Probiotic potential of β‑galactosidase‑producing lactic acid bacteria from fermented milk and their molecular characterization

Vasudha,Mahanthesh; Prashantkumar,Chakra S; Bellurkar,Mallika; Kaveeshwar,Vishwas; Gayathri,Devaraja

doi:10.3892/br.2023.1605

Probiotic potential of β‑galactosidase‑producing lactic acid bacteria from fermented milk and their molecular characterization

Authors:
- Mahanthesh Vasudha
- Chakra S Prashantkumar
- Mallika Bellurkar
- Vishwas Kaveeshwar
- Devaraja Gayathri
View Affiliations

Affiliations: Department of Studies in Microbiology, Davangere University, Davangere, Karnataka 577007, India, Central Research Laboratory, SDM College of Medical Sciences and Hospital, Shri Dharmasthala Manjunatheshwara University, Dharwad, Karnataka 580009, India, Central Research Laboratory, SDM College of Medical Sciences and Hospital, Shri Dharmasthala Manjunatheshwara University, Dharwad, Karnataka 580009, India
Published online on: February 8, 2023 https://doi.org/10.3892/br.2023.1605
Article Number: 23
Copyright: © Vasudha et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Probiotics have attained significant interest in recent years as a result of their gut microbiome modulation and gastrointestinal health benefits. Numerous fermented foods contain lactic acid bacteria (LAB) which are considered as GRAS and probiotic bacteria. The present study aimed to investigate indigenous LAB from homemade fermented milk samples collected in remote areas of Karnataka (India), in order to isolate the most potent and well‑adapted to local environmental conditions bacteria, which were then evaluated using a step‑by‑step approach focused on the evaluation of probiotic traits and β‑galactosidase‑producing ability. LAB were screened using 5‑bromo‑4‑chloro‑3‑indole‑D‑galactopyranoside (X‑Gal) and O‑nitrophenyl‑β‑D‑galactopyranoside (ONPG) as substrate, and exhibited β‑galactosidase activity ranging from 728.25 to 1,203.32 Miller units. The most promising isolates were selected for 16S rRNA gene sequence analysis and identified as Lactiplantibacillus plantarum, Limosilactobacillus fermentum, Lactiplantibacillus pentosus and Lactiplantibacillus sp. Furthermore, these isolates were evaluated by in vitro, viz., survival in gastrointestinal tract, antibiotic susceptibility, antimicrobial activity, cell surface characteristics, and haemolytic activity. All eight isolates demonstrated strong adherence and prevented pathogen penetration into HT‑29 cells, indicating potential of the bacteria to scale up industrial level production of milk products for lactose intolerants.

