Purification and characterization of fibrinolytic enzyme from local Bacillus sp.

碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 100 === This study aimed to screen local Bacillus sp. with high proteolytic activity to provide the basis for drug or health food development. The strain used in this study was isolated from local soil in Taiwan. Among 30 local Bacillus sp. isolates screened, 8 ca...

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Bibliographic Details
Main Authors: Chen-Yi Yeh, 葉振宜
Other Authors: Miao-Lin Hu
Format: Others
Language:zh-TW
Published: 2012
Online Access:http://ndltd.ncl.edu.tw/handle/74144926344045684543
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Summary:碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 100 === This study aimed to screen local Bacillus sp. with high proteolytic activity to provide the basis for drug or health food development. The strain used in this study was isolated from local soil in Taiwan. Among 30 local Bacillus sp. isolates screened, 8 candidates with higher proteolytic activity assayed with skim milk and gelatin plates methods were selected for further testing. 32732C with the highest fibrinolytic activity by fibrin plate assay was chosen among 8 candidates. The isolated 32732C was identifical by BIOLOG as Bacillus amyloliquefaciens. An extracellular fibrinolytic enzyme from B. amyloliquefaciens 32732C was purified by ammonium sulfate precipitation and DEAE sepharose FF anion exchange. The molecular weight of the purified fibrinolytic enzyme was estimated to be 50 kDa by sodium dodecyl sulphate-polyacrylamide gel electorphoresis (SDS-PAGE) and zymogram, respectively. The optimal pH and temperature of the purified enzyme were 7 and 50oC, respectively. The purified enzyme thermal and pH stability were 25-50 oC and 6.0-9.0, respectively. Results of this study demonstrate that the B. amyloliquefaciens 32732C has the potential for an antithrombotic drug or a health-care food, but we need more research to prove it.