Effect of High Hydrostatic Pressure on Activity of Restriction Endonucleases.
The effect of high hydrostatic pressure on the activities of type II restriction enzymes HindIII and XbaI in digesting plasmid pSPORT1 was studied. The endonuclease activity of HindIII and XbaI at 37 degrees were gradually inhibited by increasing pressure and completely inhibited at 200 and 180 MPa, respectively. No obvious irreversible effect was observed for HindIII after suffering high pressure, while a considerable irreversible inactivation was observed for XbaI. The standard molar volume changes for HindIII and XbaI estimated from the inhibition of endonuclease activity at different pressures were 213 and 103 ml/mol, respectively. It was also concluded that pressurization did not change the substrate sequence specificity of both HindIII and XbaI.
Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China. kcruan@sunm.shcnc.ac.cn
Shao-Min
SM
Tian
Zhen-Zhen
ZZ
Gong
Wei-Jun
WJ
Xie
Le
L
Zheng
Kang-Cheng
KC
Ruan