Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-28T01:27:26.837Z Has data issue: false hasContentIssue false

A comparative study of extraction techniques for maximum recovery of β-galactosidase from the yogurt bacterium Lactobacillus delbrueckii ssp. bulgaricus

Published online by Cambridge University Press:  03 February 2020

Rabin Gyawali*
Affiliation:
Food Microbiology and Biotechnology Laboratory, Food and Nutritional Sciences Program, North Carolina Agricultural and Technical State University, Greensboro, NC, USA
Ayowole Oyeniran
Affiliation:
Food Microbiology and Biotechnology Laboratory, Food and Nutritional Sciences Program, North Carolina Agricultural and Technical State University, Greensboro, NC, USA
Tahl Zimmerman
Affiliation:
Food Microbiology and Biotechnology Laboratory, Food and Nutritional Sciences Program, North Carolina Agricultural and Technical State University, Greensboro, NC, USA
Sulaiman O. Aljaloud
Affiliation:
College of Sport Sciences and Physical Activity, King Saud University, P.O. Box 1949, Riyadh11362, Saudi Arabia
Albert Krastanov
Affiliation:
Department of Biotechnology, University of Food Technologies, Plovdiv, Bulgaria
Salam A. Ibrahim
Affiliation:
Food Microbiology and Biotechnology Laboratory, Food and Nutritional Sciences Program, North Carolina Agricultural and Technical State University, Greensboro, NC, USA
*
Author for correspondence: Rabin Gyawali, Email: [email protected]

Abstract

The study reported in this research communication evaluates the chemical (solvents) and mechanical (sonication, bead-beater) extraction methods to determine the maximum recovery of β-galactosidase from L. bulgaricus spp. Among all extraction techniques, sonication-assisted extraction yielded the highest amounts of enzyme activity (between 1892–2156 Miller Units) in cell-free extract (supernatant). Interestingly, solvent extracted enzyme activities were found to be very low (between 83–153 Miller Units) in supernatant. SDS-polyacrylamide gel electrophoresis and the total protein determination showed that mechanical methods can completely lyse the cells. Our results thus demonstrated that the mechanical extraction method of sonication is the best one for recovering the maximum amount of lactase from L. bulgaricus strains.

Type
Research Article
Copyright
Copyright © Hannah Dairy Research Foundation 2020

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Brena, B, González-Pombo, P and Batista-Viera, F (2013) Immobilization of enzymes: a literature survey. In Guisan, JM (ed), Immobilization of enzymes and cells. Humana Press, Totowa, NJ, pp. 1531.Google Scholar
Degeest, B and De Vuyst, L (2000) Correlation of activities of the enzymes α-phosphoglucomutase, UDP-galactose 4-epimerase, and UDP-glucose pyrophosphorylase with exopolysaccharide biosynthesis by Streptococcus thermophilus LY03. Applied and Environmental Microbiology 66, 35193527.Google Scholar
Geciova, J, Bury, D and Jelen, P (2002) Methods for disruption of microbial cells for potential use in the dairy industry – a review. International Dairy Journal 12, 541553.Google Scholar
Gobinath, D and Prapulla, SG (2015) Transgalactosylating β-galactosidase from probiotic Lactobacillus plantarum MCC2156: production and permeabilization for use as whole cell biocatalyst. Journal of Food Science and Technology 52, 60036009.Google ScholarPubMed
Ibrahim, SA and O'Sullivan, DJ (2000) Use of chemical mutagenesis for the isolation of food grade β-galactosidase overproducing mutants of bifidobacteria, lactobacilli and Streptococcus thermophilus. Journal of Dairy Science 83, 923930.Google ScholarPubMed
Ibrahim, SA and Gyawali, R (2013) Lactose intolerance. In Young, WP and George, FW (eds), Milk and Dairy Products in Human Nutrition: Production, Composition and Health: John Wiley & Sons. John Wiley & Sons, New Jersey, USA pp. 246260.Google Scholar
Ismail, SA, El-Mohamady, Y, Helmy, WA, Abou-Romia, R and Hashem, AM (2010) Cultural condition affecting the growth and production of β- galactosidase by Lactobacillus acidophilus NRRL 4495. Australian Journal of Basic and Applied Sciences 4, 50515058.Google Scholar
Kreft, ME, Roth, L and Jelen, P (2001) Lactose hydrolysing ability of sonicated cultures of Lactobacillus delbrueckii ssp. bulgaricus 11842. Le Lait 81, 355364.Google Scholar
Kumari, S, Panesar, PS, Bera, MB and Singh, B (2011) Permeabilization of a yeast cells for β-galactosidase activity using mixture of organic solvents. Asian Journal of Biotechnology 3, 406414.Google Scholar
Meira, SMM, Helfer, VE, Velho, RV, Lopes, FC and Brandelli, A (2012) Probiotic potential of Lactobacillus spp. Isolated from Brazilian regional ovine cheese. Journal of Dairy Research 79, 119127.Google ScholarPubMed
Miller, JH (1992) A Short Course in Bacterial Genetics. A Labora-Tory Manual and Handbook for Escherichia Coli and Related Bacteria. Cold Spring Harbor, New York, USA: Cold Spring Harbor Laboratory Press.Google Scholar
Niven, GW and Mulholland, F (1998) Cell membrane integrity and lysis in Lactococcus lactis: the detection of a population of permeable cells in post-logarithmic phase cultures. Journal of Applied Microbiology 84, 9096.10.1046/j.1365-2672.1997.00316.xGoogle ScholarPubMed
Panesar, PS, Kumari, S and Panesar, R (2010) Potential applications of immobilized β-galactosidase in food processing industries. Enzyme Research. doi: http://dx.doi.org/10.4061/2010/473137Google ScholarPubMed
Plou, FJ, Polaina, JZ, Sanz-Aparicio, J and Fernández-Lobato, M (2017) β-Galactosidases for lactose hydrolysis and galactooligosaccharide synthesis. In Ray, RC and Rosell, CM (eds), Microbial Enzyme Technology in Food Applications. Boca Raton, FL, USA: CRC Press, pp. 307317.Google Scholar
Vasiljevic, T and Jelen, P (2001) Production of β-galactosidase for lactose hydrolysis in milk and dairy products using thermophilic lactic acid bacteria. Innovative Food Science & Emerging Technologies 2, 7585.Google Scholar
Vinderola C, G and Reinheimer, JA (2003) Lactic acid starter and probiotic bacteria: a comparative “in vitro” study of probiotic characteristics and biological barrier resistance. Food Research International 36, 895904.Google Scholar
Supplementary material: PDF

Gyawali et al. supplementary material

Figures S1-S2

Download Gyawali et al. supplementary material(PDF)
PDF 451.4 KB