Effect of Montmorillonite and Organoclay on Urease and L-Asparaginase Activities in Soil

Document Type : Research Paper


Ferdowsi University of Mashhad


The amidohydrolase play a significant role in maintaining a sustainable environment and an agriculture sector. This is mainly because it is central to biochemical reactions (e.g. organic decompositions, food cycles, and pollutant decomposition). Therefore, keeping these enzymes active and stable in soil is a crucial task. In doing so, the effect of adding montmorillonite and organoclays on activity and stability of these enzymes in soil was investigated. The organic clay was produced by modifying the Na-montmorillonite clay using the hexadecyl trimethylammonium bromide cationic surfactant. To analyze the structural and morphologic profiles of the synthesized organoclay montmorillonite, Field Emission Scanning Electron Microscope (FESEM) and the clay's cation exchange capacity were used. The effects of this organic clay and incubation period on urease and L-asparaginase activities in soil were investigated. Results from statistical analyses showed that the effects of clay type and incubation period on the activity of both enzymes (urease and L-asparaginase) were significant at the 5% level. The activity level of both enzymes in soil samples treated with organic clay (71.48 and 9.01 μgNH4+-Ng-12h-1 in urease and L-asparaginase) was significantly higher than in the control (29.12 and 4.22 μgNH4+-Ng-12h-1 in urease and L-asparaginase) and montmorillonite clay sample (39.84 and 5.26 μgNH4+-Ng-12h-1 in urease and L-asparaginase) s. The maximum declining slope in the activity of both enzymes was observed after 7 days from incubation. The organoclay prepared from the montmorillonite clay successfully maintained soil enzymes. Therefore, use of this organoclay can positively affect enzyme activity and stability.


