In my experience best way is use homogenizer, but very important point that you have to consider for choosing a protocol is next step of your test.

Is there any protocol can you suggest me?

I'd recommend you to atleast go through the following link. It will give you a brief understanding about protein purification.

https://www.embl.de/pepcore/pepcore_services/index.html

There are lots of questions when doing purification like what buffer to choose, salt concentration, additives, inhibitors, temperature, purification steps likewise. Use of homogenizer or sonicator is trivial and depends on the scale and purpose of the purification.

Thank you so much.

It was very useful for me.

Hey there

I am working on the same thing

I am trying to get proteins from Bacillus thuringiesis as well as E. coli

I am doing Sonication in order to lysis the cell and get all the protein out of the cell

I am to getting a good results and I think I have to modify my protocol

Please check the link:

https://www.researchgate.net/post/Can_anyone_suggest_a_method_to_extract_proteins_from_bacterial_cells_I_need_intracellular_as_well_as_extracellular_proteins

Hi Waeell,

Thanks for your answer.

It very much depends upon what you intend to use the protein for. If you want native protein for purification or enzyme measurements, then you need a fairly gentle protocol. Traditionally bacterial cells are broken with a French press or a sonicator, depending upon the volume of cells. French press is good for larger quantities and sonicator best for smaller. You can also lyse the cells by freeze thaw cycles after lysozyme treatment. Please note that these methods generally do NOT release membrane bound proteins, those need to be released using detergent. On the other hand, if you want total protein to run on a gel or some other procedure but don't require that the proteins be fully folded and native, then lysis by detergent is the most efficient method.

I could be wrong, but I thought homogenizers are good for larger cells but not very effective on smaller bacterial cells.

Michael J. Benedik

you are right exactly. it was my mistake. my mean was sonication.

sorry for that.

Hi Michael ,

Thank you for your answer.

I used lysis buffer containing Triton X-100 and PMSF. Also, I used two different ways to break up the bacterial cell wall which are homogenisator and sonicator.

Thanks again.