بهبود فعالیت آنزیم‌های تجزیه‌کننده سلولز با جهش‌زایی در قارچ Trichoderma reesei

نوع مقاله : علمی - پژوهشی


1 پژوهشگر، گروه گیاهپزشکی و نگهداری مواد غذایی، پژوهشکده کشاورزی هسته‌ای، پژوهشگاه علوم و فنون هسته‌ای، سازمان انرژی اتمی ایران، کرج، ایران

2 استادیار گروه گیاهپزشکی و نگهداری مواد غذایی، پژوهشکده کشاورزی هسته‌ای، پژوهشگاه علوم و فنون هسته‌ای، سازمان انرژی اتمی ایران، کرج، ایران


قارچ Trichoderma reesei یکی از گونه‌های مهم تولید­کننده آنزیم‌های تجزیه‌کننده سلولز در طبیعت است. در این پژوهش 21 جدایه جهش‌یافته پرتو گاما از قارچ T. reesei برای تولید آنزیم سلولاز روی محیط محتوی سلولز کلوئیدی غربال‌گری شد. غلظت پروتئین‌های خارج سلولی جدایه‏های وحشی و جهش‌یافته با استفاده از روش بردفورد اندازه‌گیری شد. آویسل، کربوکسی متیل سلولز و کاغذ صافی برای اندازه‌گیری فعالیت سلولازی استفاده شد. هم‌چنین خلوص و ترکیب پروتئین‌های غنی از آنزیم با استفاده از آزمون الکتروفورز ژل پلی‌آکریل‌آمید مورد ارزیابی قرار گرفت. بالاترین میزان تولید پروتئین خارج سلولی در جدایه‌های T. r M7 و T. r M17 مشاهده شد. فعالیت آنزیم‌های اندوگلوکاناز، اگزوگلوکاناز و سلولاز کل در جدایه جهش‌یافته T. r M8 بالاترین مقادیر فعالیت آنزیمی را در بین جدایه‌های جهش‌یافته و وحشی نشان داد. اختلاف وزن مولکولی باندهای آنزیمی نشان داد که آنزیم‌های EG IV، Cel 1A، Cel 12A، Cel 45A، Cel 3A، Cel 7A، Cel 6A، Cel 5A و Cel 61A سلولز کلوئیدی را به‌صورت هم‌افزایی هیدرولیز می‌کنند. این نتایج نشان می‌دهد که جهش‌زایی با پرتو گاما برای دستیابی به جهش‌یافته‌های تریکودرما با تولید بالای آنزیم سلولاز امکان‏‌پذیر است.


عنوان مقاله [English]

Improvement of Cellulose Degrading Enzymes Activity by Mutagenesis in Trichoderma reesei Fungi

نویسندگان [English]

  • Hamed Askari 1
  • Samira Shahbazi 2
1 Researcher, Plant Protection and Food Preservation Department, Nuclear Agriculture Research School, Nuclear Science and Technology Research Institute (NSTRI), Atomic Energy Organization of Iran (AEOI), Alborz, Iran
2 Assistant Professor, Plant Protection and Food Preservation Department, Nuclear Agriculture Research School, Nuclear Science and Technology Research Institute (NSTRI), Atomic Energy Organization of Iran (AEOI), Alborz, Iran
چکیده [English]

Trichoderma reesei is one of the most important fungi producing cellulose degrading enzymes in nature. In this study, 21 mutated isolate via gamma irradiation of T. reesei were screened for cellulase enzyme production on a colloidal cellulose medium. The concentration of extracellular proteins of wild and mutant isolates were measured using Bradford's method. Avicel, carboxymethyl cellulose and filter paper were used to measure cellulase activity. The purity and composition of enzyme-rich proteins were also evaluated using polyacrylamide gel electrophoresis (SDS-PAGE) test. The highest amount of extracellular protein production was observed in T. r M7 and T. r M17 isolates. Activity of endoglucanase, exoglucanase and total cellulase enzymes in mutated isolate T. r M8 showed the highest levels of enzymatic activity among mutant and wild isolates. The difference in the molecular weight of the enzyme bands showed that the EG 4, Cel 1A, Cel 12A, Cel 45A, Cel 3A, Cel 7A, Cel 6A, Cel 5A and Cel 61A enzymes hydrolyzed the colloidal cellulose, synergictly. These results indicate that, mutation induced by gamma radiation is possible to obtain high producing Trichoderma mutants of cellulase enzymes.

کلیدواژه‌ها [English]

  • Trichoderma
  • Cellulase enzyme
  • Gamma radiation
  • Mutation
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