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Three-phase partitioning and immobilization of Bacillus methylotrophicus Y37 cellulase on organo-bentonite and its kinetic and thermodynamic properties

Published online by Cambridge University Press:  25 June 2020

Yonca Avci Duman ,
A. Uğur Kaya  and
Çiğdem Yağci
Yonca Avci Duman*
Affiliation:
Faculty of Arts and Sciences, Department of Chemistry, Kocaeli University, Umuttepe Campus, 41380İzmit-Kocaeli, Turkey
A. Uğur Kaya
Affiliation:
Faculty of Arts and Sciences, Department of Physics, Kocaeli University, Umuttepe Campus, 41380İzmit-Kocaeli, Turkey
Çiğdem Yağci
Affiliation:
Faculty of Education, Department of Science Education, Kocaeli University, Umuttepe Campus, 41380İzmit-Kocaeli, Turkey
*
*E-mail: yavci@kocaeli.edu.tr
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Abstract

In this study, for the first time Bacillus methylotrophicus Y37 cellulase was purified and recovered in a single step by three-phase partitioning (TPP). The optimal purification parameters for TPP were 40% ammonium sulfate saturation (m/v) with a 1.0:1.0 (v/v) ratio of crude extract:t-butanol, which gave 5.8-fold purification with 155% recovery of cellulase. Non-covalent immobilization of the partitioned cellulase was performed using bentonite as a support material. The activity observed in the 20th experiment was 100%. The optimal pH values and temperatures determined for the free enzyme and the immobilized enzyme were 5.0 and 6.0 and 45°C and 50°C, respectively. The Arrhenius activation energy (Ea) of the immobilized enzyme was lower than that of the free enzyme, whereas the Michaelis–Menten constant (Km) and maximum velocity (Vm) of the immobilized enzyme increased. The turnover number (kcat) and the catalytic performance (kcat/Km) demonstrated the improved catalytic properties of the immobilized enzyme compared to the free enzyme. Immobilization of cellulase is thermodynamically preferred.

Keywords

bentonitecellulaseimmobilizationkineticthermodynamicsthree-phase partitioning
Type
Article
Information
Clay Minerals , Volume 55 , Issue 2 , June 2020 , pp. 120 - 131
Copyright
Copyright © The Mineralogical Society of Great Britain and Ireland, 2020

