Immobilization of Chloroperoxidase with Magnetic Graphene Oxide and Its Application of Decolorization of Acid Blue 45†

doi:10.7503/cjcu20170617

Abstract

Abstract:

Magnetic graphene oxide(MGO) was prepared using graphene oxide(GO) and Fe₃O₄. Chloro-peroxidase(CPO) was immobilized via adsorption method in MGO, and the immobilization conditions, such as the temperature, time and pH value of the buffer solution, were optimized. The operating stability of immobilized CPO was studied using CPO as catalyst for the degradation of acid blue 45. The activity stability of immobilized CPO was investigated from four aspects of acid base stability, H₂O₂ stability, thermal stability and storage stability using CPO as catalyst for the decolorization of acid blue 45. The results show that the optimal conditions are the phosphate buffer pH=3.5, the reaction time 15 min and the temperature 15 ℃. TMGO{m[NH₄Fe(SO₄)₂·12H₂O]∶m(GO)=10.7∶1} is a better immobilized carrier than FMGO{m[NH₄Fe(SO₄)₂·12H₂O]∶m(GO)=5.35∶1}. And the immobilized CPO shows a better operation stability than the free enzyme. The accumulation layer of TMGO is opened at 35—50 ℃, leading to the loss rate of immobilized enzyme activity significantly less than that of the free enzyme. TMGO-CPO has a good reusability that the activity is still over 60% after recycling 5 times.

Key words: Magnetic graphene oxide, Chloroperoxidase, Immobilization, Acid blue 45

CLC Number:

O629
Q814.3

TrendMD:

GAO Fengqin,WANG Shan,WANG Yunfang,ZHAO Danlei,CUI Ru,JIANG Yucheng. Immobilization of Chloroperoxidase with Magnetic Graphene Oxide and Its Application of Decolorization of Acid Blue 45^†[J]. Chem. J. Chinese Universities, 2018, 39(5): 904.

Figures/Tables 10

Fig.1 XRD patterns of GO(a) and TMGO(b)

Fig.2 SEM images of GO(A), TMGO(B) and TMGO-CPO(C)

Fig.3 TEM images of GO(A), FMGO(B), TMGO(C) and TMGO-CPO(D)

Fig.4 Effects of the pH values(A), time(B) and temperatures on the immobilization of CPO with FMGO

Fig.5 Effects of pH value on the catalytic activity of free(a) and immobilized CPO(b)

Fig.6 Effects of H2O2 concentration on the catalytic activity of free(a) and immobilized CPO(b)

Fig.7 Optimum temperature conditions for dye decolorization of CPO and TMGO-CPO from 15 ℃ to 40 ℃(A) and thermal stability of enzymes of CPO and TMGO-CPO from 25 ℃ to 90 ℃(B)

