2.2. Mass spectrometric analysis by protein extraction
method and intact cell method
The bacterial pellet was dissolved in 1 mL of water and
centrifuged (all centrifugation steps were performed at
13,000 g for 2 minutes) in order to wash out the residue
of BHI.
According to the protein extraction method (PEM), 1 mL
of 75% ethanol solution was added to the pellet, and after
centrifugation, the supernatant was removed by carefully
pouring it from the microtube. A second centrifugation step
was performed, and the remaining liquid was carefully
removed with a pipette. The bacterial pellets were allowed
to air-dry at room temperature for 15 minutes. A solution of
70% formic acid was added to lyse bacterial cells and to
release the inner-cell proteins, which are predominantly
ribosomal proteins that produce diagnostic ions in MALDIMS
fingerprinting. The 70% formic acid solution was added
proportionally to the size of pellet to completely dissolve it.
2.2. Mass spectrometric analysis by protein extraction
method and intact cell method
The bacterial pellet was dissolved in 1 mL of water and
centrifuged (all centrifugation steps were performed at
13,000 g for 2 minutes) in order to wash out the residue
of BHI.
According to the protein extraction method (PEM), 1 mL
of 75% ethanol solution was added to the pellet, and after
centrifugation, the supernatant was removed by carefully
pouring it from the microtube. A second centrifugation step
was performed, and the remaining liquid was carefully
removed with a pipette. The bacterial pellets were allowed
to air-dry at room temperature for 15 minutes. A solution of
70% formic acid was added to lyse bacterial cells and to
release the inner-cell proteins, which are predominantly
ribosomal proteins that produce diagnostic ions in MALDIMS
fingerprinting. The 70% formic acid solution was added
proportionally to the size of pellet to completely dissolve it.
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