تولید باکتریوسین با استفاده از باسیلوس لکینی‌فورمیس ATCC 9789 و تعیین ویژگی‌های ساختاری و ضدمیکروبی آن

نویسندگان
امین خلیلی 1
محمود رضازاد باری 2
سعید حسامی تکلو 3
صابر Amiri 4
1 دانش آموخته کارشناسی ارشد، گروه زیست شناسی، دانشکده علوم پایه، دانشگاه آزاد اسلامی واحد تهران مرکزی، تهران، ایران
2 استاد گروه علوم و صنایع غذایی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه، ایران
3 استادیار گروه زیست شناسی، دانشکده علوم پایه، دانشگاه آزاد اسلامی واحد تهران مرکزی، تهران، ایران
4 استادیار گروه علوم و صنایع غذایی، دانشکده کشاورزی، دانشگاه ارومیه، ارومیه، ایران
10.22034/FSCT.19.131.319
چکیده
باکتریوسین­ها گروهی از مولکول­های منحصر بفرد هستند که توسط برخی از میکروارگانیسم­ها تولید می­شوند و به عنوان بخشی از ایمنی ذاتی میزبان در نظر گرفته می­شوند. امروزه استفاده از نگهدارنده­های زیستی در صنایع غذایی به طور گسترده در حال افزایش است. در پژوهش حاضر برای تولید باکتریوسین از که باسیلوس لکینی فورمیس ATCC 9789 استفاده شد. فعالیت ضد میکروبی باکتریوسین تولیدی در برابر پاتوژن­های منتقله از غذا بررسی شد. در ادامه پس از خالص سازی، ویژگی­های باکتریوسین تولید شده شامل پایداری در برابر حرارت، pH، آنزیم و اشعه فرابنفش و نیز minimal inhibitory concentration (MIC) و minimal bactericidal concentration (MBC) تعیین شد. همچنین وزن مولکولی، گروه­های عاملی و ویژگی­های دمایی باکتریوسین توسط آزمون sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-Page)، Fourier-transform infrared spectroscopy (FTIR) و Differential scanning calorimetry (DSC) تعیین گردید. این باکتریوسین در حرارت­ها و pHهای مختلف، تحت تابش اشعه فرابنفش و همچنین تحت تیمار با آنزیم­های تریپسین و پپسین پایداری بسیار خوبی نشان دادند. آزمون SDS-Page نشان داد که باکتریوسین تولیدی از دو بخش با وزن مولکولی 17 و 20 کیلودالتون تشکیل شده است. نتایج آنالیز FTIR نشان دهنده ویژگی پپتیدی بود و نتایجDSC دو پیک­های اگزوترمال به ترتیب در 190 و 325 درجه سانتی­گراد داشت. به طور کلی در این پژوهش بعد از استخراج و اطمینان از وجود باکتریوسین، اثر ضد میکروبی آن با تعیین MIC و MBC بر روی پاتوژن­های منتقله از غذا مورد ارزیابی قرار گرفت. نتایج نشان داد که باکتریوسین تولیدی توسط باسیلوس لکینی‌فورمیس دارای اثر ضدمیکروبی قابل توجهی روی باکتری­های پاتوژن­های منتقله از غذا داشت.

کلیدواژه‌ها
اکتریوسین
نگهدارنده زیستی
پاتوژن‌های منتقله از غذا
باسیلوس لکینی‌فورمیس

موضوعات

عنوان مقاله English

Production of bacteriocin using Bacillus licheniformis ATCC 9789 and determination of its structural and antimicrobial properties

نویسندگان English

Amin Khalili 1
Mahmoud Rezazadeh Bari 2
Saeed Hesami Tackallou 3
Saber Amiri 4
1 MSc, Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran
2 Professor, Department of Food Science and Technology, Faculty of Agriculture, Urmia University, Urmia, Iran
3 Assistant professor, Department of Biology, Central Tehran Branch, Islamic Azad University, Tehran, Iran
4 Assistant professor, Department of Food Science and Technology, Faculty of Agriculture, Urmia University, Urmia, Iran
چکیده English

Bacteriocins are a group of unique molecules that are produced by some microorganisms and are considered as part of the host's innate immunity. Nowadays, the use of biological preservatives in the food industry is increasing widely. In the present study, Bacillus laciniformis ATCC 9789 was used to produce bacteriocin. The antimicrobial activity of produced bacteriocin was investigated against foodborne pathogens. Further, after purification, the characteristics of produced bacteriocin, including stability against heat, pH, enzyme and ultraviolet (UV) rays, as well as minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined. Also, molecular weight, functional groups and temperature characteristics of bacteriocin were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-Page), Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) tests. This bacteriocin had very good stability in different temperatures, pHs and against trypsin and pepsin enzymes and under UV rays. The SDS-Page test showed that the produced bacteriocin consists of two parts with a molecular weight of 17 and 20 kDa. The results of FTIR analysis showed peptide characteristics and DSC results had two exothermic peaks at 190 and 325 ⸰C, respectively. In general, in this research, after extracting and ensuring the existence of bacteriocin, its antimicrobial effect was evaluated by determining MIC and MBC on foodborne pathogens. The results showed that the bacteriocin produced by B. licheniformis had a significant antimicrobial effect on foodborne pathogens.

کلیدواژه‌ها English

Bacteriocin
Bio-preservative
Foodborne pathogens
Bacillus licheniformis
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