scholarly journals Proteome analysis of Mycoplasma fermentans cultured under aerobic and anaerobic conditions

2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Francesca Benedetti ◽  
Selvi Krishnan ◽  
Fiorenza Cocchi ◽  
Hervé Tettelin ◽  
Robert C. Gallo ◽  
...  
Keyword(s):  
Proteome Analysis ◽  
Anaerobic Growth ◽  
Spectrometric Analysis ◽  
Anaerobic Conditions ◽  
Two Dimensional Gel Electrophoresis ◽  
Mycoplasma Fermentans ◽  
Software Analysis ◽  
Anaerobic Microorganisms ◽  
Growing Conditions ◽  
Aerobic And Anaerobic

Abstract Background and aims Mycoplasmas are ubiquitous pathogens found not only in humans but also in animals, plants, insects and soil. Though they usually grow better in an aerobic environment, mycoplasmas are also facultative anaerobic microorganisms. Following infection, the transition of a microorganism from a normal environment into an anaerobic one (e.g. dead or dying tissue) may result in production of a higher number of bacterial toxins. The resolution of the bacterial proteome during the aerobic/anaerobic switch could thus allow the identification of potential pathogenic determinants and pathways. Methods We used two-dimensional gel electrophoresis (2-DE) coupled with matrix-assisted laser desorption/ionization time-of-flight/tandem mass spectroscopy (MALDI-TOF MS/MS) and subsequent mass spectrometric analysis to characterize the liposoluble and hydrosoluble protein fractions of a strain of Mycoplasma fermentans isolated in our lab (MFI), that was cultured under either aerobic or anaerobic conditions. Results We identified the 27 most abundant proteins in the liposoluble fraction and the 30 most abundant proteins in the hydrosoluble fraction and determined their modulation under aerobic and anaerobic growth. By using Protein ANalysis TrougH Evolutionary Relationships (PANTHER) and the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) software analysis tools, we were able to identify, define and organize the function of each protein, as well as to determine the specific interactome. Conclusions Our work provides the first proteome reference map of Mycoplasma fermentans obtained under aerobic and anaerobic growing conditions. These data may help to better understand the mechanisms of pathogenicity of this microorganism and define new diagnostic targets.

Organic Acids and Anaerobic Microorganisms in the Contents of the Cholesteatoma SAC

1983 ◽  
Vol 92 (1) ◽  
pp. 91-96 ◽  
Author(s):  
Yukiko Iino ◽  
Tomonori Takasaka ◽  
Etsuro Hoshino ◽  
Yutaka Kaneko ◽  
Sachiko Tomioka ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Organic Acids ◽  
Volatile Fatty Acids ◽  
Chromatographic Technique ◽  
Anaerobic Conditions ◽  
Clostridium Species ◽  
Anaerobic Microorganisms ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

Organic acids in the contents of the cholesteatoma sac from 28 cases were studied by gas chromatographic technique. Five volatile fatty acids (acetate, propionate, isobutyrate, butyrate and isovalerate) and lactate were detected in large amounts, which may lower the pH of the cholesteatoma content. These acids were considered to be derived from products of anaerobic microorganisms. Therefore, the contents from 12 cases were cultured anaerobically in a glove box. Obligate microorganisms were identified in 92% of the cases and Peptococcus, Bacteroides, and Clostridium species were frequently isolated. In vitro, such obligate anaerobes produced various organic acids from the cholesteatoma content. Facultatives such as Staphylococcus aureus and Proteus mirabilis produced acetate in the content under aerobic and anaerobic conditions, whereas no organic acid was produced by Pseudomonas aeruginosa. Organic acids in the cholesteatoma content could be fermentative products made by the microorganisms, anaerobes and facultatives, which use the content as a substrate for acid production.

scholarly journals The Bacillus subtilis nrdEF Genes, Encoding a Class Ib Ribonucleotide Reductase, Are Essential for Aerobic and Anaerobic Growth

2006 ◽  
Vol 72 (8) ◽  
pp. 5260-5265 ◽  
Author(s):  
Elisabeth Härtig ◽  
Anja Hartmann ◽  
Manuela Schätzle ◽  
Alessandra M. Albertini ◽  
Dieter Jahn
Keyword(s):  
Bacillus Subtilis ◽  
Regulatory System ◽  
Viable Cell ◽  
Growth Conditions ◽  
Anaerobic Growth ◽  
Cell Counting ◽  
Temperature Sensitive ◽  
Anaerobic Conditions ◽  
Transcriptional Start Sites ◽  
Aerobic And Anaerobic

