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

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.

Authors:

• Rupinder Kaur
• Parmjit S. Panesar
• Ram S. Singh

Keywords:

• β-galactosidase
• fungus
• yeast
• whey.

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.