The Hydroxymethyldihydropterin Pyrophosphokinase Domain of the Multifunctional Folic Acid Synthesis Fas Protein of Pneumocystis carinii Expressed as an Independent Enzyme in Escherichia coli: Refolding and Characterization of the Recombinant Enzyme

1994 ◽  
Vol 5 (4) ◽  
pp. 371-378 ◽  
Author(s):  
S.P. Ballantine ◽  
F. Volpe ◽  
C.J. Delves
Keyword(s):  
Escherichia Coli ◽  
Folic Acid ◽  
Pneumocystis Carinii ◽  
Acid Synthesis ◽  
Recombinant Enzyme

Identification of a halophilic α-amylase gene from Escherichia coli JM109 and characterization of the recombinant enzyme

2013 ◽  
Vol 35 (7) ◽  
pp. 1061-1065 ◽  
Author(s):  
Yutuo Wei ◽  
Xiaobo Wang ◽  
Jiayuan Liang ◽  
Xue Li ◽  
Liqin Du ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Enzyme ◽  
Amylase Gene ◽  
Escherichia Coli Jm109

Expression of the pyranose 2-oxidase from Trametes pubescens in Escherichia coli and characterization of the recombinant enzyme

2005 ◽  
Vol 120 (4) ◽  
pp. 387-395 ◽  
Author(s):  
Helena Marešová ◽  
Branislav Večerek ◽  
Marcela Hradská ◽  
Nathalie Libessart ◽  
Stanislav Bečka ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Enzyme

ucFabV Requires Functional Reductase Activity to Confer Reduced Triclosan Susceptibility in Escherichia coli

2015 ◽  
Vol 25 (6) ◽  
pp. 394-402 ◽  
Author(s):  
Taylor L. Fischer ◽  
Robert J. White ◽  
Katherine F.K. Mares ◽  
Devin E. Molnau ◽  
Justin J. Donato
Keyword(s):  
Escherichia Coli ◽  
Reductase Activity ◽  
Acid Synthesis ◽  
Temperature Sensitive ◽  
Wild Type ◽  
E Coli ◽  
Enoyl Acp Reductase ◽  
Selection For

<b><i>Background/Aims:</i></b> We previously identified the Triclo1 fosmid in a functional metagenomic selection for clones that increased triclosan tolerance in <i>Escherichia coli</i>. The active enzyme encoded by Triclo1 is ucFabV. Although ucFabV is homologous to FabV from other organisms, ucFabV contains substitutions at key positions that would predict differences in substrate binding. Therefore, a detailed characterization of ucFabV was conducted to link its biochemical activity to its ability to confer reduced triclosan sensitivity. <b><i>Methods:</i></b> ucFabV and a catalytic mutant were purified and used to reduce crotonoyl-CoA in vitro. The mutant and wild-type enzymes were introduced into <i>E. coli</i>, and their ability to confer triclosan tolerance as well as suppress a temperature-sensitive mutant of FabI were measured. <b><i>Results:</i></b> Purified ucFabV, but not the mutant, reduced crotonoyl-CoA in vitro. The wild-type enzyme confers increased triclosan tolerance when introduced into <i>E. coli</i>, whereas the mutant remained susceptible to triclosan<i>. </i>Additionally, wild-type ucFabV, but not the mutant, functionally replaced FabI within living cells. <b><i>Conclusion:</i></b> ucFabV confers increased tolerance through its function as an enoyl-ACP reductase. Furthermore, ucFabV is capable of restoring viability in the presence of compromised FabI, suggesting ucFabV is likely facilitating an alternate step within fatty acid synthesis, bypassing FabI inhibition.

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