Effect of High
Hydrostatic Pressure on Activity of Restriction Endonucleases
TIAN Shao-Min, GONG Zhen-Zhen, XIE Wei-Jun, ZHENG Le and
RUAN Kang-Cheng
( Shanghai Institute of Biochemistry, the Chinese Academy of Sciences,
Shanghai 200031, China )
Abstract 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 ℃ 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.
Key Words high pressure； restriction endonuclease； HindIII；
XbaI
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