Fig.8 Illustration of the TMGO layer dispersed due to thermal diffusion

Fig.9 Storage stability of free CPO and immobilized CPO

Fig.10 Reusability of immobilized CPO

References 33

[1]	Wang Y., Wang J.F., Li X. L., Zou H. M., Chinese Journal of Environmental Engineering, 2016, 10(8), 4177—4183
	(王艳, 汪建飞, 李孝良, 邹海明. 环境工程学报, 2016, 10(8), 4177—4183
[2]	Safavia A., Momeni S., J. Hazard. Mater., 2012, 201—202(1), 125—131
[3]	Jiang Y.Z., Shi Y. Q., Shi Y. P., Xu H. Y., Scientia Sinca Chimica, 2014, 44(10), 1528—1535
	(江银枝, 时永强, 史银瓶, 徐火英. 中国科学: 化学, 2014, 44(10), 1528—1535)
[4]	Chen B., Zhao L.L., Sun Q. F., Li Y. R., Li Y., Environmental Science & Technology, 2010, 33(11), 50—53
	(谌斌, 赵亮亮, 孙秋芳, 李燕荣, 李艳. 环境科学技术, 2010, 33(11), 50—53)
[5]	Xiao X., Cui K.P., Wang Z., Feng Y. Y., Technology of Water Treatment, 2016, 42(12), 55—59
	(肖霄, 崔康平, 王郑, 冯莹莹. 水处理技术, 2016, 42(12), 55—59)
[6]	Wang H.L., Zhao Y., Ma L. K., Fan P. H., Xu C. B., Jiao C. L., Lin A. J., Chem. [J]. Chinese Universities, 2016, 37(2), 335—341
	(王会丽, 赵越, 马乐宽, 范鹏浩, 徐从斌, 焦春磊, 林爱军. 高等学校化学学报, 2016, 37(2), 335—341)
[7]	Geng Z., Yu Y., Zhu S.Y., Yu H. B., Liu J. C., Bian D. J., Yang X., Huo H. L., Huo M. X., Chem. Res. Chinese Universities, 2017, 33(1), 36—43
[8]	Fu Y.C., Callaway Z., Lum J., Wang R. H., Lin J. H., Li Y. B., Anal. Chem., 2014, 86(4), 1965—1971
[9]	Palomo J.M., Filice M., Biotechnology Advances, 2015, 33(5), 605—613
[10]	Wu L.T., Wu S. S., Xu Z., Qiu Y. B., Li S., Xu H., Biosensors & Bioelectronics, 2016, 80, 59—66
[11]	Alneyadi A. H., Shah I., AbuQamar S. F., Ashraf S. S., Biomolecules,2017, 7(1), 31(1—18)
[12]	Morozov A. N., Chatfield D. C., Int. J. Mol. Sci., 2016, 17(8), 1297(1—12)
[13]	Lu J., Cheng L., Wang Y., Ding Y., Hu M., Li S., Zhai Q., Jiang Y., Materials & Design, 2017, 129, 219—226
[14]	Yang Q., Wang B.C., Zhang Z., Lou D. S., Tan J., Zhu L. C., RSC Adv., 2017, 7(60), 38028—38036
[15]	Rueda N., Albuquerque T. L., Bartolome-Cabrero R., Lopez L. F., Torres R., Ortiz C., Dos Santos J. C. S., Barbosa O., Lafuente R. F., Molecules,2016, 21(5), 646(1—18)
[16]	Matte C.R., Bordinhão C., Poppe J. K., Benvenutti E. V., Costa T. M. H., Rodrigues R. C., Hertz P. F., Ayub M. A. Z., [J]. Braz. Chem. Soc., 2017, 28(8), 1430—1439
[17]	Sheldon R.A., Pelt S., Chem. Soc. Rev., 2013, 42(15), 6223—6235
[18]	Jiao R.J., Tan Y., Jiang Y. C, Hu M. C, Li S. N, Zhai Q. G., Ind. Eng. Chem. Res., 2014, 53(31), 12201—12208
[19]	Guerrero E., Aburto P., Terrés E., Villegas O., González E., Zayas T., Hernández F., Torres E., [J]. Porous Mat., 2013, 20(2), 387—396
[20]	Ding Y., Cui R., Hu M.C., Li S. N, Zhai Q. G., Jiang Y. C., [J]. Mater. Sci., 2017, 52(17), 10001—10012
[21]	Chen J., Leng J., Yang X., Liao L., Liu L., Xiao A., Molecules,2017, 22(2), 221(1—11)
[22]	Yang D., Wang X., Shi J., Wang X., Zhang S., Han P., Jiang Z., Biochem. Eng. J., 2016, 105, 273—280
[23]	Patel S.K. S., Choi S. H., Kang Y. C., Lee J. K., ACS Appl. Mater. Inter., 2017, 9(3), 2213—2222
[24]	Skoronski E., Souza D.H., Ely C., Broilo F., Fernandes M., Júnior A. F., Ghislandi M. G., Int. [J]. Biol. Macromol., 2017, 99, 121—127
[25]	Hummers W.S. Jr., Offeman R. E., [J]. Am. Chem. Soc., 1958, 80(6), 1339
[26]	Mandal P. Chattopadhyay A.P., Dalton Trans., 2015, 44(25), 11444—11456
[27]	Zhou X., Xu L., Chem. Res. Chinese Universities, 2017, 33(5), 689―694
[28]	Shao Y.B., Jing T., Tian J. Z., Zheng Y. [J]., RSC Adv., 2015, 5(126), 103943—103955
[29]	Kassaee M.Z., Motamedi E., Majdi M., Chem. Eng. J., 2011, 172(1), 540—549
[30]	Zhang B., Lipase Immobilized on the Magnetic Graphene Oxide-chitosan Nanocomposite and Its Application, Lanzhou University of Technology, Lanzhou, 2013
	(张彬. 磁性氧化石墨烯-壳聚糖纳米复合材料固定化的脂肪酶及其应用, 兰州: 兰州理工大学, 2013)
[31]	Cui R., Bai C.H., Jiang Y. C., Hu M. C., Li S. N., Zhai Q. G., Chem. Eng. J., 2015, 259, 640—646
[32]	Zhou L., Deng H.P., Wan J. L., Zhang R. J., Progress in Chemistry, 2013, 25(1), 145—155
	(周丽, 邓慧萍, 万俊力, 张瑞金. 化学进展, 2013, 25(1), 145—155)
[33]	Li X.X., Zhu H., Feng J., Zhang J. W., Deng X., Zhou B. F., Zhang H. L., Xue D. S., Li F. S., Mellors N. J., Li Y. F., Peng Y., Carbon, 2013, 60, 488—497