Information on EC 2.1.1.273 - benzoate O-methyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.1.1.273
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RECOMMENDED NAME
GeneOntology No.
benzoate O-methyltransferase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + benzoate = S-adenosyl-L-homocysteine + methyl benzoate
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
volatile benzenoid biosynthesis I (ester formation)
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SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:benzoate O-methyltransferase
While the enzyme from the plant Zea mays is specific for benzoate [6], the enzymes from Arabidopsis species and Clarkia breweri also catalyse the reaction of EC 2.1.1.274, salicylate 1-O-methyltransferase [1,5]. In snapdragon (Antirrhinum majus) two isoforms are found, one specific for benzoate [2,3] and one that is also active towards salicylate [4]. The volatile product is an important scent compound in some flowering species [2].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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-
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Manually annotated by BRENDA team
Lillium hybrid cultivar
var. Yelloween, a popular OT (oriental x trumpet) hybrid, collected from the field in Xiaotang­shan, Beijing, gene LiBSMT
UniProt
Manually annotated by BRENDA team
gene PbBSMT
UniProt
Manually annotated by BRENDA team
cv. Delprim
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
Lillium hybrid cultivar
the enzyme belongs to SABATH family, a class of O-methyltransferases and N-methyltransferases
metabolism
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final enzyme in the biosynthesis of methyl benzoate
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + 3-hydroxybenzoic acid
S-adenosyl-L-homocysteine + methyl 3-hydroxybenzoate
show the reaction diagram
54% relative activity at 1 mM methyl acceptor compared to activity with benzoic acid set at 100%
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-
?
S-adenosyl-L-methionine + anthranilic acid
S-adenosyl-L-homocysteine + methyl anthranilate
show the reaction diagram
S-adenosyl-L-methionine + benzoate
S-adenosyl-L-homocysteine + methyl benzoate
show the reaction diagram
S-adenosyl-L-methionine + benzoic acid
S-adenosyl-L-homocysteine + methyl benzoate
show the reaction diagram
S-adenosyl-L-methionine + salicylate
S-adenosyl-L-homocysteine + methyl salicylate
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + benzoate
S-adenosyl-L-homocysteine + methyl benzoate
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
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5 mM, stimulates BAMT activity by a factor of 2
Mg2+
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BAMT activity not affected by the presence of 5 mM
NH4+
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5 mM, stimulates BAMT activity by a factor of 2
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
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5 mM, slight inhibitory effect, less than 10% inhibition
S-adenosyl-L-homocysteine
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competitive inhibition with respect to S-adenosyl-L-methionine and noncompetitive inhibition with respect to benzoate
additional information
presence of 5 mM Mg2+ in the assay reaction and addition of K+, NH4+ and Ca2+ does not affect activity
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alamethicin
treatment of detached Arabidopsis thaliana leaves efficiently induced methyl salicylate but not methyl benzoate emission
insect herbivory
feeding by the larvae of the diamondback moth on rosette leaves of intact plants resultes in low methyl salicylate and very low methyl benzoate emission
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.1 - 1.6
Benzoate
0.065 - 1.72
Benzoic acid
0.003 - 0.087
S-adenosyl-L-methionine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0058 - 0.3
Benzoic acid
0.006
S-adenosyl-L-methionine
Antirrhinum majus
Q8H6N2
with benzoic acid, pH 7.5, 20°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.9 - 9.6
Benzoic acid
498
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.007 - 0.014
S-adenosyl-L-homocysteine
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000054
benzoic acid, pH 7.5, 20°C
0.0004518
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recombinant protein, with benzoic acid as substrate, pH 6.9, 20°C
0.0072
purified recombinant enzyme, pH and temperature not specified in the publication
0.01962
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purified BAMT protein, pH 7.5, 20°C
additional information
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emission of methyl benzoate begins at anthesis but at a very low level, 1.8 mg per flower for 24 hr, reaches a peak between days 5 to 8, and declines thereafter, at peak, 56.5 mg is emitted per flower in 24 hr
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
with 80–90% of maximum activity
5.5 - 9.5
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with 65% of maximum activity at both pH 6.5 and 8.5 and at pH 5.5 and 9.5, the enzyme activity falls to about 50% of the optimal value
