استخراج و تخلیص آنزیم لیپاز (EC 3.1.1.3) از روده ماهی هوور مسقطی (Katsuwenus pelamis)

نوع مقاله : مقاله کامل علمی - پژوهشی

نویسندگان

1 نویسنده مسئول، پژوهشکده اکولوژی خلیج‌فارس و دریای عمان، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، بندرعباس، ایران

2 مرکز تحقیقات زیست‌شناسی سلولی و مولکولی، دانشگاه علوم پزشکی شهید بهشتی، تهران، ایران

چکیده

حدود 7 تا 8 درصد وزن بدن تون ماهیان را مجاری گوارشی تشکیل می دهد و به عنوان یک منبع اقتصادی برای استخراج آنزیم لیپاز می‌باشد. استخراج آنزیم با استفاده از بافر تریس هیدروکلرید (25 میلی مولار با pH؛ 8/7)، انجام شد. سپس عصاره خام در محدوده اشباع بین 30، 45 و 60 درصد سولفات آمونیوم رسوب کرده و دیالیز گردید. برای خالص سازی بیشتر از کروماتوگرافی تبادل یونی با استفاده از ستون DEAE-Cellulose با ابعاد (30 × 5/2 سانتی متر) و کروماتوگرافی فیلتراسیون ژل توسط ستون Sephadex G-100 (80×8/1 سانتی متر) استفاده شد. آنزیم های لیپاز خالص شده حاصل از ترسیب با سولفات آمونیوم 30، 45 و 60 درصد به ترتیب دارای وزن مولکولی (3/68، 41/38 و ND کیلودالتون)، pH (1/7، 9/6 و 4/7)، دمای بهینه (55، 42 و 38 درجه سانتی گراد) و فعالیت ویژه آنزیمی (42/3، 8/2 و U/mg 1/4) با استفاده از پارانیتروفنیل پالمیتات (pNPP) به عنوان سوبسترا بود. نتایج بیانگر امکان استخراج و تخلیص آنزیم لیپاز از دستگاه گوارش آبزیان با ویژگی های بیوشیمیایی مناسب جهت استفاده در صنایع شیمیایی و غذایی می باشد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Extraction and purification of lipase (EC 3.1.1.3) from the Skipjack intestine (Katsuwenus pelamis)

نویسندگان [English]

  • zabih alh bahmani 1
  • Bahram Kazemi 2
1 Corresponding Author, Persian Gulf and Sea of Oman Ecology Research Center, Fisheries Science Research Institute, Agricultural Research, Education and Extension Organization, Bandar Abbas, Iran
2 Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
چکیده [English]

About 7 to 8% of the body weight of tuna fish is the digestive tract and it is an economic source for lipase enzyme extraction. Enzyme extraction was done using Tris hydrochloride buffer (25 mM with pH; 7.8). Then the crude extract was precipitated and dialyzed in the range of saturation between 30, 45 and 60% ammonium sulfate. For further purification, ion exchange chromatography using a DEAE-CMC column with dimensions (2.5 × 30 cm) and gel filtration chromatography using a Sephadex G-100 column (1.8 × 80 cm) were used. Purified lipase enzymes obtained by precipitation with ammonium sulfate 30, 45 and 60%, respectively, have molecular weight (68.3, 38.41, and ND KDa), pH (7.1, 6.9, and 7.4), The optimal temperature (55, 42 and 38 °C) and specific enzyme activity (3.42, 2.8 and 1.4 U/mg) were using paranitrophenyl palmitate (pNPP) as a substrate. The results indicate the possibility of extracting and purifying lipase enzyme from the digestive system of aquatic animals with suitable biochemical characteristics for use in chemical and food industries.

کلیدواژه‌ها [English]

  • Intestinal Lipase Enzyme (ILE)
  • Ion exchange
  • Filtration
  • Molecular weight

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مجله بهره برداری و پرورش آبزیان
دوره 13، شماره 3
مهر 1403
صفحه 115-128

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