Purification of α-galactosidase by affinity precipitation with alginate

被引:2
作者
Celem, Evran Bicak [1 ]
Onal, Secil [1 ]
机构
[1] Ege Univ, Fac Sci, Dept Biochem, TR-35100 Bornova, Turkey
关键词
affinity macroligand; affinity precipitation; alginate; Vicia faba; alpha-galactosidase;
D O I
10.1080/10826060802325436
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Affinity precipitation is a technique which is known for over 20 years, but has recently received more attention due to the development of new materials for its implementation. It is a relatively simple, convenient, and reproducible technique that results in high target molecule recovery at high specificity. We describe, here, an efficient and rapid purification procedure for Vicia faba -galactosidase (EC 3.2.1.22) by using affinity precipitation with alginate. The enzyme was purified with 43% activity yield and 40-fold purification. SDS-PAGE of the purified enzyme showed a single band and a subunit weight of 44kDa. The properties of the enzyme were also searched. The results showed that the general properties of the enzyme offer potential for use of this -galactosidase in several production processes.
引用
收藏
页码:348 / 357
页数:10
相关论文
共 34 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]   Affinity chromatography matures as bioinformatic and combinatorial tools develop [J].
Clonis, YD .
JOURNAL OF CHROMATOGRAPHY A, 2006, 1101 (1-2) :1-24
[3]   THE LECTIN NATURE OF ALPHA-GALACTOSIDASES FROM VICIA-FABA SEEDS [J].
DEY, PM ;
NAIK, S ;
PRIDHAM, JB .
FEBS LETTERS, 1982, 150 (01) :233-237
[4]   Characterization of α-galactosidases from germinating soybean seed and their use for hydrolysis of oligosaccharides [J].
Guimaraes, VM ;
de Rezende, ST ;
Moreira, MA ;
de Barros, EG ;
Felix, CR .
PHYTOCHEMISTRY, 2001, 58 (01) :67-73
[5]  
Gupta MN, 1996, J MOL RECOGNIT, V9, P356, DOI 10.1002/(SICI)1099-1352(199634/12)9:5/6<356::AID-JMR328>3.0.CO
[6]  
2-X
[7]   Protein purification by affinity precipitation [J].
Hilbrig, F ;
Freitag, R .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2003, 790 (1-2) :79-90
[8]   Characteristics of an α-galactosidase associated with grape flesh [J].
Kang, HC ;
Lee, SH .
PHYTOCHEMISTRY, 2001, 58 (02) :213-219
[9]   Production, purification and characterization of a constitutive intracellular α-galactosidase from the thermophilic fungus Humicola sp [J].
Kotwal, SM ;
Gote, MM ;
Khan, MI ;
Khire, JM .
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 1999, 23 (01) :661-667
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+