Affinity partitioning of a Cellulomonas fimi beta-mannanase with a mannan-binding module in galactomannan/starch aqueous two-phase system.
J Chromatogr A
; 1123(1): 53-9, 2006 Aug 04.
Article
em En
| MEDLINE
| ID: mdl-16797561
ABSTRACT
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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cromatografia de Afinidade
/
Cellulomonas
/
Beta-Manosidase
/
Mananas
Idioma:
En
Revista:
J Chromatogr A
Ano de publicação:
2006
Tipo de documento:
Article