Crystallization and preliminary X-ray crystallographic analysis of chitinase F1 (ChiF1) from the alkaliphilic Nocardiopsis sp. strain F96.
Chitinase F1 (ChiF1) isolated from the alkaliphilic Nocardiopsis sp. strain F96 is a family 18 chitinase that hydrolyzes chitin, an insoluble beta-1,4-linked polymer of N-acetylglucosamine. Crystals of recombinant ChiF1 with molecular weight of 33 000 Da were grown to a suitable size for X-ray structure analysis using 18%(w/v) polyethylene glycol 8000, 200 mM zinc acetate dehydrate and 100 mM sodium cacodylate buffer pH 6.5. Diffraction data were collected at SPring-8 and show that the crystals belong to the trigonal space group P3(1)12 or P3(2)12, with unit-cell parameters a = 56.0, c = 179.5 A, and diffract X-rays beyond 1.2 A resolution. Crystallographic analysis was carried out using the multiwavelength anomalous diffraction (MAD) method using zinc as the anomalous scatter. The binding of Zn atoms was clarified from the Bijvoet and dispersive Patterson functions, which gave prominent zinc-zinc self-vectors on the Harker section.
Matsui
Tsutomu
T
Department of Life Science, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Kumasaka
Takashi
T
Endo
Kimiko
K
Sato
Takao
T
Nakamura
Satoshi
S
Tanaka
Nobuo
N
D000192:Actinomycetales / Q000201:enzymology*; D002688:Chitinases / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D006863:Hydrogen-Ion Concentration
D002688:Chitinases