scholarly journals Molecular Cloning cysK Gene from Escherichia coli Genome, Transferring in the Intestinal Sulfate-Reducing Bacteria and the Expression Analysis of O-acetylserine(thiol)lyase

2016 ◽  
Vol 4 (1) ◽  
pp. 19-24 ◽  
Author(s):  
Ivan V Kushkevych
Keyword(s):  
Escherichia Coli ◽  
Ulcerative Colitis ◽  
Inflammatory Bowel Disease ◽  
Sulfate Reducing Bacteria ◽  
Wild Type ◽  
Mutant Type ◽  
Sulfate Reducing ◽  
Cysteine Synthesis ◽  
Inflammatory Bowel ◽  
Reducing Bacteria

Sulfate-reducing bacteria produce hydrogen sulfide which is toxic and carcinogenic for intestinal epithelial cells and can cause the development of the inflammatory bowel disease and ulcerative colitis in the humans and animals. Enzyme O-acetylserine(thiol)lyase, localized in Escherichia coli genome, use sulfide as substrate in the cysteine synthesis pathway. In this paper, the molecular cloning cysK gene from E. coli, its genetic transferring in the intestinal sulfate-reducing bacterium Desulfovibrio piger Vib-7 and the expression analysis of the enzyme was studied. Cysteine synthesis from hydrogen sulfide as substrate in the D. piger Vib-7 strain at the first time was demonstrated and characterized. The bacterial growth, sulfate and lactate consumption, accumulation of sulfide, acetate and cysteine synthesis in both D. piger Vib-7 wild-type and mutant-type were tested. The mutant-strain consumed much faster sulfate and lactate producing cysteine in the cultivation medium. The expression of the cysK gene in the mutant-type was studied by the formation of the final reaction product (cysteine) and the activity of O-acetylserine(thiol)lyase enzyme. Cysteine level was directly proportional to consumption of sulfate in the mutant-type and accumulation of sulfide in the wild-type. The D. piger Vib-7 mutant-type completely used sulfate the 48th hour of cultivation, thereafter additional sulfite and sulfide doses from the medium were also consumed and converted to cysteine. The obtained genetically constructed mutant strain bacterium D. piger Vib-7 for therapeutic strategy could be applied as a probiotic substance for subjects with inflammatory bowel disease and ulcerative colitis. This strain can compete with other intestinal sulfate-reducing bacteria, actively growth consuming sulfate and lactate much faster, and converting the toxic sulfide to untoxic cysteine in the gut.Microbes and Health, January 2015. 4(1): 19-24

scholarly journals Analysis of pH dose-dependent growth of sulfate-reducing bacteria

Open Medicine ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. 66-74 ◽  
Author(s):  
Ivan Kushkevych ◽  
Dani Dordević ◽  
Monika Vítězová
Keyword(s):  
Ulcerative Colitis ◽  
Hydrogen Sulfide ◽  
Sulfate Reduction ◽  
Sulfate Reducing Bacteria ◽  
Alkaline Media ◽  
Dissimilatory Sulfate Reduction ◽  
Lactate Oxidation ◽  
Sulfate Reducing ◽  
Inflammatory Bowel ◽  
Reducing Bacteria

AbstractLower intraluminal colonic pH is an indication for the development of inflammatory bowel disease including active ulcerative colitis. Involvement of intestinal sulfate-reducing bacteria in decreasing bowel pH by the production of H2S and acetate as well as their sensitivity has never been reported before. The study of the relative pH and survival of Desulfovibrio piger Vib-7 by monitoring sulfate reduction parameters was the aim of this work. Monitoring was done through the measurement of bacterial growth (biomass), dissimilatory sulfate reduction parameters: sulfate consumption, lactate oxidation, hydrogen sulfide and acetate production. According to our results, we observed that lower pH (<5) significantly inhibited D. piger Vib-7 growth. This inhibition was also noticed when alkaline media (>9 pH) was used, though the reduction was not at the rate as in media with pH of 4. The research indicates that the growth of D. piger Vib-7 is inhibited at pH of 4 which is not as low as the pH found in people with severely developed inflammatory bowel diseases such as ulcerative colitis. Certainly the interaction (synergistic effect) between both hydrogen sulfide and acetate accumulation can also play an important etiological role in the development of bowel inflammation in humans and animals.

scholarly journals Kinetic properties of pyruvate ferredoxin oxidoreductase of intestinal sulfate-reducing bacteria Desulfovibrio piger Vib-7 and Desulfomicrobium sp. Rod-9

2015 ◽  
Vol 64 (2) ◽  
pp. 107-114 ◽  
Author(s):  
IVAN V. KUSHKEVYCH
Keyword(s):  
Enzyme Reaction ◽  
Sulfate Reducing Bacteria ◽  
Kinetic Properties ◽  
Bacterial Strains ◽  
Ferredoxin Oxidoreductase ◽  
Sulfate Reducing ◽  
Bowel Diseases ◽  
Inflammatory Bowel ◽  
Pyruvate Ferredoxin Oxidoreductase ◽  
Reducing Bacteria

