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Purification and characterization of α‐glucosidase in Apis cerana indica
Author(s) -
Chanchao Chanpen,
Pilalam Suwisa,
Sangvanich Polkit
Publication year - 2008
Publication title -
insect science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 45
eISSN - 1744-7917
pISSN - 1672-9609
DOI - 10.1111/j.1744-7917.2008.00203.x
Subject(s) - biology , chromatography , complementary dna , biochemistry , polyacrylamide gel electrophoresis , molecular mass , matrix assisted laser desorption/ionization , apis cerana , substrate (aquarium) , amino acid , enzyme , desorption , chemistry , botany , ecology , organic chemistry , honey bees , adsorption , gene
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 chromatographies. 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.