scholarly journals Characterization of a Group of Anaerobically Induced, fnr-Dependent Genes of Salmonella typhimurium

1999 ◽  
Vol 181 (19) ◽  
pp. 6092-6097 ◽  
Author(s):  
Yan Wei ◽  
Charles G. Miller
Keyword(s):  
Escherichia Coli ◽  
Salmonella Typhimurium ◽  
Dna Sequence ◽  
Abc Transporter ◽  
E Coli

ABSTRACT We have previously reported the isolation of a group of anaerobically regulated, fnr-dependent lacfusions in Salmonella typhimurium and have grouped theseoxd genes into classes based on map position. In order to identify these genes, we have replaced the original Mud-lacfusion in a member of each oxd class with the much smaller Mud-cam element, cloned the fusion, and determined DNA sequence sufficient to define the oxd gene. Several of the fusions correspond to previously known genes from S. typhimurium or Escherichia coli: oxd-4 = cbiA and oxd-11 = cbiK, oxd-5 = hybB, oxd-7 = dcuB, oxd-8 = moaB, oxd-12 = dmsA, and oxd-14 = napB (aeg-46.5). Two other fusions correspond to previously unknown loci: oxd-2 encodes an acetate/propionate kinase, and oxd-6 encodes a putative ABC transporter present in S. typhimurium but not in E. coli.

scholarly journals Characterization of human interleukin 2 derived from Escherichia coli

1985 ◽  
Vol 229 (2) ◽  
pp. 429-439 ◽  
Author(s):  
S M Liang ◽  
B Allet ◽  
K Rose ◽  
M Hirschi ◽  
C M Liang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Dna Sequence ◽  
Protein A ◽  
Interleukin 2 ◽  
Coding Region ◽  
Disulphide Bridge ◽  
E Coli ◽  
Escherichia Coli Cells

Interleukin 2 isolated from Escherichia coli cells expressing the human interleukin gene has been characterized. The observed properties of the protein have been compared with those properties which can be deduced from the DNA sequence alone and the published properties of natural human interleukin 2. The purified E. coli-derived interleukin 2 is a monomeric protein of Mr 15 000 with a sedimentation velocity of 1.86S. The amino acid composition of the protein and isoelectric point (7.7) are consistent with that part of the translated DNA sequence of the gene corresponding to the mature protein. A single disulphide bridge was identified between Cys-58 and Cys-105. C.d. suggested that interleukin 2 is predominantly alpha-helical in secondary structure. The E. coli-derived protein differed from natural interleukin 2 in the presence of N-terminal methionine and also in the absence of a carbohydrate moiety. Removal of the coding region for the first three amino acids of the natural interleukin 2 protein sequence (Ala-Pro-Thr) by site-specific mutagenesis resulted in a protein with N-terminal serine. The possibility that the specificity of the E. coli ribosomal methionine aminopeptidase may not recognize the sequence NH2-Met-Xaa-Pro is discussed (where Xaa is any amino acid residue).

scholarly journals Curli Fibers Are Highly Conserved between Salmonella typhimurium and Escherichia coli with Respect to Operon Structure and Regulation

1998 ◽  
Vol 180 (3) ◽  
pp. 722-731 ◽  
Author(s):  
Ute Römling ◽  
Zhao Bian ◽  
Mårten Hammar ◽  
Walter D. Sierralta ◽  
Staffan Normark
Keyword(s):  
Escherichia Coli ◽  
Salmonella Typhimurium ◽  
Transcriptional Activation ◽  
Protein Level ◽  
Amino Acid Homology ◽  
E Coli ◽  
Low Copy Number ◽  
High Level ◽  
High Degree

ABSTRACT Mouse-virulent Salmonella typhimurium strains SR-11 and ATCC 14028-1s express curli fibers, thin aggregative fibers, at ambient temperature on plates as judged by Western blot analysis and electron microscopy. Concomitantly with curli expression, cells develop a rough and dry colony morphology and bind the dye Congo red (called the rdar morphotype). Cloning and characterization of the two divergently transcribed operons required for curli biogenesis, csgBA(C)and csgDEFG, from S. typhimurium SR-11 revealed the same gene order and flanking genes as in Escherichia coli. The divergence of the curli region between S. typhimurium and E. coli at the nucleotide level is above average (22.4%). However, a high level of conservation at the protein level, which ranged from 86% amino acid homology for the fiber subunit CsgA to 99% homology for the lipoprotein CsgG, implies functional constraints on the gene products. Consequently, S. typhimurium genes on low-copy-number plasmids were able to complement respective E. coli mutants, although not always to wild-type levels. rpoS and ompR are required for transcriptional activation of (at least) the csgDpromoter. The high degree of conservation at the protein level and the identical regulation patterns in E. coli and S. typhimurium suggest similar roles of curli fibers in the same ecological niche in the two species.