ABSTRACT Ribonucleotide reductases (RNRs) are essential for the biosynthesis of the deoxyribonucleoside triphosphates of DNA. Recently, it was proposed that externally supplied deoxyribonucleosides or DNA is required for the growth of Bacillus subtilis under strict anaerobic conditions (M. J. Folmsbee, M. J. McInerney, and D. P. Nagle, Appl. Environ. Microbiol. 70:5252-5257, 2004). Cultivation of B. subtilis on minimal medium in the presence of oxygen indicators in combination with oxygen electrode measurements and viable cell counting demonstrated that growth occurred under strict anaerobic conditions in the absence of externally supplied deoxyribonucleosides. The nrdEF genes encode the only obvious RNR in B. subtilis. A temperature-sensitive nrdE mutant failed to grow under aerobic and anaerobic conditions, indicating that this oxygen-dependent class I RNR has an essential role under both growth conditions. Aerobic growth and anaerobic growth of the nrdE mutant were rescued by addition of deoxynucleotides. The nrd locus consists of an nrdI-nrdE-nrdF-ymaB operon. The 5′ end of the corresponding mRNA revealed transcriptional start sites 45 and 48 bp upstream of the translational start of nrdI. Anaerobic transcription of the operon was found to be dependent on the presence of intact genes for the ResDE two-component redox regulatory system. Two potential ResD binding sites were identified approximately 62 bp (site A) and 50 bp (site B) upstream of the transcriptional start sites by a bioinformatic approach. Only mutation of site B eliminated nrd expression. Aerobic transcription was ResDE independent but required additional promoter elements localized between 88 and 275 bp upstream of the transcriptional start.

scholarly journals Effect of Intracellular Expression of Antimicrobial Peptide LL-37 on Growth of Escherichia coli Strain TOP10 under Aerobic and Anaerobic Conditions

2013 ◽  
Vol 57 (10) ◽  
pp. 4707-4716 ◽  
Author(s):  
Wei Liu ◽  
Shi Lei Dong ◽  
Fei Xu ◽  
Xue Qin Wang ◽  
T. Ryan Withers ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Growth Conditions ◽  
Anaerobic Growth ◽  
Anaerobic Conditions ◽  
Content Type ◽  
E Coli ◽  
Intracellular Expression ◽  
Aerobic And Anaerobic Growth ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

ABSTRACTAntimicrobial peptides (AMPs) can cause lysis of target bacteria by directly inserting themselves into the lipid bilayer. This killing mechanism confounds the identification of the intracellular targets of AMPs. To circumvent this, we used a shuttle vector containing the inducible expression of a human cathelicidin-related AMP, LL-37, to examine its effect onEscherichia coliTOP10 under aerobic and anaerobic growth conditions. Induction of LL-37 caused growth inhibition and alteration in cell morphology to a filamentous phenotype. Further examination of theE. colicell division protein FtsZ revealed that LL-37 did not interact with FtsZ. Moreover, intracellular expression of LL-37 results in the enhanced production of reactive oxygen species (ROS), causing lethal membrane depolarization under aerobic conditions. Additionally, the membrane permeability was increased after intracellular expression of LL37 under both aerobic and anaerobic conditions. Transcriptomic analysis revealed that intracellular LL-37 mainly affected the expression of genes related to energy production and carbohydrate metabolism. More specifically, genes related to oxidative phosphorylation under both aerobic and anaerobic growth conditions were affected. Collectively, our current study demonstrates that intracellular expression of LL-37 inE. colican inhibit growth under aerobic and anaerobic conditions. While we confirmed that the generation of ROS is a bactericidal mechanism for LL-37 under aerobic growth conditions, we also found that the intracellular accumulation of cationic LL-37 influences the redox and ion status of the cells under both growth conditions. These data suggest that there is a new AMP-mediated bacterial killing mechanism that targets energy metabolism.

