Purification, Immobilization, and Characterization of Bovine Lactoperoxidase

Figures & data

Table 1 Purification steps of lactoperoxidase from bovine milk

Step	Activity(EU/ml)	Totalvolume(ml)	Protein(mg/ml)	Totalprotein(mg)	Totalactivity(EU)	Specificactivity(EU/mg)	Recovery(%)	Purification (fold)
Crude extract	2.8	150	1.58	237	420	1.77	100	1.00
CM Sephadex C50 column	1.36	270	0.11	29.7	367	12.35	87.3	6.97
Sephadex G100 column	2.4	120	0.11	13.2	288	21.8	68.5	12.31
Ammonium sulfate and dialysis	5.55	25	0.24	6	138.7	23	33	13

Figure 1 SDS-polyacrylamide gel electrophoresis of bovine LPO. (Color figure available online.)

Figure 2 Influence of pH on LPO immobilization efficiency on PANIG (n = 3). (Color figure available online.)

Figure 3 Relationship between the relative amounts of LPO and PANIG and the amount of immobilized LPO (n = 3). (Color figure available online.)

Figure 4 Time dependency of immobilized enzyme activities (n = 3). (Color figure available online.)

Table 2 Efficiency of PANIG on active enzyme retention

Total enzyme		Immobilized enzyme		Total protein		Retained protein
UE	%	UE	%	μg	%	μg	%	SA^a	Binding efficiency
138 ± 3.2	100	32.01 ± 1.6	23.2	3 ± 0.2	100	0.75 ± 0.03	25.31	42.68	0.91
Average ± standard derivation (n = 5).
^aSA defined by EU μg ^−1 protein.

Figure 5 AFM images of bare (a) and immobilized enzyme (b) polyaniline polymer. (Color figure available online.)

Figure 6 Storage stability of free (♦) and immobilized (▲) enzymes at 4°C (n = 3). (Color figure available online.)

Figure 7 Storage stability of free (♦) and immobilized (▲) enzymes at 30°C (n = 3). (Color figure available online.)

Figure 8 Thermal stability of free (♦) and immobilized (▲) enzymes at 60°C (n = 3). (Color figure available online.)