Isolation and identification of bile salt hydrolase-producing lactic acid bacteria with cholesterol-lowering potential probiotics in raw milk from dairy farms in Thailand
Keywords:
raw milk, lactic acid bacteria, probiotics, Lactobacillus, Lacticaseibacillus, bile salt hydrolase, cholesterolAbstract
Currently, probiotics are widely available and used in daily life, with lactic acid bacteria being the most common source found in fermented foods, the normal microbiota of humans and animals, plants, or in the environment. This study aimed to discover lactic acid bacteria with positive bile salt hydrolase (BSH) activity, an enzyme with the potential mechanism of decreasing cholesterol levels, from 21 raw milk samples collected from dairy farms or dairy cooperatives in Thailand. From these samples, 103 lactic bacteria were isolated, and 15 isolates exhibited bile salt hydrolase activity. Among them, three isolates with the highest BSH activities, namely TM1, TM7, and TM14, were selected for further screening. Molecular identification using 16S rDNA sequences revealed that TM1 was closely related to Lactobacillus gasseri ATCC33323, while TM7 and TM14 were closely related to Lacticaseibacillus rhamnosus strain NBRC 3425. The phylogenetic tree confirmed these results. Additionally, TM7 was notably distant from TM14 and other L. rhamnosus clade members, suggesting a possible new strain of L. rhamnosus. Acid tolerance tests showed that all strains were tolerant to acidity at pH 3.0 and pH 4.0 for the duration of normal gastric emptying, with TM14 even exhibiting an increase in viable cell population at pH 4.0. In the bile salt challenge test, TM1 showed an increase in growth rate with increasing bile salt concentration, indicating its preference for bile salts in its growth conditions. At a bile salt concentration of 0.3%, viable cells of TM7 and TM14 could be detected, suggesting bile salt tolerance. Moreover, all three isolates exhibited inhibition of pathogenic bacteria such as Vibrio parahaemolyticus, Klebsiella pneumoniae, Escherichia coli, and Serratia marcescens, without showing signs of antibiotic resistance to ampicillin, chloramphenicol, and tetracycline. These findings advocate further assessment and consider them as candidates for probiotic product development.
References
Martino C, Dilmore AH, Burcham ZM, et al. Microbiota succession throughout life from the cradle to the grave. Nat Rev Microbiol 2022;20:707-20. doi:10.1038/s41579-022-00768-z.
Jin M, Qian Z, Yin J, et al. The role of intestinal microbiota in cardiovascular disease. J Cell Mol Med 2019;23:2343-50. doi:10.1111/jcmm.14195.
Papa A, Santini P, De Lucia SS, et al. Gut dysbiosis-related thrombosis in inflammatory bowel disease: Potential disease mechanisms and emerging therapeutic strategies. Thromb Res 2023;232:77-88. doi:10.1016/j.thromres.2023.11.005.
Dahal RH, Kim S, Kim YK, et al. Insight into gut dysbiosis of patients with inflammatory bowel disease and ischemic colitis. Front Microbiol 2023;14:1174832. doi:10.3389/fmicb.2023.1174832.
Patra D, Banerjee D, Ramprasad P, et al. Dasgupta S. Recent insights of obesityinduced gut and adipose tissue dysbiosis in type 2 diabetes. Front Mol Biosci 2023;10:1224982. doi:10.3389/fmolb.2023.1224982.
Hamjane N, Mechita MB, Nourouti NG, et al. Gut microbiota dysbiosis –associated obesity and its involvement in cardiovascular diseases and type 2 diabetes. A systematic review. Microvasc Res 2023;151:104601. doi:10.1016/j.mvr.2023.104601.
Aslan I, Tarhan Celebi L, Kayhan H, et al. Probiotic Formulations Containing Fixed and Essential Oils Ameliorates SIBO-Induced Gut Dysbiosis in Rats. Pharmaceuticals (Basel) 2023;16. doi:10.3390/ph16071041.
