scholarly journals Deconjugation of Bile Acids with Immobilized Genetically EngineeredLactobacillus plantarum80(pCBH1)

2005 ◽  
Vol 2 (1) ◽  
pp. 31-38
Author(s):  
M. L. Jones ◽  
C. Martoni ◽  
H. Chen ◽  
W. Ouyang ◽  
T. Metz ◽  
...  
Keyword(s):  
Bile Acids ◽  
Deoxycholic Acid ◽  
Genetically Engineered ◽  
Bile Salt Hydrolase ◽  
Bacterial Cells ◽  
Normal Human ◽  
High Concentrations ◽  
Glycodeoxycholic Acid ◽  
Bsh Activity

Bile acids are important to normal human physiology. However, bile acids can be toxic when produced in pathologically high concentrations in hepatobileary and other diseases. This study shows that immobilized genetically engineeredLactobacillus plantarum80 (pCBH1) (LP80 (pCBH1)) can efficiently hydrolyze bile acids and establishes a basis for their use. Results show that immobilized LP80 (pCBH1) is able to effectively break down the conjugated bile acids into glycodeoxycholic acid (GDCA) and taurodeoxycholic acid (TDCA) with bile salt hydrolase (BSH) activities of 0.17 and 0.07 μmol DCA/mg CDW/h, respectively. The deconjugation product, deoxycholic acid (DCA), was diminished by LP80 (pCBH1) within 4 h of initial BSH activity. Thisin-vitrostudy suggests that immobilized genetically engineered bacterial cells have important potential for deconjugation of bile acids for lowering of high levels of bile acids for therapy.

scholarly journals In Vitro Bile Salt Hydrolase (BSH) Activity Screening of Different Probiotic Microorganisms

Foods ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 674
Author(s):  
Jimmy G. Hernández-Gómez ◽  
Argelia López-Bonilla ◽  
Gabriela Trejo-Tapia ◽  
Sandra V. Ávila-Reyes ◽  
Antonio R. Jiménez-Aparicio ◽  
...  
Keyword(s):  
Bile Salt ◽  
High Performance ◽  
Probiotic Strain ◽  
Saccharomyces Boulardii ◽  
Bile Salt Hydrolase ◽  
Sodium Glycocholate ◽  
Probiotic Yeast ◽  
Probiotic Strains ◽  
Bsh Activity

Bile salt hydrolase (BSH) activity in probiotic strains is usually correlated with the ability to lower serum cholesterol levels in hypercholesterolemic patients. The objective of this study was the evaluation of BSH in five probiotic strains of lactic acid bacteria (LAB) and a probiotic yeast. The activity was assessed using a qualitative direct plate test and a quantitative high-performance thin- layer chromatography assay. The six strains differed in their BSH substrate preference and activity. Lactobacillus plantarum DGIA1, a potentially probiotic strain isolated from a double cream cheese from Chiapas, Mexico, showed excellent deconjugation activities in the four tested bile acids (69, 100, 81, and 92% for sodium glycocholate, glycodeoxycholate, taurocholate, and taurodeoxycholate, respectively). In the case of the commercial probiotic yeast Saccharomyces boulardii, the deconjugation activities were good against sodium glycodeoxycholate, taurocholate, and taurodeoxycholate (100, 57, and 63%, respectively). These last two results are part of the novelty of the work. A weak deconjugative activity (5%) was observed in the case of sodium glycocholate. This is the first time that the BSH activity has been detected in this yeast.

Attachment of Salmonella to mammalian cells in vitro

1983 ◽  
Vol 29 (12) ◽  
pp. 1731-1735 ◽  
Author(s):  
Clifford S. Mintz ◽  
Dean O. Cliver ◽  
R. H. Deibel
Keyword(s):  
Cell Lines ◽  
Mammalian Cells ◽  
Culture Cell ◽  
Intestinal Cell ◽  
Tissue Culture Cell ◽  
Bacterial Cells ◽  
Intestinal Cell Line ◽  
Hela Cell Lines ◽  
High Concentrations

The attachment of Salmonella typhimurium strain PHL67342 to several mammalian tissue culture cell lines was investigated. Strain PHL67342 failed to attach in significant numbers to the Buffalo green monkey (BGM), swine testicular (ST), and HeLa cell lines. Significant attachment was observed with the Henle intestinal cell line. Log-phase cells of strain PHL67342 attached in greatest numbers to the Henle cells after 45 min of incubation at 37 °C. Attachment to the Henle cells was not affected by D-mannose or D-galactose, but was markedly inhibited by high concentrations of alpha-methyl-D-mannoside. Also, Salmonella lipopolysaccharide had no effect on the attachment of strain PHL67342 to the Henle cells. Fimbriae were not detected on the bacterial cells used in the adherence experiments. These results suggest that some bacterial factor(s) other than fimbriae and lipopolysaccharide mediate the attachment of strain PHL67342 to the Henle cells.

