نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانش آموخته کارشناسی ارشد، گروه زیست شناسی، دانشکده علوم، دانشگاه فردوسی مشهد، مشهد، ایران

2 استادیار، گروه زیست شناسی، دانشکده علوم، دانشگاه فردوسی مشهد، مشهد، ایران

3 دانشجوی دکترا، گروه بیوتکنولوژی پزشکی، دانشکده پزشکی، دانشگاه علوم پزشکی مشهد، مشهد، ایران

چکیده

آمیلاز (EC 3.2.1.1) یکی از مهم‌ترین آنزیم های صنعتی است که کاربردهای گسترده‌ای در صنایع مختلف دارد. این آنزیم پیوند گلیکوزیدی (4-1) α را در نشاسته هیدرولیز کرده و محصولاتی با وزن مولکولی کمتر مثل گلوکز، مالتوز و مالتوتریوز تولید می‌کند. هدف از انجام این پژوهش بهینه‌سازی شرایط رشد و تولید آنزیم آمیلاز در باکتری مقاوم به سرمای sp. Janthinobacterium می‌باشد. شرایط رشد و تولید آنزیم آمیلاز در مرحله اول با استفاده از روش تک متغیره برای متغیرهای دما،pH ، منبع کربن، منبع نیتروژن، درصد نشاسته و میزان تلقیح بهینه‌سازی شد. در مرحله بعدی به منظور بررسی اثر برهم کنش بین متغیرهای مختلف بهینه متغیرسازی به روش طراحی روی سطح پاسخ (Response Surface Methodology) صورت گرفت. شناسایی مولکولی جدایه با استفاده از ژن S rRNA16 نشان داد که باکتری به جنس Janthinobacterium تعلق دارند. روش تک متغیره نشان داد که متغیرهای دما، pH و درصد نشاسته بیشترین تاثیر را در تولید آنزیم دارا می‌باشند. مقایسه شرایط قبل و بعد از بهینه سازی نیز مشخص می‌سازد که میزان تولید انزیم از 094/0 به 116/0 واحد آنزیمی افزایش می‌یابد. در نتیجه اعمال شرایط بهینه شده با استفاده از روش RSM، ترشح آنزیم به میزان 434/0 واحد آنزیمی افزایش پیدا می‌کند.

کلیدواژه‌ها

عنوان مقاله [English]

Optimization of amylase enzyme production by a psychrothrophic Janthinobacterium sp. ATR812 with Response Surface Methodology

نویسندگان [English]

  • razieh ghazi birjandi 1
  • bahar shahnavaz 2
  • masome bahreini 2
  • maryam mahjoobin tehran 3

1 MSc student, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

2 Assistant Professor, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.

3 PHD student,Department of Medical Biotechnology, School of Medicine, Mashhad University of Medical Science, Mashhad, Iran

چکیده [English]

Amylases (EC 3.2.1.1) constitute a class of industrial enzymes having great potential for application in industries. Amylase hydrolyses the internal α (1, 4) glycosidase linkages in starch and produces products with lower molecular weight such as glucose, maltose and maltotriose. This research aims the optimization growth and enzyme production in a psychrotolerant bacterium. In the first step, the enzyme production was optimized by one-factor- at- a time method using different variables including temperature, pH, carbon and nitrogen sources, the level of starch, and inoculation rate. In order to examine the interaction between these variables, optimization was performed by response surface method. Molecular identification of strain using 16S rRNA gene showed that bacteria belonged to the genus Janthinobacterium. One-factor-at-a-time method showed that temperature, pH and the level of starch are the most effective factors in enzyme production, the amount of strain production increased to 0.116 U/ml by the mentioned method. The use of response surface methodology increased amylase production up to 0.434 U/ml.

کلیدواژه‌ها [English]

  • Janthinobacterium sp
  • RSM
  • Amylase
  • psychrotolerant bacteria
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