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Literature summary extracted from

  • Rohde, B.; Hans, J.; Martens, S.; Baumert, A.; Hunziker, P.; Matern, U.
    Anthranilate N-methyltransferase, a branch-point enzyme of acridone biosynthesis (2008), Plant J., 53, 541-553.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.111 expressed in Escherichia coli, BL21 cells for induction of recombinant protein expression Ruta graveolens

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.111 N298E generated by site-directed mutagenesis Ruta graveolens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.111 0.003
-
 S-adenosyl-L-methionine wild-type Ruta graveolens
2.1.1.111 0.003
-
 S-adenosyl-L-methionine N298E mutant Ruta graveolens
2.1.1.111 0.007
-
 anthranilate wild-type Ruta graveolens
2.1.1.111 0.024
-
 anthranilate N298E mutant Ruta graveolens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.111 Ca2+ 73% relative enzyme activity in presence of 1.5 mM, significantly reduced Ruta graveolens
2.1.1.111 Co2+ activity reduced in presence of 1.5 mM, 25% relative activity retained Ruta graveolens
2.1.1.111 Cu2+ activity abolished in presence of 1.5 mM of Ruta graveolens
2.1.1.111 Fe2+ activity nearly abolished in presence of 1.5 mM of Ruta graveolens
2.1.1.111 Fe3+ activity abolished in presence of 1.5 mM of Ruta graveolens
2.1.1.111 Mg2+ activity independent in presence of 1.5 mM of, 90% relative activity retained Ruta graveolens
2.1.1.111 Zn2+ activity abolished in presence of 1.5 mM of Ruta graveolens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.111 70000
-
 gel filtration, SDS-PAGE Ruta graveolens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.111 S-adenosyl-L-methionine + anthranilate Ruta graveolens first specific step in acridone alkaloid biosynthesis S-adenosyl-L-homocysteine + N-methylanthranilate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.111 Ruta graveolens A9X7L0 line R-20
-
2.3.1.159 Ruta graveolens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.111 gel filtration, SDS-PAGE, wild-type and mutant protein Ruta graveolens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.1.1.111 carpel lining the inside of Ruta graveolens
-
2.1.1.111 cell suspension culture crude extracts of total plants, elicited, 2 years old Ruta graveolens
-
2.1.1.111 flower high mRNA expression and enzyme activity Ruta graveolens
-
2.1.1.111 leaf moderate mRNA expression Ruta graveolens
-
2.1.1.111 ovary central vascular bundles of, around cup-like structures in the outer ovary wall representing secretory oil glands Ruta graveolens
-
2.1.1.111 root rhizodermis of older root parts, endodermis of younger root parts, high mRNA expression and enzyme activity Ruta graveolens
-
2.1.1.111 stem vascular region of secondary and primary shoots, moderate mRNA expression, high enzyme activity Ruta graveolens
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.1.111 additional information
-
 in-gel digestion and peptide sequencing by mass spectrometry, Western Blot and phylogenetic analysis, assays conducted at 32°C in 73.5 mM glycine-NaOH buffer at pH 8.9, HPLC separation of assay extracts, list of compounds that are not accepted as substrate in assay Ruta graveolens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.111 S-adenosyl-L-methionine + anthranilate first specific step in acridone alkaloid biosynthesis Ruta graveolens S-adenosyl-L-homocysteine + N-methylanthranilate
-
 ?
2.1.1.111 S-adenosyl-L-methionine + anthranilate first specific step in alkaloid biosynthesis, substrate specificity determines the course of acridone biosynthesis Ruta graveolens S-adenosyl-L-homocysteine + N-methylanthranilate
-
 ?
2.3.1.159 N-methylanthraniloyl-CoA + 3 malonyl-CoA anthraniloyl CoA is not accepted as starter substrate, the anthranilate N-methylation prior to CoA activation is a key step in acridone alkaloid formation, anthranilate N-methyltransferase is a branch-point enzyme of acridone biosynthesis Ruta graveolens 1,3-dihydroxy-N-methylacridone + CoA + CO2
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.1.1.111 ANMT
-
 Ruta graveolens
2.3.1.159 acridone synthase
-
 Ruta graveolens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.111 32
-
 assay at Ruta graveolens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.111 8
-
 activity optimum at Ruta graveolens