Comparative proteome analysis of Mycobacterium tuberculosis grown under aerobic and anaerobic conditions

Microbiology ◽  
2004 ◽  
Vol 150 (11) ◽  
pp. 3821-3829 ◽  
Author(s):  
Joakim Starck ◽  
Gunilla Källenius ◽  
Britt-Inger Marklund ◽  
Dan I. Andersson ◽  
Thomas Åkerlund
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Transition Point ◽  
Elongation Factor ◽  
Proteome Analysis ◽  
Mycolic Acid ◽  
Anaerobic Culture ◽  
Anaerobic Conditions ◽  
Coa Transferase ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

Data are presented from two-dimensional (2-D) PAGE analysis of Mycobacterium tuberculosis strain Harlingen grown during aerobic and anaerobic culture, according to a modified Wayne dormancy model. M. tuberculosis cultures were grown to the transition point between exponential growth and stationary phase in the presence of oxygen (7 days) and then part of the cultures was shifted to anaerobic conditions for 16 days. Growth declined similarly during aerobic and anaerobic conditions, whereas the ATP consumption rapidly decreased in the anaerobic cultures. 2-D PAGE revealed 50 protein spots that were either unique to, or more abundant during, anaerobic conditions and 16 of these were identified by MALDI-TOF. These proteins were the α-crystalline homologue (HspX), elongation factor Tu (Tuf), GroEL2, succinyl-CoA : 3-oxoacid-CoA transferase (ScoB), mycolic acid synthase (CmaA2), thioredoxin (TrxB2), β-ketoacyl-ACP synthase (KasB), l-alanine dehydrogenase (Ald), Rv2005c, Rv2629, Rv0560c, Rv2185c and Rv3866. Some protein spots were found to be proteolytic fragments, e.g. HspX and GroEL2. These data suggest that M. tuberculosis induces expression of about 1 % of its genes in response to dormancy.

scholarly journals Effect of Anaerobic Growth on Quinolone Lethality with Escherichia coli

2006 ◽  
Vol 51 (1) ◽  
pp. 28-34 ◽  
Author(s):  
Muhammad Malik ◽  
Syed Hussain ◽  
Karl Drlica
Keyword(s):  
Escherichia Coli ◽  
Nalidixic Acid ◽  
Sodium Dodecyl ◽  
Anaerobic Growth ◽  
Aerobic Growth ◽  
Anaerobic Conditions ◽  
Dna Complexes ◽  
Chromosome Fragmentation ◽  
Aerobic And Anaerobic Growth ◽  
Aerobic And Anaerobic

ABSTRACT Quinolone activity against Escherichia coli was examined during aerobic growth, aerobic treatment with chloramphenicol, and anaerobic growth. Nalidixic acid, norfloxacin, ciprofloxacin, and PD161144 were lethal for cultures growing aerobically, and the bacteriostatic activity of each quinolone was unaffected by anaerobic growth. However, lethal activity was distinct for each quinolone with cells treated aerobically with chloramphenicol or grown anaerobically. Nalidixic acid failed to kill cells under both conditions; norfloxacin killed cells when they were grown anaerobically but not when they were treated with chloramphenicol; ciprofloxacin killed cells under both conditions but required higher concentrations than those required with cells grown aerobically; and PD161144, a C-8-methoxy fluoroquinolone, was equally lethal under all conditions. Following pretreatment with nalidixic acid, a shift to anaerobic conditions or the addition of chloramphenicol rapidly blocked further cell death. Formation of quinolone-gyrase-DNA complexes, observed as a sodium dodecyl sulfate (SDS)-dependent drop in cell lysate viscosity, occurred during aerobic and anaerobic growth and in the presence and in the absence of chloramphenicol. However, lethal chromosome fragmentation, detected as a drop in viscosity in the absence of SDS, occurred with nalidixic acid treatment only under aerobic conditions in the absence of chloramphenicol. With PD161144, chromosome fragmentation was detected when the cells were grown aerobically and anaerobically and in the presence and in the absence of chloramphenicol. Thus, all quinolones tested appear to form reversible bacteriostatic complexes containing broken DNA during aerobic growth, during anaerobic growth, and when protein synthesis is blocked; however, the ability to fragment chromosomes and to rapidly kill cells under these conditions depends on quinolone structure.

scholarly journals Acid- and Base-Induced Proteins during Aerobic and Anaerobic Growth of Escherichia coli Revealed by Two-Dimensional Gel Electrophoresis

