Hello
I'm getting so many protocols for gram negative bacterial membrane protein isolation but not even single for the gram positive bacteria. Kindly help me for the same
Fractionation of bifidobacterial cell wall proteins.
The bifidobacterial cell wall
proteins were extracted from 50 ml of a bacterial culture in stationary growth
phase as reported by Hardie and Williams (13). Bifidobacterial cells were collected
by centrifugation for 10 min at 5,000 rpm at 4°C and then washed in 50
mM Tris-HCl (pH 7.6). The bacteria were resuspended in 2 ml of protoplast
buffer (50 mM Tris-HCl [pH 7.6], 1 M sucrose, 1.4 mM phenylmethylsulfonyl
fluoride, 15 mg/ml lysozyme). The suspension obtained was incubated for 90 min
at 37°C. Subsequently, the cell suspension was centrifuged for 3 min at 4,000 rpm
at 4°C, and the supernatant, containing the cell wall proteins, was collected and
stored at 20°C.
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