In Situ Encapsulation of Cellubiase Enzyme in Magnetic Mesoporous Silica via the Nonsurfactant-templated Sol-gel Method†

doi:10.7503/cjcu20150688

Abstract

Abstract:

Nonsurfactant-templated sol-gel method is an advanced technology for enzyme encapsulation. The immobilized enzyme can be reused many times, so that can reduce the cost. It has a broad application prospect in biological catalysis. D-fructose is used as non-surfactant template to generate mesopores. The average diameter of the mesoporous silica is 3.2 nm. And they significantly improve the activity of the immobilized enzyme. The best result is 70% of free enzyme. On the other hand, the mesopores are smaller than the size of cellobiase enzyme. So the enzyme will not leach out. That can help avoiding enzyme leaching. The magnetic nanoparticles can make the separation and recycle of immobilized enzyme easier and more feasible. The incorporation of magnetic nanoparticles has little effect on the enzyme activity. The magnetic encapsulated enzyme can be easily recycled by magnet force. More than 83% remained after 10 cycles. The process is simple and biocompatible as the sol-gel reaction occurs inaqueous phase and neutral pH.

Key words: Enzyme encapsulation, Cellobiase enzyme, Mesoporous silica, Magnetic nanoparticles, Sol-gel reaction, Non surfactant template

CLC Number:

O629.8

TrendMD:

ZHANG Xiang, LI Shurun, ZHAI Wentao, CAO Yingze, QI Hongxu, JI Yan, WEI Yen. In Situ Encapsulation of Cellubiase Enzyme in Magnetic Mesoporous Silica via the Nonsurfactant-templated Sol-gel Method^†[J]. Chem. J. Chinese Universities, 2015, 36(11): 2349.

Figures/Tables 10

Table 1 Information of each sample

Sample	m(SiO₂)/g	m(Fructose)/g	m(Enzyme)/mL	m(Fe₃O₄)/g
E0	9.0	0	0.5	0
M0	9.0	0	0.5	0.45
E3	9.0	3.0	0.5	0
M3	9.0	3.0	0.5	0.45
E6	9.0	6.0	0.5	0
M6	9.0	6.0	0.5	0.45
E9	9.0	9.0	0.5	0
M9	9.0	9.0	0.5	0.45

Fig.1 SEM images of immobilized enzyme with different magnifications

Table 2 Hysteresis loop tests of Fe3O4, E0, M0 and M9

Sample	Remanence/(A·m²·kg^-1)	Coercive force/(A·m^-1)
Fe₃O₄	8.50	9.0×10⁴
E0	0.0036	9.0×10⁴
M0	0.28	9.0×10⁴
M9	0.24	1.1×10⁵

Fig.2 TGA curves of E0(a), E3(b), M3(c), M6(d), E6(e), M9(f) and E9(g)

Table 3 Fructose template content, mass loss, textural properties from nitrogen adsorption-desorption and enzymatic activity of immobilized cellobiase samples

Sample	Template added(%)	Mass loss(%)	BET/(m²·g^-1)	Pore diameter/nm	Pore volume/ (cm³·g^-1)	Enzymatic activity/ (mg·dL^-1·h^-1)
Free enzyme	—	—	—	—	—	80
E0	0.0	6.9	336	2.38	0.102	2
E3	25.0	18.1	720	2.64	0.347	13
M3	24.1	20.7	703	2.71	0.353	15
E6	40.0	34.8	645	3.00	0.550	28
M6	38.8	32.1	576	2.68	0.433	31
E9	50.0	46.2	922	3.40	0.851	56
M9	48.8	45.5	730	3.32	0.632	48

Fig.3 Nitrogen adsorption-desorption isotherms of M0(a), M3(b), M6(c) and M9(d)

Fig.4 Enzyme activity plots of free enzyme(a), M3(b), M6(c) and M9(d)

Fig.5 Enzyme activity plots of free enzyme(●) and M9(■) at different temperature

Fig.6 Enzyme activity plots of E9(a) and M9(b) in recycle uses

Fig.7 Sedimentation of magnetic and non-magnetic samples at the beginning(A), 1 min(B), 10 min(C) and 1 h(D)

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