scholarly journals Cryo-EM structure of mycobacterial cytochrome bd reveals two oxygen access channels

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Weiwei Wang ◽  
Yan Gao ◽  
Yanting Tang ◽  
Xiaoting Zhou ◽  
Yuezheng Lai ◽  
...  

AbstractCytochromes bd are ubiquitous amongst prokaryotes including many human-pathogenic bacteria. Such complexes are targets for the development of antimicrobial drugs. However, an understanding of the relationship between the structure and functional mechanisms of these oxidases is incomplete. Here, we have determined the 2.8 Å structure of Mycobacterium smegmatis cytochrome bd by single-particle cryo-electron microscopy. This bd oxidase consists of two subunits CydA and CydB, that adopt a pseudo two-fold symmetrical arrangement. The structural topology of its Q-loop domain, whose function is to bind the substrate, quinol, is significantly different compared to the C-terminal region reported for cytochromes bd from Geobacillus thermodenitrificans (G. th) and Escherichia coli (E. coli). In addition, we have identified two potential oxygen access channels in the structure and shown that similar tunnels also exist in G. th and E. coli cytochromes bd. This study provides insights to develop a framework for the rational design of antituberculosis compounds that block the oxygen access channels of this oxidase.

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Melanie Radloff ◽  
Isam Elamri ◽  
Tamara N. Grund ◽  
Luca F. Witte ◽  
Katharina F. Hohmann ◽  
...  

AbstractCytochrome bd-type oxidases play a crucial role for survival of pathogenic bacteria during infection and proliferation. This role and the fact that there are no homologues in the mitochondrial respiratory chain qualify cytochrome bd as a potential antimicrobial target. However, few bd oxidase selective inhibitors have been described so far. In this report, inhibitory effects of Aurachin C (AurC-type) and new Aurachin D (AurD-type) derivatives on oxygen reductase activity of isolated terminal bd-I, bd-II and bo3 oxidases from Escherichia coli were potentiometrically measured using a Clark-type electrode. We synthesized long- (C10, decyl or longer) and short-chain (C4, butyl to C8, octyl) AurD-type compounds and tested this set of molecules towards their selectivity and potency. We confirmed strong inhibition of all three terminal oxidases for AurC-type compounds, whereas the 4(1H)-quinolone scaffold of AurD-type compounds mainly inhibits bd-type oxidases. We assessed a direct effect of chain length on inhibition activity with highest potency and selectivity observed for heptyl AurD-type derivatives. While Aurachin C and Aurachin D are widely considered as selective inhibitors for terminal oxidases, their structure–activity relationship is incompletely understood. This work fills this gap and illustrates how structural differences of Aurachin derivatives determine inhibitory potency and selectivity for bd-type oxidases of E. coli.


Science ◽  
2019 ◽  
Vol 366 (6461) ◽  
pp. 100-104 ◽  
Author(s):  
S. Safarian ◽  
A. Hahn ◽  
D. J. Mills ◽  
M. Radloff ◽  
M. L. Eisinger ◽  
...  

Cytochrome bd–type quinol oxidases catalyze the reduction of molecular oxygen to water in the respiratory chain of many human-pathogenic bacteria. They are structurally unrelated to mitochondrial cytochrome c oxidases and are therefore a prime target for the development of antimicrobial drugs. We determined the structure of the Escherichia coli cytochrome bd-I oxidase by single-particle cryo–electron microscopy to a resolution of 2.7 angstroms. Our structure contains a previously unknown accessory subunit CydH, the L-subfamily–specific Q-loop domain, a structural ubiquinone-8 cofactor, an active-site density interpreted as dioxygen, distinct water-filled proton channels, and an oxygen-conducting pathway. Comparison with another cytochrome bd oxidase reveals structural divergence in the family, including rearrangement of high-spin hemes and conformational adaption of a transmembrane helix to generate a distinct oxygen-binding site.