Intestinal sulfate-reducing bacteria reduce sulfate ions to hydrogen sulfide causing inflammatory bowel diseases of humans and animals. The bacteria consume lactate as electron donor which is oxidized to acetate via pyruvate in process of the dissimilatory sulfate reduction. Pyruvate-ferredoxin oxidoreductase activity and the kinetic properties of the enzyme from intestinal sulfate-reducing bacteria Desulfovibrio piger and Desulfomicrobium sp. have never been well-characterized and have not been yet studied. In this paper we present for the first time the specific activity of pyruvate-ferredoxin oxidoreductase and the kinetic properties of the enzyme in cell-free extracts of both D. piger Vib-7 and Desulfomicrobium sp. Rod-9 intestinal bacterial strains. Microbiological, biochemical, biophysical and statistical methods were used in this work. The optimal temperature (+35°C) and pH 8.5 for enzyme reaction were determined. The spectral analysis of the puri- fied pyruvate-ferredoxin oxidoreductase from the cell-free extracts was demonstrated. Analysis of the kinetic properties of the studied enzyme was carried out. Initial (instantaneous) reaction velocity (V0), maximum amount of the product of reaction (Pmax), the reaction time (half saturation period) and maximum velocity of the pyruvate-ferredoxin oxidoreductase reaction (V ) were defined. Michaelis constants (Km) of the enzyme reaction were calculated for both intestinal bacterial strains. The studies of the kinetic enzyme properties in the intestinal sulfate-reducing bacteria strains in detail can be prospects for clarifying the etiological role of these bacteria in the development of inflammatory bowel diseases.

scholarly journals Metabolic activity of sulfate-reducing bacteria from rodents with colitis

Open Medicine ◽  
2018 ◽  
Vol 13 (1) ◽  
pp. 344-349 ◽  
Author(s):  
Jozef Kováč ◽  
Monika Vítězová ◽  
Ivan Kushkevych
Keyword(s):  
Ulcerative Colitis ◽  
Large Intestine ◽  
Sulfate Reducing Bacteria ◽  
Dissimilatory Sulfate Reduction ◽  
Healthy Human ◽  
Sulfate Reducing ◽  
Anaerobic Microorganisms ◽  
The Difference ◽  
High Concentrations ◽  
Reducing Bacteria

AbstractSulfate-reducing bacteria (SRB) are anaerobic microorganisms, which use sulfate as an electron acceptor in the process of dissimilatory sulfate reduction. The final metabolic product of these anaerobic microorganisms is hydrogen sulfide, which is known as toxic and can lead to damage to epithelial cells of the large intestine at high concentrations. Different genera of SRB are detected in the large intestine of healthy human and animals, and with diseases like Crohn’s disease and ulcerative colitis. SRB isolated from rodents with ulcerative colitis have produced 1.14 (mice) and 1.03 (rats) times more sulfide ions than healthy rodents. The species ofDesulfovibriogenus are the most widespread among all SRB in the intestine. The object of our research was to observe and compare the difference of production of sulfide and reduction of sulfate in intestinal SRB isolated from healthy rodents and rodents with ulcerative colitis.

scholarly journals Analysis of physiological parameters of Desulfovibrio strains from individuals with colitis

2019 ◽  
Vol 13 (1) ◽  
pp. 481-488 ◽  
Author(s):  
Ivan Kushkevych ◽  
Dani Dordević ◽  
Peter Kollár
Keyword(s):  
Ulcerative Colitis ◽  
Bowel Disease ◽  
Principal Component ◽  
Biomass Accumulation ◽  
Sulfate Reducing Bacteria ◽  
Dissimilatory Sulfate Reduction ◽  
Lactate Oxidation ◽  
Sulfate Reducing ◽  
Sulfide Production ◽  
Reducing Bacteria

AbstractIntestinal sulfate-reducing bacteria are often isolated from patients with inflammatory bowel disease, including ulcerative colitis, and can be involved in the development of gut inflammation. A comparison of the metabolism of intestinal sulfate-reducing bacteria isolated from individuals with colitis and healthy controls using statistical analysis has never been studied and described before. The aim of our research was to evaluate the parameters of dissimilatory sulfate reduction inDesulfovibriospecies that were isolated from the feces of healthy objects and individuals with colitis. Principal component analysis indicates that the strains that were isolated from individuals with colitis grouped in the same cluster by biomass accumulation and sulfide production, same as the strains isolated from healthy individuals. Sulfate and lactate consumption measured over time showed negative correlation (Pearson correlations,p<0.01), healthy: -0.760; colitis: -0.770; healthy: -0.828; colitis: -0.847, respectively. The calculated linear regression (R2) was lower in biomass accumulation and hydrogen sulfide production, 0.531; 0.625 respectively. Thus, biomass accumulation and sulfide production, together with measured kinetic parameters play an important factor in bowel inflammation, including ulcerative colitis. Additionally, acetate production can also synergize with H2S, while sulfate consumption and lactate oxidation likely represent minor factors in bowel disease.

scholarly journals Sulfate-reducing bacteria in human feces and their association with inflammatory bowel diseases

2002 ◽  
Vol 40 (2) ◽  
pp. 107-112 ◽  
Author(s):  
Julien Loubinoux ◽  
Jean-Pierre Bronowicki ◽  
Ines A.C. Pereira ◽  
Jean-Louis Mougenel ◽  
Alain E. Faou
Keyword(s):  
Inflammatory Bowel Diseases ◽  
Sulfate Reducing Bacteria ◽  
Human Feces ◽  
Sulfate Reducing ◽  
Bowel Diseases ◽  
Inflammatory Bowel ◽  
Reducing Bacteria
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