2 replies to this topic

[joy123]

Hi, I am interested in an enzyme in a G(+) bacteria. It natually sits on cell membrane.

I have cloned it in E.coli. But the recombinant protein didn't have enzyme activity (I checked it by loading E coli on zymogram).

My question is: how to make a recombinant enzyme have activity? Does the procedure differs a lot for different enzymes, or there is a universal procedure?

Thanks very much! Look forward to hear from you!

[Inmost sun]

you may find out an in vitro protocol for reconstitution (f.i. Novogene)

or try coexpression with a chaperon/HSP

[joy123]

Thanks! I am working on it.