I’m testing the MBEC in Klebsiella pneumoniae and I would like to detach my biofilm to get CFUs counts. I would like to use an enzymatic method for degradation of the matrix. From the literature I know that the matrix is composed of galactose, glucose, mannose and rhamnose and extracellular DNA. Is there any commercial enzyme mixture that I could use?
Regards,
Joao
You can also use proteases such as Proteinase K etc. Its very effective in dispersing most of the biofilms.
Hi João,
Detaching for culturable microorganisms is of course different than detaching for DNA or RNA extraction. You are looking to have intact culturable organisms in the end. Therefore you can't attack the biofilm too heavily otherwise you will disrupt the cells.
Here is a paper which evaluated different methods for exactly what you are looking to do. It was done using wetland biofilm:
Weber K.P., Legge R.L. 2010. Method for the detachment of culturable bacteria from wetland gravel. Journal of Microbiological Methods. 80(3), 242-250.
alpha-glucosidase, and beta-galactosidase, in a phosphate buffer saline shaken at 100rpm for 3h was evaluated against some more simplified methods. Adding enzymes had a small positive effects. See Fig.4 specifically.
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