Synergistic Effect of Two Type Cellulase Immobilized on Chitosan Microparticle as Biocatalyst for Coconut Husk Hydrolysis

Afan Hamzah; Lidya Lorenta Sitompul; Irma Nurhanifah Fenda Putri; Soeprijanto Soeprijanto; Arief Widjaja

doi:10.22146/ijc.39714

Synergistic Effect of Two Type Cellulase Immobilized on Chitosan Microparticle as Biocatalyst for Coconut Husk Hydrolysis

https://doi.org/10.22146/ijc.39714

Afan Hamzah⁽¹⁾, Lidya Lorenta Sitompul⁽²⁾, Irma Nurhanifah Fenda Putri⁽³⁾, Soeprijanto Soeprijanto⁽⁴⁾, Arief Widjaja^(5*)

(1) Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember
(2) Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember
(3) Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember
(4) Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember
(5) Department of Chemical Engineering, Institut Teknologi Sepuluh Nopember
(*) Corresponding Author

Abstract

The effectivity of employing two types of cellulases from Aspergillus niger and Trichoderma resei covalently immobilized on chitosan microparticle was investigated. Reducing sugar from CMC yielded by immobilized cellulase from T. resei alone and A. niger alone was 0.316 g/L and 0.244 g/L, respectively. Simultaneous use of both cellulases shows a significant increase of reducing sugar produced to 1.020 g/L. The effective combination of this two types of cellulases also occurred when coconut husk was used as substrate. A very high enzyme coupling of 92.06% compared to free enzyme was obtained in the immobilization. Addition of GDA not only increased enzyme coupling to 100% but also improved sugar produced. Immobilized cellulase was successfully maintained its activity until 5 cycles

Keywords

chitosan; cellulase; immobilization; microparticle; synergy; coconut husk

Full Text:

Full Text PDF

References

[1] Li, C., Yoshimoto, M., Fukunaga, K., and Nakao, K., 2007, Characterization and immobilization of liposome-bound cellulase for hydrolysis of insoluble cellulose, Bioresour. Technol., 98 (7), 1366–1372.

[2] Ahamed, A., and Vermette, P., 2008, Enhanced enzyme production from mixed cultures of Trichoderma reesei RUT-C30 and Aspergillus niger LMA grown as fed batch in a stirred tank bioreactor, Biochem. Eng. J., 42 (1), 41–46.

[3] Martins, L.F., Kolling, D., Camassola, M., Dillon, A.J.P., and Ramos, L.P., 2008, Comparison of Penicillium echinulatum and Trichoderma reesei cellulases in relation to their activity against various cellulosic substrates, Bioresour. Technol., 99 (5), 1417–1424.

[4] Ungurean, M., Paul, C., and Peter, F., 2013, Cellulase immobilized by sol-gel entrapment for efficient hydrolysis of cellulose, Bioprocess Biosyst. Eng., 36 (10), 1327–1338.

[5] Stockton, B.C., Mitchell, D.J., Grohmann, K., and Himmel, M.E., 1991, Optimum β-D-glucosidase supplementation of cellulase for efficient conversion of cellulose to glucose, Biotechnol. Lett., 13 (1), 57–62.

[6] Juhász, T., Kozma, K., Szengyel, Z., and Réczey, K., 2003, Production of β-glucosidase in mixed culture of Aspergillus niger BKMF 1305 and Trichoderma reesei RUT C30, Food Technol. Biotechnol., 41 (1), 49–53.

[7] Liu, J., and Cao, X., 2013, Biodegradation of microcrystalline cellulose in pH-pH recyclable aqueous two-phase systems with water-soluble immobilized cellulase, Biochem. Eng. J., 79, 136–143.

[8] Cheng, C., and Chang, K.C., 2013, Development of immobilized cellulase through functionalized gold nano-particles for glucose production by continuous hydrolysis of waste bamboo chopsticks, Enzyme Microb. Technol., 53 (6-7), 444–451.

