碩士 / 國立海洋大學 / 食品科學系 / 87 / Abstract
The calpastatin activity of grass shrimp (Penaeus monodon), which eluted from DEAE-Sephacel chromatography was not significantly changed after heating for 3 min boiling at 100oC. The calpastatin was further purified to electrophoretical homogeneity by heat-treatment and DEAE-Sephacel, and Q-Sepharose. The specific activity, yield and purification were 86.9 units/mg, 129% and 52,303 fold, respectively. The MW of calpastatin was 80 and 88.7 kDa estimated from SDS-PAGE and gel filtration of Sephacryl S-200 HR. The inhibitory activity was not affected by 20 min boiling at 100 oC and at pH ranged of 4.5~10.0. However, left after 23 min boiling at 100 oC, only 50% inhibitory activity was remaining. The calpastatin could inhibit shrimp calpain, beef μ-, and m-calpains, but not papain, cathepsins B, and D, trypsin, -, and -chymotrypsins. Hence, the calpastatin might be a specific inhibitor for calpains. On the purification of shrimp calpastatin, the calpain and calpastatin were eluted at 320 and 460 mM NaCl in 20 mM Tris-HCl buffer (pH 7.5) from DEAE-Sephacel (2.6 x 10 cm) chromatography by using linear gradient elution, respectively. Likewise, shrimp muscle calpain and calpastatin were separated at 350 and 500 mM NaCl in same buffer and column by using stepwise elution and their recovery were 84.6 and 92.3 %, respectively. The results suggested that quick separation of shrimp muscle calpain and calpastatin was feasible and it can be applied on large-scale and continuous determination of proteolysis system of calpain and its inhibitor. It worth further researching if the proteolysis system would be altered with feedings and/or conditions during shrimp cultured.
| Identifer | oai:union.ndltd.org:TW/087NTOU0253017 |
| Date | January 1999 |
| Creators | Sue Mei Chu, 蘇美菊 |
| Contributors | Jiang, Shann -Tzong, Chen, Chin-San, 江善宗, 陳慶三 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 70 |
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