碩士 / 國立高雄海洋科技大學 / 水產食品科學研究所 / 97 / The objective of the present study is to purify and characterize the proteinase from Vogesella sp.7307-1 which was newly identified and isolated from fish scale. The proteinase was extracted, isolated, and purified 35.65-fold with recovery 9.96% using 20-80% saturation of ammonium sulfate fractionation, Q FF ion exchange chromatography and Superdex 200 gelfiltration. The molecular mass of the purified enzyme was 119 kDa. The Km and Vmax was 0.067 mM and 425.5 U/mg-min, respectively using azo-casein as substrate. The optimum pH of the purified enzyme was 7.5, and optimum temperature was 50℃. At temperature below 60℃ or pH ranged from 7.5 to 9.0, the enzyme was stable. Metal ion (including Cu2+、Hg2+), SDS (sodiumdodecylsulphate) and β-mercaptoethanol could inhibit enzyme activity while Triton X-100 could slight activate the enzyme activity. Moreover, the enzyme activity of the purified proteinase was completely inhibited by EDTA (ethylene diamine teraacetates), indicating the purified enzyme was probably a metalloproteinase. The crude proteinase retained more than 84.5% of enzyme activity at 4 or -20℃ for 10 weeks, and 39.9% at 25℃ for 4 weeks. Hydrolysates from fish scale treated with 7307-1 protease were found having the low MW peptides (< 3 kDa).
| Identifer | oai:union.ndltd.org:TW/097NKIM8084016 |
| Date | January 2009 |
| Creators | Yu-Ren Huang, 黃裕仁 |
| Contributors | Jen-Min Kuo, 郭建民 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 93 |
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