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Information on EC 2.3.1.183 - phosphinothricin acetyltransferase and Organism(s) Streptomyces viridochromogenes and UniProt Accession Q57146

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IUBMB Comments
The substrate phosphinothricin is used as a nonselective herbicide and is a potent inhibitor of EC 6.3.1.2, glutamine synthetase, a key enzyme of nitrogen metabolism in plants .
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Streptomyces viridochromogenes
UNIPROT: Q57146
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The taxonomic range for the selected organisms is: Streptomyces viridochromogenes
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
phosphinothricin acetyltransferase, phosphinothricin-n-acetyltransferase, phosphinothricin n-acetyltransferase, ppt acetyltransferase, gk2920, gk0593, dr_1182, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphinothricin-N-acetyltransferase
-
phosphinothricin N-acetyltransferase
-
-
phosphinothricin-N-acetyltransferase
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acyl group transfer
-
-
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:phosphinothricin N-acetyltransferase
The substrate phosphinothricin is used as a nonselective herbicide and is a potent inhibitor of EC 6.3.1.2, glutamine synthetase, a key enzyme of nitrogen metabolism in plants [2].
CAS REGISTRY NUMBER
COMMENTARY hide
111069-93-3
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + L-phosphinothricin
CoA + L-N-acetylphosphinothricin
show the reaction diagram
acetyl-CoA + phosphinothricin
CoA + N-acetylphosphinothricin
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
the enzyme confers resistance to the herbicide phosphinothricin
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + L-phosphinothricin
CoA + L-N-acetylphosphinothricin
show the reaction diagram
acetyl-CoA + phosphinothricin
CoA + N-acetylphosphinothricin
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
the enzyme confers resistance to the herbicide phosphinothricin
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
-
insertion of the gene encoding phosphinothricin acetyltransferase has resulted in cotton plants resistant to the herbicide glufosinate. Resistance to glufosinate is proportional to phosphinothricin acetyltransferase expression and activity in LibertyLink and WideStrike cotton
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20000
-
2 * 20000, about, recombinant enzyme
20621
-
x * 20621, nucleotide sequence calculation
21000
-
determined by SDS-PAGE
40000
-
recombinant enzyme, gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 20621, nucleotide sequence calculation
dimer
-
2 * 20000, about, recombinant enzyme
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
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recombinant enzyme from Escherichia coli by ammonium sulfate fractionation, anion exchange, hydrophobic interaction, and hydroxylapatite chromatography, followed by gel filtration, to about 90% purity
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recombinant enzyme from Pseudomonas fluorescens to homogeneity by ammonium sulfate fractionation, hydrophobic interaction chromatography, and gel filtration
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
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gene pat, DNA and amino acid sequence determination and analysis, translation of pat is initiated by a GTG codon, functional expression in transgenic Nicotiana tabacum plants by Agrobacterium tumefaciens strain LBA4404-mediated leaf-disc transformation replacing the GTG start codon by ATG, expression of wild-type and mutant enzymes in Streptomyces lividans strain TK23 and in Escherichia coli strains JM83 and S17.1, the gene confers resistance to L-phosphinothricin
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gene pat, expression in Nicotiana tabacum var. W38 plant leaves and roots using the root-specific pat promoter of the hemoglobin gene from Parasponia andersonii, the transgenic tobacco plants show herbicide tolerance, overview
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gene pat, expression in transgenic Nicotiana tabacum, Daucus carota, and Medicago sativa plants, the functional expression of gene pat confers resistance to the herbicide L-phosphinothricin
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gene pat, functional expression in transgenic Nicotiana tabacum and Daucus carota plant leaves using the Agrobacterium tumefaciens strain LBA4404 transfection of leaves and direct gene transfer to protoplasts, respectively, subcloning in Escherichia coli strain S 17. I, the transgenic plants are resistant to L-phosphinothricin
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gene PttR, cloning of the gene from an phosphinothricin-tripeptide-resistant mutant, DNA and amino acid sequence determination and analysis, promoter analysis, expression in Streptomyces lividans strain TK23 and Escherichia coli strain JM83 using different plasmids, overview, the gene confers resistance to L-phosphinothricin
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overexpression in Escherichia coli
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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
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pat gene is inserted into glufosinate-resistant maize hybrids (Herculex, Agrisure TL, Herculex Yieldgard, Leptra, Viptera 3, Power Core)
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
use of pat as marker gene for the screening of genetically modified crops
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
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Strauch, E.; Wohlleben, W.; Puhler, A.