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Footnotes

Associate Editor: Miroslav Pospíšil

References

Ahmad, R. & Khare, S.K. (2018) Immobilization of Aspergillus niger cellulase on multiwall carbon nanotubes for cellulose hydrolysis. Bioresource Technology, 252, 72–75.CrossRefGoogle ScholarPubMed
Alemdar, A. (2001) The Effect of Organic and Inorganic Additives on the Rheological, Viscoelastic and Colloidal Properties of Bentonite and Montmorillonite Dispersions. PhD thesis, ITU Institute of Science and Technology, Istanbul, Turkey, thesis no. 104267.Google Scholar
Andjelkovic, U., Nikolic, A.M., Jovic, N.J., Bankovic, P., Bajt, T., Mojovic, Z., Vujcic, Z. & Jovanovic, D. (2015) Efficient stabilization of Saccharomyces cerevisiae external invertase by immobilisation on modified beidellite nanoclays. Food Chemistry, 168, 262–269.CrossRefGoogle ScholarPubMed
Aytas, S., Yurtlu, M. & Donat, R. (2009) Adsorption characteristic of U(VI) ion onto thermally activated bentonite. Journal of Hazardous Materials, 172, 667–674.CrossRefGoogle ScholarPubMed
Bilal, M., Iqbal, H.M.N., Guoa, S., Hua, H., Wanga, W. & Zhanga, X. (2018) State-of-the-art protein engineering approaches using biological macromolecules: a review from immobilization to implementation view point. International Journal of Biological Macromolecules, 108, 893–901.CrossRefGoogle ScholarPubMed
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Analytical Biochemistry, 72, 248–254.CrossRefGoogle ScholarPubMed
Chen, D., Chen, J., Luan, X., Ji, H. & Xia, Z. (2011) Characterization of anion–cationic surfactants modified montmorillonite and its application for the removal of Methyl Orange. Chemical Engineering Journal, 171, 1150–1158.CrossRefGoogle Scholar
Dennison, C. & Lovrein, R. (1997) Three phase partitioning: concentration and purification of proteins. Protein Expression and Purification, 11, 149–161.CrossRefGoogle ScholarPubMed
Dong, H., Li, J., Li, Y., Hu, L. & Luo, D. (2012) Improvement of catalytic activity and stability of lipase by immobilization on organobentonite. Chemical Engineering Journal, 181, 590–596.CrossRefGoogle Scholar
Dong, H., Li, Y., Sheng, G. & Hu, L. (2013) The study on effective immobilization of lipase on functionalized bentonites and their properties. Journal of Molecular Catalysis B: Enzymatic, 95, 9–15.CrossRefGoogle Scholar
Duman, A.Y. & Kaya, E. (2013a) Purification, recovery, and characterization of chick pea (Cicer arietinum) β-galactosidase in single step by three phase partitioning as a rapid and easy technique. Protein Expression and Purification, 91, 155–160.CrossRefGoogle Scholar
Duman, A.Y. & Kaya, E. (2013b) Three-phase partitioning as a rapid and easy method for the purification and recovery of catalase from sweet potato tubers (Solanum tuberosum). Applied Biochemistry and Biotechnology, 170, 1119–1126.CrossRefGoogle Scholar
Duman, Y.A., Kazan, D., Denizci, A.A. & Erarslan, A. (2014) Water miscible mono alcohols’ effect on the proteolytic performance of Bacillus clausii serine alkaline protease. Applied Biochemistry and Biotechnology, 172, 469–486.CrossRefGoogle ScholarPubMed
Duman, Y., Yüzügüllü, Y.K., Sertel, A. & Polat, F. (2016) Production, purification and characterization of a thermo-alkali stable and metal-tolerant carboxymethylcellulase from newly isolated Bacillus methylotrophicus Y37. Turkish Journal of Chemistry, 40(5), 802–815.CrossRefGoogle Scholar
Dutta, A. & Singh, N. (2015) Surfactant-modified bentonite clays: preparation, characterization and atrazine removal. Environmental Science and Pollution Research International, 22, 3876–3885.CrossRefGoogle ScholarPubMed
Favre, H. & Lagaly, G. (1991) Organo-bentonites with quaternary alkylammonium ions. Clay Minerals, 26, 19–32.CrossRefGoogle Scholar
Gao, J., Lu, C.-L., Wang, Y., Wang, S.-S., Shen, J.-J., Zhang, J.-X. & Zhang, Y-W. (2018) Rapid immobilization of cellulase onto graphene oxide with a hydrophobic spacer. Catalysts, 8, 180–188.CrossRefGoogle Scholar