[9] Ahmad, R., and Sardar, M., 2014, Immobilization of cellulase on TiO₂ nanoparticles by physical and covalent methods: A comparative study, Indian J. Biochem. Biophys., 51 (4), 314–320.

[10] Chakrabarti, A.C., and Storey, K.B., 1988, Immobilization of cellulase using polyurethane foam, Appl. Biochem. Biotechnol., 19 (2), 189–207.

[11] Nguyen, L.T., Neo, K.R.S., and Yang, K.L., 2015, Continuous hydrolysis of carboxymethyl cellulose with cellulase aggregates trapped inside membranes, Enzyme Microb. Technol., 78, 34–39.

[12] Cherian, E., Dharmendirakumar, M., and Baskar, G., 2015, Immobilization of cellulase onto MnO₂ nanoparticles for bioethanol production by enhanced hydrolysis of agricultural waste, Chin. J. Catal., 36 (8), 1223–1229.

[13] Liao, H., Chen, D., Yuan, L., Zheng, M., Zhu, Y., and Liu, X., 2010, Immobilized cellulase by polyvinyl alcohol/Fe₂O₃ magnetic nanoparticle to degrade microcrystalline cellulose, Carbohydr. Polym., 82 (3), 600–604.

[14] El-Ghaffar, M.A.A., and Hashem, M.S., 2010, Chitosan and its amino acids condensation adducts as reactive natural polymer supports for cellulase immobilization, Carbohydr. Polym., 81 (3), 507–516.

[15] Biró, E., Németh, A.S., Sisak, C., Feczkó, T., and Gyenis, J., 2008, Preparation of chitosan particles suitable for enzyme immobilization, J. Biochem. Biophys. Methods, 70 (6), 1240–1246.

[16] Budriene, S., Gorochovceva, N., Romaskevic, T., Yugova, L.V., Miezeliene, A., Dienys, G., and Zubriene, A., 2005, β-Galactosidase from Penicillium canescens. Properties and immobilization, Cent. Eur. J. Chem., 3 (1), 95–105.

[17] Zheng, Y.G., Chen, X.L., and Wang, Z., 2005, Microbial biomass production from rice straw hydrolysate in airlift bioreactors., J. Biotechnol., 118 (4), 413–420.

[18] Zang, L., Qiu, J., Wu, X., Zhang, W., Sakai, E., and Wei, Y., 2014, Preparation of magnetic chitosan nanoparticles as support for cellulase immobilization, Ind. Eng. Chem. Res., 53 (9), 3448–3454.

[19] Mao, X., Guo, G., Huang, J., Du, Z., Huang, Z., Ma, L., Li, P., and Gu, L., 2006, A novel method to prepare chitosan powder and its application in cellulase immobilization, J. Chem. Technol. Biotechnol., 81, 189–195.

[20] Sojitra, U.V., Nadar, S.S., and Rathod, V.K., 2017, Immobilization of pectinase onto chitosan magnetic nanoparticles by macromolecular cross-linker, Carbohydr. Polym., 157, 677–685.

[21] Alftrén, J., and Hobley, T.J., 2014, Immobilization of cellulase mixtures on magnetic particles for hydrolysis of lignocellulose and ease of recycling, Biomass Bioenergy, 65, 72–78.

[22] Zheng, P., Wang, J., Lu, C., Xu, Y., and Sun, Z., 2013, Immobilized β-glucosidase on magnetic chitosan microspheres for hydrolysis of straw cellulose, Process Biochem., 48 (4), 683–687.

[23] Sanchez-Ramirez, J., Martinez-Hernandez, J.L., Segura-Ceniceros, P., Lopez, G., Saade, H., Medina-Morales, M.A., Ramos-Gonzalez, R., Aguilar, C.N., and Ilyina, A., 2017, Cellulases immobilization on chitosan-coated magnetic nanoparticles: Application for Agave atrovirens lignocellulosic biomass hydrolysis, Bioprocess Biosyst. Eng., 40 (1), 9–22.