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.67
Lillium hybrid cultivar
sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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reduced BAMT activity
Manually annotated by BRENDA team
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reduced BAMT activity
Manually annotated by BRENDA team
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the enzyme is localized predominantly in the conical cells of the inner epidermal layer and, to a much lesser extent, in the cells of the outer epidermis of flower petal lobes. The enzyme is also located in the inner epidermis of the corolla tube with little protein detected in the outer epidermis and in the yellow hairs within the tube on the bee’s way to the nectar
Manually annotated by BRENDA team
expression of mRNA about half the level in petal
Manually annotated by BRENDA team
expression of only 2fold higher than in pistil, where lowest level of mRNA expression is observed
Manually annotated by BRENDA team
lower levels of AtBSMT1 transcript
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
41000
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calculated molecular mass of the protein encoded by BAMT cDNA
43360
AlBSMT1 subunit, calculated from sequence
43370
subunit, calculated from sequence
100000
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native protein, gel filtration chromatography on Superdex 200-HR
107000
with a calibrated Sephacryl 200-HR column
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
Lillium hybrid cultivar
x * 41050, about, sequence calculation
homodimer
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 65
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BAMT is 100% stable for 30 min at 20°C and 60% stable for 30 min at 30°C, it is 20% stable for 30 min at 42°C but after 30 min incubation at 65°C, it completely loses activity
additional information
enzyme is stable at temperatures up to 42°C for 30 min but 30 min incubation at 65°C leads to a 70% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme is active in both Tris- and phosphate–citrate-based buffers, although its activity in phosphate-citrate buffer is about 20% lower than in Tris-buffer
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
the purified proteins, both from petal tissue and from Escherichia coli, are highly stable for several months when stored at -80°C. When stored in a buffer containing 50 mM Bis–Tris–HCl, pH 6.9, 10% glycerol, and 10 mM beta-mercaptoethanol at 4°C, BAMT protein is stable for 1 week
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the recombinant protein is catalytically stable for more than a year when stored at -80°C
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by nickel-based affinity chromatography
enzyme purified from upper and lower petal lobes of 5- to 10-day-old snapdragon flowers using DE53 anion exchange chromatography, hydrophobic interaction chromatography using Phenyl-Sepharose 6FF, and Mono-Q chromatography
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from 5- to 10-day-old upper and lower petal lobes by DE53 anion exchange, phenyl–Sepharose 6FF and MonoQ chromatography
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recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) Codon Plus by metal affinity chromatography
to near homogeneity in two steps, first using anion-exchange column diethylaminoethyl cellulose followed by Mono-Q anion-exchange chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
BAMT cDNA expressed in Escherichia coli
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coding regions of AlBSMT1 ligated into pCRT7/CT-TOPO TA vector for functional expression in Escherichia coli
coding regions of AtBSMT1 ligated into pCRT7/CT-TOPO TA vector for functional expression in Escherichia coli
expressed with an N-terminal His tag in Escherichia coli
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gene LiBSMT, DNA and amino acid sequence determination and analysis, sequence comparison and phylogenetic analysis and tree, quantitative real-time PCR expression analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Lillium hybrid cultivar
gene PbBSMT, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) codon plus
subcloned into expression vector pET-28a, containing an N-terminal polyhistidine 6-His-tag, and expressed in Escherichia coli
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
during flower life span, levels of methyl benzoate emission, BAMT activity, BAMT gene expression, and the amounts of BAMT protein and benzoic acid are developmentally and differentially regulated
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leaf wounding and uprooting causes AtBSMT1 transcripts to accumulate quickly within 2 h after treatment with uprooting being more effective; methyl jasmonate treatment, in which plants are exposed to methyl jasmonate vapors in a closed container, results in slow induction
under normal growing conditions AtBSMT1 promoter is only active in leaves at the base of the trichomes, in the hydathodes, and in the upper part of the sepals of flowers. Strong AtBSMT1 promoter activity is observed around lesions in the leaves caused by thrips of the genus Frankliniella