Affinity partitioning of a Cellulomonas fimi beta-mannanase with a mannan-binding module in galactomannan/starch aqueous two-phase system
Author
Summary, in English
A new approach in affinity separations was studied by partitioning of Cellulomonas fimi beta-mannanase (EC 3.2.1.78) containing a mannan-binding module in galactomannan/hydroxypropyl starch aqueous two-phase system. Comparison was made with a truncated version of C. fimi beta-mannanase which lacked the mannan-binding module. Results showed that affinity partitioning of the beta-mannanase was achieved due to biospecificity of the mannan-binding module towards the top phase containing galactomannan. Experiments were conducted at pH 8 to prevent enzyme degradation of the phase containing galactomannan. Removal of the top phase polymer was accomplished by beta-mannanase degradation allowed by shifting to the optimal pH 6. In the combination with the genetic fusion of any given protein to the mannan-binding module, the results envision a general procedure for primary affinity recovery of such fusion proteins.
Department/s
Publishing year
2006
Language
English
Pages
53-59
Publication/Series
Journal of Chromatography A
Volume
1123
Issue
1
Document type
Journal article
Publisher
Elsevier
Topic
- Biological Sciences
Keywords
- galactomannan
- affinity-partitioning
- aqueous two-phase system
- beta-mannanase
- mannan-binding module
Status
Published
ISBN/ISSN/Other
- ISSN: 0021-9673