کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10843783 | 1069293 | 2005 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Purification and characterization of fibrinolytic enzyme from cultured mycelia of Armillaria mellea
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
A fibrinolytic enzyme was purified from the cultured mycelia of Armillaria mellea by ion-exchange chromatography followed by gel filtration, and was designated A. mellea metalloprotease (AMMP). The purification protocol resulted in a 627-fold purification of the enzyme, with a final yield of 6.05%. The apparent molecular mass of the purified enzyme was estimated to be 21 kDa by SDS-PAGE, fibrin-zymography and gel filtration chromatography, which revealed a monomeric form of the enzyme. The optimal reaction pH value and temperature were, pH 6.0, and 33 °C, respectively. This protease effectively hydrolyzed fibrinogen, preferentially digesting the Aα-chain over the Bβ- and r-chains. Enzyme activity was inhibited by Cu2+ and Co2+, but enhanced by the addition of Ca2+ and Mg2+ ions. Furthermore, AMMP activity was potently inhibited by EDTA, and was found to exhibit a higher specificity for the substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsin-like metalloprotease. The first 24 amino acid residues of the N-terminal sequence were MFSLSSRFFLYTLCL SAVAVSAAP, which is extremely similar to the 24 amino acid residues of the N-terminal sequence of the fruiting body of A. mellea. These data suggest that the fibrinolytic enzyme AMMP, obtained from the A. mellea exhibits a profound fibrinolytic activity. The mycelia of A. mellea may thus represent a potential source of new therapeutic agents to treat thrombosis.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 43, Issue 1, September 2005, Pages 10-17
Journal: Protein Expression and Purification - Volume 43, Issue 1, September 2005, Pages 10-17
نویسندگان
Sook-Young Lee, Jae-Sung Kim, Ji-Eun Kim, Kumar Sapkota, Ming-Hua Shen, Seung Kim, Hong-Sung Chun, Jin-Cheol Yoo, Han-Suk Choi, Myung-Kon Kim, Sung-Jun Kim,