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Signal peptides E. coli and Methionin


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#1 pDNA

pDNA

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Posted 28 January 2022 - 06:31 AM

Dear people,

 

   long time no see :)

 

I have a long standing issue that troubles me all the time i'm designing constructs for the periplasmic expression of heterologous proteins in E. coli.

 

Do you guys keep the original methionin of the gene of interest or do you work with particular amino acids that you put after the signal peptide cleavage site.

It is said that the +1 position for secreted proteins in E. coli is mainly A or polar amino acids like D and E.

 

The give some sort of motif here:

Flanking signal and mature peptide residues influence signal peptide cleavage | BMC Bioinformatics | Full Text (biomedcentral.com)

 

I would be interested how the crowd is approaching this and if you have seen a huge experience on the cleavage efficiency of the signal peptidase depending on the downstream amino acids.

 

Or some people just skip the M and start with the sequence of the GOI.

 

What do you think is the best option?

 

Many thanks for your time and help in advance!

 

Best,

pDNA






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