Purification and characterization of α-glucosidase in Apis cerana indica  

作　　者：Chanpen Chanchao Suwisa Pilalam Polkit Sangvanich 

机构地区：[1]Department of Biology Faculty of Science, Chulalongkom University, Bangkok, Thailand [2]Program of Biotechnology Faculty of Science, Chulalongkom University, Bangkok, Thailand [3]Department of Chemistry, Faculty of Science, Chulalongkom University, Bangkok, Thailand

出　　处：《Insect Science》2008年第3期217-224,共8页昆虫科学（英文版）

摘　　要：Apis cerana indica foragers were used for the isolation of a full-length α- glucosidase cDNA, and for purification of the active nascent protein by low salt extraction of bee homogenates, ammonium sulphate precipitation and diethylaminoethyl-cellulose and Superdex 200 c hromatographies. The molecular mass of the purified protein was estimated by polyacrylamide gel electrophoresis resolution, and the pH, temperature, incubation, and substrate optima for enzymic activity were determined. Conformation of the purified enzyme as α-glucosidase was performed by BLAST software homology comparisons between matrix assisted laser desorption ionization time of flight mass spectroscopy analysed partial tryptic peptide digests of the purified protein with the predicted amino acid sequences deduced from the α-glucosidase cDNA sequence.Apis cerana indica foragers were used for the isolation of a full-length α- glucosidase cDNA, and for purification of the active nascent protein by low salt extraction of bee homogenates, ammonium sulphate precipitation and diethylaminoethyl-cellulose and Superdex 200 c hromatographies. The molecular mass of the purified protein was estimated by polyacrylamide gel electrophoresis resolution, and the pH, temperature, incubation, and substrate optima for enzymic activity were determined. Conformation of the purified enzyme as α-glucosidase was performed by BLAST software homology comparisons between matrix assisted laser desorption ionization time of flight mass spectroscopy analysed partial tryptic peptide digests of the purified protein with the predicted amino acid sequences deduced from the α-glucosidase cDNA sequence.

关 键 词：Α-GLUCOSIDASE Apis cerana indica CDNA CHROMATOGRAPHY PURIFICATION 

分 类 号：Q969[生物学—昆虫学]