On-column refolding of recombinant fungal endoglucanase
Endoglucanase is an industrially important enzyme involved synergistically in degradation of cellulosic materials into constitutive monomers with other cellulase enzymes. In this study, overexpression of (His)6-tagged endoglucanase from Fusarium oxysporum sp. in E. coli BL21(DE3) resulted to high...
| Main Authors: | , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
INSI Publications
2012
|
| Subjects: | |
| Online Access: | http://irep.iium.edu.my/23411/ http://irep.iium.edu.my/23411/ http://irep.iium.edu.my/23411/1/jamil_on_column_refolding.pdf |
| id |
iium-23411 |
|---|---|
| recordtype |
eprints |
| spelling |
iium-234112012-06-15T02:12:08Z http://irep.iium.edu.my/23411/ On-column refolding of recombinant fungal endoglucanase Jamaluddin, Mohd Jamil Aizat Mohd. Salleh, Hamzah TA164 Bioengineering TP248.13 Biotechnology Endoglucanase is an industrially important enzyme involved synergistically in degradation of cellulosic materials into constitutive monomers with other cellulase enzymes. In this study, overexpression of (His)6-tagged endoglucanase from Fusarium oxysporum sp. in E. coli BL21(DE3) resulted to high tendency of endoglucanase in the form of inactive inclusion bodies when the host cells were cultured between at 37°C and induced by 0.20 mM (final concentration) IPTG. A simple and time-efficient on-column refolding scheme using immobilized metal-chelate affinity chromatography (IMAC) aided by AKTA purifier, an automated fast protein liquid chromatography (FPLC) system afforded highly purified refolded recombinant endoglucanase 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. INSI Publications 2012-01 Article PeerReviewed application/pdf en http://irep.iium.edu.my/23411/1/jamil_on_column_refolding.pdf Jamaluddin, Mohd Jamil Aizat and Mohd. Salleh, Hamzah (2012) On-column refolding of recombinant fungal endoglucanase. Australian Journal of Basic and Applied Sciences, 6 (1). pp. 128-133. ISSN 1991-8178 http://www.insipub.com/ajbas/2012/January/128-133.pdf |
| repository_type |
Digital Repository |
| institution_category |
Local University |
| institution |
International Islamic University Malaysia |
| building |
IIUM Repository |
| collection |
Online Access |
| language |
English |
| topic |
TA164 Bioengineering TP248.13 Biotechnology |
| spellingShingle |
TA164 Bioengineering TP248.13 Biotechnology Jamaluddin, Mohd Jamil Aizat Mohd. Salleh, Hamzah On-column refolding of recombinant fungal endoglucanase |
| description |
Endoglucanase is an industrially important enzyme involved synergistically in degradation
of cellulosic materials into constitutive monomers with other cellulase enzymes. In this study,
overexpression of (His)6-tagged endoglucanase from Fusarium oxysporum sp. in E. coli BL21(DE3)
resulted to high tendency of endoglucanase in the form of inactive inclusion bodies when the host cells
were cultured between at 37°C and induced by 0.20 mM (final concentration) IPTG. A simple and
time-efficient on-column refolding scheme using immobilized metal-chelate affinity chromatography
(IMAC) aided by AKTA purifier, an automated fast protein liquid chromatography (FPLC) system
afforded highly purified refolded recombinant endoglucanase 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.
|
| format |
Article |
| author |
Jamaluddin, Mohd Jamil Aizat Mohd. Salleh, Hamzah |
| author_facet |
Jamaluddin, Mohd Jamil Aizat Mohd. Salleh, Hamzah |
| author_sort |
Jamaluddin, Mohd Jamil Aizat |
| title |
On-column refolding of recombinant fungal endoglucanase |
| title_short |
On-column refolding of recombinant fungal endoglucanase |
| title_full |
On-column refolding of recombinant fungal endoglucanase |
| title_fullStr |
On-column refolding of recombinant fungal endoglucanase |
| title_full_unstemmed |
On-column refolding of recombinant fungal endoglucanase |
| title_sort |
on-column refolding of recombinant fungal endoglucanase |
| publisher |
INSI Publications |
| publishDate |
2012 |
| url |
http://irep.iium.edu.my/23411/ http://irep.iium.edu.my/23411/ http://irep.iium.edu.my/23411/1/jamil_on_column_refolding.pdf |
| first_indexed |
2023-09-18T20:35:25Z |
| last_indexed |
2023-09-18T20:35:25Z |
| _version_ |
1777409013568241664 |