Rapid affinity purification processes for cyclodextrin glycosyltranferase from Bacillus circulans.

ROSSO, A..; FERRAROTTI, S.; MIRANDA, M.V.; KRYMKIEWICZ, N.; NUDEL, B.C.; CASCONE, O.

BIOTECHNOLOGY LETTERS

SPRINGER

Año: 2005 vol. 27 p. 1171 - 1175

0141-5492

Bacillus circulans DF9R cyclodextrin glycosyltransferase (CGTase) purification were developed: affinity precipitation with starch and aqueous two-phase partition. The first method was optimised by a factorial design, allowing an 80% CGTase adsorption at 11% starch and 1.6% ammonium sulphate, and a 64.5% recovery after elution with 10 mM cyclodextrin. The purification factor was 17.1. Aqueous two-phase partition yielded a 72% CGTase recovery in a two-step procedure: in the first step, the enzyme accumulated in the top phase of a poly(ethyleneglycol)/phosphates system, pH 7.0, and then, in the second step, another aqueous two-phase system was built with the top phase of the first system and magnesium sulphate and Reppal. CGTase was obtained in the bottom phase with a purification factor of 36.7.