[24] Sangian, H.F., Kristian, J., Rahma, S., Agnesty, S.Y., Gunawan, S., and Widjaja, A., 2015, Comparative Study of the preparation of reducing sugars hydrolyzed from high-lignin lignocellulose pretreated with ionic liquid, alkaline solution and their combination, J. Eng. Technol. Sci., 47 (2), 137–148.

[25] Safarik, I., Horska, K., Pospiskova, K., and Safarikova, M., 2012, One-step preparation of magnetically responsive materials from non-magnetic powders, Powder Technol., 229, 285–289.

[26] Anwar, N., Widjaja, A., and Winardi, S., 2012, Study of the enzymatic hydrolysis of alkaline-pretreated rice straw using cellulase of various sources and compositions, Int. Rev. Biophys. Chem., 3 (1), 272–278.

[27] Bradford, M.M., 1976, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem., 72 (1-2), 248–254.

[28] Miller, G.L., 1959, Use of Dinitrosalicylic acid reagent for determination of reducing sugar, Anal. Chem., 31 (3), 426–428.

[29] Wang, J., Zhao, G., Li, Y., Liu, X., and Hou, P., 2013, Reversible immobilization of glucoamylase onto magnetic chitosan nanocarriers, Appl. Microbiol. Biotechnol., 97 (2), 681–692.

[30] Al-Remawi, M.M.A., 2012, Properties of chitosan nanoparticles formed using sulfate anions as crosslinking bridges, Am. J. Appl. Sci., 9 (7), 1091–1100.

[31] Abdel-Naby, M.A., Ismail, A.M.S., Abdel-Fattah, A.M., and Abdel-Fattah, A.F., 1999, Preparation and some properties of immobilized Penicillium funiculosum 258 dextranase, Process Biochem., 34 (4), 391–398.

[32] Chang, M.Y., and Juang, R.S., 2005, Activities, stabilities, and reaction kinetics of three free and chitosan-clay composite immobilized enzymes, Enzyme Microb. Technol., 36 (1), 75–82.

[33] Çetinus, Ş.A., and Öztop, H.N., 2003, Immobilization of catalase into chemically crosslinked chitosan beads, Enzyme Microb. Technol., 32 (7), 889–894.

[34] Mistry, B.D., 2009, A Handbook of Spectroscopic Data: UV, IR, PMR, CNMR and Mass Spectroscopy, Oxford Book Company.

[35] Xu, J., Huo, S., Yuan, Z., Zhang, Y., Xu, H., Guo, Y., Liang, C., and Zhuang, X., 2011, Characterization of direct cellulase immobilization with superparamagnetic nanoparticles, Biocatal. Biotransform., 29 (2-3), 71–76.

[36] Jordan, J., Kumar, C.S.S.R., and Theegala, C., 2011, Preparation and characterization of cellulase-bound magnetite nanoparticles, J. Mol. Catal. B: Enzym., 68 (2), 139–146.

[37] Sangian, H.F., Ranggina, D., Ginting, G.M., Purba, A.A., Gunawan, S., and Widjaja, A., 2015, Study of the preparation of sugar from high-lignin lignocellulose applying subcritical water and enzymatic hydrolysis: Synthesis and consumable cost evaluation, St. Cerc. St. CICBIA, 16, 13–27.

[38] Chen, H., Zhang, Q., Dang, Y., and Shu, G., 2013, The Effect of glutaraldehyde cross-linking on the enzyme activity of immobilized β-galactosidase on chitosan bead, Adv. J. Food Sci. Technol., 5 (7), 932–935.

[39] Vieira, D.C., Lima, L.N., Mendes, A.A., Adriano, W.S., Giordano, R.C., Giordano, R.L.C., and Tardioli, P.W., 2013, Hydrolysis of lactose in whole milk catalyzed by β-galactosidase from Kluyveromyces fragilis immobilized on chitosan-based matrix, Biochem. Eng. J., 81, 54–64.

DOI: https://doi.org/10.22146/ijc.39714