INTRODUCTION:

Lactic acid bacteria (LAB) are living microorganisms capable of creating, among other substances, lactic acid, hydrogen peroxide, and bacteriocins. Lactic acid bacteria are typically considered safe and can assist the host by producing antibacterial compounds, adherence to the intestinal epithelium, and reducing pathogenic microbe adhesion.¹ Probiotic bacteria have positive health effects, including lowering cholesterol levels.^2,3,4Probiotic bacteria' primary value lies in their ability to keep the host's gut flora in check.¹

In addition to cholesterol absorption and enhanced disease resistance and immunology, a number of probiotic microorganisms have been identified to give other advantages.^5,6 Several LAB, particularly those belonging to the genera Bifidobacterium and Lactobacillus, have been found to exhibit potent probiotic properties.^7,8 Lactic acid bacteria, particularly Lactobacillus, are the most common probiotic bacteria.^9,10Multiple lactic acid bacterial species have shown high cholesterol-cholesterol activity in both animal and human studies.^11,12,13,14

The amount of food consumed may affect the blood's overall cholesterol levels. The main risk factors for cardiovascular and cerebrovascular disease include high levels of triglycerides and cholesterol in the blood and diet.^15,16Because of the expenses and adverse effects that are still present, pharmacological therapy is not yet regarded an optimal solution for this condition. Therefore, employing natural foods and harmless microbes to lower dietary and blood cholesterol and triglyceride levels is an effective technique. Recently, many LAB with possible cholesterol-lowering activities have been found.^15,16,17Probiotics have been demonstrated to have a positive effect on cholesterol levels via many mechanisms. These mechanisms include the absorption of probiotic cholesterol, the deconjugation of bile acids through enzymatic activity, and the deconjugation of bile acids. bile, including cholesterol binding to probiotic cell walls, cholesterol absorption into probiotic cell membranes, cholesterol conversion to coprostanol, and short chain fatty acid (SCFA) synthesis by probiotic fermentation in the presence of prebiotics.¹⁸ The ability of probiotic bacteria to deconjugate bile acids is an important factor that allows these bacteria to tolerate bile acids in the digestive tract. Lactic acid bacteria (LAB) are a type of bacteria that is often used as a probiotic.

Most Probiotic lactic acid bacteria strains that reduce cholesterol in humans are found in fermented product; vegetables¹⁹, dairy products, meat products²⁰, seafood²¹ and human or animal waste (feces)²². Dadiah is a fermented food made from buffalo milk that has a probiotic-potential microbiome of lactic acid bacteria. One of them is Lactobacillus plantarum 2DA which was isolated from Payakumbuhdadiah which has the potential as a source of probiotics.²³Several factors influence the cholesterol-lowering ability of lactic acid bacteria; the removal of intestinal cholesterol by probiotic cells^24,25, inhibition of small intestinal cholesterol absorption by regulation of intestinal NPC1L1 protein levels²⁶, and increased faecal bile acid excretion by bile salt deconjugation catalysed by bile salt hydrolase (BSH) from probiotic cells are the three main proposed mechanisms.^14,27

The particular mechanism responsible for the cholesterol-lowering activity is still unknown, according to experts.Using a bioinformatics method, it is therefore necessary to examine how well LAB can reduce cholesterol in terms of secondary metabolites. Biionformatics approach can be used to determine the ability of Lactobacillus plantarum 2DA as an anticholesterol.

Methods:

Bacteria used were Lactobacillus plantarum 2DA culture isolated from the Payakumbuhdadiah collection from the Animal Products Technology Laboratory, Andalas University.

Search of Secondary Metabolite Search:

Search for bacterial secondary metabolites was carried out by studying literature from published journals. Each compound that has been determined is then searched for its canonical structure and isomeric SMILE (simplified molecular-input line-entry system) in the PubChem database (https://pubchem.ncbi.nlm.nih.gov/).

