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Literature summary extracted from

  • Suzuki, H.; Sawada, S.; Watanabe, K.; Nagae, S.; Yamaguchi, M.A.; Nakayama, T.; Nishino, T.
    Identification and characterization of a novel anthocyanin malonyltransferase from scarlet sage (Salvia splendens) flowers: an enzyme that is phylogenetically separated from other anthocyanin acyltransferases (2004), Plant J., 38, 994-1003.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.214 Ss5MaT2 or SsAT412, DNA and amino acid sequence determination and analysis, phylogenetic analysis, soluble N-terminally His-tagged SsAT412 expression in Escherichia coli strain JM109 Salvia splendens
2.3.1.224 expressed in Escherichia coli JM109 cells Salvia splendens

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.1.224 acetyl-CoA 32% residual activity at 0.1 mM Salvia splendens
2.3.1.224 bisdemalonylsalvianin 31% residual activity at 0.05 mM Salvia splendens
2.3.1.224 caffeic acid 38% residual activity at 1 mM Salvia splendens
2.3.1.224 Cd2+ complete inhibition at 0.1 mM Salvia splendens
2.3.1.224 CoA-SH complete inhibition at 0.1 mM Salvia splendens
2.3.1.224 Cu2+ complete inhibition at 0.1 mM Salvia splendens
2.3.1.224 diethyldicarbonate 69% residual activity at 0.1 mM Salvia splendens
2.3.1.224 Fe2+ complete inhibition at 0.1 mM Salvia splendens
2.3.1.224 Hg+ complete inhibition at 0.1 mM Salvia splendens
2.3.1.224 N-ethylmaleimide complete inhibition at 5 mM Salvia splendens
2.3.1.224 Zn2+ complete inhibition at 0.1 mM Salvia splendens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.214 0.0069
-
 malonyl-CoA pH 8.5, temperature not specified in the publication Salvia splendens
2.3.1.214 0.0513
-
 4'''-demalonylsalvianin pH 8.5, temperature not specified in the publication Salvia splendens
2.3.1.224 0.0046
-
 malonyl-CoA recombinant enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 0.0069
-
 malonyl-CoA native enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 0.0513
-
 monodemalonylsalvianin native enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 0.0563
-
 monodemalonylsalvianin recombinant enzyme, at pH 7.0 and 30°C Salvia splendens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.3.1.224 additional information not influenced by Ca2+, Co2+, Mg2+, Mn2+, Ni2+, and Sn2+ Salvia splendens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.1.214 43643
-
 1 * 46000, SDS-PAGE, 1 * 43643, sequence calculation Salvia splendens
2.3.1.214 45000
-
 gel filtration Salvia splendens
2.3.1.214 46000
-
 1 * 46000, SDS-PAGE, 1 * 43643, sequence calculation Salvia splendens
2.3.1.224 46000
-
 gel filtration Salvia splendens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.224 malonyl-CoA + cyanidin 3-O-(6''-O-4-coumarylglucoside)-5-O-(6'''-O-malonylglucoside) Salvia splendens
-
 CoA + ?
-
 ?
2.3.1.224 malonyl-CoA + monodemalonylsalvianin Salvia splendens
-
 CoA + salvianin
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.214 Salvia splendens
-
 scarlet sage
-
2.3.1.214 Salvia splendens Q6TXD2
-
-
2.3.1.224 Salvia splendens Q6TXD2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.214 native Ss5MaT2 510fold by ammonium sulfate fractionation, a first anion exchange chromatographic step, hydrophobic interaction and ceramic hydroxyapatite chromatography, gel filtration, and a second, different anion exchange chromatograpic step to over 95% purity. Recombinant N-terminally His-tagged SsAT412 from Escherichia coli strain JM109 to homogeneity Salvia splendens
2.3.1.224 ammonium sulfate fractionation, DEAE-Sepharose column chromatography, phenyl Sepharose column chromatography, Mono Q column chromatography, and Superdex 200 gel filtration Salvia splendens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.3.1.214 flower recently opened, fully pigmented flowers Salvia splendens
-
2.3.1.214 additional information quantitative real-time PCR expression analysis in tissues, ovverview Salvia splendens
-
2.3.1.224 flower highest expression Salvia splendens
-
2.3.1.224 root
-
 Salvia splendens
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.1.214 0.144
-
 purified native enzyme, pH 8.5, temperature not specified in the publication Salvia splendens
2.3.1.224 0.0000047
-
 crude extract, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 0.0024
-
 after 510fold purification, at pH 7.0 and 30°C Salvia splendens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.214 malonyl-CoA + 4'''-demalonylsalvianin
-
 Salvia splendens CoA + salvianin
-
 ?
2.3.1.214 malonyl-CoA + monodemalonylsalvianin Ss5MaT2 activity with monodemalonylsalvianin Salvia splendens CoA + salvianin
-
 ?
2.3.1.214 additional information the enzyme does not show any 5MaT1 activity with bisdemalonylsalvianin as substrate Salvia splendens ?
-
 ?
2.3.1.224 malonyl-CoA + cyanidin 3-O-(6''-O-4-coumarylglucoside)-5-O-(6'''-O-malonylglucoside)
-
 Salvia splendens CoA + ?
-
 ?
2.3.1.224 malonyl-CoA + monodemalonylsalvianin
-
 Salvia splendens CoA + salvianin
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.1.214 monomer 1 * 46000, SDS-PAGE, 1 * 43643, sequence calculation Salvia splendens
2.3.1.224 monomer 1 * 46000, SDS-PAGE Salvia splendens

