Cloning, expression, purification and crystallization of saccharopine reductase from Magnaporthe grisea.
The gene coding for saccharopine reductase (E.C. 1.5.1.10), an enzyme of the alpha-aminoadipic pathway of lysine biosynthesis in the pathogenic fungus Magnaporthe grisea, was cloned and expressed in Escherichia coli. The purified enzyme was crystallized in space groups C2 and C222(1) using ammonium sulfate pH 4.8 or PEG 6000 pH 4. 1 as precipitants. The unit-cell parameters are a = 115.0, b = 56.6, c = 74.3 A, beta = 111.1 degrees for space group C2, and a = 89.3, b = 119.0, c = 195.9 A for space group C222(1). The crystals diffract to resolutions of 2.0 A (C2) and 2.4 A (C222(1)) at synchrotron sources.
Johansson
E
E
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden.
Steffens
J J
JJ
Emptage
M
M
Lindqvist
Y
Y
Schneider
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D004926:Escherichia coli; D020082:Magnaporthe / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012443:Saccharopine Dehydrogenases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification
D011994:Recombinant Proteins; D012443:Saccharopine Dehydrogenases; C041897:saccharopine dehydrogenase (NADP, L-glutamate-forming)