Prediction of bacterial content's Structure-activity Relationship (SAR) as a cholesterol-lowering treatment:

Secondary metabolites in bacteria from the literature study were analyzed for their potential using WAY2DRUG PASS prediction (http://www.pharmaexpert.ru/passonline/predict.php) as a cholesterol-lowering treatment. Previously, each compound needed to find the SMILE structure obtained from the PubChem database. Then the potency of the compound was analyzed using WAY2DRUG PASS prediction to determine its potential for cholesterol reduction treatment. We recommend using a score of 0.5 as the cut off score. The Pa value means the accuracy of the prediction function obtained, the higher the Pa value of a function, the better the accuracy level.²⁸

RESULT AND DISCUSSION:

The following table shows the results of SAR predictions.

Table 1: Prediction Results of SAR Way2 Drug Pass

Compound	Anti hyper Cholesterol -emic	Cholesterol Antagonist	Cholesterol oxidase inhibitor
Butyric acid	0.58	0.71	0.52
Propionic acid	0.61	0.64	0.48
Lactic acid	0.5	0.62	0.49
Valeric acid	0.61	0.72	0.53
Caproic acid	0.61	0.72	0.53
Rata-rata	0.45	0.49	0.37

The value of Pa (Probability to be Active) reflects the potential of a compound being examined. This value is determined by comparing the structure of the input chemical to a compound that has been demonstrated to cure a certain condition. Potential analysis is done using Way2Drug Pass Server. If the Pa value is more than 0.7, the compound is expected to have a high potential as an antidiabetic owing to its high similarity to a proven therapeutic molecule in the database.

Table 1 can be seen based on the Prediction of SAR Way2 Drug Pass Server secondary metabolites of Lactobacillus plantarum 2DA on antihypercholesterolemic 0.45, Cholesterol antagonist 0.49 and Cholesterol oxidase inhibitor 0.37. Where the best result in the SAR Way2 Drug Pass Server prediction is 0.7, where Lactobacillus plantarum 2DA is the best as a Cholesterol antagonist. This is because L. plantarum 2DA is included as a probiotic lactic acid bacteria so that the prediction results can be applied in vivo in experimental animals and in vitro with application to food products. Probiotic bile salt hydrolases' enzymatic deconjugation of bile acids is one of the methods that have been suggested.When food is consumed, the gallbladder retains and concentrates bile, a consequence of the liver's production of water-soluble cholesterol, before releasing it into the duodenum.²⁹This concoction includes electrolytes, conjugated bile acids, bile pigments, phospholipids, and cholesterol. Deconjugated bile acids are absorbed by the intestines and passed through the faeces because they are less soluble than conjugated bile acids. In a process known as homeostasis, cholesterol is needed to create new bile acids, which lowers blood cholesterol.³⁰ Jones et al.³¹ used Lactobacillus plantarum to test the effect of bile salt hydrolase in decreasing cholesterol in an in vitro investigation. According to the researchers, BSH activity may hydrolyze conjugated glycodeoxycholic acid and taurodeoxycholic acid, leading to the deconjugation of glycodeoxycholic acid and taurobylic acid. Probiotics' hypocholesterolemic effects are also related to their capacity to bind cholesterol in the small intestine.

Amino acids butyric acid, valric acid and caproic acid have high values for cholesterol antagonists with Pa values of 0.71, 0.72 and 0.72 meaning they have the highest values in the SAR prediction results so that they can be recommended as therapeutic media to lower cholesterol. Usman³²a particular strain of Lactobacillus gasseri has previously been shown to absorb cholesterol from cellular surfaces in laboratory systems. It seems that the capacity to bind cholesterol is development and strain dependent. The concept was then supported by Kimoto et al.³¹ who tested cholesterol elimination by probiotic cells under various growing conditions. Cells that were both alive and not growing (suspended in phosphate buffer) were contrasted with those that were both alive and not growing. Cholesterol could still be extracted from the media by heat-killed cells, showing that some cholesterol remained attached to the cell surface. By permeating cell membranes during growth, probiotics also remove cholesterol. Kimoto et al.³¹ investigated how different Lactococcus strains removed cholesterol from the medium. Cells developing with and without cholesterol showed different patterns of fatty acid distribution.