2016 ◽  
Vol 1 (2) ◽  
Author(s):  
Bertoka Fajar S.P. Negara ◽  
. Riyanti ◽  
Bintang Marhaeni ◽  
Aradea B. Kusuma

Circulation of synthetic antibacterial chemicals which is used continuously can cause resistance to bacteria. Alternative synthetic antibiotics research then comes up as an urgent need. Recently, the discovery of new antibacterial agents is looking for Actinomycetes. Actinomycetes are one of the active marine bacteria which produce secondary metabolites that could be used as antibacterial. The objective of this study was to evaluate the antibacterial activities of Actinomycetes symbiont with Halimeda macroloba, Gelidiella acerosa, and Turbinaria ornata in Pangandaran beach, Indonesia. A total of 41 Actinomycetes were isolated from 3 species of seaweed. Rep PCR amplification method was used to characterize and identify the relationship between samples. Primer BOX A1R (5’-CTACGGCAAGGCGACGCTGACGCTGACG-3’), 27F (5'-AGTTTGATCMTGGCTCAG-3') and 1492R (5'-TACGGYTACCTTGTTACGACTT-3') was used in amplification 16s DNA. Six isolates from the representative cluster dendogram were selected for secondary metabolites extraction using ethyl acetate solvent, meanwhile agar diffusion method was used as Antibacterial activity assay using E. coli, Proteus sp., and Enterobacter sp. as the bacterial test. The results showed that the activity of secondary metabolite extracts of Actinomycetes symbiont with Halimeda macroloba presented the highest inhibition zone on 3 bacterial tests. In the future, it could be a potential antibacterial agent against animal pathogenic bacteria.


2020 ◽  
Vol 13 (6) ◽  
Author(s):  
Mohamad Hossein Rezaei ◽  
Farshad Kakian ◽  
Abolfazl Gholipour ◽  
Behnam Zamanzad

Background: Nosocomial infections are acquired during hospital treatment or in a hospital environment. One such infecting agent is uropathogenic Escherichia coli and many virulence genes enable it to become pathogenic, thereby causing damage to the host. Objectives: This study aimed to identify aer, traT, and PAI genes in E. coli isolates collected from fecal and urinary tract infection (UTI) specimens and determine the relationship between them in both populations studied in a center in Iran by multiplex polymerase chain reaction (PCR) assay. Methods: Seventy-five isolates of E. coli from the urine of inpatients and 75 isolates from commensal fecal without UTI and diarrhea were collected. The E. coli bacteria were detected and isolated, using biochemical techniques and supplementary tests in the Microbiology Laboratory of Shahrekord University of Medical Sciences. Antibiotic susceptibility pattern for 14 antibiotics was done utilizing the disc diffusion method. The existence of aer, traT, and PAI virulence genes among all isolates was investigated by multiplex PCR. Results: Among the urinary pathogenic E. coli isolates, the highest antibiotic resistance was observed in cefazolin, ampicillin, and cotrimoxazole antibiotics. The prevalence rates of aer, traT, and PAI genes in the fecal isolates were 92%, 90.6%, and 46.6%, respectively. Further, their prevalence rates in urine isolates were 96%, 97.3%, and 41.3%, in that order. Conclusions: The presence of the high frequency of pathogenic islands (PAIs), especially in fecal samples, is important because these genes are easily transmitted and convert a commensal bacterium into a pathogen. Because only the genome of pathogenic bacteria has been unwrapped, little attention has been paid to PAIs in commensal bacteria.


2018 ◽  
Vol 22 (2) ◽  
pp. 285-288
Author(s):  
A.P. Prevar ◽  
A.V. Kryzshanovskaya ◽  
V.A. Radionov ◽  
V.M. Mrug

The main factor in the treatment of suppurative and inflammatory processes is the timely optimization of treatment measures taking into account the nature of the microflora and its susceptibility to antimicrobial drugs. The purpose of the study is to monitor the spectrum of microorganisms – pathogens of purulent-inflammatory processes of soft tissues in surgical patients; study of the sensitivity of isolated strains to antibiotics. The material was collected in accordance with aseptic rules. The identification of a pure culture of bacteria was carried out according to morphological, culture, biochemical properties, and the presence of virulence enzymes. Sensitivity of bacteria to antibiotics was determined by the standard disks method (by Kirby-Bauer’s). 255 patients with purulent-inflammatory processes of soft tissues were examined for the period from 2014 to 2017. 229 strains of isolated bacteria were included to Escherichia coli, Citrobacer freundii, Enterobacter cloacae, E.aerogenes, Proteus vulgaris, Staphylococcus aureus, S.epidermidis, Streptococcus pyogenes, S.viridians, S.agalactiae, Pseudomonas aeruginosa. The main cause of purulent-inflammatory processes of soft tissues is Staphylococci (67,2%). Compared to previous studies, the number of P.aeruginosa isolated cultures increased (7.9%). In monoculture and in association with other microorganisms, E. coli (9.6% of cases), E.cloacae et aerogenes (3.9% of cases), P.vulgaris (3.9% of cases), C.freundi (2.5% of cases), S.agalactiae, S.pyogenes, S.viridans (3.5%). The number of associated sows reaches 12%. Clinical strains of microorganisms remain most sensitive to fluoroquinolones, cephalosporins, and also retains high sensitivity to gentamicin, lincomycin, rifampicin, which is important for empirical antibiotic therapy. To increase the effectiveness of antibacterial therapy, strict adherence to the mode of appointment of antibiotics, justification of indications, a combination of antibiotics of different spectrum of action, mandatory correction after determining the sensitivity of the pathogen.


