A simple preparation method of crystals of soybean hull peroxidase.
Soybean hull peroxidase (SHP) was crystallised from an enzyme solution with low purity by a simple method. The enzyme solution was purified by cooperation salting out of acetone and ammonium sulphate, and lumpy crystals were obtained with the size of about 40 x 30 mum when ammonium sulphate was quickly added to the enzyme solution. The crystal was examined and confirmed to be an SHP crystal by the method of activity test. The result shows that, though the purity of the enzyme solution was not high, crystals could be formed when the enzyme solution rapidly reached to a degree of supersaturation, which was different from the traditional methods of protein crystallisation. Additionally, a purification method of acetone and ammonium sulphate fractional salting out was also studied, in which the procedure was simplified, and a satisfactory purification effect was obtained.
Liu
Junhong
J
Qingdao University of Science and Technology, P.O. Box 70, Qingdao 266042, People's Republic of China. liujh@qust.edu.cn
Liu
Haizhou
H
Zhang
Yuanyuan
Y
Qiu
Longhui
L
Su
Feng
F
Li
Fengmei
F
Su
Zhongliang
Z
Li
Junfeng
J
D000096:Acetone / Q000737:chemistry; D000645:Ammonium Sulfate / Q000737:chemistry; D001709:Biotechnology / Q000379:methods*; D003460:Crystallization; D009195:Peroxidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013025:Glycine max / Q000201:enzymology*
D000096:Acetone; D009195:Peroxidase; D000645:Ammonium Sulfate