Overexpression of Escherichia coli Genes Encoding Nucleoside Phosphorylases in the pET/Bl21(DE3) System Yields Active Recombinant Enzymes

2002 ◽  
Vol 24 (1) ◽  
pp. 56-60 ◽  
Author(s):  
Roman S. Esipov ◽  
Alexandr I. Gurevich ◽  
Dmitry V. Chuvikovsky ◽  
Larisa A. Chupova ◽  
Tatyana I. Muravyova ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Enzymes ◽  
Genes Encoding ◽  
Nucleoside Phosphorylases

scholarly journals Products Released from Structurally Different Dextrans by Bacterial and Fungal Dextranases

Foods ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 244
Author(s):  
Silke L. Pittrof ◽  
Larissa Kaufhold ◽  
Anja Fischer ◽  
Daniel Wefers
Keyword(s):  
Escherichia Coli ◽  
Enzyme Activity ◽  
Sugar Industry ◽  
Streptococcus Salivarius ◽  
Bacteroides Thetaiotaomicron ◽  
Recombinant Enzymes ◽  
Penicillium Sp ◽  
Hydrolysis Products ◽  
Medical Sector ◽  
Genes Encoding

Dextran hydrolysis by dextranases is applied in the sugar industry and the medical sector, but it also has a high potential for use in structural analysis of dextrans. However, dextranases are produced by several organisms and thus differ in their properties. The aim of this study was to comparatively investigate the product patterns obtained from the incubation of linear as well as O3- and O4-branched dextrans with different dextranases. For this purpose, genes encoding for dextranases from Bacteroides thetaiotaomicron and Streptococcus salivarius were cloned and heterologously expressed in Escherichia coli. The two recombinant enzymes as well as two commercial dextranases from Chaetomium sp. and Penicillium sp. were subsequently used to hydrolyze structurally different dextrans. The hydrolysis products were investigated in detail by HPAEC-PAD. For dextranases from Chaetomium sp., Penicillium sp., and Bacteroides thetaiotaomicron, isomaltose was the end product of the hydrolysis from linear dextrans, whereas Penicillium sp. dextranase led to isomaltose and isomaltotetraose. In addition, the latter enzyme also catalyzed a disproportionation reaction when incubated with isomaltotriose. For O3- and O4-branched dextrans, the fungal dextranases yielded significantly different oligosaccharide patterns than the bacterial enzymes. Overall, the product patterns can be adjusted by choosing the correct enzyme as well as a defined enzyme activity.

scholarly journals Molecular Epidemiology of NDM-1-Producing Enterobacteriaceae and Acinetobacter baumannii Isolates from Pakistan

2014 ◽  
Vol 58 (9) ◽  
pp. 5589-5593 ◽  
Author(s):  
Anna L. Sartor ◽  
Muhammad W. Raza ◽  
Shahid A. Abbasi ◽  
Kathryn M. Day ◽  
John D. Perry ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Molecular Epidemiology ◽  
Genetic Relatedness ◽  
Antibiotic Resistance Genes ◽  
Enterobacter Cloacae ◽  
Content Type ◽  
Plasmid Replicon ◽  
Genes Encoding ◽  
Clonal Spread ◽  
Encoding Genes

ABSTRACTThe molecular epidemiology of 66 NDM-producing isolates from 2 Pakistani hospitals was investigated, with their genetic relatedness determined using repetitive sequence-based PCR (Rep-PCR). PCR-based replicon typing and screening for antibiotic resistance genes encoding carbapenemases, other β-lactamases, and 16S methylases were also performed. Rep-PCR suggested a clonal spread ofEnterobacter cloacaeandEscherichia coli. A number of plasmid replicon types were identified, with the incompatibility A/C group (IncA/C) being the most common (78%). 16S methylase-encoding genes were coharbored in 81% of NDM-producingEnterobacteriaceae.

scholarly journals Shiga Toxins, and the Genes Encoding Them, in Fecal Samples from Native Idaho Ungulates

2008 ◽  
Vol 75 (3) ◽  
pp. 862-865 ◽  
Author(s):  
Jeremy J. Gilbreath ◽  
Malcolm S. Shields ◽  
Rebekah L. Smith ◽  
Larry D. Farrell ◽  
Peter P. Sheridan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Wild Ungulates ◽  
Fecal Samples ◽  
Shiga Toxins ◽  
Toxin Genes ◽  
Genes Encoding ◽  
Shiga Toxin Genes

ABSTRACT Cattle are a known reservoir of Shiga toxin-producing Escherichia coli. The prevalence and stability of Shiga toxin and/or Shiga toxin genes among native wild ungulates in Idaho were investigated. The frequency of both Shiga genes and toxin was similar to that reported for Idaho cattle (∼19%).