The Effect of Conjugated and Unconjugated Bile Acids on Water and Electrolyte Absorption in the Human Jejunum

1973 ◽  
Vol 45 (3) ◽  
pp. 301-311 ◽  
Author(s):  
R. I. Russell ◽  
J. G. Allan ◽  
V. P. Gerskowitch ◽  
K. M. Cochran
Keyword(s):  
Bile Acids ◽  
Deoxycholic Acid ◽  
Proximal Jejunum ◽  
Acid Solutions ◽  
Perfusion System ◽  
High Concentration ◽  
Electrolyte Absorption ◽  
Occluding Balloon ◽  
Glycodeoxycholic Acid ◽  
Inhibited Water

1. The effect of conjugated and unconjugated bile acids in various concentrations on the absorption of water, sodium and chloride in the proximal jejunum in man has been studied using a perfusion system incorporating a proximal occluding balloon. 2. Unconjugated cholic acid and its glycine and taurine conjugates had no significant effect on water and electrolyte absorption. Unconjugated deoxycholic acid significantly inhibited water and electrolyte absorption at 3 mm concentration and net secretion occurred at concentrations of 5 mm, 7 mm and 10 mm. Similar results were found when taurodeoxycholic and glycodeoxycholic acid solutions were used. A greater effect was noted when higher concentrations of bile acids were used. 3. Unconjugated chenodeoxycholic acid caused net water and electrolyte secretion at 3 mm concentration and this effect became more marked with higher bile acid concentrations. Inhibition of absorption also occurred with taurochenodeoxycholic acid in a concentration of 3 mm and net secretion occurred at a concentration of 10 mm. 4. The results show that dihydroxy bile acids, conjugated or unconjugated, inhibit water and electrolyte absorption in the proximal jejunum in low concentration and cause net secretion in high concentration.

scholarly journals Ketamine used in the therapy of depressive disorders impacts protein profile, proliferation rate and phagocytosis resistance of enterococci

2021 ◽  
Author(s):  
Tomasz Artur Jarzembowski ◽  
Agnieszka Daca ◽  
Wiesław J. Cubała ◽  
Marek Bronk ◽  
Łukasz Naumiuk
Keyword(s):  
Depressive Disorders ◽  
Early Stage ◽  
Protein Profile ◽  
Proliferation Rate ◽  
Bacterial Suspension ◽  
Bacterial Cells ◽  
High Concentrations ◽  
Etiological Agents ◽  
Different Response

Abstract Background: Ketamine is known to cause rapid anti-depressive effect. Additionally, it has been also proved that at high concentrations ketamine inhibits bacterial growth. It is also widely known that even sub-inhibitory concentration of chemicals, as concentration of ketamine used in therapy of depression, may change bacterial properties, including their virulence. The knowledge about possible influence of ketamine on bacterial commensals seems to be essential, as the mechanism of ketamine’s action in depression is believed to result also from its’ anti-inflammatory activity. In the current study we aimed to evaluate the in vitro influence of ketamine on proliferation rate, phagocytosis resistance and toxicity of enterococci. Results: The studied enterococcal strains were isolated as etiological agents of infection and collected in the Department of Medical Microbiology, Medical University of Gdansk. To measure metabolic activity of Enterococcus faecalis 10µM of CFDA-SE was added to bacterial suspension. The number of bacterial cells and fluorescence of particles were determined using FACSVerse flow cytometer. Additionally, for the determination of phagocytosis resistance, THP-1 human monocytes cell line from ATCC was used. Suspension of monocytes which engulfed bacteria was then stained with propidium iodide to determine cells’ membrane permeability and to evaluate cytotoxicity of enterococci. The result of the study proved diverse influence of therapeutic concentration of ketamine on Enterococci. In 23.1% of strains both proliferation rate and metabolism activity were inhibited. This group of strains was more susceptible to phagocytosis and had lower cytotoxicity than in culture without ketamine. Different response of isolates to ketamine was also visible in changes of protein profile determined by MALDI TOF. Conclusions: The analysis of bacteria at early stage of growth curve demonstrate the bacterial diversity in response to ketamine and let us set the hypothesis that microbiome susceptibility to ketamine may be one of the elements which should be taken into consideration when planning the successful pharmacotherapy of depression.

