ ﻪﺠ ﯿﺮ ي
ﻔﻠﺘﺨﻣ عاﻮﻧا ﯽ
ﺮﺘﮐﺎﺑ زا ﯾ ﺳﻮ ﯿ ﺮﺘﮐﺎﺑ زا ﺎﻫ ﻦ ي
ﺎﻫ ي ﺳا ﯿ ﺘﮐﻻ ﺪ ﯿ ﻪﮐ ﮏ
ﺎﻫ اﺬﻏ رد ي ﺎﺳﺎﻨﺷ ،ﺪﻧراد رﻮﻀﺣ ﻒﻠﺘﺨﻣ ﯾ
ﯽ ﺻﻮﺼﺧ و ﯿ
ﺖ ﯾﺑﺎ ﯽ ﺪﻧا هﺪﺷ
18] - 17 .[ ﻟد ﻪﺑ ﯿ اﺰـﻓا ﻞ ﯾ اﺮـﺑ ﺎـﺿﺎﻘﺗ ﺶ ي
تﻻﻮﺼـﺤﻣ
ﺒﻃ ﯿﻌ ﯽ داﻮﻣ و ﺮﺗ ي
ﻣ ظﺎﺤﻟ زا ﻪﮐ ﯿ
ﺑوﺮﮑ ﯿ ژﻮﻟﻮ ﯾ ﮑ ﯽ ا ﯾ ،ﺪﻨـﺷﺎﺑ ﻦـﻤ
ﻧﯿ ﺎﻫ شور ﻪﺑ زﺎ ي
راﺪﻬﮕﻧ ي ز ﯾ ﺘﺴ ﯽ ﻣ سﺎﺴﺣا اﺬﻏ ﺖﻌﻨﺻ رد ﯽ
دﻮﺷ . ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺴﻧﺎﺘﭘ ﺎﻫ ﻦ ﯿ
ﻬﺟﻮﺗ ﻞﺑﺎﻗ ﻞ ﯽ
اﺮﺑ ي راﺪﻬﮕﻧ ي ،اﺬـﻏ
ﺎﻫ دﺮﺑرﺎﮐ ي ﻧﺎﻣرد ﯽ ﺎﻫ ﻞﻤﮑﻣ ناﻮﻨﻋ ﻪﺑ نﺎﺴﻧا رد ـﯾ
ﺎـﺟ ﺎ ﯾ ﺰﮕ ﯾﻨ ﯽ
ﺮﺑ ا ي ﺘﻧآ ﯽ ﺑ ﯿ ﺗﻮ ﯿ ﺎﻫ ﮏ ي ار ﯾ ﺪﻧراد ،هدﺎﻔﺘﺳا درﻮﻣ ﺞ 20]
- 19 .[
ﺎﺘﻧ ﻪﺟﻮﺗ ﺎﺑ ﺮﺿﺎﺣ ﺶﻫوﮋﭘ رد ﯾ
ﻌﺗ زا ﻞﺻﺎﺣ ﺞ ﯿﯿ
ﻃ ﻦ ﯿ رﺎﻬﻣ ﻒ ي
ﺮﺘﮐﺎﺑ ﻪﮐ ﺪﺷ هﺪﻫﺎﺸﻣ ﯾ
ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ ﻮﺳ ود ﺮﻫ ﻂﺳﻮﺗ هﺪﺷ ﺪ ﯾ
ﻪ
ﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﻠﻋ ﺮﺑ ،ﻪﻌﻟﺎﻄﻣ درﻮﻣ
ﯿ ﺮﺘﮐﺎﺑ ﻪ ي ﺎﻫ ي مﺮﮔ
ﻔﻨﻣ مﺮﮔ و ﺖﺒﺜﻣ ﯽ
ﺶﻫوﮋﭘ رد ﻪﺘﻓر رﺎﮐ ﻪﺑ ا ،ﺪﻧدﻮﺑ لﺎﻌﻓ
ـﯾ رد ﻦ
ﻟﺎﺣ ﯽ ﺮﺘﮐﺎﺑ ﻪﮐ ﺖﺳا ﯾ
ﺳﻮ ﯿ ﺮﺘﮐﺎﺑ ﻦ ي ﺎﻫ ي ﺳا ﯿ ﺘﮐﻻ ﺪ ﯿ ﺮﺑ ﻻﻮﻤﻌﻣ ﮏ
