Rapid Affinity Purification Processes for Cyclodextrin Glycosyltransferase from Bacillus circulans

• Published in: Biotechnology letters Vol. 27; no. 16; pp. 1171 - 1175
• Main Authors: Rosso, A, Ferrarotti, S, Miranda, M.V, Krymkiewicz, N, Nudel, B.C, Cascone, O
• Format: Journal Article
• Language: English
• Published: Dordrecht Kluwer Academic Publishers 01.08.2005; Springer; Springer Nature B.V
• Subjects: adsorption; alpha-cyclodextrin; Ammonium; ammonium sulfate; ammonium sulphate; Bacillus - classification; Bacillus - enzymology; Bacillus circulans; Bacteria; Biological and medical sciences; Biotechnology; Chromatography, Affinity - methods; cyclomaltodextrin glucanotransferase; Fundamental and applied biological sciences. Psychology; Glucosyltransferases - biosynthesis; Glucosyltransferases - chemistry; Glucosyltransferases - isolation & purification; purification; purification methods; Species Specificity; Starch; Starch - chemistry; Cyclodextrin; Purification; Bacillales; Bacillus circulans; Enzyme; Transferases; Glycosyltransferases; Bacteria; Affinity; cyclodextrin glycosyltransferase; Bacillaceae; Operating process
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-005-8654-6

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 α-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.