差向異構酶包涵體復性最適化研究
碩士 === 國立中興大學 === 化學工程學系 === 91 === GlcNAc 2-epimerase was expressed in Escherichia coli in the form of inlcusion bodies. The epimerase from the purified inclusion bodies was solubilized in 100 mM Tris-HCl buffer at pH 12.5 and > 0.3 % SDS had significatly solubilization effect. The optimal condi...
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| Format: | Others |
| Language: | zh-TW |
| Published: |
2003
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| Online Access: | http://ndltd.ncl.edu.tw/handle/32201443936545409469 |
| Summary: | 碩士 === 國立中興大學 === 化學工程學系 === 91 === GlcNAc 2-epimerase was expressed in Escherichia coli in the form of inlcusion bodies. The epimerase from the purified inclusion bodies was solubilized in 100 mM Tris-HCl buffer at pH 12.5 and > 0.3 % SDS had significatly solubilization effect. The optimal conditions for denaturation of epimerase were when inclusion bodies were solubilized in 50mM Tris-HCl buffer. The refolding method is that the solubilized protein was adjusted to neutral pH. Optimal soluble protein recovery is about 65% when the solubilized proteins (8mg/ml) were refolded at pH 8 .In this study,we added GSH and GSSG to promoted activity of refolded epimerase utilizing an oxido-shuffling system .The dimer fractions of refolded epimerase had 30% relative specific activity .
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