نوع مقاله : مقاله پژوهشی

نویسندگان

1 استادیار گروه علوم دامی، مؤسسه آموزش عالی علمی کاربردی جهاد کشاورزی، تهران، ایران

2 دانشیار گروه علوم دامی، دانشکده کشاورزی، دانشگاه آزاد اسلامی واحد علوم و تحقیقات، تهران، ایران

3 کارشناس، گروه علوم دامی، مؤسسه آموزش عالی علمی- کاربردی جهاد کشاورزی، تهران، ایران

چکیده

این تحقیق به منظور بررسی تولید آنزیم‌های هضم‌کنندة الیاف در شکمبه و روش‌های مختلف استخراج و تغلیظ آن‌ها از محتویات شکمبة دام‌های کشتارشده انجام شد. به همین منظور، از دو روش هوموژن‌سازی و سونیکیشن برای آزادسازی آنزیم‌های متصل به مواد جامد محتویات شکمبه و از روش‌های رسوب دادن پروتئین‌ها با اتانل، تری‌کلرو استیک اسید و سولفات آمونیم، همچنین خشک کردن با برودت به منظور تغلیظ آنزیم‌های هضم‌کنندة الیاف استفاده شد. به منظور بررسی اثر دما و مدت زمان نگهداری بر فعالیت آنزیم‌ها از روش‌های نگهداری در نیتروژن مایع، فریزر 70- و 20- درجة سانتی‌گراد استفاده شد. میانگین فعالیت ویژة آنزیم‌های سلولاز و زایلاناز به ترتیب 5/7 و 5/16 واحد به ازای هر میلی‌گرم پروتئین موجود در محلول بود. میزان آزادسازی و استخراج آنزیم از محتویات شکمبه در روش هوموژن‌سازی بیشتر از روش سونیکیشن بود. فعالیت و بازیافت آنزیم‌ها در روش‌های تغلیظ با برودت و رسوب دادن با سولفات آمونیم بیشتر بود. روش نیتروژن مایع برای نگهداری طولانی‌مدت و روش فریزر 70- برای نگهداری میان‌مدت مناسب‌تر بود. نتایج این تحقیق نشان داد که محتویات شکمبة دام‌های کشتارشده منبع مناسبی از آنزیم‌های هضم‌کنندة الیاف گیاهی است و می‌توان با به‌کارگیری روش‌های استخراج و تغلیظ مناسب از آن‌ها استفاده کرد.

کلیدواژه‌ها

عنوان مقاله [English]

Investigation of methods of extraction, concentration and conservation of rumen content fibrolytic enzymes

نویسندگان [English]

  • Hossein Manafi Rasi 1
  • Mohammad Chamani 2
  • Salman Afshar 3

1 Assistant Professor, Department of Animal Science, Institute of Scientific-Applied Higher Education of Jihad-e-Agriculture, Tehran, Iran

2 Association Professor, Department of Animal Science, Agriculture Faculty, Science and Research Branch, Islamic Azad University, Tehran, Iran

3 Research Expert, Department of Animal Science, Institute of Scientific-Applied Higher Education of Jihad-e-Agriculture, Tehran, Iran

چکیده [English]

This study was carried out to determine the potential of rumen microorganisms in samples taken from slaughterhouse to produce fibrolytic enzymes and to compare different methods for extraction, concentration and conservation of them.  According to the results, the average specific activity of cellulase and xylanase were 7.5 and 16.5 U/mg protein respectively. The application of homogenizing and sonication methods to extract rumen content enzymes showed that the former had better performance in the liberation of enzymes. To determine optimum time length for the maximum release of enzymes from solid phases, various time lengths including one, 1.5 and two min were examined. Based on this experiment, one minute appeared the best. To attain a high level of concentration of enzymes obtained from rumen liquor, various procedures were applied. The results indicate that freeze drying and precipitation of enzyme using ammonium sulphate were the best methods, while trichloroacetic acid precipitation turned out to be the most inappropriate method due to improper effect on enzymatic activity. In order to examine the effect of temperature and time length on enzymes activities, some methods including liquid nitrogen, freezing in -70° and freezing in -20° were applied. The comparison of different methods of enzymes conservation indicated that applying liquid nitrogen for long term as well as keeping in the freezer (-70°) for medium term purposes were the best.

کلیدواژه‌ها [English]

  • Cellulase
  • concentration
  • Conservation
  • extraction
  • rumen
  • Xylanase

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