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Literature summary for 3.1.3.2 extracted from

  • Leelapon, O.; Sarath, G.; Staswick, P.E.
    A single amino acid substitution in soybean VSPalpha increases its acid phosphatase activity nearly 20-fold (2004), Planta, 219, 1071-1079.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of GST-tagged wild-type and mutant APase in Escherichia coli strain BL21(DE3) Glycine max
DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of GST-tagged wild-type and mutant VSPalpha in Escherichia coli strain BL21(DE3) Glycine max

Protein Variants

Protein Variants Comment Organism
S106D site-directed mutagenesis of VSPalpha, the mutation increases the enzyme activity about 20fold with substrate GMP compared to the wild-type enzyme Glycine max

Inhibitors

Inhibitors Comment Organism Structure
cAMP
-
Glycine max
GMP
-
Glycine max

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.9
-
GMP pH 6.0, 37°C, recombinant enzyme Glycine max
8.8
-
4-nitrophenol phosphate pH 6.0, 37°C, recombinant enzyme Glycine max

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Glycine max

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
27000
-
x * 27000, recombinant enzyme, SDS-PAGE Glycine max

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Glycine max VSPalpha functions as temporary storage reserve and as plant acid phosphatase ?
-
?

Organism

Organism UniProt Comment Textmining
Glycine max O49855 APase
-
Glycine max P10742 VSPalpha, i.e. VSP25, fragment
-

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged wild-type and mutant APase 3.0fold from Escherichia coli strain BL21(DE3) by ammonium sulfate precipitation and ion exchange chromatography Glycine max
recombinant GST-tagged wild-type and mutant VSPalpha from Escherichia coli strain BL21(DE3) by ammonium sulfate precipitation and ion exchange chromatography, or by glutathione affinity chromatography Glycine max

Source Tissue

Source Tissue Comment Organism Textmining
root nodule APase Glycine max
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.5
-
purified recombinant enzyme, substrate phospho-L-tyrosine Glycine max
3.2
-
purified recombinant enzyme, substrate 4-nitrophenyl phosphate Glycine max
20
-
purified recombinant S106D mutant enzyme, substrate phospho-L-tyrosine Glycine max
22.6
-
purified recombinant enzyme, substrate GMP Glycine max
35
-
purified recombinant S106D mutant enzyme, substrate 4-nitrophenyl phosphate Glycine max
72.1
-
purified recombinant GST-tagged enzyme, substrate 4-nitrophenyl phosphate Glycine max
439
-
purified recombinant S106D mutant enzyme, substrate GMP Glycine max
845
-
purified recombinant detagged enzyme, substrate GMP Glycine max

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl phosphate + H2O
-
Glycine max 4-nitrophenol + phosphate
-
?
ADP + H2O
-
Glycine max AMP + phosphate
-
?
FMN + H2O
-
Glycine max flavin nucleoside + phosphate
-
?
GMP + H2O
-
Glycine max guanosine + phosphate
-
?
GMP + H2O best substrate, recombinant enzyme Glycine max guanosine + phosphate
-
?
additional information VSPalpha functions as temporary storage reserve and as plant acid phosphatase Glycine max ?
-
?
additional information ADP, FMN, and triphosphate are no substrates Glycine max ?
-
?
additional information no activity with triphosphate Glycine max ?
-
?
phospho-L-tyrosine + H2O
-
Glycine max L-tyrosine + phosphate
-
?

Subunits

Subunits Comment Organism
? x * 27000, recombinant enzyme, SDS-PAGE Glycine max

Synonyms

Synonyms Comment Organism
APase
-
Glycine max
More VSPalpha belongs to the haloacid dehalogenase superfamily Glycine max
vegetative storage protein alpha
-
Glycine max
VSP25
-
Glycine max
VSPalpha
-
Glycine max

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Glycine max

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
-
Glycine max
6
-
assay at Glycine max

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.015
-
GMP pH 6.0, 37°C, recombinant GST-tagged enzyme Glycine max