1999 ◽  
Vol 181 (7) ◽  
pp. 2209-2216 ◽  
Author(s):  
Darcy Blankenhorn ◽  
Judith Phillips ◽  
Joan L. Slonczewski
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Low Ph ◽  
Anaerobic Growth ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
High Ph ◽  
Significant Difference ◽  
Aerobic And Anaerobic Growth ◽  
Aerobic And Anaerobic

ABSTRACT Proteins induced by acid or base, during long-term aerobic or anaerobic growth in complex medium, were identified inEscherichia coli. Two-dimensional gel electrophoresis revealed pH-dependent induction of 18 proteins, nine of which were identified by N-terminal sequencing. At pH 9, tryptophan deaminase (TnaA) was induced to a high level, becoming one of the most abundant proteins observed. TnaA may reverse alkalinization by metabolizing amino acids to produce acidic products. Also induced at high pH, but only in anaerobiosis, was glutamate decarboxylase (GadA). Thegad system (GadA/GadBC) neutralizes acidity and enhances survival in extreme acid; its induction during anaerobic growth may help protect alkaline-grown cells from the acidification resulting from anaerobic fermentation. To investigate possible responses to internal acidification, cultures were grown in propionate, a membrane-permeant weak acid which acidifies the cytoplasm. YfiD, a homologue of pyruvate formate lyase, was induced to high levels at pH 4.4 and induced twofold more by propionate at pH 6; both of these conditions cause internal acidification. At neutral or alkaline pH, YfiD was virtually absent. YfiD is therefore a strong candidate for response to internal acidification. Acid or propionate also increased the expression of alkyl hydroperoxide reductase (AhpC) but only during aerobic growth. At neutral or high pH, AhpC showed no significant difference between aerobic and anaerobic growth. The increase of AhpC in acid may help protect the cell from the greater concentrations of oxidizing intermediates at low pH. Isocitrate lyase (AceA) was induced by oxygen across the pH range but showed substantially greater induction in acid or in base than at pH 7. Additional responses observed included the induction of MalE at high pH and induction of several enzymes of sugar metabolism at low pH: the phosphotransferase system components ManX and PtsH and the galactitol fermentation enzyme GatY. Overall, our results indicate complex relationships between pH and oxygen and a novel permeant acid-inducible gene, YfiD.

Proteome Analysis of CD4+ T Cells Reveals Differentially Expressed Proteins in Infertile Polycystic Ovary Syndrome Patients

2020 ◽  
Vol 20 ◽  
Author(s):  
Fatemeh Nasri ◽  
Maryam Zare ◽  
Mehrnoosh Doroudchi ◽  
Behrouz Gharesi-Fard
Keyword(s):  
Mass Spectrometry ◽  
T Cells ◽  
T Lymphocytes ◽  
Gel Electrophoresis ◽  
Cd4 T Cells ◽  
Polycystic Ovary ◽  
Proteome Analysis ◽  
Two Dimensional ◽  
Ovary Syndrome ◽  
Two Dimensional Gel Electrophoresis

Background: Polycystic ovary syndrome (PCOS) is the most frequent endocrine disorder affecting 6–7% of premenopausal women. Recent studies revealed that the immune system especially CD4+ T helper cells are important in the context PCOS. Proteome analysis of CD4+ T lymphocytes can provide valuable information regarding the biology of these cells in the context of PCOS. Objective: To investigate immune dysregulation in CD4+ T lymphocytes at the protein level in the context of PCOS using two-dimensional gel electrophoresis (2DE) and mass spectrometry (MS). Methods: In the present study, we applied two-dimensional gel electrophoresis / mass spectrometry to identify proteins differentially expressed by peripheral blood CD4+ T cells in ten PCOS women compared with ten healthy women. Western blot technique was used to confirm the identified proteins. Results: Despite the overall proteome similarities, there were significant differences in the expression of seven spots between two groups (P <0.05). Three proteins, namely phosphatidylethanolamine-binding protein 1, proteasome activator complex subunit 1 and triosephosphate isomerase 1 were successfully identified by Mass technique and confirmed by western blot. All characterized proteins were over-expressed in CD4+ T cells from patients compared to CD4+ T cells from controls (P <0.05). In-silico analysis suggested that the over-expressed proteins interact with other proteins involved in cellular metabolism especially glycolysis and ferroptosis pathway. Conclusion: These findings suggest that metabolic adjustments in CD4+ T lymphocytes, which is in favor of increased glycolysis and Th2 differentiation are important in the context of PCOS.

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