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.
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