Crystallization and preliminary X-ray analysis of Thermoactinomyces vulgaris R-47 maltooligosaccharide-metabolizing enzyme homologous to glucoamylase.
A maltooligosaccharide-metabolizing enzyme from Thermoactinomyces vulgaris R-47 (TGA) homologous to glucoamylase degrades maltooligosaccharides more efficiently than starch, unlike fungal glucoamylases. TGA was crystallized and the state of the protein in solution was analyzed by gel-filtration chromatography. Diffraction data were collected to 3.31 A resolution. The TGA crystal belongs to the orthorhombic space group P2(1)2(1)2(1) or P2(1)2(1)2, with unit-cell parameters a = 110.2, b = 317.6, c = 144.9 A, and is expected to contain five to eight TGA molecules per asymmetric unit. Gel-filtration and native PAGE analyses indicated that TGA exists as a dimer in solution. This is the first report of the crystallization of an oligomeric glucoamylase.
Ichikawa
Kazuhiro
K
Department of Applied Biological Science, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509, Japan.
Tonozuka
Takashi
T
Mizuno
Masahiro
M
Tanabe
Yoshihiro
Y
Kamitori
Shigehiro
S
Nishikawa
Atsushi
A
Sakano
Yoshiyuki
Y
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D002850:Chromatography, Gel; D003460:Crystallization; D008848:Micromonosporaceae / Q000201:enzymology*; D009844:Oligosaccharides / Q000378:metabolism*; D014961:X-Ray Diffraction; D000520:alpha-Glucosidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*
D001426:Bacterial Proteins; D009844:Oligosaccharides; C021705:maltooligosaccharides; D000520:alpha-Glucosidases