Ghiaci, M., Aghaei, H., Soleimanian, S. & Sedaghat, M.E. (2009a) Enzyme immobilization: part 1. Modified bentonite as a new and efficient support for immobilization of Candida rugosa lipase. Applied Clay Science, 43, 289–295.CrossRefGoogle Scholar
Ghiaci, M., Aghaei, H., Soleimanian-Zad, S., Sedaghat, M.E. (2009b) Enzyme immobilization: part 2. Immobilization of alkaline phosphatase on Na-bentonite and modified bentonite. Applied Clay Science, 43, 308–316.CrossRefGoogle Scholar
Ghiaci, M., Kalbasi, R.J., Khani, H., Abbaspur, A. & Shariatmadari, H. (2004) Free-energy of adsorption of a cationic of adsorption layer by X-ray spectroscopy. Journal of Chemical Thermodynamics, 36, 707–713.CrossRefGoogle Scholar
Grim, R.E. (1988) The history of the development of clay mineralogy. Clay Minerals, 36, 97–101.CrossRefGoogle Scholar
Hasan, A., Shah, A.A. & Hameed, A. (2006) Industrial applications of microbial lipases. Enzyme and Microbial Technology, 39(2), 235–251.CrossRefGoogle Scholar
Homaei, A. (2015) Enzyme immobilization and its application in the food industry. Advances in Food Biotechnology, 9, 145–164.CrossRefGoogle Scholar
Ingle, A.P., Rathod, J., Pandit, R., Silva, S. & Rai, M. (2017) Comparative evaluation of free and immobilized cellulose for enzymatic hydrolysis of lignocellulosic biomass for sustainable bioethanol production. Cellulose, 24, 5529–5540.CrossRefGoogle Scholar
Jović-Jovičić, N., Milutinović-Nikolić, A., Gržetić, I. & Jovanović, D. (2008) Organobentonite as efficient textile dye sorbent. Chemical Engineering and Technology, 31, 567–574.CrossRefGoogle Scholar
Karakuş, Y., Özler, A. & Pekyardimci, Ş. (2008) Noncovalent immobilization of pectinesterase (Prunus Armeniaca L.) onto bentonite. Artificial Cells, Blood Substitutes, and Immobilization Biotechnology, 36, 535–550.CrossRefGoogle ScholarPubMed
Khenifi, A., Bouberka, Z., Sekrane, F., Kamech, M. & Derriche, Z. (2007) Adsorption study of an industrial dye by an organic clay. Adsorption, 13, 149–158.CrossRefGoogle Scholar
Kumar, A.G., Swarnalatha, S., Kamatchi, P. & Sekaran, G. (2009) Immobilization of high catalytic acid protease on functionalized mesoporous activated carbon particles. Biochemical Engineering Journal, 43, 185–190.CrossRefGoogle Scholar
Kumararaja, P., Manjaiaha, K.M., Datta, S.C. & Sarkar, B. (2017) Remediation of metal contaminated soil by aluminium pillared bentonite: synthesis, characterisation, equilibrium study and plant growth experiment. Applied Clay Science, 137, 115–122.CrossRefGoogle Scholar
Kumari, A., Kaur, B., Srivastava, R. & Sangwan, R.S. (2015) Isolation and immobilization of cellulase on mesoporous silica and mesoporous ZSM-5 zeolite materials for improved catalytic properties. Biochemistry and Biophysics Reports, 2, 108–114.CrossRefGoogle Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.CrossRefGoogle ScholarPubMed
Li, J., Cai, J., Zhongc, L. & Du, Y. (2012) Immobilization of a protease on modified chitosan beads for the depolymerization of chitosan. Carbohydrate Polymers, 87, 2697–2705.CrossRefGoogle Scholar
Liaoa, H., Chenb, D., Yuanb, L., Zhenga, M., Zhua, Y. & Liu, X., (2010) Immobilized cellulase by polyvinyl alcohol/Fe2O3 magnetic nanoparticle to degrade microcrystalline cellulose. Carbohydrate Polymers, 82, 600–604.CrossRefGoogle Scholar
Mateo, C., Palomo, J.M., Fernandez-Lorente, G., Guisan, J.M. & Fernandez-Lafuente, R. (2007) Improvement of enzyme activity, stability and selectivity via immobilization techniques. Enzyme and Microbial Techology, 40, 1451–1463.CrossRefGoogle Scholar
Miller, G.L. (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 426–428.CrossRefGoogle Scholar
Mohapatra, B.R., Gould, W.D., Dinardo, O., Papavinasam, S., Koren, D. & Revie, R.W. (2008) Effect of immobilization on kinetic and thermodynamic characteristics of sulfide oxidase from Arthrobacter species. Preparative Biochemistry and Biotechnology, 38, 61–73.CrossRefGoogle ScholarPubMed
