A new study of cell disruption to release recombinant thermostable enzyme from Escherichia coli by thermolysis.
Extraction of intracellular protein from Escherichia coli is traditionally achieved by mechanical, chemical or enzymatic disruption technology. In this study, a novel thermolysis method was used to disrupt E. coli cells to release a recombinant thermostable esterase. We found that heat treatment of E. coli was highly effective to destroy the integrity of bacterial cell walls and release the recombinant hyperthermophilic esterase at temperatures above 60 degrees C. The effects of temperature, pH and cell concentration on the efficiency of cell disruption were examined. The most effective temperature for cell disruption was at 80 degrees C. The pH and cell concentration had only minor effect on the release of the hyperthermophilic esterase. In addition, we found that the hyperthermophilic esterase could be purified at the early stage of the thermolysis, which is a major advantage of the thermolysis method. Finally, a comparison between thermolysis and traditional methods for the disruption of cells and the release of the thermostable enzyme was made.
Ren
Xiaodong
X
Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun, PR China.
Yu
Dawei
D
Yu
Lei
L
Gao
Gui
G
Han
Siping
S
Feng
Yan
Y
D002463:Cell Membrane Permeability; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology*; D004950:Esterases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D006358:Hot Temperature; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013816:Thermodynamics; D014465:Ultrasonics
D011994:Recombinant Proteins; D004950:Esterases