Immobilization of Candida rugosa lipase on Magnetized Dacron: Kinetic Study

Figures & data

Table 1. Immobilization of lipase on various supports

Lipase	Support	Specific activity retention (%)	References
Candida cylindracea	Spherosyl-carbodiimide beads (covalent)	12	Villeneuve et al. [[2]]
Candida cylindracea	Agarose by tosylation method (covalent)	16	Sánchez et al. [[4]]
Candida cylindracea	Magnetic microspheres (adsorption)	[80% of protein absorbed]	Guo et al. [[8]]
Candida cylindracea	Magnetic poly (methacrylate-divinylbenzene) microspheres (covalent)	72	Liu et al. [[11]]
Candida rugosa	Superparamagnetic nanoparticles via carbodiimide activation (covalent)	[no activity retention indicated]	Bai et al. [[9]]
Fusarium solani FS1	Magnetized Dacron (covalent)	71	Knight et al. [[26]]
Mucor meihei	Magnetic polysiloxane-polyvinyl alcohol (PVA) (covalent)	10	Bruno et al. [[13]]

Figure 1 Thermal stabilities of soluble (▪) and immobilized (▴) on ferromagnetic dacron-azide lipases. The initial specific activities for soluble and immobilized lipases were 0.81 U/mg protein and 0.13 U/g support, respectively.

Figure 2 Storage stability of immobilized lipase on ferromagnetic dacron-azide (0.13 U/g support).

Figure 3 Effect of solvent (isopropanol) on the activity of on soluble (▪) and immobilized (▴) lipase activities. The initial specific activities for soluble and immobilized lipases were 0.81 U/mg protein and 0.13 U/g support, respectively.

Figure 4 Effect of substrate concentration (4-NPP) on soluble (▪) and immobilized (▴) lipase activities. The initial specific activities for soluble and immobilized lipases were 0.81 U/mg protein and 0.13 U/g support, respectively.

Figure 5 Effect of various fatty acid 4-nitrophenyl esters, 4-NPB (▴), 4-NPL (▪), 4-NPP (•), concentration on the activity of soluble lipase.

Figure 6 Effect of various fatty acid 4-nitrophenyl esters, 4-NPB (▴), 4-NPL (▪), 4-NPP (•), concentration on the activity of immobilized lipase. Immobilized derivative retained 16% of soluble specific activity.

Table 2. The apparent Michaelis-Menten kinetics and efficiency for the hydrolysis of 4-nitrophenyl esters by soluble lipase

Substrate (4-nitrophenyl esters)	App KM (mM)	VMAX (µmol/min)	App Kcat (µmol ⋅ min^−1/mg protein)	Efficiency AppKcat/AppKM(µmol ⋅ min^−1/mg protein ⋅ mM)
4-NPB (C4)	0.206±0.043	0.675±0.009	1.654	8.03
4-NPL (C12)	0.193±0.035	0.428±0.004	1.049	5.43
4-NPP (C16)	0.119±0.017	0.568±0.004	1.392	11.69

E_T = 0.408 mg/mL.

Table 3. The apparent Michaelis-Menten kinetics and efficiency for the hydrolysis of 4-nitrophenyl esters by immobilized lipase

Substrate (4-nitrophenyl esters)	App KM (mM)	VMAX (µmol/min)	App Kcat (µmol ⋅ min^−1/mg protein)	Efficiency AppKcat/AppKM(µmol ⋅ min^−1/mg protein ⋅ mM)
4-NPB (C4)	0.119±0.020	0.0562±0.004	0.045	0.38
4-NPL (C12)	0.235±0.035	0.0715±0.006	0.057	0.24
4-NPP (C16)	0.133±0.079	0.0275±0.008	0.022	0.16

E_T = 1.2 mg/0.1 g support.

Immobilized derivative retained 16% of soluble specific activity (0.13 U/g support).

Figure 7 Synthesis of triglyceride (oleic acid + glycerol) catalysed by immobilized lipase on ferromagnetic dacron-azide (0.13 U/g support).

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