Cloning of a phosphinothricin N-acetyltransferase gene from Streptomyces viridochromogenes Tu494 and its expression in Streptomyces lividans and Escherichia coli
Gene
63
65-74
1988
Streptomyces viridochromogenes, Streptomyces viridochromogenes Tue494
Manually annotated by BRENDA team
Wohllehen, W.; Arnold, W.; Broer, L.; Hillemaan, D.; Strauch, E.; Ptihler, A.
Nucleotide sequence of the phosphinothricin N-acetyltransferase gene from Streptomyces viridochromogenes T494 and its expression in Nicotiana tabacum
Gene
70
25-37
1988
Streptomyces viridochromogenes, Streptomyces viridochromogenes Tue494
Manually annotated by BRENDA team
van der Hoeven, C.; Dietz, A.; Landsmann, J.
Expression of phosphinothricin acetyltransferase from the root specific pat promoter in transgenic tobacco plants is sufficient for herbicide tolerance
Plant Cell Rep.
14
165-170
1994
Streptomyces viridochromogenes
Manually annotated by BRENDA team
Droege-Laser, W.; Siemeling, U.; Puhler, A.; Broer, I.
The metabolites of the herbicide L-phosphinothricin (glufosinate). Identification, stability, and mobility in transgenic, herbicide-resistant, and untransformed plants
Plant Physiol.
105
159-166
1994
Streptomyces viridochromogenes, Streptomyces viridochromogenes DSM 40736 / JCM 4977 / BCRC 1201 / Tue 494 / Tu 494
Manually annotated by BRENDA team
Droege, W.; Broer, I.; Puehler, A.
Transgenic plants containing the phosphinothricin-N-acetyltransferase gene metabolize the herbicide L-phosphinothricin (glufosinate) differently from untransformed plants
Planta
187
142-151
1992
Streptomyces viridochromogenes, Streptomyces viridochromogenes Tue494
Manually annotated by BRENDA team
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
Protein Expr. Purif.
55
352-360
2007
Streptomyces viridochromogenes
Manually annotated by BRENDA team
Vinnemeier, J.; Droege-Laser, W.; Pistorius, E.K.; Broer, I.
Purification and partial characterization of the Streptomyces viridochromogenes Tue494 phosphinothricin-N-acetyltransferase mediating resistance to the herbicide phosphinothricin in transgenic plants
Z. Naturforsch. C
50
796-805
1995
Streptomyces viridochromogenes, Streptomyces viridochromogenes Tue494
-
Manually annotated by BRENDA team
Randhawa, G.J.; Chhabra, R.; Singh, M.
Multiplex PCR-based simultaneous amplification of selectable marker and reporter genes for the screening of genetically modified crops
J. Agric. Food Chem.
57
5167-5172
2009
Streptomyces hygroscopicus (P16426), Streptomyces viridochromogenes (Q57146)
Manually annotated by BRENDA team
Carbonari, C.; Latorre, D.; Gomes, G.; Velini, E.; Owens, D.; Pan, Z.; Dayan, F.
Resistance to glufosinate is proportional to phosphinothricin acetyltransferase expression and activity in LibertyLink and WideStrike cotton
Planta
243
925-33
2016
Streptomyces hygroscopicus (P16426), Streptomyces hygroscopicus ATCC 21705 (P16426), Streptomyces viridochromogenes
Manually annotated by BRENDA team
Krenchinski, F.H.; Carbonari, C.A.; S Cesco, V.J.; P Albrecht, A.J.; Campos Arcuri, M.L.; de Godoy Maia, I.; Velini, E.D.
Glufosinate resistance level is proportional to phosphinothricin acetyltransferase gene expression in glufosinate-resistant maize
J. Agric. Food Chem.
66
12641-12650
2018
Streptomyces viridochromogenes
Manually annotated by BRENDA team