جداسازی و شناسایی باکتری تولید کننده آنزیم ال-آسپاراژیناز برون سلولی فاقد خاصیت گلوتامینازی از خلیج فارس

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشکده علوم و فناوری زیستی، دانشگاه شهید بهشتی، اوین، تهران

2 میکروبیولوژی و زیست فناوری میکروبی دانشگاه شهید بهشتی

3 مرکز تحقیقات پروتئین، دانشگاه شهید بهشتی، اوین،

4 ژوهشگاه ملی مهندسی ژنتیک و زیست فناوری، بزرگراه کرج، تهران

چکیده

آنزیم ال-آسپاراژیناز کاربردهای مختلفی در پزشکی و صنایع غذایی دارد. به دلیل ایجاد برخی عوارض جانبی  به دلیل فعالیت نامطلوب گلوتامینازی؛ تلاش برای یافتن منابع میکروبی جدید تولید کننده آنزیم فاقد فعالیت گلوتامینازی ادامه دارد. هدف مطالعه حاضر جداسازی باکتری تولید کننده آنزیم ال-آسپاراژیناز نوع دوم، فاقد خاصیت گلوتامینازی از آب خلیج فارس و اندازه گیری میزان آنزیم  و شرایط افزایش آن میباشد. جهت بررسی تولید آنزیم آسپاراژیناز برون سلولی، باکتریهای خالص جداسازی شده  از آب  دریا در محیط  M9 حاوی فنل رد و آسپاراژین کشت داده شد . به منظور بررسی خاصیت گلوتامینازی باکتریهایی که تست آسپاراژینازی مثبت داشتند در محیط M9  دارای گلوتامین و فنل رد کشت داده شد. جهت شناسایی جدایه باکتریایی دارای خاصیت آسپاراژینازی مثبت و خاصیت گلوتامینازی منفی از روش‌های مورفولوژی، بیوشیمیایی و توالی یابی  S rDNA 16 استفاده شد. ارزیابی  فعالیت آنزیمی ال- آسپاراژیناز طبق روش رنگ سنجی انجام گرفت. تاثیر القای بی هوازی در میزان فعالیت آنزیم ال- آسپاراژینازی II با کشت در دو شرایط هوادهی و بدون هوادهی بررسی شد.  نتایج مطالعه نشان داد که جدایه باکتری، Rhizobium nepotum strain SHN1 می باشد. فعالیت آنزیم آسپاراژینازی این باکتری معادل  0.467 U/mL و فعالیت ویژه آنزیمی آن  IU/mg 0.015  بدست آمد. این فعالیت در شرایط القا بی هوازی بیش از 50 درصد افزایش داشت. نتایج حاکی از آن بود که فلور میکروبی  آبهای خلیج فارس میتواند منبع مناسبی برای تولید آنزیم ال – آسپاراژیناز دارای خاصیت ضد سرطانی و فاقد فعالیت گلوتامینازی باشد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Isolation and characterization of a bacterium with free glutaminase L-Asparaginase II from the Persian Gulf

نویسندگان [English]

  • Saloomeh Shoaei Naeeni 1
  • Gholamhossein Ebrahimipour 2
  • Seyed Omid Ranaei Siada 3
  • Bijan Bambai 4

1 Faculty of Biological Science and technology, Shahid Beheshti University, Evin, Tehran

2 Microbiology and microbial Biotechnology in Shahid Beheshti University

3 protein Research Center, Shahid Beheshti University, Evin,

4 National Gohshgah of Genetics Engineering and Biotechnology, Karaj Highway, Tehran

چکیده [English]

L-asparaginase enzyme has various applications, particularly in medicine and food industry. Given some side effects and adverse possession of a minor amount of glutaminase activity, the search for new sources of microbes producing glutaminase-free L-asparaginase type II is underway. The present study aimed to isolate a bacterium from the Persian Gulf that produces glutaminase-free L-asparaginase type II. It is also aimed to measure the amount of this enzyme and the condition under which it increases. In order to assess the capability of extracellular asparagine production, the isolated bacteria from the seawater were cultured in M9 medium containing phenol red and asparagine. Those bacteria with positive asparaginase test were cultured in M9 medium containing glutamine and phenol red to assess their glutamine activity. The bacterium isolate producing asparaginase was identified using morphological and biochemical means and 16 SrDNA. L-asparaginase enzyme activity of the isolate was explored with a colorimetric method. The effect of anaerobic condition on the amount of L-asparaginase type II activity was explored by culturing under aeration condition and with no aeration. The result showed that the isolated bacterium was Rhizobium nepotum strain SHN1. The asparaginase enzyme activity and the specific activity of this bacterium were 0.467 IU/mL and 0.015 IU/mg, respectively. These characteristics increased to more than 50% in anaerobic condition. The results indicated that microbial flora from the Persian Gulf flora could be a remarkable source of glutaminase-free L-asparaginase enzyme.

کلیدواژه‌ها [English]

  • L-asparaginase enzyme
  • Rhizobium nepotum
  • glutamination

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