Utilization of Whey for the Production of β-Galactosidase Using Yeast and Fungal Culture

Authors: Rupinder Kaur, Parmjit S. Panesar, Ram S. Singh

Abstract:

Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050 was observed to be the maximum extracellular enzyme producer.

Keywords: β-galactosidase, fungus, yeast, whey.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1107195

References:

[1] A. C. Adam, M. Rubio-Texeira, J. Polaina, “Lactose: the milk sugar from a biotechnological perspective,” Crit. Rev. Food Sci. Nutr., vol. 44, pp. 553- 557, Dec. 2010
[2] P. S Panesar, S. Kumari, R. Panesar, “Potential applications of immobilized β-galactosidase in food processing industries”. Enzyme Res.,vol, 2010, pp. 1-16, Dec. 2010.
[3] S. Kumari, P. S. Panesar, M. B. Bera, B. Singh, “Permeabilization of yeast cells for β-galactosidase activity using a mixture of organic solvents: A response surface methodology approach,” Asian J. Biotechnol., vol. 3, pp. 406-414, June 2011.
[4] A. O. Johnson, J. G. Semenya, M. S. Buchowski, C. O. Enwonwu, N. S. Scrimshaw, “Correlation of lactose maldigestion, lactose intolerance and milk intolerance,” Am. J. Clin. Nutr., vol. 57, pp. 199-401, March 1993.
[5] M. Becerra, B. Barolib, A. M. Fadda, J. B. Méndez, M. I. González- Siso, “Lactose bioconversion by calcium-alginate immobilization of Kluyveromyceslactis cells,” Enzyme Microb. Technol., vol. 29, pp. 506- 512, Nov. 2001.
[6] T. Haider, Q. Husain, “Calcium alginate entrapped preparations of Aspergillusoryzae beta galactosidase: its stability and applications in the hydrolysis of lactose,” Int. J. Biol. Macromol., vol. 41, pp. 72-80, June 2007b.
[7] A. Santos, M. Ladero, F. Garcia-Ochoa, “Kinetic modelling of lactose hydrolysis by a β-galactosidase from Kluyveromyces fragilis,”Enzyme Microb. Technol., vol. 22, pp. 558-567, May 1998.
[8] S. A. Shaikh, J. M. Khire, M. I. Khan, “Production of β-galactosidase from thermophilic fungus Rhizomucor sp,” J. Ind. Microbiol. Biotechnol., vol. 19, pp. 239-245, Oct. 1997.
[9] Q. Z. K. Zhou, X. D. Chen, “Effects of temperature and pH on the catalytic activity of the immobilized β-galactosidase from Kluyveromyceslactis,”Biochem. Eng. J., vol. 9, pp. 33-40, Nov. 2001.
[10] R. E. Huber, M. N. Gupta, S. K. Khare, “The active site and mechanism of the β-galactosidase from Escherichia coli,” Int. J. Biochem., vol. 26, pp. 309-318, March 1994.
[11] F.Zhao, J. Yu, “L-Asparginase release from Escherichia coli cells with K2HPO4 and triton X-100,” Biotechnol. Progress, vol. 17, pp. 490-494, June 2001.
[12] C. Spălățelu, “Biotechnological valorisation of whey,” Innov. Rom. Food Biotechnol.,vol. 10, pp. 1-8, March 2012.
[13] F. V. Kosikowski, “Whey utilization and whey products,” J. Dairy Sci., vol. 62, pp. 1149-1160, July 1979.
[14] S. S. Marwaha, J. F. Kennedy, “Review: whey-pollution problem and potential utilization,” Int. J. Food Sci. Technol., vol. 23, pp. 323-336, Aug. 1988.
[15] H. M. Sonawat, A. Agrawal, S. M. Dutta, “Production of β-galactosidase from Kluyveromyces fragilis grown on whey,” Folia Microbiol., vol. 26, pp. 370-376, Feb. 1981.
[16] J. H. Miller, “Experiments in molecular genetics,” Cold Spring Harbor Laboratory, New York, 1972, pp. 352.
[17] K. Reczey, H. Stalbrand, B. Hahn-Hegerdal, F.Tijernal, “Myceliaassociated β-galactosidase activity in microbial pellets of Aspergillus and Penicillium strains,” Appl. Microbiol.Biotechnol., vol. 38, pp. 393- 397, Dec. 1992.
[18] M. Puri, S. Gupta, P. Pahuja, A. Kaur, J. R. Kanwar, J. F. Kennedy, “Cell disruption optimization and covalent immobilization of β- galactosidase from Kluyveromyces marxianus YW-1 for lactose hydrolysis in milk,” Appl. Biochem. Biotechnol.,vol. 160, pp. 98-108, Jan. 2010.
[19] A. M. Gupte, J. S. Nair, “β-galactosidase production and ethanol fermentation from whey using Kluyveromyces marxianus NCIM 3551,” J. Sci. Ind. Res.,vol, 69, pp, 855-859, Nov. 2010.
[20] S. Bansal, H. S. Oberoi,G. S. Dhillon, R. T. Patil, “Production ofβ- galactosidase by Kluyveromyces marxianus MTCC 1388 using whey and effect of four different methods of enzyme extraction on β- galactosidase activity,” Indian J. Microbiol., vol. 48, pp. 337-341, Sep. 2008.
[21] P. S. Panesar, R. Panesar, R. S. Singh, J. F. Kennedy, H. Kumar, “Microbial production, immobilization and applications ofβ- galactosidase,” J. Chem. Technol. Biotechnol., vol.81, pp. 530-543, Apr. 2006.
[22] R. R. Saad, “Purification and some properties of β-galactosidase from Aspergillus japonicas,” Ann. Microbiol., vol. 54, pp. 299-306, 2004.