ﻠﻋ ﯿ ﺮﺘﮐﺎﺑ ﻪ ي ﺎﻫ ي ﻔﻨﻣ مﺮﮔ ﯽ ﻧ لﺎﻌﻓ ﯿ ﮐﺪﻧا تﺎﺷراﺰﮔ ﺎﻬﻨﺗ و ﺪﻨﺘﺴ ﯽ
ﻟﺎﻌﻓ زا ﯿ ﻠﻋ ﺮﺑ ﺖ ﯿ ﺎﻫ ﻪﻧﻮﮔ ﻪ ي ﺖـﺳا هﺪـﺷ ﻪـﺋارا ﻼﻧﻮﻤﻟﺎﺳ 21]
.[
ﺮﺘﮐﺎﺑ رﺎﻬﻣ ي ﺎﻫ ي ﻔﻨﻣ مﺮﮔ ﯽ ﺮﺘﮐﺎﺑ ﻂﺳﻮﺗ ﯾ
ـﺳﻮ ﯿ ﺮﺘﮐﺎـﺑ ﻦ ي ﺎـﻫ ي
ﺳا ﯿ ﺘﮐﻻ ﺪ ﯿ ﮏ ﯾ و ﮏ ﯾ ﮔﮋ ﯽ ﻣ دﺮﻓ ﻪﺑ ﺮﺼﺤﻨﻣ ﯽ
دﺮﺑرﺎـﮐ ﻪـﮐ ﺪـﺷﺎﺑ
ﺎﻫ ي وراد ﯾ ﯽ ﻧ ار نآ ـﯿ ـﻣ شﺮﺘﺴـﮔ ﺰ ﯽ
ﺪـﻫد . ﺧا ـﯿ تﺎـﺷراﺰﮔ اﺮ
دﺪﻌﺘﻣ ي ﺎﻫﺎﺸﻏ رﺎﻬﻣ درﻮﻣ رد ي
ـﻔﻨﻣ مﺮﮔ ﯽ ﺎـﻣا هﺪـﺷ ﻪـﺋارا ﺎـﻫ
ﺋﺰﺟ ﯿ ﻧﺎﮑﻣ تﺎ ـﯿ ﺎﺑ ﺎـﻫ ﻢﺴ ـﯾ ﺑ ﺪ ـﯿ ﻘﺤﺗ درﻮـﻣ ﺮﺘﺸ ـﯿ
راﺮـﻗ ﻖ ﮔ ـﯿ دﺮ
23] - 22 .[ ﺧﺮﺑ ﯽ ﻘﺤﺗ ﯿ ﻟﺎﻌﻓ تﺎﻘ ﯿ ﺮﺘﮐﺎﺑ ﺖ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻟﻮﺗ ﯿ هﺪﺷ ﺪ
ﻂــﺳﻮﺗ ــﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎــﺘﻧﻼﭘ سﻮﻠ LR14]
24 ،[ ــﺳﺎﺑﻮﺘﮐﻻ ﯿ سﻮﻠ
مورﺎﺘﻧﻼﭘ 25ZJ5]
ﻧ ﯿ[ ﻠﻋ ﺮﺑ ﺰ ﯿ ﺮﺘﮐﺎﺑ ﻪ ي ﺎﻫ ي مﺮـﮔ و ﺖـﺒﺜﻣ مﺮﮔ
ﻔﻨﻣ ﯽ ﺪﻧا هداد شراﺰﮔ ار .ا
ﯾ ﻟﺎﺣ رد ﻦ ﯽ ﺮﺘﮐﺎﺑ ﺖﺳا ﯾ
ـﺳﻮ ﯿ ﺎـﻫ ﻦ ي
ﻟﻮﺗ ــﯿ ﻂــﺳﻮﺗ هﺪــﺷ ﺪ ــﺳﺎﺑﻮﺘﮐﻻ
ﯿ مورﺎــﺘﻧﻼﭘ سﻮﻠ SIK-8]
26 ،[
ﺳﺎﺑﻮﺘﮐﻻ ﯿ ﺳا سﻮﻠ ﯿ ﻓوﺪ ﯿ سﻮﻠ 27] ﻠﻋ ﺮﺑ ﺎﻬﻨﺗ [ ﯿ
ﺮﺘﮐﺎﺑ ﻪ ي ﺎﻫ ي مﺮﮔ
ﻓ ﺖﺒﺜﻣ ﻟﺎﻌ ﯿ ﺪﻧا هداد نﺎﺸﻧ ﺖ .