scholarly journals In vivo capture of bacterial cells by remote guiding

2021 ◽  
Author(s):  
Iaroslav A. Rybkin ◽  
Sergey I. Pinyaev ◽  
Olga A. Sindeeva ◽  
Sergey V. German ◽  
Maja Koblar ◽  
...  
Keyword(s):  
Genetic Material ◽  
Local Concentration ◽  
Bacterial Cells ◽  
Bacterial Strains ◽  
Toxic Side Effect ◽  
High Concentrations ◽  
Strong Barrier ◽  
Growth Distribution

Recently, it has been shown that several bacterial strains can be very efficient in cancer treatment since they possess many important properties such as self-targeting, ease of detection, sensing and toxicity against tumors. However, there are only a few relevant candidates for such an approach, as targeting and detection one of the biggest challenges as well as there are many limitations in the use of genetic approaches. Here, it is proposed the solution that enables surface modification of alive bacterial cells without interfering with their genetic material and potentially reduces their toxic side effect. By the electrostatic interaction fluorescently labeled polyelectrolytes (PEs) and magnetite nanoparticles (NPs) were deposited on the bacterial cell surface to control the cell growth, distribution and detection of bacteria. According to the results obtained in vivo, by the magnet entrapment of the modified bacteria the local concentration of the cells was increased more than 5 times, keeping the high concentrations even when the magnet is removed. Since the PEs create a strong barrier, in vitro it was shown that the division time of the cells can be regulated for better immune presentation.

scholarly journals Dissection of Catalytic Site in Crucial Gut Microbiome Enzyme: Bile Salt Hydrolase

2019 ◽  
Author(s):  
Yashpal Yadav ◽  
Mrityunjay K. Tiwari ◽  
Deepak Chand ◽  
Debjoyati Boral ◽  
Archana Pundle ◽  
...  
Keyword(s):  
Bile Salt ◽  
Bile Acids ◽  
Active Sites ◽  
Electrostatic Force ◽  
Enteric Bacteria ◽  
Significant Loss ◽  
Bile Salt Hydrolase ◽  
Hydrolysis Of ◽  
Determining Factor ◽  
Bsh Activity

AbstractBile Salt Hydrolases (BSHs) are enzymes from enteric bacteria that catalyze the hydrolysis of Bile Acids and consequently promote the reduction of cholesterol level in the mammalian body. Out of several reported BSHs, the Enterococcus faecalis BSH (EfBSH) has been reported to have the highest enzymatic activity. Herein, we have investigated the mechanistic details of the EfBSH activity. The study was carried out employing two mutants of EfBSH: E269A and R207A, which shows differential catalytic activity. The mutant E269A exhibits significant loss in the BSH activity with an increased affinity towards the substrate as compared to R207A mutant. Further, R207A was found to be involved in allostery with an increased EfBSH activity towards tauro-conjugated bile acids. The structural and electrostatic force analyses of the active sites of the E269A mutant and the wild type EfBSH (wt EfBSH) revealed that the interaction between Glu21 and Arg207 is the determining factor in maintaining the dynamic allostery and high activity of EfBSH.

scholarly journals Development of a Covalent Inhibitor of Gut Bacterial Bile Salt Hydrolases

2019 ◽  
Author(s):  
Arijit A. Adhikari ◽  
Tom C. Seegar ◽  
Scott B. Ficarro ◽  
Megan D. McCurry ◽  
Deepti Ramachandran ◽  
...  
Keyword(s):  
Bile Acid ◽  
Bile Salt ◽  
Bile Acids ◽  
Unmet Need ◽  
Cysteine Residue ◽  
Gut Bacteria ◽  
Bile Salt Hydrolase ◽  
Secondary Bile Acid ◽  
Bsh Activity

AbstractBile salt hydrolase (BSH) enzymes are widely expressed by human gut bacteria and catalyze the gateway reaction leading to secondary bile acid formation. Bile acids regulate key metabolic and immune processes by binding to host receptors. There is an unmet need for a potent tool to inhibit BSHs across all gut bacteria in order to study the effects of bile acids on host physiology. Here, we report the development of a covalent pan-inhibitor of gut bacterial BSH. From a rationally designed candidate library, we identified a lead compound bearing an alpha-fluoromethyl ketone warhead that modifies BSH at the catalytic cysteine residue. Strikingly, this inhibitor abolished BSH activity in conventional mouse feces. Mice gavaged with a single dose of this compound displayed decreased BSH activity and decreased deconjugated bile acid levels in feces. Our studies demonstrate the potential of a covalent BSH inhibitor to modulate bile acid composition in vivo.

