Supplemental fig 1. Separation of α-glucosidase from mice intestine by HSCCC using
reverse micelle solvent system. Separation conditions: stationary phase was pH 4.5
Tris–HCl buffer phase containing 50 mM Tris–HCl and 50 mM KCl; the mobile
phase A was isooctane containing 30 mM AOT, while, 50 mM Tris–HCl buffer
containing 500 mM KCl (pH 8.0) was used as mobile phase B; The retention ratio of
the stationary phase is 50 % .α-glucosidase detained in stationary phase was eluted in
fractions collected at 130-156 min. A: bands of fraction 4; B: bands of protein
marker ;
Supplemental fig 2. Separation of α-glucosidase from mice intestine by HSCCC using
reverse micelle solvent system. Separation conditions: stationary phase was pH 4.5
Tris–HCl buffer phase containing 50 mM Tris–HCl and 50 mM KCl; the mobile
phase A was isooctane containing 50 mM AOT, while, 50 mM Tris–HCl buffer
containing 500 mM KCl (pH 8.0) was used as mobile phase B; The retention ratio of
the stationary phase is 48 % .α-glucosidase detained in stationary phase was eluted in
fractions collected at 246-268 min. A: bands of fraction 10; B: bands of protein
marker ;
AB CD
4
8
Supplemental fig 3. Separation of α-glucosidase from mice intestine by HSCCC using
reverse micelle solvent system. Separation conditions: stationary phase was pH 4.0
Tris–HCl buffer phase containing 50 mM Tris–HCl and 50 mM KCl; the mobile
phase A was isooctane containing 30 mM AOT, while, 50 mM Tris–HCl buffer
containing 500 mM KCl (pH 8.0) was used as mobile phase B; Rotation speed is 700
rpm; 25 ml (23.9 mg) crude enzyme was injected through the injection valve. The
separation temperature was set at 25 ℃. The retention ratio of the stationary phase is
49 % .SDS-PAGE of separated sample is placed in the upper right corner. A: bands of
fraction 4; B: bands of fraction 8; C: bands of crude enzyme ; D: bands of protein
marker ;
0.9
Crude enzyme
Purified enzyme
0.8
OD value
Y = 0.0703x + 0.3589
2
R = 0.9988
0.7
Y = 0.0106x + 0.4329
2
R = 0.9871
0.6
0.5
0.4
2
4
6
8
10
12
14
16
18
20
22
Time/min
Supplemental fig 2.The comparison of α-glucosidase catalytic activity of fraction 8
and the crude enzyme (0.0703 vs 0.0106).