Immobilization of paraoxonase onto chitosan and its characterization

Figures & data

Table I. Purification of PON 1 enzyme isolated from human serum

Purification Step	Volume (ml)	Total Activity (unit)	Total Protein (mg)	Spesific Activity (unit/mg)	Yield (%)	Purification-fold
Serum	55	743,3	317,7	2,34	100	–
60 - 80 % (NH4)SO4 Precipitation	34	493,9	201,9	2,44	66,44	1,04
Hydrophobic interaction chromatography	1,5	109,6	0,089	1231	14,74	526

Figure 1. Immobilization of PON1 on chitosan.

Table II. Immobilization of PON1 from human serum on chitosan beads

Free Enzyme	Immobilized Enzyme
Protein (mg/10 chitosan beads) (x)	Specific Activity (unit/mg)	Protein in washing (mg) (y)	Bound Protein (mg) (x–y = A)	Total Activity (unit)	Specific Activity (unit/mg)	Protein Yield (%) ((A/x) × 100)
47	2,34	15	32	93,97	2,93	68

Table III. Catalytic efficiency values for free and immobilized PON1

Derivative	Vmax (EU)	Km (mM)	Catalytic Efficiency Vmax/Km (10³ EU/M)	Kcat (s^−1)	Kcat/Km (s^−1 M^−1)
Free Enzyme	125	1,067	117,15	2,65	2,4925x10³
Immobilized Enzyme	181	1,755	103,13	5,65	3,2229x10³

Figure 2. The effect of pH on the activity of free and immobilized PON1.

Figure 3. The effect of temperature on the activity of free and immobilized PON1.

Figure 4. The thermal stability profile for free PON1.

Figure 5. The thermal stability profile for immobilized PON1.

Figure 6. Storage stability of free and immobilized PON1 from human serum.