scholarly journals Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Masuzu Kikuchi ◽  
Keiichi Kojima ◽  
Shin Nakao ◽  
Susumu Yoshizawa ◽  
Shiho Kawanishi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Proton Pump ◽  
Expression System ◽  
Spectroscopic Characterization ◽  
Functional Expression ◽  
Proton Pumping ◽  
E Coli ◽  
Oxyrrhis Marina ◽  
Domains Of Life

AbstractMicrobial rhodopsins are photoswitchable seven-transmembrane proteins that are widely distributed in three domains of life, archaea, bacteria and eukarya. Rhodopsins allow the transport of protons outwardly across the membrane and are indispensable for light-energy conversion in microorganisms. Archaeal and bacterial proton pump rhodopsins have been characterized using an Escherichia coli expression system because that enables the rapid production of large amounts of recombinant proteins, whereas no success has been reported for eukaryotic rhodopsins. Here, we report a phylogenetically distinct eukaryotic rhodopsin from the dinoflagellate Oxyrrhis marina (O. marina rhodopsin-2, OmR2) that can be expressed in E. coli cells. E. coli cells harboring the OmR2 gene showed an outward proton-pumping activity, indicating its functional expression. Spectroscopic characterization of the purified OmR2 protein revealed several features as follows: (1) an absorption maximum at 533 nm with all-trans retinal chromophore, (2) the possession of the deprotonated counterion (pKa = 3.0) of the protonated Schiff base and (3) a rapid photocycle through several distinct photointermediates. Those features are similar to those of known eukaryotic proton pump rhodopsins. Our successful characterization of OmR2 expressed in E. coli cells could build a basis for understanding and utilizing eukaryotic rhodopsins.

scholarly journals Enhancement of the Efficiency of Secretion of Heterologous Lipase in Escherichia coli by Directed Evolution of the ABC Transporter System

2005 ◽  
Vol 71 (7) ◽  
pp. 3468-3474 ◽  
Author(s):  
Gyeong Tae Eom ◽  
Jae Kwang Song ◽  
Jung Hoon Ahn ◽  
Yeon Soo Seo ◽  
Joon Shick Rhee
Keyword(s):  
Escherichia Coli ◽  
Directed Evolution ◽  
Abc Transporter ◽  
Microtiter Plate ◽  
Single Amino Acid ◽  
Wild Type ◽  
E Coli ◽  
Single Amino Acid Change ◽  
Secretion Efficiency

ABSTRACT The ABC transporter (TliDEF) from Pseudomonas fluorescens SIK W1, which mediated the secretion of a thermostable lipase (TliA) into the extracellular space in Escherichia coli, was engineered using directed evolution (error-prone PCR) to improve its secretion efficiency. TliD mutants with increased secretion efficiency were identified by coexpressing the mutated tliD library with the wild-type tliA lipase in E. coli and by screening the library with a tributyrin-emulsified indicator plate assay and a microtiter plate-based assay. Four selected mutants from one round of error-prone PCR mutagenesis, T6, T8, T24, and T35, showed 3.2-, 2.6-, 2.9-, and 3.0-fold increases in the level of secretion of TliA lipase, respectively, but had almost the same level of expression of TliD in the membrane as the strain with the wild-type TliDEF transporter. These results indicated that the improved secretion of TliA lipase was mediated by the transporter mutations. Each mutant had a single amino acid change in the predicted cytoplasmic regions in the membrane domain of TliD, implying that the corresponding region of TliD was important for the improved and successful secretion of the target protein. We therefore concluded that the efficiency of secretion of a heterologous protein in E. coli can be enhanced by in vitro engineering of the ABC transporter.

scholarly journals ATIVIDADE ANTIMICROBIANA DE MICRORGANISMOS ISOLADOS DE GRÃOS DE KEFIR

2018 ◽  
Vol 19 (0) ◽  
Author(s):  
Priscila Alves Dias ◽  
Daiani Teixeira Silva ◽  
Cláudio Dias Timm
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Salmonella Enterica ◽  
Escherichia Coli O157 ◽  
E Coli

