Ekta Arya, Mundhe Shyam Vasantrao and Duni Chand*

Department of Biotechnology, Gyanpath, Himachal Pradesh University, Shimla, India

*Corresponding Author: Duni Chand, Department of Biotechnology, Gyanpath, Himachal Pradesh University, Shimla, India.

Received: November 06, 2023; Published: November 21, 2023

Abstract

β-galactosidase, also referred to as lactase (EC.3.2.1.23), is extensively utilized in the pharmaceutical sector for the management of lactose intolerance, a condition that impacts around 75% of the global population. The present study is centered on the improvement of lactase production derived from the yeast Kluyveromyces lactis through the optimization of production and reaction parameters using the one variable at a time (OVAT) approach. The initial screening process for lactase-producing microbes involved cultivating the organisms on an agar plate supplemented with X-gal. The lactase-producing positive isolate exhibits a blue coloration on agar plates. Following the initial selection process, the hyperproducer strain was chosen for subsequent investigations. The cultivation and investigation of Kluyveromyces lactis pure cultures were conducted in order to optimize the synthesis of lactase. The study focused on examining specific production characteristics, like the starting pH and incubation temperature, in isolation. The highest level of lactase synthesis (402.74 U/mg dcw) was achieved under the following conditions: an initial pH of 7.0, a temperature of 35℃, the use of galactose and peptone as carbon and nitrogen sources, and the utilization of oNPG (Ortho-nitrophenyl- β-D-galactopyranoside) as the substrate. The application of optimization techniques led to a significant increase in the total activity of the enzyme, with a seven-fold boost seen.

Keywords: Lactose Intolerance; Kluyveromyces lactis; Cultivation

References

1. Vandenplas Y. “Lactose intolerance”. Asia Pacific Journal of Clinical Nutrition 24 (2015): S9-13.
2. Storhaug CL., et al. “Country, regional, and global estimates for lactose malabsorption in adults: a systematic review and meta-analysis”. Lancet Gastroenterology and Hepatology 2 (2017): 738-746.
3. Heyman MB. “Lactose intolerance in infants, children, and adolescents”. Pediatrics 3 (2006): 1279-1286.
4. Panesar PS., et al. “Potential applications of immobilized βgalactosidase in food processing industries”. Enzyme Research (2010): 1-16.
5. Bayramoglu G., et al. “Immobilization of beta-galactosidase onto magnetic poly (GMA-MMA) beads for hydrolysis of lactose in bed reactor”. Catalysis Communications 8 (2007): 1094-1101.
6. Haider T and Husain Q. “Preparation of lactose-free milk by using ammonium sulfate fractionated proteins from almonds”. Journal of the Science of Food and Agriculture 87 (2007): 1278-1283.
7. Picard C., et al. “Review article: bifidobacteria as probiotic agents-physiological effects and clinical benefits”. Alimentary Pharmacology and Therapeutics 22 (2005): 495-512.
8. Nathalie V., et al. “Assessment of lactase activity in humans by measurement of galactitol and galactonate in serum and urine after milk intake”. The American Journal of Clinical Nutrition2 (2019): 470-477.
9. Scopes RK. “Protein Purification”. Principles and Practice. Springer-Verlag New York Heidelberg Berlin, 356 p.
10. Laemmli UK., et al. “A factor preventing the major head protein of bacteriophage T4 from random aggregation”. Journal of Molecular Biology1 (1970b): 69- 85.
11. Alazzeh AY., et al. “Carbohydrate and protein sources influence the induction of β-galactosidases in Lactobacillus reuteri”. Food Chemistry 4 (2009): 654-659.
12. Hsu CA., et al. “Production of beta-galactosidase by Bifidobacteria as influenced by various culture conditions”. International Journal of Food Microbiology 104 (2005): 197-206.
13. Kumari S., et al. “Production of beta-galactosidase using novel yeast isolate from Whey”. International Journal of Dairy Science 6 (2011): 150-157.
14. Domingues L., et al. “Aspergillus niger -galactosidase production by yeast in a continuous high-density reactor”. Process Biochemistry 40 (2005): 1151-1154.
15. Soni S and Timande S. “Biotechnological utilization of dairy waste to solve environmental problems”. Journal of Environmental Research and Development3A (2012).
16. Ramesh MV and Lonsane BK. “Solid state fermentation for production of αamylase by Bacillus megaterium 16M”. Biotechnology Letter5 (1987): 323-328.
17. Akcan N. “High-level production of extracellular galactosidase from Bacillus licheniformis ATCC12759 in submerged fermentation”. African Journal of Microbiology Research 5 (2011): 4615-4621.
18. Mozumder NHMR., et al. “Study on isolation and partial purification of lactase Enzyme from Lactobacillus bacteria isolated from Yogurt” (2012).

Citation

Citation: Duni Chand., et al. “Studies on Isolation and Optimization of Lactase from Kluyveromyces lactis LPY-08".Acta Scientific Microbiology 6.12 (2023): 57-62.

Copyright

Copyright: © 2023 Duni Chand., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.