A facile technique to prepare cross-linked enzyme aggregates using p-benzoquinone as cross-linking agent

Anming Wang; Fangkai Zhang; Feifei Chen; Meizhang Wang; Haifeng Li; Zhaowu Zeng; Tian Xie; Zhenming Chen

Korean Journal of Chemical Engineering, Vol.28, No.4, 1090-1095, April, 2011

DOI10.1007/s11814-010-0476-0 Export Citation

A facile technique to prepare cross-linked enzyme aggregates using p-benzoquinone as cross-linking agent

Anming Wang^{1, †}, Fangkai Zhang^{1, 2}, Feifei Chen^{1, 3}, Meizhang Wang¹, Haifeng Li¹, Zhaowu Zeng¹, Tian Xie¹, and Zhenming Chen¹

¹Research Center for Biomedicine and Health, Hangzhou Normal University, Hangzhou 310012, P. R. China
²College of Biological and Environmental Sciences, Hangzhou Normal University, Hangzhou 310012, P. R. China
³College of Materials, Chemistry and Chemical Engineering, Hangzhou Normal University, Hangzhou 310012, P. R. China

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To obtain robust and thermo-stable enzyme aggregates, p-benzoquinone was used as cross-linker and bovine serum albumin (BSA) as crowding macromolecules to prepare cross-linked enzyme aggregates (CLEAs) of lipase. Effects of cross-linking time and cross-linker content on the activity, thermal stability and characteristics of enzyme aggregates were examined carefully. It was observed that when the content of p-benzoquinone was 5 mM and amount of BSA was 125% of that of lipase (w/w), the specific activity of cross-linked co-aggregates of lipase and BSA was 79.8 U mg.1, 2.44-fold of that of cross-linked enzyme aggregates of lipase without BSA. Moreover, after heat treatment for 96 h at 50 oC, the CLEAs prepared with this facile routine kept 75.18% of their initial activity, 5.01-fold more than that of the just CLEAs using glutaraldehyde. Furthermore, BSA macromolecules in lipase CLEAs enhanced the catalytic efficiency of free and just lipase CLEAs without BSA by 1.45 and 2.83 times, respectively. The proposed crosslinking technique would rank among the potential strategies for efficiently preparing robust and thermo-stable enzyme aggregates.

Keywords:Cross-linked Enzyme Aggregates;p-Benzoquinine;Macromolecule Crowding;Lipase;Bovine Serum Albumin

[References]

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[Referenced By]

[1] Cao L and Schmid RD, Carrier-bound immobilized enzymes: Principles, application and design, WILEY-VCH Verlag GmbH & Co. Weinheim (2005)

[2] Jiang YY, Guo C, Xia HS, Mahmood I, Liu CZ, Liu HZ, J. Mol. Catal., B Enzym., 58, 103 (2009)

[3] Zhou C, Wang A, Du Z, Zhu S, Shen S, Korean J. Chem. Eng., 26(4), 1065 (2009)

[4] Blanco RM, Terreros P, Munoz N, Serra E, J. Mol. Catal., B Enzym., 47, 13 (2007)

[5] Roessl U, Nahalka J, Nidetzky B, Biotechnol. Lett., 32(3), 341 (2010)

[6] Cao LQ, van Rantwijk F, Sheldon RA, Org. Lett., 2, 1361 (2000)

[7] Sheldon RA, Adv. Synth. Catal., 349, 1289 (2007)

[8] Xie T, Wang A, Huang L, Li H, Chen Z, Wang Q, Yin X, Afr. J. Biotechnol., 8, 4724 (2009)

[9] Kaul P, Stolz A, Banerjee UC, Adv. Synth. Catal., 349, 2167 (2007)

[10] Mateo C, Palomo JM, van Langen LM, van Rantwijk F, Sheldon RA, Biotechnol. Bioeng., 86(3), 273 (2004)

[11] Wilson L, Fernandez-Lorente G, Fernandez-Lafuente R, Illanes A, Guisan JM, Palomo JM, Enzyme Microb. Technol., 39(4), 750 (2006)

[12] Gupta P, Dutt K, Misra S, Raghuwanshi S, Saxena RK, Bioresour. Technol., 100, 4074 (2009)

[13] Lee J, Lee D, Oh E, Kim J, Kim YP, Jin S, Kim HS, Hwang Y, Kwak JH, Park JG, Shin CH, Hyeon T, Angew. Chem. Int. Ed. Engl., 44, 7427 (2005)

[14] Sheldon RA, Biochem. Soc. Trans., 35, 1583 (2007)

[15] Lopez-Gallego F, Betancor L, Hidalgo A, Alonso N, Fernandez-Lafuente R, Guisan JM, Biomacromolecules, 6(4), 1839 (2005)

[16] Migneault I, Dartiguenave C, Bertrand MJ, Waldron KC, BioTechniques., 37, 790 (2004)

[17] Wang A, Wang H, Zhu S, Zhou C, Du Z, Shen S, Bioprocess Biosystems Eng., 31, 509 (2008)

[18] Ihara F, Kageyama Y, Hirata M, Nihira T, Yamada Y, J. Biol. Chem., 266, 18135 (1991)

[19] Wang AM, Liu MQ, Wang H, Zhou C, Du ZQ, Zhu SM, Shen SB, Ouyang PK, J. Biosci. Bioeng., 106(3), 286 (2008)

[20] Jiang M, Guo ZH, J. Am. Chem. Soc., 129(4), 730 (2007)

[21] Dalal S, Kapoor M, Gupta MN, J. Mol. Catal., B Enzym., 44, 128 (2007)

[22] Wang AM, Zhou C, Du ZQ, Liu MQ, Zhu SM, Shen SB, Ouyang PK, J. Biosci. Bioeng., 107(3), 219 (2009)

[23] Martinek K, Mozhaev VV, Pure Appl. Chem., 63, 1533 (1991)

[24] Ellis RJ, Minton AP, Nature., 425, 27 (2003)

[25] Wang XQ, Lu DN, Austin R, Agarwal A, Mueller LJ, Liu Z, Wu JZ, Feng PY, Langmuir, 23(10), 5735 (2007)

[26] Du F, Zhou Z, Mo ZY, Shi JZ, Chen J, Liang Y, J. Mol. Biol., 364, 469 (2006)

[27] Moran-Zorzano MT, Viale A, Munoz F, Alonso-Casajus N, Eydallin G, Zugasti B, Baroja-Fernandez E, Pozueta-Romero J, Febs Letters., 581, 1035 (2007)

[28] Shah S, Sharma A, Gupta MN, Anal. Biochem., 351, 207 (2006)

[29] Torchilin VP, Maksimenko AV, Smirnov VN, Berezin IV, Klibanov AM, Martinek K, Biochim. Biophys. Acta., 522, 277 (1978)