Information on EC 2.3.1.165 - 6-methylsalicylic-acid synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
2.3.1.165
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RECOMMENDED NAME
GeneOntology No.
6-methylsalicylic-acid synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + 3 malonyl-CoA + NADPH + H+ = 6-methylsalicylate + 4 CoA + 3 CO2 + NADP+ + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Claisen condensation
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-
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reduction
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-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
patulin biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
acyl-CoA:malonyl-CoA C-acyltransferase (decarboxylating, oxoacyl-reducing, thioester-hydrolysing and cyclizing)
A multienzyme complex with a 4'-phosphopantetheine prosthetic group on the acyl carrier protein. It has a similar sequence to vertebrate type I fatty acid synthase. Acetoacetyl-CoA can also act as a starter molecule.
CAS REGISTRY NUMBER
COMMENTARY hide
9045-37-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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UniProt
Manually annotated by BRENDA team
strain NRRL 2615, NRRL 5253, NRRL 1678, NRRL 5254, IHEM 3076, MUCL 39714 CBS100.11, NRRL 3849, ATCC 24008
UniProt
Manually annotated by BRENDA team
strain NRRL 2615, NRRL 5253, NRRL 1678, NRRL 5254, IHEM 3076, MUCL 39714 CBS100.11, NRRL 3849, ATCC 24008
UniProt
Manually annotated by BRENDA team
expression in Aspergillus nidulans
SwissProt
Manually annotated by BRENDA team
toxigenic Penicillium expansum strain 7015, genome of non-toxigenic strain FV 268 does not contain msas gene
TREMBL
Manually annotated by BRENDA team
strain DSM 40725
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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6-methylsalicylic acid synthase belongs to the iterative type I polyketide synthases
metabolism
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
acetoacetyl-CoA + 3 malonyl-CoA + NADPH + H+
6-methyl triacetic acid lactone + CoA + NADP+
show the reaction diagram
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-
-
-
?
acetoacetyl-CoA + malonyl-CoA
6-methyl triacetic acid lactone + CoA
show the reaction diagram
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5% of the rate of 6-methylsalicylic acid synthesis
-
?
acetoacetyl-CoA + malonyl-CoA + NADPH
?
show the reaction diagram
acetyl-CoA + 3 malonyl-CoA + NADPH + H+
6-methylsalicylate + 4 CoA + 3 CO2 + NADP+ + H2O
show the reaction diagram
acetyl-CoA + malonyl-CoA
triacetic lactone
show the reaction diagram
acetyl-CoA + malonyl-CoA + NADPH
6-methylsalicylate + CoA + CO2 + NADP+
show the reaction diagram
acetyl-CoA + malonyl-CoA + NADPH + H+
6-methylsalicylate + CoA + CO2 + NADP+ + H2O
show the reaction diagram
-
-
-
?
acetyl-CoA + N-acetylcysteamine + NADPH
?
show the reaction diagram
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-
-
-
?
acetyl-CoA + pantetheine
acetyl pantetheine + CoA
show the reaction diagram
butyryl-CoA + malonyl-CoA
6-propyl triacetic acid lactone + CoA
show the reaction diagram
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5% of the rate of 6-methylsalicylic acid synthesis
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?
crotonyl-CoA + malonyl-CoA
6-(2-propene) triacetic acid lactone + CoA
show the reaction diagram
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5% of the rate of 6-methylsalicylic acid synthesis
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?
heptanoyl-CoA + malonyl-CoA
6-hexyl triacetic acid lactone + CoA
show the reaction diagram
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5% of the rate of 6-methylsalicylic acid synthesis
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?
hexanoyl-CoA + malonyl-CoA
6-pentyl triacetic acid lactone + CoA
show the reaction diagram
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5% of the rate of 6-methylsalicylic acid synthesis
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?
propionyl-CoA + malonyl-CoA
6-ethyl triacetic acid lactone + CoA
show the reaction diagram
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5% of the rate of 6-methylsalicylic acid synthesis
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?
propionyl-CoA + malonyl-CoA + NADPH
6-ethylsalicylate + CoA + CO2 + NADP+
show the reaction diagram
propionyl-CoA + pantetheine
propionyl pantetheine + CoA
show the reaction diagram
valeryl-CoA + malonyl-CoA
6-butyl triacetic acid lactone + CoA
show the reaction diagram
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5% of the rate of 6-methylsalicylic acid synthesis
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?
