On-column refolding of recombinant fungal endoglucanase I
Endoglucanase is an industrially important enzyme involved in degradation of cellulosic materials into constitutive monomers synergistically with other cellulase enzymes. In this study, overexpression of (His)6-tagged endoglucanase I from Fusarium oxysporum sp. in E. coli BL21(DE3) resulted to high...
| Main Authors: | , |
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| Format: | Conference or Workshop Item |
| Language: | English |
| Published: |
2011
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/16211/ http://irep.iium.edu.my/16211/ http://irep.iium.edu.my/16211/1/On-column_Refolding_of_Recombinant_Fungal.pdf |
| Summary: | Endoglucanase is an industrially important enzyme involved in degradation of cellulosic materials into constitutive monomers synergistically with other cellulase enzymes. In this study, overexpression of (His)6-tagged endoglucanase I from Fusarium oxysporum sp. in E. coli BL21(DE3) resulted to high tendency of endoglucanase I in the form of inactive inclusion bodies when the host cells were cultured between 25-37°C and induced by 0.05-0.20 mM (final concentration) IPTG. A simple and time-efficient on-column refolding scheme using IMAC aided by AKTA purifier, an automated fast protein liquid chromatography (FPLC) system afforded highly purified refolded recombinant endoglucanase I that was active in carboxymethyl cellulose (CMC) assay. The on-column refolding scheme is reliable and can be applied for efficient and robust method for lab-scale production of recombinant endoglucanase I. |
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