Synonyms

EC Number Synonyms Comment Organism
2.3.1.214 malonyl-CoA:anthocyanin 5-glucoside 4'''-O-malonyltransferase
-
 Salvia splendens
2.3.1.214 Ss5MaT2
-
 Salvia splendens
2.3.1.214 SsAT412
-
 Salvia splendens
2.3.1.224 5MaT2
-
 Salvia splendens
2.3.1.224 anthocyanin malonyltransferase
-
 Salvia splendens
2.3.1.224 malonyl-CoA:anthocyanin 5-glucoside 4'''-O-malonyltransferase
-
 Salvia splendens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.214 0.55
-
 malonyl-CoA pH 8.5, temperature not specified in the publication Salvia splendens
2.3.1.214 0.55
-
 4'''-demalonylsalvianin pH 8.5, temperature not specified in the publication Salvia splendens
2.3.1.224 0.55
-
 malonyl-CoA native enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 0.55
-
 monodemalonylsalvianin native enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 0.76
-
 malonyl-CoA recombinant enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 0.76
-
 monodemalonylsalvianin recombinant enzyme, at pH 7.0 and 30°C Salvia splendens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.214 8.5
-
-
 Salvia splendens

General Information

EC Number General Information Comment Organism
2.3.1.214 evolution the enzyme belongs to the BAHD family of acyltrasferases, and phylogenetic analysis shows, that it is more related to the acetyl-CoA:benzylalcohol acetyltransferases than to the anthocyanin acyltransferases. Ss5MaT2 contains the BAHD conserved motifs 1 HXXXD and 3 DFGWG, but lacks motif 2 YFGNC Salvia splendens

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.3.1.214 10.7
-
 4'''-demalonylsalvianin pH 8.5, temperature not specified in the publication Salvia splendens
2.3.1.214 79.7
-
 malonyl-CoA pH 8.5, temperature not specified in the publication Salvia splendens
2.3.1.224 10.7
-
 monodemalonylsalvianin native enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 13.5
-
 monodemalonylsalvianin recombinant enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 79.7
-
 malonyl-CoA native enzyme, at pH 7.0 and 30°C Salvia splendens
2.3.1.224 165
-
 malonyl-CoA recombinant enzyme, at pH 7.0 and 30°C Salvia splendens