Resumo Kefir é o produto da fermentação do leite pelos grãos de kefir. Esses grãos contêm uma mistura simbiótica de bactérias e leveduras imersas em uma matriz composta de polissacarídeos e proteínas. Muitos benefícios à saúde humana têm sido atribuídos ao kefir, incluindo atividade antimicrobiana contra bactérias Gram positivas e Gram negativas. A atividade antimicrobiana de 60 microrganismos isolados de grãos de kefir, frente à Escherichia coli O157:H7, Salmonella enterica subsp. enterica sorotipos Typhimurium e Enteritidis, Staphylococcus aureus e Listeria monocytogenes, foi estudada através do teste do antagonismo. A ação antimicrobiana dos sobrenadantes das bactérias ácido-lácticas que apresentaram atividade no teste do antagonismo foi testada. O experimento foi repetido usando sobrenadantes com pH neutralizado. Salmonella Typhimurium e Enteritidis sobreviveram por 24 horas no kefir em fermentação. E. coli O157:H7, S. aureus e L. monocytogenes foram recuperados até 72 horas após o início da fermentação. Todos os isolados apresentaram atividade antimicrobiana contra pelo menos um dos patógenos usados no teste do antagonismo. Sobrenadantes de 25 isolados apresentaram atividade inibitória e três mantiveram essa atividade com pH neutralizado. As bactérias patogênicas estudadas sobreviveram por tempo superior àquele normalmente utilizado para a fermentação do kefir artesanal, o que caracteriza perigo em potencial para o consumidor quando a matéria-prima não apresentar segurança sanitária. Lactobacillus isolados de grãos de kefir apresentam atividade antimicrobiana contra cepas de E. coli O157:H7, Salmonella sorotipos Typhimurium e Enteritidis, S. aureus e L. monocytogenes além daquela exercida pela diminuição do pH.

Antimicrobial Effects of Novel H2O2-Ag+ Complex on Membrane Damage to Staphylococcus aureus,Escherichia coli and Salmonella typhimurium

2021 ◽  
Author(s):  
Bing Han ◽  
Xiaoyu Han ◽  
Mengmeng Ren ◽  
Yilin You ◽  
Jicheng Zhan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Hydrogen Peroxide ◽  
Staphylococcus Aureus ◽  
Salmonella Typhimurium ◽  
Membrane Damage ◽  
Bactericidal Effect ◽  
E Coli ◽  
Bacterial Cell Membrane ◽  
Time Kill ◽  
Environmental Disinfection

Diseases caused by harmful microorganisms pose a serious threat to human health. Safe and environment-friendly disinfectants are, therefore, essential in preventing and controlling such pathogens. This study aimed to investigate the antimicrobial activity and mechanism of a novel hydrogen peroxide and silver (H 2 O 2 -Ag + ) complex (HSC) in combatting Staphylococcus aureus ATCC 29213, Escherichia coli O157:H7 NCTC 12900 and Salmonella typhimurium SL 1344. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values against S. aureus were found to be 0.014 % H 2 O 2 -3.125 mg/L Ag + , while 0.028 % H 2 O 2 -6.25 mg/L Ag + for both E. coli and S. typhimurium . Results of the growth curve assay and time-kill trial suggest that the HSC could inhibit the growth of the tested bacteria, as 99.9 % of viable cells were killed following treatment at the 1 MIC for 3 h. Compared with Oxytech D10 disinfectant (0.25 % H 2 O 2 -5 mg/L Ag + ), the HSC exhibited better antibacterial efficacy at a lower concentration (0.045 % H 2 O 2 -10 mg/L Ag + ). The mechanism of antibacterial action of HSC was found including the disruption of the bacterial cell membrane, followed by entry into the bacteria cell to reduce intracellular adenosine triphosphate (ATP) concentration, and inhibit the activity of antioxidases, superoxide dismutase (SOD) and catalase (CAT). The enhanced bactericidal effect of hydrogen peroxide combined with silver indicates a potential for its application in environmental disinfection, particularly in the food industry.

In Vitro Inhibition of the Growth of Salmonella typhimurium and Escherichia coli 0157:H7 by Bacteria Isolated from the Cecal Contents of Adult Chickens

1991 ◽  
Vol 54 (7) ◽  
pp. 496-501 ◽  
Author(s):  
ARTHUR HINTON ◽  
GEORGE E. SPATES ◽  
DONALD E. CORRIER ◽  
MICHAEL E. HUME ◽  
JOHN R. DELOACH ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Salmonella Typhimurium ◽  
Mixed Cultures ◽  
Enterococcus Durans ◽  
E Coli ◽  
Pure Cultures ◽  
In Vitro Inhibition ◽  
Lactic Acids

A Veillonella species and Enterococcus durans were isolated from the cecal contents of adult broilers. Mixed cultures of Veillonella and E. durans inhibited the growth of Salmonella typhimurium and Escherichia coli 0157:H7 on media containing 2.5% lactose (w/v). The growth of S. typhimurium or E. coli 0157:H7 was not inhibited by mixed cultures containing Veillonella and E. durans on media containing only 0.25% lactose or by pure cultures of Veillonella or E. durans on media containing either 0.25% or 2.5% lactose. The mixed cultures of Veillonella and E. durans produced significantly (P<0.05) more acetic, propionic, and lactic acids in media containing 2.5% lactose than in media containing 0.25% lactose. The inhibition of the enteropathogens was related to the production of lactic acid from lactose by the E. durans and the production of acetic and propionic acids from lactic acid by the Veillonella.

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