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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.

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