Generation of catalytic protein particles in Escherichia coli cells using the cellulose-binding domain from Cellulomonas fimi as a fusion partner = 대장균에서 활성형 단백질입자의 제조

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Title
Generation of catalytic protein particles in Escherichia coli cells using the cellulose-binding domain from Cellulomonas fimi as a fusion partner = 대장균에서 활성형 단백질입자의 제조
Author(s)
Su Lim Choi; Sang Jun Lee; J S Ha; Jae Jun Song; Y H Rhee; Seung Goo Lee
Bibliographic Citation
Biotechnology and Bioprocess Engineering, vol. 16, no. 6, pp. 1173-1179
Publication Year
2011
Abstract
The cellulose-binding domain (CBD) of a Cellulomonas fimi exo-glucanase was translationally fused with β-glucuronidase (GusA) from Escherichia coli and β-glycosidase (BglA) from Thermus caldophilus, respectively. Two fusion proteins (GusA-CBD and BglA-CBD) were expressed as insoluble aggregates in cells and isolated by centrifugation of the cell lysates. Interestingly, activity assays revealed that > 90% of the catalytic activity of both proteins was localized in the insoluble fractions. For example, the GusA-CBD particles exhibited 21 units per mg protein, which corresponded to 19% specific activity of the highly purified soluble GusA. The specific activity increased further up to 42 units per mg protein when treated with either sonication or chaotropic L-arginine. These results demonstrate that fusion with CBD family II may activate catalytic protein particles in E. coli cells, and that internal proteins of the particles are also active. Finally, the protein particles were tested in repeated batch operations after being cross-linked with chemicals, indicating that they have potential as a new preparation for immobilized biocatalysts.
Keyword
β-glucuronidaseβ-glycosidaseCellulose-binding domain (CBD)Escherichia coliImmobilizationProtein particles
ISSN
1226-8372
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s12257-011-0336-8
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
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