Functional expression in Escherichia coli of the Haemophilus influenzae gene coding for selenocysteine-containing selenophosphate synthetase

1997 ◽  
Vol 169 (1) ◽  
pp. 71-75 ◽  
Author(s):  
Reinhard Wilting ◽  
Kalliopi Vamvakidou ◽  
A. Böck
Keyword(s):  
Escherichia Coli ◽  
Haemophilus Influenzae ◽  
Functional Expression ◽  
Expression In Escherichia Coli ◽  
Gene Coding ◽  
Selenophosphate Synthetase

Arabidopsis homologs of the shaggy and GSK-3 protein kinases: molecular cloning and functional expression in Escherichia coli

1994 ◽  
Vol 242 (3) ◽  
pp. 337-345 ◽  
Author(s):  
Michele W. Bianchi ◽  
Dominique Guivarc'h ◽  
Martine Thomas ◽  
James R. Woodgett ◽  
Martin Kreis
Keyword(s):  
Escherichia Coli ◽  
Molecular Cloning ◽  
Protein Kinases ◽  
Functional Expression ◽  
Expression In Escherichia Coli

scholarly journals Cloning of a Novel Aldo-Keto Reductase Gene from Klebsiella sp. Strain F51-1-2 and Its Functional Expression in Escherichia coli

2007 ◽  
Vol 73 (15) ◽  
pp. 4959-4965 ◽  
Author(s):  
Hong Jiang ◽  
Chao Yang ◽  
Hong Qu ◽  
Zheng Liu ◽  
Q. S. Fu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Organophosphorus Compounds ◽  
Functional Expression ◽  
Nitrilotriacetic Acid ◽  
Coding Region ◽  
Reductive Transformation ◽  
Cofactor Specificity ◽  
Expression In Escherichia Coli ◽  
Reductase Gene ◽  
One Step

ABSTRACT A soil bacterium capable of metabolizing organophosphorus compounds by reducing the P═S group in the molecules was taxonomically identified as Klebsiella sp. strain F51-1-2. The gene involved in the reduction of organophosphorus compounds was cloned from this strain by the shotgun technique, and the deduced protein (named AKR5F1) showed homology to members of the aldo-keto reductase (AKR) superfamily. The intact coding region for AKR5F1 was subcloned into vector pET28a and overexpressed in Escherichia coli BL21(DE3). Recombinant His6-tagged AKR5F1 was purified in one step using Ni-nitrilotriacetic acid affinity chromatography. Assays for cofactor specificity indicated that reductive transformation of organophosphorus compounds by the recombinant AKR5F1 specifically required NADH. The kinetic constants of the purified recombinant AKR5F1 toward six thion organophosphorus compounds were determined. For example, the Km and k cat values of reductive transformation of malathion by the purified recombinant AKR5F1 are 269.5 ± 47.0 μΜ and 25.7 ± 1.7 min−1, respectively. Furthermore, the reductive transformation of organophosphorus compounds can be largely explained by structural modeling.

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