Construction of CPO Immobilized Enzyme Reactor Based on Center-radial Dendritic Mesoporous Silica and Its Application †

doi:10.7503/cjcu20190191

Abstract

Abstract:

The mesoporous silica particles(DSP) with open three-dimensional center-radial dendrimers and uniform particle size were first prepared. Then CPO@DSP immobilized enzyme reactor was prepared by electrostatic interaction with chloroperoxidase(CPO) entrapped into the pore channel. The pore size was regulated by changing the concentrations of tetraethyl orthosilicate(TEOS) and the template agent cetyltrimethylammo-nium chloride(CTAC). The effect of pore size on the catalytic activity of immobilized enzyme reactor was studied. At the same time, the limiting effect of pore channel was discussed based on the kinetic analysis of enzymatic reaction. In addition, sodium alginate(SA) hydrogel film coated on the surface of CPO@DSP was used to improve the leakage of enzyme molecules in the enzyme reactor, and the reusability of the obtained SA-CPO@DSP enzyme reactor was significantly improved. After 10 times of reuse, more than 90% of the catalytic activity was still maintained. When SA-CPO@DSP was applied to the degradation of residual antibiotics in environmental water, the degradation rate of 100 μg/mL substrate in 25 min was over 88%. SA-CPO@DSP was also used for visual colorimetric detection of phenol and 5 μmol/L phenol could be detected by naked eyes. It showed the application prospect of SA-CPO@DSP in environmental protection.

Key words: Immobilized enzyme reactor, Mesoporous silica, Chloroperoxidase, Environmental application

CLC Number:

TrendMD:

SONG Yichao, HU Mancheng, LI Shuni, ZHAI Quanguo, JIANG Yucheng. Construction of CPO Immobilized Enzyme Reactor Based on Center-radial Dendritic Mesoporous Silica and Its Application ^†[J]. Chem. J. Chinese Universities, 2019, 40(9): 1805.

Figures/Tables 8

Fig.1 TEM images of DSP(A) and SA-CPO@DSP(B)

Fig.2 CLSM images of DSP(A), CPO@DSP(B) and overlay of images(A) and (B) (C)

Sample	CPO@DSP-1	CPO@DSP-2	CPO@DSP-3	CPO@DSP-4	CPO@DSP-5
Pore size/nm	9.78	7.43	6.16	4.41	11.13
Catalytic activity(%)	98.36	77.95	73.19	23.87	98.08

Sample	Absorbance
Sample	1st	3rd	5th	8th	10th
CPO@DSP	0.044	0.096	0.148	0.178	0.216
SA-CPO@DSP	0.020	0.032	0.046	0.065	0.084

Fig.3 Reusabilities of CPO@DSP(A) and SA-CPO@DSP(A)

Fig.4 Absorbance of MCD(a), MCD after conversion by CPO@DSPs(b) and SA-CPO@DSPs(c)(A) and comparison of activity of free CPO, CPO@DSPs and SA-CPO@DSPs(B)

Catalyst	v_max/(mmol·L^-1·s^-1)	K_m/(mmol·L^-1)	k_cat/s^-1	(k_cat/K_m)/(L·s^-1·mmol^-1)
Free CPO	1.17	1.62	5.86×10⁴	3.62×10⁴
CPO@DSP	0.19	0.24	9.31×10³	3.91×10⁴
SA-CPO@DSP	0.10	0.13	5.19×10³	4.17×10⁴

Fig.5 Colorimetric change of the solutions containing SA-CPO@DSP, H2O2 and phenol(A) and plots of A505 vs. phenol concentration(B) The concentration unit in image (A) is μmol/L.

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