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GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins
Published on: October 29, 2013
Rapid affinity purification processes for cyclodextrin glycosyltransferase from Bacillus circulans
A Rosso1, S Ferrarotti, M V Miranda
1Departamento de Ciencias Básicas, Universidad Nacional de Luján, Rutas 5 y 7, 6700, Luján, Buenos Aires, Argentina.
Abstract:
Two rapid and easy-to-scale-up methods for the purification of cyclodextrin glycosyltransferase (CGTase) from Bacillus circulans were developed: affinity precipitation with starch and aqueous two-phase partition. The first method, optimised by a factorial design, gave an 80% CGTase adsorption at 11% starch and 1.6% ammonium sulphate, and a 65% recovery after elution with 10 mM alpha-cyclodextrin. The purification factor was 17. Aqueous two-phase partition yielded a 72% CGTase recovery in a two-step procedure; CGTase was obtained in the bottom phase with a purification factor of 37.

