scholarly journals Cloning and sequencing of a cellobiose phosphotransferase system operon from Bacillus stearothermophilus XL-65-6 and functional expression in Escherichia coli.

1993 ◽  
Vol 175 (20) ◽  
pp. 6441-6450 ◽  
Author(s):  
X Lai ◽  
L O Ingram
Keyword(s):  
Escherichia Coli ◽  
Bacillus Stearothermophilus ◽  
Functional Expression ◽  
Phosphotransferase System ◽  
Cloning And Sequencing ◽  
Expression In Escherichia Coli

scholarly journals Cloning and Sequencing of a Poly(dl-Lactic Acid) Depolymerase Gene from Paenibacillus amylolyticus Strain TB-13 and Its Functional Expression in Escherichia coli

2003 ◽  
Vol 69 (5) ◽  
pp. 2498-2504 ◽  
Author(s):  
Yukie Akutsu-Shigeno ◽  
Teerawat Teeraphatpornchai ◽  
Kamonluck Teamtisong ◽  
Nobuhiko Nomura ◽  
Hiroo Uchiyama ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Amino Acid ◽  
Functional Expression ◽  
Amino Acid Residues ◽  
Cloning And Sequencing ◽  
Gene Encoding ◽  
Butylene Succinate ◽  
Expression In Escherichia Coli ◽  
Poly Ethylene

ABSTRACT The gene encoding a poly(dl-lactic acid) (PLA) depolymerase from Paenibacillus amylolyticus strain TB-13 was cloned and overexpressed in Escherichia coli. The purified recombinant PLA depolymerase, PlaA, exhibited degradation activities toward various biodegradable polyesters, such as poly(butylene succinate), poly(butylene succinate-co-adipate), poly(ethylene succinate), and poly(ε-caprolactone), as well as PLA. The monomeric lactic acid was detected as the degradation product of PLA. The substrate specificity toward triglycerides and p-nitrophenyl esters indicated that PlaA is a type of lipase. The gene encoded 201 amino acid residues, including the conserved pentapeptide Ala-His-Ser-Met-Gly, present in the lipases of mesophilic Bacillus species. The identity of the amino acid sequence of PlaA with Bacillus lipases was no more than 45 to 50%, and some of its properties were different from those of these lipases.

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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