For the function of cell on contaminant resistance or degradation, including cell wall, cell membrane and extracellular secretion, as well as functional groups, such as carboxyl and amino.
I think that any process subcellular fractionation has two stages: cell disruption and separation of the desired fraction from the remaining components of the cell by a criterion as can be a difference in density (gradient centrifugation or differential centrifugation), the presence of any antigen (immunoaffinity chromatography, immunoprecipitation, etc.).
To break you can use various techniques: homogenization, sonication, osmotic shock...
For separation you can use centrifugation. Using centrifugal force and time variables. According to Stokes' law, you can also separate the different components according to their relative densities with respect to a density gradient.
To measure you can do plate counts prior to the steps of breakdown and separation. knowing the number of CFU there for each separate cell component.
I am not an expert on the subject, I've deduced using my limited knowledge.
luck.
To examine extracellular compounds, you just grow cells in a minimal medium, remove cells by centrifugation and use supernatants. To fractionate Gram negative bacteria you can easily separate periplasmic, cytoplasmic and membrane proteins using one of the following protocols. Depending on if you need to work with proteins under native or denaturing conditions you can follow the methods described by M G Klotz and S W Hutcheson (1992; Appl. Environ. Microbiol. 58(8):2468) or Quan et al (2013; Methods Mol Biol. 966:359-66. doi: 10.1007/978-1-62703-245-2_22.), respectivelly. These methodologies are very simple and you do not need expensive reagents or instruments. I hope this is helpful for you,
Good luck!
Thank Santander!
These methods may be ok, but for Gram positive bacteria, I'll check them.
Try as in - Yeasts are single-celled, their progeny,or daughter cells, will sometimes not separate
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