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افزایش تولید سورفکتین در باکتری Bacillus subtilis UTB96 با استفاده از بهینه‌سازی فرایند فرمانتاسیون | ||
| مهار زیستی در گیاه‌پزشکی | ||
| مقاله 2، دوره 5، شماره 2 - شماره پیاپی 10، اسفند 1396، صفحه 13-26 اصل مقاله (341.18 K) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/bcpp.2018.117885 | ||
| نویسندگان | ||
| سحر کشاورزی1؛ مسعود احمدزاده1؛ سعید میرزایی2؛ کیوان بهبودی1؛ مژگان بنده‌پور بنده‌پور3 | ||
| 1گروه گیاه‌پزشکی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج، ایران | ||
| 2گروه بیوتکنولوژی کشاورزی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته کرمان، کرمان، ایران | ||
| 3دانشکده فناوری‌های نوین پزشکی، دانشگاه علوم پزشکی شهید بهشتی، تهران، ایران | ||
| چکیده | ||
| در میان باکتری‌های زیست مهارگر، باکتری Bacillus subtilis در تولید طیف وسیعی از ترکیبات ضدمیکروبی مطرح می‌باشد که از این میان، بیوسورفکتانت‌ها از جمله سورفکتین از اهمیت ویژه‌ای برخورد است. سورفکتانت‌هایی که بهوسیلۀ میکروارگانیسم‌ها تولید می‌شوند به‌دلیل سمیّت پایین و خواص منحصر به فرد، کاربردهای متعددی دارند اما به‌دلیل هزینه بالای تولید و خالصسازی، تولید در بازده بالا هنوز میسر نشده است. در این تحقیق بهمنظور ارزیابی تولید سورفکتین در جدایه مذکور، شناسایی ژن تولید کننده آن (srf) و تعیین مقدار سورفکتین تولید شده به‌ترتیب از روش‌های متلاشی شدن قطره (Drop collapse)، روش مولکولی واکنش زنجیرهای پلیمراز (PCR) و آنالیز کروماتوگرافی مایع با کارایی بالا (HPLC) استفاده شد. بررسی مولکولی با آغازگر اختصاصی نشان داد که این جدایه دارای ژنsrf می‌باشد. برای بهینه‌سازی شرایط عملیاتی، اثر شرایط فیزیک وشیمیایی و القا کننده‌های مختلف بر میزان زیست توده و سورفکتین خام تولیدی بررسی شد. شرایط بهینه فیزیکی و شیمیایی به‌دست آمده شامل: مدّت زمان نگهداری کشت 96 ساعت، دمای 30 درجۀ سلسیوس، اسیدیته 7 و دور شیکر rpm 250 می‌باشد. از میان القاکننده‌ها که شامل منابع کربن، منابع نیتروژن و اسیدهای آمینه، بیشترین مقدار تولید سورفکتین پس از تیمار محیط کشت با گلوکز به‌مقدار 243/1 گرم بر لیتر محیط کشت می‌باشد. پس از آن تأثیر عناصر القاکننده‌ای که بیشترین تأثیر را در افزایش تولید سورفکتین داشته‌اند، بهصورت توأم بررسی شد. نتایج نشان داد بیشترین مقدار تولید سورفکتین mg/l 055/5 در حضور یون‌های آهن، منیزیم و منگنز بهصورت توأم می‌باشد. همچنین مقدار و نوع منابع غذایی تأثیر بهسزایی در بازده تولید سورفکتین دارد. بنابر این تغییر در مقدار و ترکیب القا کننده‌ها روشی اقتصادی برای تولید تجاری این ماده محسوب می‌شود. | ||
| کلیدواژه‌ها | ||
| سورفکتانت؛ فرمانتاسیون؛ ژن srf؛ کروماتوگرافی مایع با کارایی بالا | ||
| عنوان مقاله [English] | ||
| Enhancing surfactant production in Bacillus subtilis UTB96 by fermentation optimization | ||
| نویسندگان [English] | ||
| Sahar Keshavarzi1؛ Masoud Ahmad zadeh1؛ Saeid Mirzaei2؛ Keivan Behboudi1؛ Mojgan bandeh pour3 | ||
| 2Department of Biotechnology, Graduate University of Advanced Technology, Kerman, Iran | ||
| 3Biotechnology Department, School of Advacned Technologies in Medicine, Shahid Beheshti,University of Medical Sciences, Tehran, Iran | ||
| چکیده [English] | ||
| A biocontrol bacterium, Bacillus subtilis is well known for producing wide spectrum of antimicrobial compounds. Among these products, biosurfactant compounds such as surfactin have high importance. Biosurfactants that are produced by the microorganisms are applied in the various aspects of industry, medicine and agriculture because of their low toxicity and individual properties. However, these compounds are not produced in high quantity due to their low production yield and high purification cost. In the current study for investigating the production of surfactin in the isolate (UTB96), recognition of srf gene and determination of the amount of surfactin production, we used Drop collapse, PCR and HPLC methods. Molecular analysis with specific primer indicated that the isolate had srf gene. To optimize the fermentation process, influence of various inducers and physiochemical factors were studied on the biomass and surfactin production. The results indicated that maximum surfactin production occurred after amending the culture medium with glucose (1.243 g/l) solution. After that influence of the combined inducer elements were studied in increasing the amount of surfactin production. Results revealed that maximum production (5.055 g/l) was done in the presence of Fe2+, Mn2+ and Mg2+ mixture in the culture medium. The results also showed that the amount and the type of the food sources had remarkable impact on the surfactin production efficiency. The changes in the amount and composition of the food sources therefore, could be economically considered for the commercial production of this compound. | ||
| کلیدواژه‌ها [English] | ||
| biosurfactant, fermentation, srf gene, HPLC | ||
| مراجع | ||
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