Characteristics of Shiga Toxin–Producing Escherichia coli Isolated from Swiss Raw Milk Cheese within a 3-Year Monitoring Program

2010 ◽  
Vol 73 (1) ◽  
pp. 88-91 ◽  
Author(s):  
C. ZWEIFEL ◽  
N. GIEZENDANNER ◽  
S. CORTI ◽  
G. KRAUSE ◽  
L. BEUTIN ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Monitoring Program ◽  
Raw Milk ◽  
Health Impact ◽  
Toxin Gene ◽  
Genes Encoding ◽  
Raw Milk Cheese ◽  
Human Infections ◽  
Hard Cheese

Food is an important vehicle for transmission of Shiga toxin–producing Escherichia coli (STEC). To assess the potential public health impact of STEC in Swiss raw milk cheese produced from cow's, goat's, and ewe's milk, 1,422 samples from semihard or hard cheese and 80 samples from soft cheese were examined for STEC, and isolated strains were further characterized. By PCR, STEC was detected after enrichment in 5.7% of the 1,502 raw milk cheese samples collected at the producer level. STEC-positive samples comprised 76 semihard, 8 soft, and 1 hard cheese. By colony hybridization, 29 STEC strains were isolated from 24 semihard and 5 soft cheeses. Thirteen of the 24 strains typeable with O antisera belonged to the serogroups O2, O22, and O91. More than half (58.6%) of the 29 strains belonged to O:H serotypes previously isolated from humans, and STEC O22:H8, O91:H10, O91:H21, and O174:H21 have also been identified as agents of hemolytic uremic syndrome. Typing of Shiga toxin genes showed that stx1 was only found in 2 strains, whereas 27 strains carried genes encoding for the Stx2 group, mainly stx2 and stx2vh-a/b. Production of Stx2 and Stx2vh-a/b subtypes might be an indicator for a severe outcome in patients. Nine strains harbored hlyA (enterohemorrhagic E. coli hemolysin), whereas none tested positive for eae (intimin). Consequently, semihard and hard raw milk cheese may be a potential source of STEC, and a notable proportion of the isolated non-O157 STEC strains belonged to serotypes or harbored Shiga toxin gene variants associated with human infections.

scholarly journals Ferric uptake regulator (Fur) reversibly binds a [2Fe-2S] cluster to sense intracellular iron homeostasis in Escherichia coli

2020 ◽  
Vol 295 (46) ◽  
pp. 15454-15463 ◽  
Author(s):  
Chelsey R. Fontenot ◽  
Homyra Tasnim ◽  
Kathryn A. Valdes ◽  
Codrina V. Popescu ◽  
Huangen Ding
Keyword(s):  
Escherichia Coli ◽  
Iron Content ◽  
Iron Homeostasis ◽  
Ferric Uptake Regulator ◽  
Intracellular Iron ◽  
Free Iron ◽  
E Coli ◽  
Genes Encoding ◽  
Fur Protein ◽  
Mutant Cells

The ferric uptake regulator (Fur) is a global transcription factor that regulates intracellular iron homeostasis in bacteria. The current hypothesis states that when the intracellular “free” iron concentration is elevated, Fur binds ferrous iron, and the iron-bound Fur represses the genes encoding for iron uptake systems and stimulates the genes encoding for iron storage proteins. However, the “iron-bound” Fur has never been isolated from any bacteria. Here we report that the Escherichia coli Fur has a bright red color when expressed in E. coli mutant cells containing an elevated intracellular free iron content because of deletion of the iron–sulfur cluster assembly proteins IscA and SufA. The acid-labile iron and sulfide content analyses in conjunction with the EPR and Mössbauer spectroscopy measurements and the site-directed mutagenesis studies show that the red Fur protein binds a [2Fe-2S] cluster via conserved cysteine residues. The occupancy of the [2Fe-2S] cluster in Fur protein is ∼31% in the E. coli iscA/sufA mutant cells and is decreased to ∼4% in WT E. coli cells. Depletion of the intracellular free iron content using the membrane-permeable iron chelator 2,2´-dipyridyl effectively removes the [2Fe-2S] cluster from Fur in E. coli cells, suggesting that Fur senses the intracellular free iron content via reversible binding of a [2Fe-2S] cluster. The binding of the [2Fe-2S] cluster in Fur appears to be highly conserved, because the Fur homolog from Hemophilus influenzae expressed in E. coli cells also reversibly binds a [2Fe-2S] cluster to sense intracellular iron homeostasis.

Detection of genes encoding for virulence and adherence factors in Escherichia coli isolated in slaughtered Sarda breed sheep

2014 ◽  
Vol 168 (1) ◽  
pp. 234-239 ◽  
Author(s):  
Busia Gianluca ◽  
Mureddu Anna ◽  
Mazza Roberta ◽  
Meloni Domenico ◽  
Consolati Simonetta G. ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genes Encoding ◽  
Breed Sheep
Sign in / Sign up

Export Citation Format

Share Document