View Figures

View References

1	Lomer MC, Parkes GC and Sanderson JD: Review Article: Lactose intolerance in clinical practice-myths and realities. Aliment Pharmacol Ther. 27:93–103. 2008.PubMed/NCBI View Article : Google Scholar
2	Gayathri D and Vasudha M: Lactose Intolerance with Special Emphasis on Probiotics for Management. EC Nutrition. 13:325–332. 2018.
3	Ingram CJ, Mulcare CA, Itan Y, Thomas MG and Swallow DM: Lactose digestion and the evolutionary genetics of lactase persistence. Human Genet. 124:579–591. 2009.PubMed/NCBI View Article : Google Scholar
4	Babu J, Kumar S, Babu P, Prasad JH and Ghoshal UC: Frequency of lactose malabsorption among healthy Southern and Northern Indian populations by genetic analysis and lactose hydrogen breath and tolerance tests. Am J Clin Nutr. 91:140–146. 2010.PubMed/NCBI View Article : Google Scholar
5	Sriphannam W, Lumyong S, Niumsap P, Ashida H, Yamamoto K and 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. 50:119–126. 2012.PubMed/NCBI View Article : Google Scholar
6	Harrington LK and Mayberry JF: A re-appraisal of lactose intolerance. Int J Clin Pract. 62:1541–1546. 2008.PubMed/NCBI View Article : Google Scholar
7	Heyman MB: Committee on Nutrition. Lactose intolerance in infants, children, and adolescents. Pediatrics. 118:1279–1286. 2006.PubMed/NCBI View Article : Google Scholar
8	FAO/WHO: Working Group on Drafting Guidelines for the Evaluation of Probiotics in Food, London Ontario, Canada, 2002.
9	Patel RM and Lin PW: Developmental biology of gut-probiotic interaction. Gut Microbes. 1:186–195. 2010.PubMed/NCBI View Article : Google Scholar
10	Pan Q, Zhu J, Liu L, Cong Y, Hu F, Li J and Yu X: Functional identification of a putative β-galactosidase gene in the special lac gene cluster of Lactobacillus acidophilus. Curr Microbiol. 60:172–178. 2010.PubMed/NCBI View Article : Google Scholar
11	Pagnini C, Saeed R, Bamias G, Arseneau KO, Pizarro TT and Cominelli F: Probiotics promote gut health through stimulation of epithelial innate immunity. Proc Natl Acad Sci USA. 107:454–459. 2010.PubMed/NCBI View Article : Google Scholar
12	Swamy CT, Gayathri D, Devaraja TN, Bandekar M, D'Souza SE, Meena RM and Ramaiah N: Plant growth promoting potential and phylogenetic characteristics of a lichenized nitrogen fixing bacterium, Enterobacter cloacae. J Basic Microbiol. 56:1369–1379. 2016.PubMed/NCBI View Article : Google Scholar
13	Rashmi BS and Gayathri D: Molecular characterization of gluten hydrolyzing Bacillus sp. and their efficacy and biotherapeutic potential as probiotics using Caco-2 cell line. J Appl Microbiol. 123:759–772. 2017.PubMed/NCBI View Article : Google Scholar
14	Gheytanchi E, Heshmati F, Shargh BK, Nowroozi J and Movahedzadeh F: Study on β-galactosidase enzyme produced by isolated lactobacilli from milk and cheese. Afr J Microbiol Res. 4:454–458. 2010.
15	Vinderola CG and Reinheimer JA: Lactic acid starter and probiotic bacteria: A comparative ‘in vitro’ study of probiotic characteristics and biological barrier resistance. Food Res Int. 36:895–904. 2003.
16	Li J, Zhang W, Wang C, Yu Q, Dai R and Pei X: Lactococcus lactis expressing food-grade β-galactosidase alleviates lactose intolerance symptoms in post-weaning Balb/c mice. Appl Microbiol Biotechnol. 96:1499–1506. 2012.PubMed/NCBI View Article : Google Scholar
17	Vos P, Garrity G, Jones D, Krieg NR, Ludwig W, Rainey FA, et al: Bergey's manual of systematic bacteriology. The Firmicutes (Vol. 3). SSBM, 2011.
18	Asha and Gayathri D: Antagonistic potential of Lactobacillus against enteropathogenic bacteria; purification and characterization of their bacteriocins. Adv J Food Sci Technol. 4:265–269. 2012.
19	Princely S, Basha NS, Kirubakaran JJ and Dhanaraju MD: Biochemical characterization, partial purification and production of an intracellular beta-galactosidase from Streptococcus thermophilus grown in whey. Euro J Exp Bio. 3:242–251. 2013.
20	William S, Feil H and Copeland A: Bacterial genomic DNA isolation using CTAB. Sigma. (50 (6876))2012.
21	Shokryazdan P, Sieo CC, Kalavathy R, Liang JB, Alitheen NB, Faseleh Jahromi M and Ho YW: Probiotic potential of Lactobacillus strains with antimicrobial activity against some human pathogenic strains. Biomed Res Int. 2014(927268)2014.PubMed/NCBI View Article : Google Scholar
