Extracting enzymes from E. coli

Baltimoir baltimoir at home.com
Mon Dec 4 16:11:40 EST 2000


On 4 Dec 2000 19:08:57 -0000, arsphys at cc.usu.edu (Phil Harrison)
wrote:


>2.  Would freeze-thaw cycles in liquid nitrogen disrupt the cells?

An interesting effect can sometimes be obtained by freeze thawing and
resuspension. A single freeze thaw cycle frequently only disrupts the
periplasmic space leaving the cell intact. If your protein is
overexpressed and some makes it to the periplasmic space (no quick way
to tell this so just assume for the moment it does), then a freeze
thaw cycle of the pelleted Ecoli followed by resuspension will often
extract your protein of interest in a relatively purified state
without bursting the Ecoli.


good luck







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