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Biomedicine

Volume: 43 Issue: Supplementary 1

  • Open Access
  • Original Article

Fourier transform infrared spectroscopy analysis of Lactose hydrolysis by beta-galactosidase from Lactiplantibacillus plantarum GV54 and Lactiplantibacillus sp. GV66

Mahanthesh Vasudha1, M. N. Kalasad2, S. C. Sharath2, Devaraja Gayathri1

1Department of Studies in Microbiology,
2Department of Studies in Physics, Davangere University, Shivagangothri, SH-76, Davangere, 577 007, Karnataka, India

*Corresponding author: Devaraja Gayathri. Email: [email protected]

Year: 2023, Page: 146-150, Doi: https://doi.org/10.51248/.v43i1.2577

Abstract

Introduction and Aim: The enzyme beta-galactosidase is extremely important in the food and pharmaceutical industries. The goal of this work was to improve the lactose hydrolysis process employing beta-galactosidase to convert glucose and galactose for alleviating lactose intolerance.
Methods: Intracellular beta-galactosidase was extracted from Lactiplantibacillus spp. (GV54 and GV66) by cell permeabilized method using toluene/acetone solvents. Lactose hydrolysis was performed using 50 g/L of lactose with partially purified beta-galactosidase enzyme at different time intervals 2, 4, 6 and 12 h. Fourier transforms infrared spectroscopy (FTIR) was used to evaluate the degree of hydrolysis at different time intervals and lactose hydrolysis was high at 6 h of incubation.
Results: The FTIR peak of lactose molecules hydrolysis by beta-galactosidase (GV54 and GV66) with different time intervals (2h, 4h, 6h, and 12h) indicates similarities in bond lengths and vibrational modes. The amide I group of beta-galactosidase is uniformly bound to lactose at 1634 per cm and C-H group of glucose and galactose was observed at 1386 per cm. In the range of 2000-800 per cm, we could identify variations in the shape and strength of these peaks, compared with control (lactose solution).
Conclusion: The precise and direct measurement of individual carbohydrates such as glucose and galactose in lactose solution hydrolyzed by beta-galactosidase enzyme was successfully accomplished using FTIR spectrometry. Comparing FTIR analysis to traditional procedures, the time required is drastically decreased. These findings have the analytical power to follow the dynamics of in vitro enzymatic activity of naturally occurring substrates rich in natural carbohydrate polymers.

Keywords: FTIR; lactose; beta-galactosidase; Lactiplantibacillus plantarum; lactose intolerance

References

1. Lara-Mota, E. E., Nicolás–Vázquez, M. I., López-Martínez, L. A., Espinosa-Solis, V., Cruz-Alcantar, P., Toxqui-Teran, A., et al., Phenomenological study of the synthesis of pure anhydrous β-lactose in alcoholic solution. Food Chem. 2021;340: 128054.

2. Gayathri, D., Vasudha, M. Lactose intolerance with special emphasis on probiotics for management. EC Nutrition, 2018; 13(5): 325-332.

3. Gänzle, M. G., Haase, G., Jelen, P. Lactose: crystallization, hydrolysis, and value-added derivatives. Int Dairy J, 2008; 18(7): 685-694.

4. Vasudha, M., Gayathri, D. Kinetic and modeling analyses of lactose-hydrolyzing β-galactosidase from Lactiplantibacillus plantarum GV54. World Acad Sci J. 2023; 5: 11.

5. Gayathri, D., Vasudha, M., Prashantkumar, C. S. Gut-Brain Axis: Probiotic Interactions and Implications for Human Mental Health. In Microbiome-Gut-Brain Axis (2022). (pp. 261-280). Springer, Singapore.

6. Gayathri, D., Prashantkumar, C. S., Vasudha, M. Current Insights on the Modulation of Gut Microbiome, and Its Effect on Human Health. Microbiome-Gut-Brain Axis, 2022; 29-51.

7. Gayathri, D. Probiotics for total health: Better today and tomorrow. J Matern Pediatr Nutr. 2016; 2(107):2.

8. Asha, A., Gayathri, D. Synergistic impact of Lactobacillus fermentum, Lactobacillus plantarum and vincristine on 1, 2-dimethylhydrazine-induced colorectal carcinogenesis in mice. Exp Therap Med. 2012; 3(6): 1049-1054.

9. Kumara, S. S., Bashisht, A., Venkateswaran, G., Hariprasad, P., Gayathri, D. Characterization of novel Lactobacillus fermentum from curd samples of indigenous cows from Malnad region, Karnataka, for their aflatoxin B1 binding and probiotic properties. Prob Antimicrob Prot; 2019; 11(4): 1100-1109.

