Hi Guys, Lactose is a metabolisable sugar molecule and affected by inducer exclusion and catabolite repression. Any experience of using lactose as inducer instead of IPTG to improve recombinant protein expression in E.coli BL21(DE3).Please advise me the usage of lactose and its advantages over IPTG for large scale production of therapeutics.Thanks !
Hi there,
The use of lactose instead of IPTG is the goal of auto-induction medium from Studier. It implies the medium is first totally depleted with glucose otherwise repression by glucose prevents any induction by lactose. For information, the recipe indicates 2g/L for lactose.
I think this should interest you as it compares auto-induction and IPTG induction:
http://www.nature.com/app_notes/nmeth/2005/051122/full/nmeth0305-233.html
Hope it helps
Hi Dominique Thanks for the information. My aim is to express protein in cytoplasm as soluble with high yield using lactose/glucose system. Growing cells in very low conc of glucose in the pool along with lactose as inducer is possible to induce expression at low temperature(25/30C)?
Yes!
That's the purpose of autoinduction medium : biomass forms on glucose then glycerol is used and finally lactose. With this mode of culture, you don't need to monitor the growth : typical experiment is run overnight at 20-25°C.
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