Glutathion in Bacterial Media
Dr. Yossef Av-Gay
yossi at unixg.ubc.ca
Sun Jun 16 20:47:33 EST 1996
Achim Recktenwald wrote:
> corboy at utdallas.edu wrote:
> > Seeking knowledge of the effects of glutathion in bacterial media. The
> > purpose is analagous to adding 2-ME or DTT to solutions when purifying a
> > protein by standard methods. My protein is secreted into the medium, and
> > I am trying to maximize the availability of a single free Cysteine SH
> > group by protecting it (anti-oxidant). To my knowledge, you cannot use
> > 2-ME or DTT in culture because they are membrane permeable and toxic. As
> > far as I know, glutathion is not membrane permeable, but can be imported
> > by bacteria. What I want to know is how much glutathion can I add to
> > bacteria growing in liquid culture to provide SH protection, but not
> > affect growth or reduce existing disulfide bonds, like you would use 2-ME
> > for in purification. Thanks in advance for any help I may get.
> > Mike Corboy
> > University of Texas at Dallas
> Ever thought of adding cysteine to the medium?
Some Bacteria mainly gram positive do not have or produve glutathion.
Thus they do not need it. Any hoe the amount requiered to keep it
reduced is so low you do not need to worry about. Grow the bacteria.
HAve fun purifing/manipulating and when you do the enzymatic assay add
DTT/DTE. Even if you want to use affinity chromatography you can use
DTT prior to the purification
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