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Fwd: Slimy recombinant e.coli pellet

Megha Goyal via methods%40net.bio.net (by mgbiotec from gmail.com)
Thu Nov 16 02:57:04 EST 2017


dear DK sir,

yes we also assume the same. so any suggestion how can we handle it. Our
protein is in form of inclusion bodies and we get our protein but slimy
cell mass makes it difficult to handle the cell paste. Also our induction
is with IPTG for only 3 hours. so can anyone suggest how do we prevent the
cell lysis if its occurring as we maintain DO, temperature etc during the
process and we do feeding with glucose at intervals during the induction
phase.

On Fri, Nov 10, 2017 at 5:51 AM, DK <dk from no.email.thankstospam.net> wrote:

> In article <mailman.110.1510233337.15473.methods from net.bio.net>, Megha
> Goyal <mgbiotec from gmail.com> wrote:
> >Dear All,
> >
> >We are doing fermentation for recombinant protein using E.coli culture.
> >Earlier we used to obtain compact cell pellet on harvesting (3 Hrs after
> >IPTG Induction). But recently we observed that the cell pellet harvested
> >are of slimy consistency and they do not homogenise in the washing buffer
> >(tris-nacl-edta). We observe thread like consistency and no matter of
> >stirring it do we get a homogeneous suspension.
> >
> >Can someone guide me what is the reason for such phenomenon.
>
> Partial cell lysis. "Slime" = chromosomal DNA.
>
> DK
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