Naik, S., Scholin, J. & Goss, B. (2016) Stabilization of phytase enzyme on montmorillonite clay. Journal of Porous Materials, 23, 401–406.CrossRefGoogle Scholar
Parolo, M.E., Pettinari, G.R., Musso, T.B., Sánchez-Izquierdo, M.P. & Fernández, L.G. (2014) Characterization of organo-modified bentonite sorbents: the effect of modification conditions on adsorption performance. Applied Surface Science, 320, 356–363.CrossRefGoogle Scholar
Patel, A.K., Singhania, R.R., Sima, S.J. & Pandey, A. (2019) Thermostable cellulases: review and perspectives. Bioresource Technology, 279, 385–392.CrossRefGoogle Scholar
Pei, H.Y., Hu, W.R. & Liu, Q.H. (2010) Effect of protease and cellulase on the characteristic of activated sludge. Journal of Hazardous Materials, 178, 397–403.CrossRefGoogle ScholarPubMed
Pike, R.N. & Dennison, C. (1989) Protein fractionation by three-phase partitioning in aqueous/t-butanol mixtures. Biotechnology and Bioengineering, 33, 221–228.CrossRefGoogle ScholarPubMed
Rao, M.B., Tanksale, A.M., Ghatge, M.S. & Deshpande, V.V. (1998) Molecular and biotechnological aspects of microbial proteases. Microbiology and Molecular Biology Reviews, 62(3), 597–635.CrossRefGoogle ScholarPubMed
Roy, I., Gupta, A., Khare, S.K., Bisaria, V.S. & Gupta, M.N. (2003) Immobilization of xylan-degrading enzymes from Melanocarpus albomyces IIS 68 on the smart polymer Eudragit L-100. Applied Microbiology and Biotechnology, 61, 309–313.CrossRefGoogle ScholarPubMed
Roy, I. & Gupta, M.N. (2002) Three-phase affinity partitioning of proteins. Analytical Biochemistry, 1, 11–14.CrossRefGoogle Scholar
Saleem, M., Rashid, M.H., Jabbar, A., Perveen, R., Khalid, A.M. & Rajoka, M.I. (2005) Kinetic and thermodynamic properties of an immobilized endoglucanase from Arachniotus citrinus. Process Biochemistry, 40, 849–855.CrossRefGoogle Scholar
Sedaghat, M.E., Ghiaci, M., Aghaei, H. & Soleimanian-Zad, S. (2009) Immobilization of α-amylase on Na-bentonite and modified bentonite. Applied Clay Science, 46, 125–130.CrossRefGoogle Scholar
Segel, J. (1975) Enzyme Kinetics, 1st edition. Willey Classics Library, New York, NY, USA.Google Scholar
Tao, Q., Li, Y., Shi, Y., Liu, R., Zhang, Y. & Guo, J. (2016) Application of molecular imprinted magnetic Fe3O4@SiO2 nanoparticles for selective immobilization of cellulase. Journal of Nanoscience and Nanotechnology, 16, 6055–6060.CrossRefGoogle ScholarPubMed
Tavano, O.L., Berenguer-Murcia, A., Secundo, F. and Fernandez-Lafuente, R. (2018) Biotechnological applications of proteases in food technology. Comprehensive Reviews in Food Science and Food Safety, 17, 412–431.CrossRefGoogle Scholar
Tu, M., Zhang, X., Kurabi, A., Gilkes, N., Mabee, W. & Saddler, J. (2006) Immobilization of β-glucosidase on Eupergit C for lignocellulose hydrolysis. Biotechnology Letters, 28, 151–156.CrossRefGoogle ScholarPubMed
Vitola, G., Büning, D., Schumacher, J., Mazzei, R., Giorno, L. & Ulbricht, M. (2017) Development of a novel immobilization method by using microgels to keep enzyme in hydrated microenvironment in porous hydrophobic membranes. Macromolecular Science, 17(5), 1600381.Google ScholarPubMed
Worsfold, P.J. (1995) Classification and chemical characteristics of immobilized enzymes. Pure and Applied Chemistry, 67(4), 597–600.CrossRefGoogle Scholar
Zhang, D., Hegab, H.E., Lvov, Y., Snow, L.D. & Palmer, J. (2016) Immobilization of cellulase on a silica gel substrate modified using a 3-APTES self-assembled monolayer. SpringerPlus, 5, 48–68.CrossRefGoogle ScholarPubMed
Zhao, Y., Wang, R., Fang, K., Tan, Y., Chen, S., Guan, Y. & Hao, L. (2019) Investigating the synergetic dispersing effect of hydrolyzed biomacromolecule cellulase and SDS on CuPc pigment. Colloids and Surfaces B: Biointerfaces, 184, 110568.CrossRefGoogle ScholarPubMed