ﺳرﺮﺑ رد ﯽ ﺎﭘ ﯾ راﺪ ي ﺗراﺮﺣ ﯽ ﺎﺘﻧ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ﯾ
،هﺪـﻣآ ﺖـﺳد ﻪـﺑ ﺞ
ﺮﺘﮐﺎﺑ ﻪــﮐ ﺪــﺷ ﺺﺨﺸــﻣ ﯾ
ــﺳﻮ ﯿ ﺎــﻫ ﻦ ي ﻟﻮﺗ ــﯿ ﻂــﺳﻮﺗ هﺪــﺷ ﺪ
ــــﺳﺎﺑﻮﺘﮐﻻ ﯿ
مورﺎــــﺘﻧﻼﭘ سﻮﻠ LP31]
28
، [ ــــﺳﺎﺑﻮﺘﮐﻻ ﯿ
سﻮﻠ
مورﺎـــــــــــــﺘﻧﻼﭘ TF711]
29 ،[ ـــــــــــــﺳﺎﺑﻮﺘﮐﻻ ﯿ
سﻮﻠ
مورﺎﺘﻧﻼﭘ NCIM2084]
1 ﻧﯿ[ ﺮﺘﮐﺎﺑ ﺪﻨﻧﺎﻤﻫ ﺰ ﯾ
ﺳﻮ ﯿ ﺎـﻫ ﻦ ي ﻟﻮﺗ ـﯿ ﺪ
ﻮﺳ ود ﻂﺳﻮﺗ هﺪﺷ ﯾ
ﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ رد ﻪـﻌﻟﺎﻄﻣ درﻮﻣ
اﯾ ﺎﻫ تراﺮﺣ ﻪﺑ ،ﺶﻫوﮋﭘ ﻦ ي
ﺎﺗ ﻻﺎﺑ C 121° ﺪـﻧدﻮﺑ موﺎـﻘﻣ .
رد
ﺮﺘﮐﺎﺑ ﻞـــﺑﺎﻘﻣ ﯾ
ـــﺳﻮ ﯿ ﻟﻮﺗ ﻦ ـــﯿ ﻂـــﺳﻮﺗ هﺪـــﺷ ﺪ ـــﺳﺎﺑﻮﺘﮐﻻ
ﯿ سﻮﻠ
مورﺎﺘﻧﻼﭘ رد و هدﻮـﺑ سﺎﺴـﺣ ﻻﺎـﺑ تراﺮـﺣ ﻪﺑ LC74
C 95°
ﻟﺎﻌﻓ ﯿ ﺮﺘﮐﺎﺑ ﺪﺿ ﺖ ﯾﺎﯾ
ﯽ داﺪﻧ نﺎﺸﻧ 5]
.[
ﺎﺘﻧ سﺎﺳا ﺮﺑ ﯾ
ﻪـﺑ ﺞ ﺳﺎﺴـﺣ زا هﺪـﻣآ ﺖـﺳد ﯿ
ﺰـﻧآ ﻪـﺑ ﺖ ﯾ
ﺎـﻫ ﻢ ي
زﺎـﺌﺗوﺮﭘ ي ــﺳرﺮﺑ و ﯽ ــﺻﺎﺧ ﯿ ﺮﺘﮐﺎــﺑ ﺖ ي ﺸــﮐ ﯽ ــﯾ ﺪــﺷر رﺎــﻬﻣ ﺎ
ﺮﺘﮐﺎﺑ ــﯾ ﺎﯾ ﯽ ﺮﺘﮐﺎﺑ ﯾ ــﺳﻮ ﯿ ﺎــﻫ ﻦ ي ﻟﻮﺗ ــﯿ ﻪــﮐ ﺪــﺷ هﺪﻫﺎﺸــﻣ ،هﺪــﺷ ﺪ
ﺮﺘﮐﺎﺑ ﯾ ـــﺳﻮ ﯿ ﺎـــﻫ ﻦ ي ﻟﻮﺗ ـــﯿ ﻂـــﺳﻮﺗ هﺪـــﺷ ﺪ ـــﺳﺎﺑﻮﺘﮐﻻ
ﯿ سﻮﻠ
مورﺎﺘﻧﻼﭘ 23C19]
ﺗ ﻪﺑ سﺎﺴﺣ [ ﯿ
ﺰـﻧآ رﺎﻤ ﯾ زﺎـﺌﺗوﺮﭘ ﻢ ي
و هدﻮـﺑ
ﺪــﺷر رﺎــﻬﻣ دﺮــﮑﻠﻤﻋ ي
