Run Fang, Yin Lin, Xian-Su Cheng

Abstract: Synthesis of two lignin derivatives, lignophenol and lignin-aminophenol, were presented in this article. The chemical structure and the func-tional groups of lignin derivatives were charac-terized through FT-IR analysis. The immobiliza-tion of three proteases (papain, trypsin and pepsin) on lignin and lignin derivatives was carried out using adsorption technique. The influence of contact time and pH on the enzyme adsorption by different adsorbents was inves-tigated. Furthermore, enzyme activity recovery was also evaluated. Results showed that lignin and lignin derivatives could adsorb proteases effectively and the adsorption capacity of lingo- phenol and lignin-aminophenol was higher than that of pure lignin. Meanwhile, the activity re-covery of papain and pepsin immobilized on lignin derivatives was very high. This pheno- menon suggested that there is a supramole- cular interaction between enzymes and lignin derivatives which do not inhibit enzyme activity. Therefore, lignophenol and lignin-aminophenol are both promising adsorbents for enzyme im-mobilization under acid and neutral conditions.

Keywords: Lignin Derivative; Papain; Trypsin; Pepsin; Adsorption

Cite this paper: nullFang, R. , Lin, Y. and Cheng, X. (2009) Preparation of lignin derivatives and their application as protease adsorbents. Natural Science, 1, 185-190. doi: 10.4236/ns.2009.13024.

References

[1]   F. Hildebrand and S. Lutz, (2006) Immobilization of alcohol dehydrogenase from Lactobacillus brevis and its application in a plugflow reactor. Tetrahedron Asymme-try, 17, 3219-3225.

[2]   C. Mateo, O. Abian, L. R. Fernandez, and J. M. Guisan, (2000) Increase in conformational stability of enzymes immobilized on epoxy-activated supports by favoring additional multipoint covalent attachment. Enzyme Mi-crob. Technol., 26, 509–515.

[3]   R. Schoevaart, M. W. Wolbers, M. Golubovic et al., (2004) Preparation, optimization and structures of cross-linked enzyme aggregates (CLEAs). Biotechnol. Bioeng., 87, 754-762.

[4]   R. A. Sheldon, R. Schoevaart, and L. M. Van Langen, (2005) Cross-linked enzyme aggregates (CLEAs): A novel and versatile method for enzyme immobilization. Biocatal. Biotransform., 23, 141-147.

[5]   A. C. Pierre, (2004)The sol-gel encapsulation of enzymes. Biocatal. Biotransform., 22, 145-170.

[6]   L. Veum, U. Hanefeld, and A. Pierre, (2004) The first encapsulation of hydroxynitrile lyase from Hevea brasil-iensis in a sol-gel matrix. Tetrahedron, 60, 10419-10425.

[7]   X. S. Cheng and X. L. Liu, (2006) Separation of lignin from cornstalks residue by enzymatic hydrolysis and its properties. Morden Chemical Industry, Supplement, 26, 99-102.

[8]   X. H. Zhang, X. S. Cheng, and J. Tang. (2006) Adsorp-tion of bromelain with HBS lignin and its derivatives. The Chinese Journal of Process Engineering, 6, 87-90.

[9]   Y. Q. Jin and X. S. Cheng, (2006) Study on the adsorp-tion of endotoxin by HBS lignin and its derivatives. Journal of Xi’an University of Engineering Science and Technology, 20, 723-726.

[10]   M. J. Li and X. S. Cheng, (2004) Research on the ad-sorption of endotoxin by lignophenol. Journal of Cellu-lose Science and Technology, 12, 1-6.

[11]   Y. X. Luo, (2000) Studies on the determination of enzy-matic active for papain. Chinese Pharmaceutical Journal, 35, 556-558.

[12]   R. Fang, X. S. Cheng., J. Fu., and Z. B. Zheng, (2009) Research on the graft copolymerization of EH-lignin with acrylamide. Natural Science, 1, 17-22.

[13]   S. Solis, J. Paniagua, J. C. Mart?nez, and M. Asomoza, (2006) Immobilization of papain on mesoporous silica: pH effect. J. Sol-Gel Sci. Technol., 37, 125-127.

[14]   L. Qiao and Y. Liu, (2008) A nanoporous reactor for effi-cient proteolysis. Chem. Eur. J., 14, 151-157.

[15]   M. Casas1 and J. Mi?ones1, (1992) Effect of polymer-ized silicic acid on mixed lipid-protein monolayers used as cell-membrane models II pepsin-sphingomyelin films. Colloid. Polym. Sci., 270, 485-491.

[16]   S. Dumitriu, P. Magny, D. Montane, P. F. Vidal, and E. Chornet, (1994) Polyionic hydrogels obtained by com-plexation between xanthan and chitosan: their properties as supports for enzyme immobilization. J. Bioact. Com-pat. Polym., 9, 184-209.

[17]   N. Dizge, C. Aydiner, E. Demirbas, M. Kobya, and S. Kara, (2008) Adsorption of reactive dyes from aqueous solutions by fly ash: Kinetic and equilibrium studies. J. Hazard. Mater., 150, 737–746.

[18]   B. Al-Duri and Y. P. Yong, (2000) Lipase immobilisation: An equilibrium study of lipases immobilised on hydro-phobic and hydrophilic/hydrophobic supports. Biochem. Eng. J., 4, 207–215.

[19]   P. Kauper, (2004) From production to product Part 1: Solution states of alkaline bagasse lignin solution. Indus-trial Crops and Products, 20, 151-157.