scholarly journals Microencapsulated Genetically EngineeredLactobacillus plantarum80 (pCBH1) for Bile Acid Deconjugation and Its Implication in Lowering Cholesterol

2004 ◽  
Vol 2004 (1) ◽  
pp. 61-69 ◽  
Author(s):  
Mitchell Lawrence Jones ◽  
Hongmei Chen ◽  
Wei Ouyang ◽  
Terrence Metz ◽  
Satya Prakash
Keyword(s):  
Bile Acid ◽  
Bile Acids ◽  
Lactobacillus Plantarum ◽  
Blood Cholesterol ◽  
Bile Salt Hydrolase ◽  
Regular Exercise ◽  
Bile Acid Sequestrants ◽  
Oral Doses ◽  
Glycodeoxycholic Acid ◽  
Conjugated Bile Acids

Cholesterol is known to be a major risk factor for coronary heart disease (CHD). Current treatments for elevated blood cholesterol include dietary management, regular exercise, and drug therapy with fibrates, bile acid sequestrants, and statins. Such therapies, however, are often suboptimal and carry a risk for serious side effects. This study shows that microencapsulatedLactobacillus plantarum80 (pCBH1) cells can efficiently break down and remove bile acids, and establishes a basis for their use in lowering blood serum cholesterol. Results show that microencapsulated LP80 (pCBH1) is able to effectively break down the conjugated bile acids glycodeoxycholic acid (GDCA) and taurodeoxycholic acid (TDCA) with bile salt hydrolase (BSH) activities of 0.19 and 0.08μmol DCA/mg CDW/h respectively. This article also summarizes the physiological interrelationship between bile acids and cholesterol and predicts the oral doses of microencapsulatedLactobacillus plantarum80 (pCBH1) cells required for lowering cholesterol.

scholarly journals Deoxycholic acid activates colonic afferent nerves via 5-HT3 receptor-dependent and -independent mechanisms

2019 ◽  
Vol 317 (3) ◽  
pp. G275-G284 ◽  
Author(s):  
Yang Yu ◽  
Egina C. Villalobos-Hernandez ◽  
Sabindra Pradhananga ◽  
Corey C. Baker ◽  
Christopher Keating ◽  
...  
Keyword(s):  
Bile Acids ◽  
Deoxycholic Acid ◽  
Neuronal Cell ◽  
Distal Colon ◽  
Proximal Colon ◽  
High Threshold ◽  
Secondary Bile Acid ◽  
Afferent Nerves ◽  
Spinal Afferents

Increased bile acids in the colon can evoke increased epithelial secretion resulting in diarrhea, but little is known about whether colonic bile acids contribute to abdominal pain. This study aimed to investigate the mechanisms underlying activation of colonic extrinsic afferent nerves and their neuronal cell bodies by a major secondary bile acid, deoxycholic acid (DCA). All experiments were performed on male C57BL/6 mice. Afferent sensitivity was evaluated using in vitro extracellular recordings from mesenteric nerves in the proximal colon (innervated by vagal and spinal afferents) and distal colon (spinal afferents only). Neuronal excitability of cultured dorsal root ganglion (DRG) and nodose ganglion (NG) neurons was examined with perforated patch clamp. Colonic 5-HT release was assessed using ELISA, and 5-HT immunoreactive enterochromaffin (EC) cells were quantified. Intraluminal DCA increased afferent nerve firing rate concentration dependently in both proximal and distal colon. This DCA-elicited increase was significantly inhibited by a 5-HT3 antagonist in the proximal colon but not in the distal colon, which may be in part due to lower 5-HT immunoreactive EC cell density and lower 5-HT levels in the distal colon following DCA stimulation. DCA increased the excitability of DRG neurons, whereas it decreased the excitability of NG neurons. DCA potentiated mechanosensitivity of high-threshold spinal afferents independent of 5-HT release. Together, this study suggests that DCA can excite colonic afferents via direct and indirect mechanisms but the predominant mechanism may differ between vagal and spinal afferents. Furthermore, DCA increased mechanosensitivity of high-threshold spinal afferents and may be a mechanism of visceral hypersensitivity. NEW & NOTEWORTHY Deoxycholic acid (DCA) directly excites spinal afferents and, to a lesser extent, indirectly via mucosal 5-HT release. DCA potentiates mechanosensitivity of high-threshold spinal afferents independent of 5-HT release. DCA increases vagal afferent firing in proximal colon via 5-HT release but directly inhibits the excitability of their cell bodies.

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