Purification and characterization of chitinase from shoots of Green Bamboo Bambusa oldhamii
| Autor: | Chung-Min Lee, 李宗明 |
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| Rok vydání: | 2003 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 91 This study is to investigate the effect of endogenous factor(ethylene) and exogenous factor(chitosan) on the chitinase activity of green bamboo shoots and to purify and characterize chitinases from bamboo shoots. Endo- and exochitinases were purified from base parts of green bamboo shoots by sequential steps of ammonium sulfate fractionation, Sephacryl S-100 HR gel filtration, DEAE-Sephacel anion -exchange chromatography and MonoQ fast protein liquid chromatography. There are four or five endochitinases and their molecular masses were 90 kDa, 42 kDa, 40 kDa, 30 kDa, and 28 kDa as estimated by SDS-PAGE, named chitinase Ⅰ. And two exochitinases that their molecular masses were 90 kDa and 30 kDa. The 90 kDa exochitinase was named chitinaseⅡ. For ethylene glycol chitin hydrolysis, chitinaseⅠhad an optimal pH of 5 and an optimal temperature of 60℃. For 4-MU-(GlcNAc)3 hydrolysis, chitinaseⅡ had an optimal pH of 3 and an optimal temperature of 60℃.ChitinaseⅠhad an isoelectric point of 4 to 6 and chitinaseⅡ had an isoelectric point of 7 to 9, as estimated by ion-exchange chromatography. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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