title: Affinity partitioning of a Cellulomonas fimi beta-mannanase with a mannan-binding module in galactomannan/starch aqueous two-phase system.
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.
authors.affiliation: Department of Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, S-22100 Lund, Sweden.
authors.forename: Mirjana
authors.initials: M
authors.lastname: Antov
authors.forename: Lars
authors.initials: L
authors.lastname: Anderson
authors.forename: Alexandra
authors.initials: A
authors.lastname: Andersson
authors.forename: Folke
authors.initials: F
authors.lastname: Tjerneld
authors.forename: Henrik
authors.initials: H
authors.lastname: Stålbrand
mesh_terms: D040143:Cellulomonas / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008351:Mannans / Q000378:metabolism*; D011485:Protein Binding; D044902:beta-Mannosidase / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D008351:Mannans; D044902:beta-Mannosidase