Easy Synthesis of Porous Polyurea and Its Application in Enzyme Immobilization and Kinetic Resolution of Racemic Phenylethanol†

doi:10.7503/cjcu20160827

Abstract

Abstract:

Porous polyurea(PPU) was prepared through one step precipitation polymerization in a mixed solvent of water-acetone with toluene diisocyanate(TDI) as the only monomer through step polymerization of TDI with its amine derivatives in-situ produced by TDI reaction with water. The morphology, pore size and size distribution of the PPU were characterized by scanning electron microscope and mercury intrusion. The results demonstrate that PPU is of irregular and granular form with the size of the granules varied from 30 μm to 50 μm. PPU is featured by a continuous pore size distribution from 2 nm to 100 μm with two main regions of pore size distribution, one from 2 nm to 100 nm and another from 5 μm to 100 μm. Amine groups on PPU surface are converted to aldehyde via a treatment with glutaraldehyde, followed by immobilization of lipase from pseudomonas fluorescens(PFL) onto the surface of activated PPU by covalent bonding of the aldehyde groups with the primary amine of the enzyme. Impact of GA and PFL concentration in the process was studied. It is found that the optimized results are obtained when PPU is activated by GA at pH=8 with a concentration of 0.17 mol/L followed by PFL immobilization with PFL concentration of 2.56 mg/mL. Under the optimized conditions, a maximal PFL immobilization of 95.2 mg/g was observed with a high activity of the immobilized PFL of 375 U/mg and the relative activity of 76% relative to free PFL. The immobilized PFL is then used as the catalyst in the kinetic resolution of 1-phenylethanol, the reactivity and the enantioselectivity compared with the free PFL. The results reveal that the activity and the enantioselectivity of the immobilized PFL are greatly enhanced in comparison with the free enzyme. The kinetic resolution of the racemic PEOH is achieved with pure enantiomers. Therefore, this PPU material prepared through simple precipitation is a good potential candidate for enzyme immobilization, the immobilized enzyme is efficient for kinetic resolution of racemic molecules.

Key words: Precipitation polymerization, Porous polyurea, Enzyme immobilization, Activity and enantioselectivity, Kinetic resolution

CLC Number:

O631

TrendMD:

ZHOU Yamei, KONG Xiangzheng, HAN Hui, JIANG Xubao, ZHU Xiaoli. Easy Synthesis of Porous Polyurea and Its Application in Enzyme Immobilization and Kinetic Resolution of Racemic Phenylethanol^†[J]. Chem. J. Chinese Universities, 2017, 38(3): 495.

