title: Purification and characterization of alpha-glucosidase I from Japanese honeybee (Apis cerana japonica) and molecular cloning of its cDNA.
abstract: alpha-Glucosidase (JHGase I) was purified from a Japanese subspecies of eastern honeybee (Apis cerana japonica) as an electrophoretically homogeneous protein. Enzyme activity of the crude extract was mainly separated into two fractions (component I and II) by salting-out chromatography. JHGase I was isolated from component I by further purification procedure using CM-Toyopearl 650M and Sephacryl S-100. JHGase I was a monomeric glycoprotein (containing 15% carbohydrate), of which the molecular weight was 82,000. Enzyme displayed the highest activity at pH 5.0, and was stable up to 40 degrees C and in a pH-range of 4.5-10.5. JHGase I showed unusual kinetic features: the negative cooperative behavior on the intrinsic reaction on cleavage of sucrose, maltose, and p-nitrophenyl alpha-glucoside, and the positive cooperative behavior on turanose. We isolated cDNA (1,930 bp) of JHGase I, of which the deduced amino-acid sequence (577 residues) confirmed that JHGase I was a member of alpha-amylase family enzymes. Western honeybees (Apis mellifera) had three alpha-glucosidase isoenzymes (WHGase I, II, and III), in which JHGase I was considered to correspond to WHGase I.
authors.affiliation: Graduate School of Agriculture, Hokkaido University, Sapporo.
authors.forename: Jintanart
authors.initials: J
authors.lastname: Wongchawalit
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nakai
authors.forename: Young-Min
authors.initials: YM
authors.lastname: Kim
authors.forename: Natsuko
authors.initials: N
authors.lastname: Sato
authors.forename: Mamoru
authors.initials: M
authors.lastname: Nishimoto
authors.forename: Masayuki
authors.initials: M
authors.lastname: Okuyama
authors.forename: Haruhide
authors.initials: H
authors.lastname: Mori
authors.forename: Osamu
authors.initials: O
authors.lastname: Saji
authors.forename: Chanpen
authors.initials: C
authors.lastname: Chanchao
authors.forename: Siriwat
authors.initials: S
authors.lastname: Wongsiri
authors.forename: Rudee
authors.initials: R
authors.lastname: Surarit
authors.forename: Jisnuson
authors.initials: J
authors.lastname: Svasti
authors.forename: Seiya
authors.initials: S
authors.lastname: Chiba
authors.forename: Atsuo
authors.initials: A
authors.lastname: Kimura
mesh_terms: D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000201:enzymology*; D050260:Carbohydrate Metabolism; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D013696:Temperature; D000520:alpha-Glucosidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; C042059:glucosidase I; D000520:alpha-Glucosidases