ﻠﻋ ﺮــﺑ ــﯿ نژﻮﺗﺎــﭘ ﻪ ﺮﮕﻧﺎﺸــﻧ
ﻟ ــﯿ ﺮﺘﺴ ﯾ ﺎ
ــﺳﻮﻧﻮﻣ ﯿ ﺰﻧژﻮﺘ IP6818 ﺖــﺷاد . ﺸــﻫوﮋﭘ رد ﯽ
د ﯾ ور ﻪــﮐ ﺮــﮕ ي
ﺳﺎﺑﻮﺘﮐﻻ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ ﻪـﮐ ﺪـﺷ ﺺﺨﺸـﻣ ،ﺪـﺷ مﺎﺠﻧا LP84
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻟﻮﺗ ﻦ ﯿ ﺪ ي ﻠﻋ ﺮﺑ نآ ـﯿ ـﺳﺎﺑ ﻪ ﯿ سﻮﺋﺮـﺳ سﻮﻠ F4810
و
ﺮﺷا ﯾ ﺸ ﯿ ﻠﮐﺎ ﯽ ﻟﺎﻌﻓ D21 ﯿ ﺮﺘﮐﺎﺑ ﺖ ي ﺸﮐ ﯽ ﻧ و ﻪﺘﺷاد ـﯿ ﺰـﻧآ ﻪـﺑ ﺰ ﯾ ﻢ
ﺎــﻫ ي زﺎــﺌﺗوﺮﭘ ي دﻮــﺑ سﺎﺴــﺣ 29]
.[ ــﺷ ﻪــﺘﻔﮔ ﻪــﮐ رﻮﻄﻧﺎــﻤﻫ ﺪ
ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻫﺎﻣ ﻦ ﯿﺘ ﯽ ﺌﺗوﺮﭘ ﯿﻨ ﯽ ﺳﺎﺴﺣ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ و دراد ﯿ
ﻪﺑ نآ ﺖ
ﺰﻧآ ﯾ ﺎﻫ ﻢ ي زﺎﺌﺗوﺮﭘ ي ﺰﻧآ ﻂﺳﻮﺗ ﯾ
ﺎﻫ ﻢ ي ـﺷراﻮﮔ ﯽ ـﻣ ﻪﺘﺴـﮑﺷ ﯽ
دﻮﺷ . اﺮﺑﺎﻨﺑ ﯾ ﻣ ﻦ ﯽ اﺮﺑ ار ﺎﻫ نآ ناﻮﺗ ي
تﻻﻮﺼـﺤﻣ رد فﺮﺼـﻣ
ﻤﺨﺗ ﯿﺮ ي ا ، ﯾ ﺖﻓﺮﮔ ﺮﻈﻧ رد ﻦﻤ 10]
.[ ﺎﻧاﻮﺗ ﯾ ﯽ ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ اﺮـﺑ ﻦ ي
ﺎﻫ نژﻮﺗﺎﭘ ﻦﺘﺸﮐ ي
ﻣﻮﻤﺴـﻣ ﻞـﻣﺎﻋ ﯿ
ﺬـﻏ دﺎﺴـﻓ و ﺖ اﯾ
ﯽ ﺎـﻧاﻮﺗ ﯾ ﯽ
ﻣز رد ﺪﻨﻤﺷزرا ﯿ
اﺬﻏ تﻻﻮﺼﺤﻣ و ﺖﻌﻨﺻ ﻪﻨ ﯾ
ﯽ ﻣ ﯽ ز ﺪﺷﺎﺑ ﯾ ﺎﺑ اﺮ
ﺑ زا لﺎﻤﺘﺣا ﯿ
ﺮﺘﮐﺎﺑ ﻦﺘﻓر ﻦ ﯾ
ﺳﻮ ﯿ ﻃ ﻦ ﯽ آﺮﻓ ﯾ ﺎﻫ ﺪﻨ ي راﺪﻬﮕﻧ ي ﺛﺎﺗ ، ﯿ ﺮ
ﺷ داﻮﻣ ﯿﻤ ﯿﺎ ﯾ ﯽ و ....
