scholarly journals Isolation and characterization of the flavin-binding domain of flavocytochrome b2 expressed independently in Escherichia coli

1995 ◽  
Vol 309 (2) ◽  
pp. 601-605 ◽  
Author(s):  
A Balme ◽  
C E Brunt ◽  
R L Pallister ◽  
S K Chapman ◽  
G A Reid
Keyword(s):  
Escherichia Coli ◽  
Binding Domain ◽  
Flavin Mononucleotide ◽  
Exponential Process ◽  
Flavocytochrome B2 ◽  
Terminal Domain ◽  
Isolation And Characterization ◽  
Flavin Binding ◽  
Single Exponential

Flavocytochrome b2 consists of two distinct domains. The N-terminal domain contains protohaem IX and the larger, C-terminal domain contains flavin mononucleotide (FMN). We describe here the isolation of the flavin-binding domain expressed in Escherichia coli independent of the cytochrome domain. The isolated domain is an efficient lactate dehydrogenase with ferricyanide as electron acceptor but reduces cytochrome c, the physiological oxidant for flavocytochrome b2, extremely poorly; electron transfer from the flavin-binding domain to the separately expressed cytochrome domain is undetectable. FMN reduction by lactate occurs as a single exponential process in the isolated flavin-binding domain, in contrast to the biphasic kinetics observed with native flavocytochrome b2.

scholarly journals Isolation and characterization of the cytochrome domain of flavocytochrome b2 expressed independently in Escherichia coli

1992 ◽  
Vol 283 (1) ◽  
pp. 87-90 ◽  
Author(s):  
C E Brunt ◽  
M C Cox ◽  
A G P Thurgood ◽  
G R Moore ◽  
G A Reid ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Field ◽  
Cytochrome B5 ◽  
Microsomal Cytochrome ◽  
E Coli ◽  
Flavocytochrome B2 ◽  
Isolation And Characterization ◽  
Exchange Rate Constant ◽  
Microsomal Cytochrome B5

The cytochrome domain of flavocytochrome b2 (L-lactate dehydrogenase) was expressed in the bacterium Escherichia coli and a purification procedure was developed. When expressed in E. coli, the b2-cytochrome domain contains protohaem IX and has an electronic absorption spectrum identical with that of the cytochrome b2 ‘core’ produced by proteolytic cleavage of the enzyme isolated from yeast. The b2-cytochrome domain isolated from E. coli has an Mr of 10,500 and a redox potential of -31 +/- 2 mV. High-field n.m.r. studies indicate pKa values for the haem propionate groups to be 4.8 and 4.6, consistent with these groups being exposed to solvent rather than buried inside the protein. Using n.m.r. spectroscopy, we have determined an electron self-exchange rate constant for the b2-cytochrome domain of 2.3 x 10(6) M-1.s-1, which is more than two orders of magnitude larger than the value obtained for microsomal cytochrome b5, a homologue of b2-cytochrome domain.

Isolation and characterization of a novel Escherichia coli Kayfunavirus phage DY1

2021 ◽  
Vol 293 ◽  
pp. 198274
Author(s):  
Xiaoming Yuan ◽  
Shuhong Zhang ◽  
Juan Wang ◽  
Chun Li ◽  
Na Li ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Isolation And Characterization

scholarly journals Isolation and characterization of pathogenic Escherichia coli bacteriophages from chicken and beef offal

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Celosia Lukman ◽  
Christopher Yonathan ◽  
Stella Magdalena ◽  
Diana Elizabeth Waturangi
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
High Efficiency ◽  
Multiplicity Of Infection ◽  
Isolation And Characterization ◽  
Transmission Electron ◽  
Pathogenic Escherichia Coli ◽  
Family Based

Abstract Objective This study was conducted to isolate and characterize lytic bacteriophages for pathogenic Escherichia coli from chicken and beef offal, and analyze their capability as biocontrol for several foodborne pathogens. Methods done in this research are bacteriophage isolation, purification, titer determination, application, determination of host range and minimum multiplicity of infection (miMOI), and bacteriophage morphology. Results Six bacteriophages successfully isolated from chicken and beef offal using EPEC and EHEC as host strain. Bacteriophage titers observed between 109 and 1010 PFU mL−1. CS EPEC and BL EHEC bacteriophage showed high efficiency in reduction of EPEC or EHEC contamination in meat about 99.20% and 99.04%. The lowest miMOI was 0.01 showed by CS EPEC bacteriophage. CI EPEC and BL EPEC bacteriophage suspected as Myoviridae family based on its micrograph from Transmission Electron Microscopy (TEM). Refers to their activity, bacteriophages isolated in this study have a great potential to be used as biocontrol against several foodborne pathogens.

Isolation and characterization of the heat stable enterotoxin from a pathogenic bovine strain of Escherichia coli

1984 ◽  
Vol 9 (4) ◽  
pp. 399-414 ◽  
Author(s):  
Charles Gerday ◽  
Marianne Herman ◽  
Jacques Olivy ◽  
Nicole Gerardin-Otthiers ◽  
Dominique Art ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Heat Stable ◽  
Isolation And Characterization ◽  
Bovine Strain
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