Acosta-Martinez, V. and Tabatabai M. A. (2000). Arylamidase activity of soils. Soil Science of Society of America Journal, 64(1), 215-21.
Acosta-Martinez, V., Cruz, L., Sotomayor-Ramirez, D. and Perez-Alegria, L. (2007). Enzyme activities as affected by soil properties and land use in tropical watershed. Applied Soil Ecology, 35(1), 35-45.
An, J. H. and Stefan, D. (2007). Adsorption of tannic acid on chitosan-montmorillonite as a function of pH and surface charge properties. Applied Clay Science, 36(4), 256–264.
Bashour, I. I. and Sayegh, A. H. (2007). Methods of analysis for soils of arid and semi arid regions: Food and agriculture organization of the united nations.
Bastida, F. Z. A., Hernandez, H. and Garcia, C. (2008). Past, present and future of soil quality indices: A biological perspective. Geoderma, 147(3-4), 159-171.
Bors, J., Dultz, S. and Riebe, B. (2000). Organophilic bentonites as adsorbents for radionuclides: II. Chemical and mineralogical properties of HDPy-montmorillonite. Applied Clay Science, 16(1-2), 15–29.
Burns, R. G., Pukite, A. H. and Mclaren, A. (1972). Concerning the location and persistence of soil urease. Soil Science Society of America Journal, 30(2), 308-311.
Cantu, M. B. A., Bedano, C. and Schiavo, H. (2007). Evaluaion de la calidad de suelos mediante el uso de indicadores e indices. Ci Suelo, 25(2), 173-8.
Cengiz, S., Cavas, L. and Yurdokoc, K. (2012). Bentonite and sepiolite as supporting media: immobilization of catalase. Applied Clay Science, 65-66, 114-120.
Dick, R. P. (1997). Soil enzyme activities as integrative indicators of soil health. In C. E. Pankhurst, B. M. Doube, and V. V. S. R. Gupta (Eds.), Biological indicators of soil health (pp. 121-156). Cab international, UK.
Dick, R. P., Sandeno, J., Taylor, A., Wagner, R., Pascoe, N. and Knight, T. (2002). Soil enzyme activity as a sensitive indicator of ecosystem disturbance. In: Proceedings of 17th World Congress of Soil Science, 14-21 Aug., Bangkok, Thailand, p. 1056.
Frankenberger, W. T. and Tabatabai, M. A. (1980). Amidase Activity in Soils: II. Kinetic parameters. Soil Science Society of America Journal, 44(3), 532-536.
Galindo-Gonzalez, C., Vicente, J. D., Ramos-Tejada, M. M., Lopez-Lopez, M. T., Gonzalez-Caballero, F. and Duran, J. D. G. (2005). Preparation and sedimentation behavior in magnetic fields of magnetite-covered clay particles. Langmuir, 21(10), 4410–4419.
Gianfreda, L., Rao, M. A. and Violante, A. (1992). Adsorption activity and kinetic properties of urease and Al(OH)x montmorillonite, aluminium hydroxide and montmorillonite complexes. Journal of Soil Biology and Biochemistry, 24, 51-58.
Huang, P. M. and Schnitzer, M. (1986). Intractions of soil minerals whith natural organics and microbes. Madison: Soil Science Society of America. USA: Madison, Wisconsin
Janssens, J., Deng, Z., Sonwa, D., Torrico, J. C., Mulindabigwi, V. and Pohlan, J. (2006). Relating agro-climax of orchards to eco-climax of natural vegetation. Acta Horticulturae, 707, 181-186.
Jung, L. C., Wang, C. C., Lee, C. and Hsu, T. (2007). Dyes adsorption onto organoclay and MCM-41. Journal of Environmental Enginering and  Management, 17(1), 29-38.
Kandeler, E., Poll, C., Frankenberger, W. T. and Tabatabai, M. A. (2011). Nitrogen cycle enzymes. In R. P. Dick (Ed.), Methods of soil enzymology (pp 211-245). Soil Science Society of America, Madison.
Leinweber, P., Jandl, G., Baum, C., Eckhardt, K. U. and Kandeler, E. (2008). Stability and composition of soil organic matter control respiration and soil enzyme activities. Siol Biology and Biochemistry, 40(6), 1496-1505.
Li, S., Wu, P., Li, H., Zhu, N., Li, P., Wu, J., Wang, X. and Dang, Z. (2010). Synthesis and characterization of organo montmorillonite supported iron nanoparticles. Applied Clay Sciences, 50, 330-336.
Nannipieri, P. (1994). The potential use of soil enzymes as indicators of productivity, sustainability and pollution. In C. E. Pankhurst, M. Doube, G. S. R. and P. R. Grace (Eds.), Soil Biota, Management in Sustainable Farming Systems (pp. 238-244). CSIRO Publications, Australia.
Nannipieri, P., Ceccanti, B., Cervelli, S. and Sequi, P. (1978). Stability and kinetic properties of humus-urease complexes. Soil Biology and Biochemistry, 10(2), 359-362.
Patil, A. J., Muthusamy, E. and Mann, S. (2004). Synthesis and self-assembly of organoclay-wrapped biomolecules. Angewandte Chemie, 116(37), 5036–5041.
Pettit, N. m., Smith, A. R. J., Freedman, R. B. and Burns, R. G. (1976). Soil urease: activity, stability and kinetic properties. Journal of Soil Biology and Biochemistry, 8, 479-484.
Sarioglan, S., Gurbuz, S., Ipeksac, T., Sedan, M. G. and Erol, M. (2014). Pararosaniline and crystal violet tagged montmorillonite for latent fi ngerprint investigation. Applied Clay Science, 87, 235–244.
Shrigadi, N. B., Shinde, A. B. and Samant, S. D. (2003). Study of catalytic activity of free and K10-supported iron oxyhydroxides and oxides in the Friedel-Crafts benzylation reaction using benzyl chloride/alcohol to understand their role in the catalysis by the Fe-exchanged/impregnated K10 catalysts. Applied Catalysis A: General, 252(1), 23–35.
Wang, C. C., Juang, L. C., Lee, C. K., Hsu, T. C., Lee, J. F. and Chao, H. P. (2004). Effects of exchanged surfactant cations on the pore structure and adsorption characteristics of montmorillonite. Journal of Colloid and Interface Science, 280(1), 27–35.
Weil, R. and Magdoff, F. (2004). Significance of soil organic matter to soil quality and health: Advances in Agroecology. In F. Magdoff, and R. Weil (Eds.), Soil organic matter in sustainable agriculture (pp. 1-43). CRC Press, Boca Raton.
Wu, L., Liao, L., Lv, G., Qin, F. and Li, Z. (2014). Microstructure and process of intercalation of imidazolium ionic liquids into montmorillonite. Chemical Engineering Journal, 236, 306-313.
Yilmaz, N. and Yapar, S. (2004). Adsorption properties of tetradecyl- and hexadecyltrimethy-lammonium bentonites. Applied Clay Science, 27(3), 223–228.
Zantua, M. I. and Bremner, J. M. (1977). Stability of urease in soils. Soil Biology and Biochemistry, 9(2), 135-140.
Zhai, R., Zhang, B., Liu, L., Xie, Y., Zhang, H. and Liu, J. (2010). Immobilization of enzyme biocatalyst on natural halloysite nanotubes. Catalysis Communications, 12(4), 259-263.
Zhang, Z., Zhang, J., Liao, L. and Xia, Z. (2013). Synrgistic effect of cationic and anionic surfactant for the modification of Ca-montmorillonite. Material Research Bulletin, 48(5), 1811-1816.
Zhu, L. Z., Zhu, R. L., Xu, L.H. and Ruan, X. X. (2007). Influence of clay charge densities and surfactant loading amount on the microstructure of CTMA-montmorillonite hybrids. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 304(1), 41–48.
Zhuang, G., Zhang, Z., Guo, J., Liao, L. and Zhao, J. (2015). A new ball milling method to produce organo-montmorillonite from anionic and nonionic surfactants. Applied clay science, 104, 18-26.