Membranes were shown to contain the majority of probiotic lipids, implying that cholesterol delivered into cell membranes changed the fatty acid makeup of cells. The content of saturated and unsaturated fatty acids rises as a consequence of cholesterol absorption into cell membranes, increasing membrane strength and cellular resistance to lysis.^32,33 The site of cholesterol incorporation inside the phospholipid bilayer of probiotic cell membranes was investigated further.³² The fluorescent probe was introduced into the bilayer membrane of probiotic cells growing with or without cholesterol. The polar head, top phospholipid region, and tail area of the phospholipid bilayer cell membrane all exhibited cholesterol incorporation when cells developing in the presence of cholesterol were contrasted with control cells. Cholesterol may be converted to coprostanol in the stomach, which is then expelled directly in the faeces. This reduces the quantity of cholesterol absorbed, hence lowering the concentration of cholesterol in the physiological pool. Chiang et al.³⁴investigated if microorganisms could convert cholesterol to coprostanol. If seen in table 1, the amino acid compounds produced by L. plantarum 2DA such as butyric acid, propionic acid, lactic acid, valeric acid and caproic acid are also capable of producing metabolites that can be used as a source of therapy to lower cholesterol. Where in addition to these amino acid compounds, probiotics generally also produce peptides which are also able to lower cholesterol. Exopolysaccharides released by probiotics, as well as the amount of amino acids and peptidoglycan in probiotic cell walls, may explain the strong contact force between living and dead probiotic cells that lower cholesterol in culture media^35,36 The decrease of cholesterol by this pathway is, however, strain and growth dependent.³¹Anti hypercholesterolemic in L. plantarum was highest in amino propionic acid, valeric acid and caproic acid where the Pa value was close to the highest level, namely 0.7. This of course can be recommended as hypercholesterolemic therapy because it is able to assimilate cholesterol. The risk of cardiovascular disease is thought to be reduced by cholesterol assimilation, which is thought to enhance cholesterol excretion in the stool and decrease cholesterol absorption by enterocytes. Although animal studies are necessary to demonstrate its anti-hypercholesterolemic efficacy, this might suggest that this isolate could be used as a probiotic to lower cholesterol levels. Cholesterol-lowering action of probiotics In vitro tests, such as those employed in this study, may be used to predict cholesterol-lowering efficacy in vivo.³⁷Since high cholesterol levels are directly linked to cardiovascular disease and are a leading cause of death, the search for probiotic candidates with potential anti-hypercholesterolemic effect is imperative. A blood cholesterol level of greater than 240mg/dL is referred to medically as hypercholesterolemia.³⁸

Probiotics may help regulate cholesterol levels in the blood. Therefore, the ability of probiotics to decrease cholesterol is now thought to be a key factor in the selection of bacterial strains for use in human probiotics. A number of processes, including the synthesis of BSH, have been connected to the hypocholesterolemic effects of probiotics.^39,2 the ability to assimilate cholesterol and incorporate it into cellular membranes^40,41 and the production of ferulic acid^32,42 which can inhibit the activity of hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase.⁴³Probiotics' is a functional food that contains a living microbe that, when given to the host in sufficient quantities, has a beneficial effect on the host's health.^44-50Probiotics' ability to bind cholesterol in the small intestine is linked to their hypocholesterolemic impact. The enzyme bile salt hydrolase (BSH), which catalyses the deconjugation of bile salts throughout their enterohepatic circulation, is one of the most crucial processes. Glyco- and tauro-bile acid deconjugation occurred due to BSH activity hydrolyzing conjugated forms of glycodeoxycholic acid and taurodeoxycholic acid.

CONCLUSION:

Based on the Pa (Probability to Be Active) value on anti-hypercholesterolemic, cholesterol antagonist, and Cholesterol oxidase inhibitors, Lactobacillus plantarum 2DA bacteria isolated from dadiahpayakumbuh can be recommended for cholesterol lowering therapy in terms of the Pa (Probability to Be Active) value on anti-hypercholesterolemic, cholesterol antagonist, and Cholesterol oxidase inhibitors, so further research

ACKNOWLEDGEMENT:

Authors are acknowledging the help and supports byBaiturrahmah University.

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Received on 29.09.2022 Modified on 10.12.2022

Research J. Pharm. and Tech 2023; 16(12):5915-5919.

DOI: 10.52711/0974-360X.2023.00959