Table 1.
Purification step | Total protein (mg) b | Volume (ml) | Purity (%) c | Amount of HP-NAP (mg) d | Step recovery of HP-NAP (%) e | Total recovery of HP-NAP (%) f |
---|---|---|---|---|---|---|
Whole cell lysate | 26.70 | 20.00 | 69.45 | 18.54 | 100 | 100 |
Supernatant | 28.60 | 20.00 | 73.34 | 20.98 | 113 | 113 |
DEAE Sephadex chromatography | 19.78 | 59.95 | 95.97 | 18.98 | 90.5 | 102 |
Dialysis | 18.54 | 57.95 | 97.12 | 18.01 | 94.9 | 97.1 |
Acrodisc syringe filtration | 15.33 | 56.79 | 99.32 | 15.23 | 84.6 | 82.1 |
aFrom 0.115 g of E.coli cell paste obtained from 50 ml culture.
bProtein concentration determined by the Bradford method with bovine serum albumin (BSA) as the reference.
cValues determined from densitometry measurement as described in Methods.
dValues determined by multiplying the values in the column of “Total protein” and “Purity”.
eValues determined by dividing the amount of HP-NAP from each purification step by that from the previous step.
fValues determined by dividing the amount of HP-NAP from each purification step by that from the whole cell lysate.