2019 ◽  
Vol 14 (1) ◽  
pp. 28-31 ◽  
Author(s):  
Rowles H. L.

Probiotics are live microorganisms, which when ingested in sufficient amounts, confer health benefits to the host by improving the gut microflora balance. The purpose of this research was to determine whether commercial probiotic products containing multitude of commensal bacteria would reduce the growth rate of pathogenic bacteria, specifically Escherichia coli and Salmonella typhimurium. Growth curves were established, and the growth rates were compared for samples of E. coli, S. typhimurium, Nature’s Bounty Controlled Delivery probiotic, Sundown Naturals Probiotic Balance probiotic, and cocultures of the pathogenic bacteria mixed with the probiotics. The findings of this research were that the commercial probiotics significantly reduced the growth rate of E. coli and S. typhimurium when combined in cocultures. Probiotics containing multiple strains may be taken prophylactically to reduce the risk of bacterial infections caused by E. coli and S. typhimurium. Probiotics could be used to reduce the high global morbidity and mortality rates of diarrheal disease.


Author(s):  
Pramod Dhakal ◽  
Ankit a Achary ◽  
Vedamurthy Joshi

Bioenhancers are drug facilitator which do not show the typical drug activity but in combination to enhance the activity of other molecule in several way including increase the bioavailability of drug across the membrane, potentiating the drug molecules by conformational interaction, acting as receptor for drug molecules and making target cell more receptive to drugs and promote and increase the bioactivity or bioavailability or the uptake of drugs in combination therapy. The objective of the present study was to evaluate the antibacterial and activity of combination in Azadirachta indica extract with cow urine distillate and pepper extract against common pathogenic bacteria, a causative agent of watery diarrhea. It has been found that Indian indigenous cow urine and its distillate also possess bioenhancing ability. Bioenhancing role of cow urine distillate (CUD) and pepper extract was investigated on antibacterial activity of ethanol extract of Azadirachta indica. Antibacterial activity of ethanol extract neem alone and in combination with CUD and pepper extract were determined the ATCC strains against Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and E-coli by cup plate diffusion method. Ethanol extract of neem has showed more effect on P. aeruginosa, E-coli than S. aureus and K. pneumonia with combination of CUD and pepper extract. CUD and pepper did not show any inhibition of test bacteria in low concentration. The antibacterial effect of combination of extract and CUD was higher than the inhibition caused by extract alone and is suggestive of the bioenhancing role of cow urine distillate and pepper. Moreover, inhibition of test bacteria was observed with less concentration of extract on combining with CUD


1995 ◽  
Vol 31 (5-6) ◽  
pp. 19-26 ◽  
Author(s):  
G. J. Medema ◽  
I. A. van Asperen ◽  
J. M. Klokman-Houweling ◽  
A. Nooitgedagt ◽  
M. J. W. van de Laar ◽  
...  