دﯾ ﺜﮑﺗ ﻪﺑ ردﺎﻗ هﺪﺷ ﻪﺘﺸﮐ نژﻮﺗﺎﭘ ﺮﮕ ﯿ
دﺪﺠﻣ ﺮ
دﻮﺑ ﺪﻫاﻮﺨﻧ .
ﻪــﺟﻮﺗ ﺎـﺑ ﺎــﺘﻧ ﻪــﺑ
ﯾ لژ ﺞ SDS-PAGE ﮑﺸــﺗ و
ﯿ ﺪــﻧﺎﺑ ﮏــﺗ ﻞ
ﺌﺗوﺮﭘ ﯿﻨ ﯽ ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎﻫ ﻦ ي ﻮﺳ ود ﺮﻫ ﯾ
ﺳﺎﺑﻮﺘﮐﻻ ﻪ ﯿ مورﺎﺘﻧﻼﭘ سﻮﻠ
هدوﺪــﺤﻣ رد ﻪـﻌﻟﺎﻄﻣ درﻮـﻣ 10kDa
- 5 نارﺎــﮑﻤﻫ و ورودﻮـﺗ ،
) 2004 ( ﻧﯿ ﻟﻮﮑﻟﻮﻣ نزو ﺰ ﯽ
10kDa اﺮﺑ ار ي ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻟﻮﺗ ﻦ ـﯿ ﺪ
ﻮﺳ ﻂﺳﻮﺗ هﺪﺷ ﺳﺎﺑﻮﺘﮐﻻ ﯾﻪ
ﯿ مورﺎـﺘﻧﻼﭘ سﻮﻠ ST13BR
شراﺰـﮔ
ﺪﻧدﺮﮐ . ﻟآ ﯽ نارﺎﮑﻤﻫ و )
2007 ( ﺮﺘﮐﺎﺑ ﻪﮐ ﺪﻧدﺮﮐ شراﺰﮔ ﯾ
ـﺳﻮ ﯿ ﻦ
ﻟﻮﺗ ــﯿ ــﺳﺎﺑﻮﺘﮐﻻ ﻂــﺳﻮﺗ هﺪــﺷ ﺪ ﯿ
مورﺎــﺘﻧﻼﭘ سﻮﻠ IIA-1A5
رد
هدوﺪﺤﻣ 5kDa /6 ﺗوﺮﭘ ﺪﻧﺎﺑ ﮏﺗ ﺌﯿﻨ
ﯽ ﮑﺸﺗ ﯿ داد ﻞ . ﻟﻮﮑﻟﻮﻣ نزو ﯽ
5kDa /9 اﺮﺑ ي ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﻟﻮﺗ ﻦ ﯿ ﻂـﺳﻮﺗ هﺪﺷ ﺪ ـﺳﺎﺑﻮﺘﮐﻻ
ﯿ سﻮﻠ
مورﺎﺘﻧﻼﭘ ﺷ زا هﺪﺷ اﺪﺟ ﯿ
ا ﻪﮐ ﺪﺷ شراﺰﮔوﺎﮔ مﺎﺧ ﺮ ـﯾ
ﺘﻧ ﻦ ﯿ ﻪـﺠ
ﭘﯿ ﻣ دﺎﻬﻨﺸ ﯽ ا ﺖﺳا ﻦﮑﻤﻣ ﻪﮐ ﺪﻫد ﯾ
ﺮﺘﮐﺎﺑ ﻦ ﯾ ﺳﻮ ﯿ ﻧ ﻦ ﯿ ﻪﺘﺳد ءﺰﺟ ﺰ
ﺮﺘﮐﺎﺑ II ﯾ ﺳﻮ ﯿ ﺪﺷﺎﺑ ﺎﻫ ﻦ 10]
.[
ﺎﺘﻧ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ﯾ
ﻨﺒﻣ هﺪﻣآ ﺖﺳد ﻪﺑ ﺞ ﯽ
ﻃ ﺮـﺑ ـﯿ رﺎـﻬﻣ ﻒ ي ـﺳو ﯿ ،ﻊ
ﺎﭘ ــﯾ راﺪ ي ــﺗراﺮﺣ ﯽ ﺳﺎﺴــﺣ و ﯿ
ﺰــﻧآ ﻪــﺑ ﺖ ﯾ
ﺎــﻫ ﻢ ي زﺎــﺌﺗوﺮﭘ ي و
ﺻﺎﺧ ﯿ ﺮﺘﮐﺎﺑ ﺖ ي ﺸﮐ ﯽ ﺮﺘﮐﺎﺑ ﺪﺷر رﺎﻬﻣ و ﯾﺎﯾ
ﯽ ﺮﺘﮐﺎﺑ ﯾ ﺳﻮ ﯿ ﺎـﻫ ﻦ ي
ﻟﻮﺗ ﯿ ا رد هﺪﺷ ﺪ ﯾ
ﻣ ،ﺶﻫوﮋﭘ ﻦ ﯽ
ﺘﻧ ناﻮﺗ ﯿ ﻪﺑ ﻪﺟﻮﺗ ﺎﺑ ﻪﮐ ﺖﻓﺮﮔ ﻪﺠ
ﺷﺎﻧ ضراﻮﻋ ﯽ
ﮐﺮﺗ فﺮﺼﻣ زا ﯿ
ﻣ ﺪـﺿ تﺎﺒ ﯿ
ـﺑوﺮﮑ ﯽ ـﺷ ﯿﻤ ﯿﺎ ﯾ ﯽ و
ﻋﻮﻨﺼﻣ ﯽ ا ، ﯾ ﻟﻮﺑﺎﺘﻣ ﻦ ﯿ ﻣ ﺎﻫ ﺖ ﯽ ﺑ ﺪﻨﻧاﻮﺗ ﮐﺮﺗ ناﻮﻨﻋ ﻪ ﯿ
ﺎﻫ ﺐ ي ﺪﺿ
ﻣﯿ ﺑوﺮﮑ ﯽ ﺒﻃ ﯿﻌ ﯽ اﺬﻏ دﺮﺑرﺎﮐ ﯾ
ﯽ وراد و ﯾ ﯽ ﻪﺘﺷاد ﺪﻨﺷﺎﺑ .
5 - ﻊﺑﺎﻨﻣ
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Partial Purification and Characterization of produced Bacteriocins by Two East-Azarbayjan Native Isolates of
Lactobacillus plantarum
Khaledzade, Sh.1, Hejazi, M. A.2, Masoomi Jahandizi, R. 3
1. MSc student of Microbial Biotechnology, Faculty of Basic Science, University of Maragheh 2. Associate Professor, Agriculture Biotechnology Research Institute of North and North-West (ABRII) 3. Assistant Professor, Department of Microbial Biotechnology, Faculty of Basic science, University of
