شناسایی انتروسین‌ها و بررسی برون-تنی فعالیت ضدمیکروبی و ضدسرطانی آن‌ها در سویه‌های انتروکوک جداشده از محصولات تخمیری سنتی

محورهای موضوعی : بهداشت مواد غذایی

فائزه سالک ¹ , حمید میرزایی ² , جلیل خندقی ³ , افشین جوادی ⁴ , یوسف نامی ⁵

1 - گروه بهداشت مواد غذایی، دانشکده دامپزشکی، علوم پزشکی تبریز، دانشگاه آزاد اسلامی، تبریز، ایران.
2 - گروه بهداشت مواد غذایی، دانشکده دامپزشکی، واحد علوم پزشکی تبریز، دانشگاه آزاد اسلامی، تبریز، ایران
3 - گروه علوم و صنایع غذایی، واحد سراب، دانشگاه آزاد اسلامی، سراب، ایران
4 - گروه بهداشت مواد غذایی، دانشکده دامپزشکی، واحد علوم پزشکی تبریز، دانشگاه آزاد اسلامی، تبریز، ایران
5 - گروه بیوتکنولوژی مواد غذایی، واحد شمال غرب و غرب پژوهشکده بیوتکنولوژی کشاورزی، سازمان تحقیقات، آموزش و ترویج کشاورزی (AREEO)، تبریز، ایران

تاریخ دریافت : 1402/02/26 تاریخ پذیرش : 1402/05/22 تاریخ انتشار : 1402/04/01

کلید واژه: باکتریوسین, ضدمیکروبی, انتروکوکوس, ضدتوموری,

چکیده مقاله :

جایگزینی ترکیبات زیست‌فعال به‌دست آمده از منابع طبیعی مانند متابولیت‌های میکروبی با ترکیبات نگهدارنده سنتزی و بهره‌مندی از خواص فراسودمند این متابولیت‌ها ازجمله اثرات درمانی آن‌ها روز به‌روز در حال گسترش است. در این میان، باکتریوسین‌ها که دارای فعالیت فیزیولوژیـک متعـدد از جمله خواص ضـدمیکروبی و ضدسرطانی هستند، کانون توجه بسیاری از محققین بوده‌اند. در این مطالعه ژن‌های تولید انتروسین جدایه‌های انتروکوکوس دو محصول لبنی تخمیری (پنیر موتال و ترخینه) به‌روش ملکولی ردیابی شده و کینتیک تولید ترکیبات شبه باکتریوسینی و پایداری آن‌ها در دمای بالا، pH‌های مختلف و تحت تاثیر آنزیم‌های گوناگون بررسی شد. همچنین خواص آنتاگونیستی (به‌روش انتشار در چاهک) و ضدسرطانی (با تکنیک رنگ‌سنجی MTT) انتروسین‌های تولید شده توسط این جدایه‌ها بر روی دو رده سلولی سرطانی انسان مورد امطالعه قرار گرفت. نتایج آزمونPCR نشان داد که دو جدایه انتروکوکوس فاسیوم AUT-7KB و انتروکوکوس فکالیس KUMS-T48 حاوی هر سه ژن‌ تولید انتروسین مورد مطالعه بوده و متابولیت‌های شبه باکتریوسینی این سویه‌ها اثرات آنتاگونیستی قوی بر روی باکتری‌های لیستریا مونوسایتوجنز و باسیلوس سرئوس داشتند. این متابولیت‌ها از پایداری بالایی در pHهای مختلف برخوردار بودند و تحت تاثیر آنزیم‌های پروتئاز (تریپسین و پپسین) بی‌اثر شدند که نشانگر ماهیت پروتئینی آن‌ها بود. به‌علاوه، متابولیت‌های جدایه KUMS-T48 اثرات سایتوتوکسیک قوی روی دو رده سلولی سرطانی AGS و HT-29 نشان داد. با توجه به یافته‌های تحقیق حاضر، انتروسین‌های سویه انتروکوکوس فکالیس KUMS-T48 به‌عنوان گزینه مناسبی برای مطالعات ضدتوموری بیشتر مانند بررسی‌های درون-تنی پس از بررسی‌های کامل جنبه‌های ایمنی پیشنهاد می‌شوند.

چکیده انگلیسی:

Replacing naturally occurring bioactive substances, such as microbial metabolites, with synthetic preservative compounds, and getting access to their beneficial properties, is expanding every day. Meanwhile, the focus of many researchers' attention has been on bacteriocins, which have multiple physiological functions, including antibacterial and anticancer properties. In this study, the enterocin production genes of Enterococcus isolates from two fermented dairy products (Motal cheese and Tarkhineh) were tracked using a molecular method. The kinetics of bacteriocin-like substance production, as well as its stability (at high temperatures, different pH levels, and under different enzyme influences), were investigated. The antagonistic (well diffusion method) and anticancer (with MTT colorimetric technique) characteristics of enterocins generated by these isolates were also investigated in two human cancer cell lines. The results of the PCR test showed that isolates of Enterococcus faecium AUT-7KB and Enterococcus faecalis KUMS-T48 contained all three studied enterocin production genes, and the bacteriocin-like metabolites of these strains had strong antagonistic effects on Listeria monocytogenes and Bacillus cereus. These metabolites had high stability at different pH and became ineffective under the influence of protease enzymes (trypsin and pepsin), which indicated their protein nature. Additionally, metabolites of the KUMS-T48 isolate showed strong cytotoxic effects on two cancer cell lines, AGS and HT-29. According to the findings of the current research, enterocins of the E. faecalis KUMS-T48 strain are suggested as a suitable candidate for further anti-tumor studies, such as in vivo studies, after complete investigations of safety aspects.

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