Figures/Tables 7

References 40

[1]	Yousefi M., Mohammadi M., Habibi Z., J. Mol. Catal. B: Enzym.,2014, 104, 87—94
[2]	Marszall M. P, Siódmiak T., Catal. Commun., 2012, 24, 80—84
[3]	Silva M. R., Mattos M. C., Oliveira M. C. F., Tetrahedron,2014, 70, 2264—2271
[4]	Bonduelle C., Martin-Vaca B., Bourissou D., Biomacromolecules,2009, 10, 3069—3073
[5]	Sobiech M., ski P., Maciejewska D., Talanta, 2015, 156, 556—567
[6]	Yu S. C., Yu F., Li Y. P., Liu L., Zhang H. Q., Qu L. B., Food Control, 2015, 60, 500—504
[7]	Jamwal S., Dharela R., Gupta R., Ahn J. H., Chem. Eng. Res. Des., 2015, 95, 159—164
[8]	Han H., Zhou Y., Li S., Wang Y., Kong X. Z., ACS Appl. Mater. Interfaces, 2016, 8, 25714—25724
[9]	Mouad A. M., Taupin D., Lehr L., J. Mol. Catal. B: Enzym. ,2015, 126, 64—68
[10]	Choi J. M., Han S. S., Kim H. S., Biotechnol. Adv., 2015, 33, 1443—1454
[11]	B. M., Biochem, Eng. J., 2015, 93, 73—83
[12]	Kim S. H., Kim S., Park S., Kim H. K., J. Mol. Catal. B, Enzym.,2013, 85, 10—16
[13]	Bayramoglu G., Akbulut A., Ozalp V. C., Arica M. Y., Chem. Eng. Res. Des., 2015, 95, 12—21
[14]	Hu Y., Yang J., Tang S. S., Chu X. M., Zou B., Huang H., Chem. J. Chinese Universities, 2013, 34(5), 1195—1202
	(胡燚, 杨姣, 唐苏苏, 初旭明, 邹彬, 黄和. 高等学校化学学报, 2013, 34(5), 1195—1202)
[15]	Monsan P., J. Mol. Catal., 1978, 3, 371—384
[16]	Yang G., Wu J., Xu G., Yang L., Colloids. Surf. B, 2010, 78, 351—356
[17]	Bornscheuer U. T., Angew Chem. Int. Ed., 2003, 42, 3336—3337
[18]	Han H., Li S. S., Jiang X. B., Kong X. Z., RSC Adv., 2014, 4, 33520—33529
[19]	Li S. S., Jiang X. B., Kong X. Z., Chem. J. Chinese Universities, 2013, 34(4), 992—999
	(李树生, 姜绪宝, 孔祥正. 高等学校化学学报, 2013, 34(4), 992—999)
[20]	Li S. S., Kong X. Z., Jiang X. B., Zhu X., Chinese Chem. Lett., 2013, 24, 287—290
[21]	Ye P., Xu Z. K., Wu J., Innocent C., Seta P., Macromolecules,2006, 39, 1041—1045
[22]	Migneault I., Dartiguenave C., Bertrand M. J., Waldron K. C., Biotechniques,2004, 37, 790—802
[23]	Li S.Y., Zhu X. L., Kong X. Z., Jiang X. B.Acta Polym. Sin., 2016, (3), 391—398
	(李世彦, 朱晓丽, 孔祥正, 姜绪宝. 高分子学报, 2016, (3), 391—398)
[24]	Chen C. S., Fujimoto Y., Girdaukas G., Sih C. J., J. Am. Chem. Soc. ,1982, 104, 1294—1299
[25]	Pangon A., Dillon G. P., Runt J., Polymer,2014, 55, 1837—1844
[26]	Zhou Q., Cao L., Li Q., Yao Y. L., Ouyang Z. F., Su Z. Q., Chen X. N., J. Appl. Polym. Sci., 2012, 125, 3695—3701
[27]	Jiang X. B., Li X. M., Zhu X. L., Kong X. Z., Ind. Eng. Chem. Res., 2016, 55, 11528—11535
[28]	Betancor L., López-Gallego F., Hidalgo A., Alonso-Morales N., Mateo G. D. C., Fernández-Lafuente R., Guisán J. M., Enzym. Microb. Technol., 2006, 39, 877—882
[29]	Klein M. P., Nunes M. R., Rodrigues R. C., Benvenutti E. V., Costa T. M., Hertz P. F., Ninow J. L., Biomacromolecules,2012, 13, 2456—2464
[30]	Meng X., Xu G., Zhou Q. L., Wu J. P., Yang L. R., J. Mol. Catal. B: Enzym.,2013, 89, 86—92
[31]	Feng X., Patterson D. A., Balaban M., Emanuelsson E. A. C., Colloids. Surf. B: Biointerfaces, 2013, 102, 526—533
[32]	Bellusci M., Francolini I., Martinelli A., Ilario L. D., Piozzi A., Biomacromolecules,2012, 13, 805—813
[33]	Kojima Y., Kobayashi M., Shimizu S., J Biosci. Bioeng., 2003, 96, 242—249
[34]	Lima L. N., Oliveira G. C., Rojas M. J., Castro H. F., Da Rós P. C. M., Mendes A. A., Giordano R. L. C., Tardioli P. W., J. Ind. Microbiol. Biotechnol., 2015, 42, 523—535
[35]	Salis A., Bhattacharyya M. S., Monduzzi M., Solinas V., J. Mol. Catal. B: Enzym.,2009, 57, 262—269
[36]	De Lima. L. N., Aragon C. C., Mateo C., Palomo J. M., Giordano R. L. C., Tardioli P. W., Guisan J. M., Fernandez-Lorente G., Process Biochem., 2013, 48, 118—123
[37]	El-Aassar M. R., J. Mol. Catal. B: Enzym., 2013, 85, 140—148
[38]	Wang C. T., Liang J. R., Yin S., Zhou D., Sun B. G., Zhao J. X., J. Chin. Inst. Food Sci. Technol.,2016, 16, 78—86
	(王成涛, 梁婧如, 尹胜, 周娣, 孙宝国, 赵吉兴. 中国食品学报, 2016, 16, 78—86)
[39]	Chen Y. Z., Yang C. T., Ching C. B., Xu R., Langmuir,2008, 24, 8877—8884
[40]	Machado A. C. O., Da Silva A. A . T., Borges C. P., Simas A. B. C., Freire D. M. G., J. Mol. Catal. B: Enzym. ,2011, 69, 42—46
	(Ed.:D, Z)

Concentration of GA/(mol·L^-1)	Immobilized PFL/(mg·g^-1)	Immobilized PFL ratio(%)	Activity/(U·mg^-1)	Activity retention(%)
0.0334	72.2	46.5	125.3	25.4
0.1002	92.3	61.0	242.5	49.2
0.1669	95.2	62.0	375.0	76.0
0.3339	93.8	60.1	274.6	55.7
0.5008	71.4	50.0	142.2	28.8

Concentration of GA/(mol·L^-1)	Immobilized PFL/(mg·g^-1)	Immobilized PFL ratio(%)	Activity/(U·mg^-1)	Activity retention(%)
0.0334	72.2	46.5	125.3	25.4
0.1002	92.3	61.0	242.5	49.2
0.1669	95.2	62.0	375.0	76.0
0.3339	93.8	60.1	274.6	55.7
0.5008	71.4	50.0	142.2	28.8

PFL concentration/(mg·mL^-1)	Immobilized PFL/(mg·g^-1)	Immobilized PFL ratio(%)	Activity/(U·mg^-1)	Activity retention(%)
0.64	38.4	100.0	203.8	41.3
1.28	67.6	88.0	328.8	66.7
1.71	83.7	81.7	362.5	73.5
2.56	95.2	62.0	375.0	76.0
3.36	95.0	45.5	372.5	75.6

PFL concentration/(mg·mL^-1)	Immobilized PFL/(mg·g^-1)	Immobilized PFL ratio(%)	Activity/(U·mg^-1)	Activity retention(%)
0.64	38.4	100.0	203.8	41.3
1.28	67.6	88.0	328.8	66.7
1.71	83.7	81.7	362.5	73.5
2.56	95.2	62.0	375.0	76.0
3.36	95.0	45.5	372.5	75.6

Reaction time/h	Free enzyme
Reaction time/h	c(%)	ee_p(%)	E	c(%)	ee_p(%)	E
2	6.4	>99.5	>200	9.9	>99.5	>200
4	14.5	>99.5	>200	19.0	>99.5	>200
8	20.6	>99.5	>200	33.3	>99.5	>200
10	25.1	94.1	44.6	38.0	98.3	>200
12	28.1	93.6	43.4	43.3	97.5	181