|
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.
