Rapid affinity purification processes for cyclodextrin glycosyltransferase from Bacillus circulans

被引:7
|
作者
Rosso, A
Ferrarotti, S
Miranda, MV
Krymkiewicz, N
Nudel, BC
Cascone, O
机构
[1] Univ Buenos Aires, Fac Farm & Bioquim, Catedra Microbiol Ind & Biotecnol, RA-1113 Buenos Aires, DF, Argentina
[2] Univ Nacl Lujan, Dept Ciencias Basicas, RA-6700 Buenos Aires, DF, Argentina
关键词
affinity; Bacillus circulans; cyclodextrin glycosyltransferase; purification;
D O I
10.1007/s10529-005-8654-6
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:1171 / 1175
页数:5
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