Rapid affinity purification processes for cyclodextrin glycosyltransferase from Bacillus circulans.
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.
Rosso
A
A
Departamento de Ciencias Básicas, Universidad Nacional de Luján, Rutas 5 y 7, 6700, Luján, Buenos Aires, Argentina.
Ferrarotti
S
S
Miranda
M V
MV
Krymkiewicz
N
N
Nudel
B C
BC
Cascone
O
O
D001407:Bacillus / Q000145:classification / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods*; D005964:Glucosyltransferases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification*; D013045:Species Specificity; D013213:Starch / Q000737:chemistry*
D013213:Starch; D005964:Glucosyltransferases; C019963:cyclomaltodextrin glucanotransferase