Statistical Modeling for Permeabilization of a Novel Yeast Isolate for β-Galactosidase Activity Using Organic Solvents

Authors: Shweta Kumari, Parmjit S. Panesar, Manab B. Bera

Abstract:

The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 ^oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.

Keywords: β-galactosidase, optimization, permeabilization, response surface methodology, yeast.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1092924

References:

[1] Y. Chisti and M. Moo-Young, "Disruption of microbial cells for intracellular products,” Enz. Microb. Technol., vol. 8, pp.194-204, Apr. 1986.
[2] P.S. Panesar, S. Kumari and R. Panesar, "Potential applications of immobilized β-galactosidase in food processing industries,” Enz. Res., vol. 2010, pp. 1-16, Dec. 2010.
[3] S. Dagbagli and Y. Goksungur, "Optimization of β-galactosidase production using Kluyveromyces lactis NRRL Y-8279-8279 by response surface methodology,” Elec. J. Biotechnol., vol. 2008, pp. 1-12, Oct. 2008.
[4] L.M. Huerta, C. Vera, C. Guerrero, L. Wilson and A. Illanes, "Synthesis of galacto-oligosaccharides at very high lactose concentrations with immobilized β-galactosidases from Aspergillus oryzae,” Process Biochem., vol. 46, pp. 245-252, Jan. 2011.
[5] W. Li, X. Xiang, S. Tang, B. Hu, L. Tian, Y. Sun, H. Ye and X. Zeng, "Effective enzymatic synthesis of lactosucrose and its analogues by β-D-galactosidase from Bacillus circulans,” J. Agric. Food Chem., vol. 57, pp. 3927-3933, May 2009.
[6] X. Hua, R. Yang, W. Zhang, Y. Fei, Z. Jin and B. Jiang, "Dual-enzymatic synthesis of lactulose in organic-aqueous two-phase media,” Food Res. Inter., vol. 43, pp. 716-722, Apr. 2010.
[7] K. Adinarayana, P. Ellaiah, B. Srinivasulu, R.B. Devi and G. Adinarayana, "Response surface methodological approach to optimize the nutritional parameters for neomycin production by Streptomyces marinensis under solid-state fermentation,” Process Biochem., vol. 38, pp. 565-572, June 2003.
[8] M. Harju, "Milk sugars and minerals as ingredients,” Int. J. Dairy Technol., vol. 54, pp. 61-63, May 2001.
[9] I.H. Boyaci, "A new approach for determination enzyme kinetic constants using response surface methodology,” Biochem. Engg. J., vol. 25, pp. 55-62, Aug. 2005.
[10] J.H. Miller, "Experiments in molecular genetics,” Cold Spring Harbor Laboratory, New York, 1972, pp. 352.
[11] R. Panesar, P.S. Panesar, R.S. Singh, J.F. Kennedy and M.B. Bera, "Production of lactose hydrolyzed milk using ethanol permeabilized yeast cells,” Food Chem., vol. 101, pp. 786-790, 2007.
[12] R.C. Dickson and J.S. Markin, "Physiological studies of betagalactosidase induction in K. lactis,” J. Bacteriol., vol. 142, pp. 777-785, June 1980.
[13] G. Kaur, P.S. Panesar, M.B. Bera and B. Singh, "Optimization of permeabilization process for lactose hydrolysis in whey using response surface methodology,” J. Food Process Engg., vol. 32, pp. 355-368, June 2009.
[14] M.S. Flores, C.E. Voget and R.J.J. Ertola, "Permeabilization of yeast cells (Kluyveromyces lactis) with organic solvents,” Enz. Microb. Technol., vol. 16, pp. 340-346, Apr. 1994.
[15] M. Decleire, W. De-Cat and N. Van-Huynh, "Comparison of various permeabilization treatments on Kluyveromyces by determining in situ β-galactosidase activity,” Enz. Microb. Technol., vol. 9, pp. 300-302, May 1987.
[16] P.S. Panesar, "Application of response surface methodology in the permeabilization of yeast cells for lactose hydrolysis,” Biochem. Engg. J., vol. 39, pp. 91-96, Apr. 2008.