This pilot study was carried out to determine the relationship between microbiological water quality parameters and the occurrence of health complaints among triathletes. Data were collected at an Olympic distance triathlon (n=314) and a run-bike-run (n=81; controls for exposure to fresh water). At the time of the triathlon, the concentrations of Escherichia coli , thermotolerant coliforms, faecal streptococci, entero- and reoviruses, F-specific RNA phages, Salmonella, Campylobacter, Aeromonas, Plesiomonas shigelloides, Pseudomonas aeruginosa and Staphylococcus aureus were examined over the swimming course. Information on the occurrence of health complaints during the competition and in the week thereafter was collected through a written questionnaire. The results show that triathletes and run-bike-runners are comparable with respect to factors other than water exposure (age, sex, training history, physical stress, lower intestinal health complaints during the competition) that may influence the occurrence of health complaints in the week after the competition. Triathletes and run-bike-runners reported gastro-intestinal (7.7% vs 2.5%), respiratory (5.5% vs 3.7%), skin/mucosal (2.6% vs 1.2%), general (3.5% vs 1.2%) and total symptoms (14.8% vs 7.4%) in the week after the event. The health risks for triathletes for all symptom groups are not significantly higher than for run-bike-runners. The geometric mean concentration of faecal indicator bacteria is relatively low: E. coli 170/100 ml; faecal streptococci 13/100 ml, enteroviruses were present at concentrations of 0.1/l. The group of triathletes was homogeneusly and relatively intensely exposed to water; they all swam in the same body of water at the same time and 75% reported to have swallowed freshwater. It was concluded that this study design is suitable to study the relationship between health complaints and microbiological water quality. In the summers of 1993 and 1994, a study will be carried out concerning several run-bike-runs and triathlons in freshwaters of different quality.


1997 ◽  
Vol 35 (11-12) ◽  
pp. 35-40 ◽  
Author(s):  
B. Genthe ◽  
N. Strauss ◽  
J. Seager ◽  
C. Vundule ◽  
F. Maforah ◽  
...  

Efforts to provide water to developing communities in South Africa have resulted in various types of water supplies being used. This study examined the relationship between the type of water supply and the quality of water used. Source (communal taps, private outdoor and indoor taps) and point-of-use water samples were examined for heterotrophic plate counts (HPC), total and faecal coliforms, E. coli, and coliphages. Ten percent of samples were also analysed for enteric viruses, Giardia and Cryptosporidium. Approximately 320 households were included in a case-control study. In addition, a cross-sectional study was conducted. Both studies examined the relationship between different types of water facilities and diarrhoea among pre-school children. The source water was of good microbial quality, but water quality was found to have deteriorated significantly after handling and storage in both case and control households, exceeding drinking water quality guideline values by 1-6 orders of magnitude. Coliphage counts were low for all water samples tested. Enteric viruses and Cryptosporidium oocysts were not detected. Giardia cysts were detected on one occasion in case and control in-house samples. Comparisons of whether in-house water, after handling and storage, complied with water quality guideline values demonstrated households using communal taps to have significantly poorer quality than households using private outdoor or indoor taps for HPC and E. coli (χ2 = 14.9, P = 0.001; χ2 = 6.6, P = 0.04 respectively). A similar trend (although not statistically significant) was observed for the other microbial indicators. The cross-sectional study demonstrated an apparent decrease in health risk associated with private outdoor taps in comparison to communal taps. This study suggests that a private outdoor tap is the minimum level of water supply in order to ensure the supply of safe water to developing communities.


2019 ◽  
Vol 25 (34) ◽  
pp. 3645-3663 ◽  
Author(s):  
Muhammad Ismail ◽  
Kalsoom Akhtar ◽  
M.I. Khan ◽  
Tahseen Kamal ◽  
Murad A. Khan ◽  
...  

: Water pollution due to waste effluents of the textile industry is seriously causing various health problems in humans. Water pollution with pathogenic bacteria, especially Escherichia coli (E. coli) and other microbes is due to the mixing of fecal material with drinking water, industrial and domestic sewage, pasture and agricultural runoff. Among the chemical pollutants, organic dyes due to toxic nature, are one of the major contaminants of industrial wastewater. Adequate sanitation services and drinking quality water would eliminate 200 million cases of diarrhea, which results in 2.1 million less deaths caused by diarrheal disease due to E. coli each year. Nanotechnology is an excellent platform as compared to conventional treatment methods of water treatment and remediation from microorganisms and organic dyes. In the current study, toxicity and carcinogenicity of the organic dyes have been studied as well as the remediation/inactivation of dyes and microorganism has been discussed. Remediation by biological, physical and chemical methods has been reviewed critically. A physical process like adsorption is cost-effective, but can’t degrade dyes. Biological methods were considered to be ecofriendly and cost-effective. Microbiological degradation of dyes is cost-effective, eco-friendly and alternative to the chemical reduction. Besides, certain enzymes especially horseradish peroxidase are used as versatile catalysts in a number of industrial processes. Moreover, this document has been prepared by gathering recent research works related to the dyes and microbial pollution elimination from water sources by using heterogeneous photocatalysts, metal nanoparticles catalysts, metal oxides and enzymes.


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