Maragheh
(Received: 2016/08/09 Accepted:2018/10/21)
In this study, some characters of produced bacteriocins by two isolates of L.plantarum were studied.
At fiest, two native isolates of L.plantarum BL1 and L.plantarum EL3 were selected from microbial collection of ABRII, and then the antibacterial trait and inhibitory spectrum of produced bacteriocins were determined. Therefore, bacteriocidal or bacteriostatic activity of produced bacteriocins, and thermal stability of them in different tempretures were studied. Molecular weight of bacteriocins were estimated by SDS-PAGE technique. According to achieved results, produced bacteriocins by two isolates were active against all used pathogens and had wide inhibitory spectrum, and even after different thermal treatments they retain their antibacterial activity. Also, it was observed that both produced bacteriocins were sensitive to proteinase-K and destroyed completely. In contrast, alpha- chymotrypsin was not able to destroy bacteriocins properly. On the other hand, it was revealed that L.plantarum BL1 bacteriocin had bacteriocidal effect on Eschrichia coli (ATCC 1399), but bacteriostatic activity against Yerssinia entrocolitica (ATCC 35669) as indicator pathogens.
L.plantarum EL3 bacteriocin showed bacteriostatic activity on both noted indicator pathogens.
Produced bacteriocins by both studied L.plantarum strains, had molecular weight lower than 10 kDa ( 5 kDa < produced bacteriocins < 10 kDa) and taked place in specified range of class IIb bacteriocins.
Keywords: Lactic acid bacteria, Lactobacillus plantarum, Bacteriocin, Partial purification, Antibacterial activities