Kim S, Lee S, Kim TY, et al. Newly isolated Lactobacillus paracasei strain modulates lung immunity and improves the capacity to cope with influenza virus infection. Microbiome 2023;11:260. doi:10.1186/s40168-023-01687-8.
Kleniewska P, Kopa-Stojak PN, Hoffmann A, et al. The potential immunomodulatory role of the gut microbiota in the pathogenesis of asthma: An in vitro study. Sci Rep 2023;13:19721. doi:10.1038/s41598-023-47003-0.
Takeuchi T, Kubota T, Nakanishi Y, et al. Gut microbial carbohydrate metabolism contributes to insulin resistance. Nature 2023;621:389-95. doi:10.1038/s41586-023-06466-x.
Gominak SC. Vitamin D deficiency changes the intestinal microbiome reducing B vitamin production in the gut. The resulting lack of pantothenic acid adversely affects the immune system, producing a “proinflammatory” state associated with atherosclerosis and autoimmunity. Med Hypotheses 2016;94:103-7. doi:10.1016/j.mehy.2016.07.007.
Jia B, Zou Y, Han X, et al. Gut microbiomemediated mechanisms for reducing cholesterol levels: implications for ameliorating cardiovascular disease. Trends Microbiol 2023;31:76-91. doi:10.1016/j.tim.2022.08.003.
Hernandez-Gomez JG, Lopez-Bonilla A, Trejo-Tapia G, et al. In Vitro Bile Salt Hydrolase (BSH) Activity Screening of Different Probiotic Microorganisms. Foods 2021;10. doi:10.3390/foods10030674.
Jittapranerat J, Saetiew K, Limtrakul A, et al. Isolation and characterization of lactic acid bacteria with bile salt hydrolase activity. J Med Health Sci 2020;27:77-91.
Kaya Y, Kök MŞ, Öztürk M. Molecular cloning, expression and characterization of bile salt hydrolase from Lactobacillus rhamnosus E9 strain. Food Biotechnology 2017;31:128-40. doi:10.1080/08905436.2017.1303778.
Shehata MG, El Sohaimy SA, El-Sahn MA, et al. Screening of isolated potential probiotic lactic acid bacteria for cholesterol lowering property and bile salt hydrolase activity. Annals of Agricultural Sciences 2016;61:65-75. doi:10.1016/j.aoas.2016.03.001.
Jamjuree P, Kasorn A, Vitheejongjaroen P, et al. Characterization and product formulation of cholesterol-lowering probiotic strains for prevention of metabolic syndrome. J Med Health Sci 2023;30:114-30.
S AF, G Z, O SOA, et al. Antimicrobial activity and Probiotic Properties of Lactic Acid Bacteria Isolated From Traditional Fermented Dairy Products. Journal of Modern Research 2020;2:40-8. doi:10.21608/jmr.2020.22931.1015.
B S, J M, L OM, et al. Is the mucosal route of administration essential for probiotic function? Subcutaneous administration is associated with attenuation of murine colitis and arthritis. Gut. Gut 2004;53:694-700. doi:10.1136/gut.2003.027789.
Yang Z, Li C, Wang T, et al. Novel gellan gum-based probiotic film with enhanced biological activity and probiotic viability: Application for fresh-cut apples and potatoes. Int J Biol Macromol 2023;239:124128. doi:10.1016/j.ijbiomac.2023.124128.
Hamdy SM, Danial AW, Halawani EM, et al. Biofabrication strategy of silvernanodrug conjugated polyhydroxybutyrate degrading probiotic and its application as a novel wound dressing. Int J Biol Macromol 2023;250:126219. doi:10.1016/j.ijbiomac.2023.126219.
Kamel DG, Hammam ARA, Nagm El-Diin MAH, et al. Abdel-Rahman AM. Nutritional, antioxidant, and antimicrobial assessment of carrot powder and its application as a functional ingredient in probiotic soft cheese. J Dairy Sci 2023;106:1672-86. doi:10.3168/jds.2022-22090.
Haghani S, Hadidi M, Pouramin S, et al. Application of Cornelian Cherry (Cornus mas L.) Peel in Probiotic Ice Cream: Functionality and Viability during Storage. Antioxidants (Basel) 2021;10. doi:10.3390/antiox10111777.