22	Armas F, Camperio C and Marianelli C: In vitro assessment of the probiotic potential of Lactococcus lactis LMG 7930 against ruminant mastitis-causing pathogens. PLoS One. 12(e0169543)2017.PubMed/NCBI View Article : Google Scholar
23	Kumara SS, Bashisht A, Venkateswaran G, Hariprasad P and 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. Probiotics Antimicrob Proteins. 11:1100–1109. 2019.PubMed/NCBI View Article : Google Scholar
24	Aissi EA, Lecocq M, Brassart C and Bouquelet S: Adhesion of some Bifidobacterial strains to human enterocyte-like cells and binding to mucosal glycoproteins. Microb Ecol Health Dis. 13:32–39. 2001.
25	Duary RK, Rajput YS, Batish VK and Grover S: Assessing the adhesion of putative indigenous probiotic lactobacilli to human colonic epithelial cells. Indian J Med Res. 134:664–671. 2011.PubMed/NCBI View Article : Google Scholar
26	Botes M, Loos B, van Reenen CA and Dicks LM: Adhesion of the probiotic strains Enterococcus mundtii ST4SA and Lactiplantibacillus plantarum 423 to Caco-2 cells under conditions simulating the intestinal tract, and in the presence of antibiotics and anti-inflammatory medicaments. Arch Microbiol. 190:573–584. 2008.PubMed/NCBI View Article : Google Scholar
27	Maleki Kakelar H, Barzegari A, Hanifian S, Barar J and Omidi Y: Isolation and molecular identification of Lactobacillus with probiotic potential from abomasums driven rennet. Food Chem. 272:709–714. 2019.PubMed/NCBI View Article : Google Scholar
28	Reuben RC, Roy PC, Sarkar SL, Rubayet Ul, Alam ASM and Jahid IK: Characterization and evaluation of lactic acid bacteria from indigenous raw milk for potential probiotic properties. J Dairy Sci. 103:1223–1237. 2020.PubMed/NCBI View Article : Google Scholar
29	Bin Masalam MS, Bahieldin A, Alharbi MG, Al-Masaudi S, Al-Jaouni SK, Harakeh SM and Al-Hindi RR: Isolation, molecular characterization and probiotic potential of lactic acid bacteria in Saudi raw and fermented milk. Evid Based Complement Alternat Med. 2018(7970463)2018.PubMed/NCBI View Article : Google Scholar
30	Wang CY, Lin PR, Ng CC and Shyu YT: Probiotic properties of Lactobacillus strains isolated from the feces of breast-fed infants and Taiwanese pickled cabbage. Anaerobe. 16:578–585. 2010.PubMed/NCBI View Article : Google Scholar
31	Shokryazdan P, Faseleh Jahromi M, Liang JB and Ho YW: Probiotics: From isolation to application. J Am Coll Nutr. 36:666–676. 2017.PubMed/NCBI View Article : Google Scholar
32	Hsieh PS, Chen CW, Kuo YW and Ho HH: Lactobacillus spp. reduces ethanol-induced liver oxidative stress and inflammation in a mouse model of alcoholic steatohepatitis. Exp Ther Med. 21(188)2021.PubMed/NCBI View Article : Google Scholar
33	Zago M, Fornasari ME, Carminati D, Burns P, Suàrez V, Vinderola G, Reinheimer J and Giraffa G: Characterization and probiotic potential of Lactiplantibacillus plantarum strains isolated from cheeses. Food Microbiol. 28:1033–1040. 2011.PubMed/NCBI View Article : Google Scholar
34	Succi M, Tremonte P, Reale A, Sorrentino E, Grazia L, Pacifico S and Coppola R: Bile salt and acid tolerance of Lactobacillus rhamnosus strains isolated from Parmigiano Reggiano cheese. FEMS Microbiol Lett. 244:129–137. 2005.PubMed/NCBI View Article : Google Scholar
35	Byakika S, Mukisa IM, Byaruhanga YB and Muyanja C: Probiotic potential of lactic acid starter cultures isolated from a traditional fermented sorghum-millet beverage. Int J Microbiol. 2020(7825943)2020.PubMed/NCBI View Article : Google Scholar
36	Fonseca HC, de Sousa Melo D, Ramos CL, Dias DR and Schwan RF: Probiotic properties of lactobacilli and their ability to inhibit the adhesion of enteropathogenic bacteria to Caco-2 and HT-29 cells. Probiotics Antimicrob Proteins. 13:102–112. 2021.PubMed/NCBI View Article : Google Scholar
37	Behbahani BA, Noshad M and Falah F: Inhibition of Escherichia coli adhesion to human intestinal Caco-2 cells by probiotic candidate Lactiplantibacillus plantarum strain L15. Microb Pathog. 136(103677)2019.PubMed/NCBI View Article : Google Scholar
38	Goh YJ and Klaenhammer TR: Functional roles of aggregation-promoting factor in stress tolerance and adherence of Lactiplantibacillus acidophilus NCFM. Appl Environ Microbiol. 76:5005–5012. 2010.PubMed/NCBI View Article : Google Scholar
39	Del Piano M, Morelli L, Strozzi GP, Allesina S, Barba M, Deidda F, Lorenzini P, Ballaré M, Montino F, Orsello M, et al: Probiotics: From research to consumer. Dig Liver Dis. 38 (Suppl 2):S248–S255. 2006.PubMed/NCBI View Article : Google Scholar