افزایش تولید آنتی‌بیوتیک تایلوزین در باکتری Streptomycess fradiae با استفاده از روش موتاسیون

نویسندگان

عضو هیئت علمی پژوهشکده بیوتکنولوژی

چکیده

در این بررسی از دو موتاژن اشعه uv با طول موج nm 280 و (EMS (Ethylmethanesulfonate با غلظت‌های متفاوت برای ایجاد موتاسیون و افزایش تولید تایلوزین در باکتری Streptomycess fradiae استفاده شد. منحنی مرگ برای هر دو موتاژن رسم گردید و شرایط بهینه برای ایجاد موتاسیون توسط اشعه  uvبا فاصله 20 سانتی‌متری و مدت زمان 40 تا 60 ثانیه و برای EMS مدت زمان 30 دقیقه و غلظت (w/v) 4 درصد تشخیص داده شد. در مرحله اول، موتاسیون با uv انجام شد و سویه‌های دارای تولید بالا به روش پلاک آگار ارزیابی شدند. در غربال‌گری ثانویه سویه 24U با تولید mg/ml 892/0 تایلوزین و 28/1 برابر بیشتر از سویه وحشی به عنوان سویه مناسب انتخاب شد و در مرحله دوم تحت تیمار با EMS قرار گرفت. در نهایت سویه موتانتی با تولید mg/ml 18/2 تایلوزین و 12/3 برابر افزایش تولید نسبت به سویه وحشی بدست آمد. 

عنوان مقاله [English]

Enhancement of tylosin production in Streptomyces fradiae by Mutation

نویسندگان [English]

  • D Zareh
  • M Azin
  • H Rasi
  • S.S Mirdamadi
Scientific Member of Biotechnology Institute
چکیده [English]

In order to increase tylosin production by Streptomyces fradiae, two mutagens, i. e. uv and EMS (Ethylmethanesulfonate) were used. Death curves were plotted for these mutagens. Optimum condition for obtaining mutants by uv radiation was determined by using a uv source lamp at a distance of 20 cm for 40-60 seconds. When EMS was used at 4% (w/v) concentration, the best mutants were isolated after 30 minutes of exposure to the mutagen. In the first mutation step, which was done by uv radiation, high tylosin producing strains were isolated by Plaque Agar method as the first screening step. For the second step of screening, a mutant strain (U24) with a production of 0.892 mg tylosin per ml was isolated, which was 1.27 times more productive than the wild type. The secondary mutation step on this mutant strain was performed by EMS. Finally, a mutant strain (UE1) with a production of 2.18 mg tylosin per ml was obtained, which was 3.12 times more productive than the wild type.

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