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
acetyl-CoA + 3 malonyl-CoA + NADPH + H+
6-methylsalicylate + 4 CoA + 3 CO2 + NADP+ + H2O
show the reaction diagram
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-
-
-
?
acetyl-CoA + malonyl-CoA + NADPH
6-methylsalicylate + CoA + CO2 + NADP+
show the reaction diagram
acetyl-CoA + malonyl-CoA + NADPH + H+
6-methylsalicylate + CoA + CO2 + NADP+ + H2O
show the reaction diagram
P22367
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-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S,3R)-2,3-epoxy-4-oxo-7,10-dodecadienoylamide
1,3-dibromo-propane-2-one
5,5'-dithiobis(2-nitrobenzoic acid)
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0.01 mM, rapid inactivation, inactivation half-life: approx. 15 s, both acetyl-CoA and malonyl-CoA protect
iodoacetamide
NAD+
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0.3 mM, 21% inhibition in crude extracts
NADH
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0.3 mM, 26% inhibition in crude extracts
NADP+
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0.3 mM, 21% inhibition in crude extracts
nicotinamide hypoxanthine dinucleotide phosphate
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reduced form, 0.3 mM, 32% inhibition in crude extracts, oxidized form, 0.3 mM, 24% inhibition in crude extracts
additional information
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no inhibition by phenylmethylsulfonyl fluoride, a serine protease-type thioesterase inhibitor at 1 mM
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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in order to activate 6-MSAS a 4'-phosphopantetheinyl transferase, PPTase, is required, therefor a Bacillus subtilis or a Aspergillus nidulans PPTase is coexpressed in yeast
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.76 - 12.3
6-Methylsalicylate
0.065
acetoacetyl-CoA
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0.01
acetyl-CoA
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0.007
malonyl-CoA
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0125 - 0.023
6-Methylsalicylate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
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assay at
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
750000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homotetramer
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every subunit of 6-methylsalicylic acid synthase from Aspergillus terreus consist of ketosynthase (KS), acyltransferase (AT), dehydratase (DH), ketoreductase (KR), and acyl carrier protein (ACP) domain, proposed from deletion mutant analyses
tetramer
additional information
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.6
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more than 60% activity remains after 120 min at 30C in the presence of PMSF, NADPH, acetyl-CoA and dithiothreitol
486446
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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half-life in crude extracts: 16 min; half-life in crude extracts in the presence of 1.5 mM NAD+: 27 min; half-life in crude extracts in the presence of 1.5 mM NADH: 27 min; half-life in crude extracts in the presence of 1.5 mM NADP+: 36 min; half-life in crude extracts in the presence of 1.5 mM NADPH, 1 mM PMSF, 0.25 mM acetyl-CoA, 0.75 mM malonyl-CoA and 10 mM dithiothreitol: 188 min; half-life in crude extracts in the presence of 1.5 mM NADPH: 38 min; half-life in crude extracts in the presence of 1 mM PMSF: 53 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
activity is lost upon storage
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PMSF and benzamidine in disruption buffer extend half-life from 20 min to 6 h, 15% glycerol and proteinase inhibitors further improve stability
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very sensitive to proteolysis, significant stabilization by a combination of NADPH, acetyl-CoA, malonyl-CoA, PMSF, and dithitothreitol
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, ammonium sulfate precipitate, loss of 5% activity per month
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, poly(ethylene glycol) 6000, DEAE-Sepharose, hydroxyapatite, Mono Q
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recombinant N-terminally His6-tagged ATX and its thioester hydrolase domain, and the truncated mutant enzyme form, THID, from Escherichia coli strain BL21(DE3)-CodonPlus RIPL, by nickel affinity chromatography, and THID further by anion exchange chromatography, followed by gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Aspergillus nidulans
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expressed in Saccharomyces cerevisiae
expression in Aspergillus nidulans
expression in Escherichia coli
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expression in Escherichia coli and Saccharomyces cerevisiae can be used for 4-hydroxy-6-methyl-2-pyrone production
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for expression in Saccharomyces cerevisiae
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functional expression in Aspergillus nidulans
functional expression of ATX in Escherichia coli strain BL21(DE3)-CodonPlus RIPL, expression of N-terminally His6-tagged ATX and its thioester hydrolase domain, and the truncated mutant enzyme form, THID
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into the PCR2.1 Topo cloning kit
paart of Chl gene cluster of chlorothricin producer Streptomyces antibioticus
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
5 mg/ml of both the phenolic compounds quercetin and umbelliferone decrease the expression of msas
5 mg/ml of the phenolic compounds quercetin or umbelliferone slightly increases the expression of msas
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C216A
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mutant without catalytic activity
H939A
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mutant with catalytic activity
H972A
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site-directed mutagenesis, the reaction intermediate is bound to the thioester hydrolase domain of ATX formed in the presence of NADPH, and is released as 6-methylsalicylic acid by both the intact ATX and by truncated THID mutant in trans
S1761A
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mutant without catalytic activity
S667A
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mutant without catalytic activity
H958A
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mutation in the THID active site
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
food industry
medicine
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production of 6-methylsalicylic acid which is used as human pharmaceutical