10. Jurado, E., Camacho, F., Luzon, G., Vicaria, J.M. A new kinetic model proposed for enzymatic hydrolysis of lactose by a β-galactosidase from Kluyveromyces fragilis. Enzyme Microb Technol. 2002; 31:300-309.

11. Sriphannam, W., Lumyong, S., Niumsap, P., Ashida, H., Yamamoto, K., Khanongnuch, C. A selected probiotic strain of Lactobacillus fermentum CM33 isolated from breast-fed infants as a potential source of β-galactosidase for prebiotic oligosaccharide synthesis. J Microbiol. 2012; 50:119-126.

12. Oliveira, C., Guimarães, P.M., Domingues, L. Recombinant microbial systems for improved β-galactosidase production and biotechnological applications. Biotechnol Adv. 2011;29(6):600-609.

13. Verma, M. L., Barrow, C. J., Kennedy, J. F., Puri, M. Immobilization of β-d-galactosidase from Kluyveromyces lactis on functionalized silicon dioxide nanoparticles: characterization and lactose hydrolysis. Int J Biol Macromol. 2012; 50(2): 432-437.

14. Staiano, M., Bazzicalupo, P., Rossi, M., D’Auria, S. Glucose biosensors as models for the development of advanced protein based biosensors. Mol BioSyst. 2005; 354(1).

15. Mlichova, Z., Rosenberg., M. Current trends of β-galactosidase application in food technology. J Food Nutr Res. 2006; 2(45):47-54.

16. Lei, Y., Zhou, Q., Zhang, Y. L., Chen, J. B., Sun, S. Q., Noda, I. Analysis of crystallized lactose in milk powder by Fourier-transform infrared spectroscopy combined with two-dimensional correlation infrared spectroscopy. J Mol Struct. 2010; 974(1-3): 88-93

17. Ibrahim, M., Alaam, M., El-Haes, H., Jalbout, A. F., Leon, A. D. Analysis of the structure and vibrational spectra of glucose and fructose. Eclet Quim. 2006;31: 15-21.

18. Vinderola, C.G., Reinheimer, J.A. Lactic acid starter and probiotic bacteria: a comparative “in vitro” study of probiotic characteristics and biological barrier resistance. Food Res Int. 2003; 36:895-904.

19. Selvarajan, E., Mohanasrinivasan, V. Kinetic studies on exploring lactose hydrolysis potential of β galactosidase extracted from Lactobacillus plantarum HF571129. J Food Sci Technol. 2015; 52(10): 6206-6217.

20. Nguyen, T.H., Splechtna, B., Steinböck, M., Kneifel, W., Lettner, H.P., Kulbe, K.D., et al., Purification and characterization of two novel β-galactosidases from Lactobacillus reuteri. J Agri Food Chem. 2006; 54: 4989-4998.

21. Das, B., Roy, A. P., Bhattacharjee, S., Chakraborty, S., Bhattacharjee, C. Lactose hydrolysis by β-galactosidase enzyme: optimization using response surface methodology. Ecotoxicol Environ Saf. 2015; 121: 244-252.

22. Białkowska, A.M., Cieśliński, H., Nowakowska, K.M., Kur, J., Turkiewicz, M. A new β-galactosidase with a low temperature optimum isolated from the Antarctic Arthrobacter sp. 20B: gene cloning, purification and characterization. Arch Microbiol. 2009; 191: 825-835.

23. Husain, Q., Ansari, S. A., Alam, F., Azam, A. Immobilization of Aspergillus oryzae β galactosidase on zinc oxide nanoparticles via simple adsorption mechanism. Inter J Biol Macromol. 2011; 49(1): 37-43.

24. Adina, C., Florinela, F., Abdelmoumen, T., Carmen, S. Application of FTIR spectroscopy for a rapid determination of some hydrolytic enzymes’ activity on sea buckthorn substrate. Rom Biotechnol Letters. 2010; 15(6): 5738-5744.

25. Wang, J., Kliks, M. M., Jun, S., Jackson, M., Li, Q. X. Rapid analysis of glucose, fructose, sucrose, and maltose in honeys from different geographic regions using Fourier transform infrared spectroscopy and multivariate analysis. J Food Sci. 2010; 75(2): C208-C214.

Cite this article

Mahanthesh Vasudha, M. N. Kalasad, S. C. Sharath, Devaraja Gayathri. Fourier transform infrared spectroscopy analysis of Lactose hydrolysis by beta-galactosidase from Lactiplantibacillus plantarum GV54 and Lactiplantibacillus sp. GV66. Biomedicine: 2023; 43(1) Supplementary issue: 146-150

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