Labenz J, Borkenstein DP, Heil FJ, et al. Application of a multispecies probiotic reduces gastro-intestinal discomfort and induces microbial changes after colonoscopy. Front Oncol 2022;12: 1078315. doi:10.3389/fonc.2022.1078315.
Park MK, Ngo V, Kwon YM, et al. Lactobacillus plantarum DK119 as a probiotic confers protection against influenza virus by modulating innate immunity. PLoS One 2013;8:e75368. doi:10.1371/journal.pone.0075368.
Maruscakova IC, Schusterova P, Bielik B, et al. Effect of Application of Probiotic Pollen Suspension on Immune Response and Gut Microbiota of Honey Bees (Apis mellifera). Probiotics Antimicrob Proteins 2020;12:929-36. doi:10.1007/s12602-019-09626-6.
Javed S, Munir A, Javed GA, et al. Genetic diversity, cholesterol reduction, and presence of conserved bile salt hydrolase gene in probiotic strains from human milk. Lett Appl Microbiol 2023;76. doi:10.1093/lambio/ovad024.
Jones ML, Tomaro-Duchesneau C, Martoni CJ, et al. Cholesterol lowering with bile salt hydrolase-active probiotic bacteria, mechanism of action, clinical evidence, and future direction for heart health applications. Expert Opin Biol Ther 2013;13:631-42. doi:10.1517/14712598.2013.758706.
Solana MJ, Lopez-Herce J, Sanchez C, et al. Comparison of continuous pH-meter and intermittent pH paper monitoring of gastric pH in critically ill children. Eur J Gastroenterol Hepatol 2012;24:33-6. doi:10.1097/MEG.0b013e32834d4bac.
Ahmed S, Sharif MK, Butt MS, et al. Development of nutritious probiotic sportsman drink. Pak J Pharm Sci 2020;33:191-7. Available from: https://pubmed.ncbi.nlm.nih.gov/32122848/.
Vitale K, Getzin A. Nutrition and Supplement Update for the Endurance Athlete: Review and Recommendations. Nutrients 2019;11. doi:10.3390nu11061289.
Goodman C, Keating G, Georgousopoulou E, et al. Probiotics for the prevention of antibiotic-associated diarrhoea: A systematic review and meta-analysis. BMJ Open 2021;11:e043054. doi:10.1136/bmjopen-2020-043054.
Zhang C, Gao X, Ren X, et al. BacteriaInduced Colloidal Encapsulation for Probiotic Oral Delivery. ACS Nano 2023;17:6886-98. doi:10.1021acsnano.3c00600.
Gan D, Chen J, Tang X, et al. Impact of a probiotic chewable tablet on stool habits and microbial profile in children with functional constipation: A randomized controlled clinical trial. Front Microbiol 2022;13:985308. doi:10.3389/fmicb.2022.985308.
Azhar MA, Munaim MSA. Design and optimization of a probiotic tablet for gastrointestinal tolerance by a simplexcentroid mixture. Drug Dev Ind Pharm 2021;47:189-96. doi:10.1080/03639045.2020.1862176.
Wang L, Fang M, Hu Y, et al. Characterization of the most abundant Lactobacillus species in chicken gastrointestinal tract and potential use as probiotics for genetic engineering Acta Biochimica et Biophysica Sinica 2014;46:8. doi:https://doi.org/10.1093/abbs/gmu037.
Farhangfar A, Gandomi H, Akhondzadeh Basti A, et al. Noori N. Study of growth kinetic and gastrointestinal stability of acid-bile resistant Lactobacillus plantarum strains isolated from Siahmazgi traditional cheese. Vet Res Forum 2021;12:235-40. doi:10.30466/vrf.2019.101880.2428.
Liu W, Chen M, Duo L, et al. Characterization of potentially probiotic lactic acid bacteria and bifidobacteria isolated from human colostrum. J Dairy Sci 2020;103:4013-25. doi:10.3168/jds.2019-17602.
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