In vitro effects of some anaesthetic drugs on lactoperoxidase enzyme activity

Figures & data

Figure 1 SDS-Polyacrylamide gel electrophoresis of purified LPO. (lane 1; Serum albumin (M.W 66.000) ovalbumine (M.W 45.000), carbonic anhydrase (M.W 30.000.) (Lane 2; Crude extract), (Lane 3–4: Ammonium sulfate precipitation), (Lane 5–6: purified bovine LPO in CM sephadex C-50 column), (Lane 7–8: purified bovine LPO in sephadex–G100 column).

Figure 2 % Activity-[drug] graphs for LPO for two drugs; (A) Bupivacaine and (B) Ketamine.

Figure 3 Lineweaver-Burk graph for 6 different substrate (ABTS) concentration and 3 different ketamine concentrations for determination of K_i constant.

Figure 4 Lineweaver-Burk graph for 6 different substrate (ABTS) concentration and 3 different Bupivacaine concentrations for determination of K_i constant.

Table I.  Purification steps of lactoperoxidase from bovine milk.

Step	Activity (EÜ/ mL)	Total Volume (mL)	Protein (mg/ml)	Total Protein (mg)	Total Activity (EU)	Specific Activity (EU/mg)	Recovery (%)	Purification (Fold)
Crude homogenate	3.33	150	1.85	277	499	1.80	100	1.00
Ammonium sulfate	15.6	26	1.97	50.8	405	7.97	82	4.42
CM-Sephadex C50 column	1.29	300	0.09	27.0	387	14.33	77.4	7.96
Ammonium sulfate	12.2	26	0.70	18.2	317	17.41	64	9.67
Sephadex G100 column	1.59	140	0.08	11.2	222	19.8	44	11
Ammonium sulfate and dialyse	5.81	24	0.28	6.7	139.4	20.7	28	11.5

Table II.  I₅₀ values, K_i constant, and inhibition types of Ketamine and Bupivacaine for bovine LPO.

Inhibitors	[drugs] (mM)	Ki Constant (mM)	Mean Ki constant(mM)	Inhibition Type	I50values (mM)
	0.006	0.018
Ketamine	0.012	0.019	0.019 ± 0.031	Non-Competitive	0.29
	0.018	0.019
	0.001	0.014
Bupivacaine	0.003	0.015	0.015 ± 0.021	Non-Competitive	0.155
	0.007	0.015