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Literature summary for 2.3.1.183 extracted from

  • Madduri, K.M.; Snodderley, E.M.
    Expression of phosphinothricin N-acetyltransferase in Escherichia coli and Pseudomonas fluorescens: Influence of mRNA secondary structure, host, and other physiological conditions (2007), Protein Expr. Purif., 55, 352-360.
    View publication on PubMed
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Application

Application Comment Organism
agriculture since the presence of the PAT gene confers resistance to the host harboring it, this gene has been used extensively as a selectable marker for the transformation of a wide range of plants Streptomyces viridochromogenes

Cloned(Commentary)

Cloned (Comment) Organism
a linear template for in vitro transcription and translation is created, the PAT gene is cloned into the vector pCR2.1TOPO, verified and subcloned into pET280 and pDOW1169 for expression in Escherichia coli or Pseudomonas fluorescens, respectively Streptomyces viridochromogenes
overexpression in Escherichia coli Bl21(DE3) and Pseudomonas fluorescens strain DC454 requiring modification of the 5' end of the pat ORF, in vitro transcription and translation, optimization of the expression method, Pseudomonas fluorescens is the better host, the secondary structures in the 5' coding region of pat gene influences the expression efficiency, overview Streptomyces viridochromogenes

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
21000
-
 determined by SDS-PAGE Streptomyces viridochromogenes

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acetyl-CoA + phosphinothricin Streptomyces viridochromogenes
-
 CoA + N-acetylphosphinothricin
-
 ?

Organism

Organism UniProt Comment Textmining
Streptomyces viridochromogenes
-
-
-

Purification (Commentary)

Purification (Comment) Organism
for the purification of PAT from Pseudomonas fluorescens ammonium sulfate precipitation, a Source 15 Phenyl XK hydrophobic interaction column, and a Superdex 75 XK gel filtration column are used Streptomyces viridochromogenes
recombinant enzyme from Pseudomonas fluorescens to homogeneity by ammonium sulfate fractionation, hydrophobic interaction chromatography, and gel filtration Streptomyces viridochromogenes

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + phosphinothricin
-
 Streptomyces viridochromogenes CoA + N-acetylphosphinothricin
-
 ?

Synonyms

Synonyms Comment Organism
Pat
-
 Streptomyces viridochromogenes
phosphinothricin N-acetyltransferase
-
 Streptomyces viridochromogenes