Information on EC 2.3.1.161 - lovastatin nonaketide synthase

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The expected taxonomic range for this enzyme is: Aspergillus terreus

EC NUMBER
COMMENTARY hide
2.3.1.161
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RECOMMENDED NAME
GeneOntology No.
lovastatin nonaketide synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
9 malonyl-CoA + 11 NADPH + 10 H+ + S-adenosyl-L-methionine + holo-[lovastatin nonaketide synthase] = dihydromonacolin L-[lovastatin nonaketide synthase] + 9 CoA + 9 CO2 + 11 NADP+ + S-adenosyl-L-homocysteine + 6 H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cyclization
decarboxylation
dehydration
reduction
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
lovastatin biosynthesis
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-
SYSTEMATIC NAME
IUBMB Comments
acyl-CoA:malonyl-CoA C-acyltransferase (dihydromonacolin L acid-forming)
This fungal enzyme system comprises a multi-functional polyketide synthase (PKS) and an enoyl reductase. The PKS catalyses many of the chain building reactions of EC 2.3.1.85, fatty-acid synthase, as well as a reductive methylation and a Diels-Alder reaction, while the reductase is responsible for three enoyl reductions that are necessary for dihydromonacolin L acid production.
CAS REGISTRY NUMBER
COMMENTARY hide
235426-97-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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release of reaction product from enzyme LovB is mediated by multifunctional esterase LovG. LovG is also involved in the clearance of aberrant intermediates from LovB
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(4E,6E)-2-methyl-3-oxoocta-4,6-dienoyl-S-ACP + NAPDH + H+
(4E,6E)-2-methyl-3-hydroxyocta-4,6-dienoyl-S-ACP + NAPDH
show the reaction diagram
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reaction of ketoreductase subunit, natural substrate. ACP = acyl-carrier protein
-
-
?
(4E,6E)-3-oxoocta-4,6-dienoyl-N-acetylcysteamine + S-adenosyl-L-methionine
(4E,6E)-2-methyl-3-oxoocta-4,6-dienoyl-N-acetylcysteamine + S-adenosyl-L-homocysteine
show the reaction diagram
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reaction of methyltransferase subunit, natural substrate analogue. ACP = acyl-carrier protein
-
-
?
(4E,6E)-3-oxoocta-4,6-dienoyl-S-ACP + S-adenosyl-L-methionine
(4E,6E)-2-methyl-3-oxoocta-4,6-dienoyl-S-ACP + S-adenosyl-L-homocysteine
show the reaction diagram
-
reaction of methyltransferase subunit, natural substrate. ACP = acyl-carrier protein
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-
?
2-methyl-3-oxohexanoyl-N-acetylcysteamine + NAPDH + H+
3-hydroxy-2-methyl-hexanoyl-N-acetylcysteamine + NAPDH
show the reaction diagram
-
reaction of ketoreductase subunit, natural substrate
-
-
?
3-oxohexanoyl-N-acetylcysteamine + NAPDH + H+
3-hydroxyhexanoyl-N-acetylcysteamine + NAPDH
show the reaction diagram
-
reaction of ketoreductase subunit, natural substrate
-
-
?
3-oxooctanoyl-N-acetylcysteamine + S-adenosyl-L-methionine
2-methyl-3-oxooctanoyl-N-acetylcysteamine + S-adenosyl-L-homocysteine
show the reaction diagram
-
reaction of methyltransferase subunit, artificial substrate. 0.3% of the activity with (4E,6E)-3-oxoocta-4,6-dienoyl-N-acetylcysteamine
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-
?
acetyl-CoA + 8 malonyl-CoA + 11 NADPH + S-adenosyl-L-methionine + 11 H+
dihydromonacolin L + 9 CoA + 8 CO2 + 11 NADP+ + S-adenosyl-L-homocysteine + 6 H2O
show the reaction diagram
acetyl-CoA + malonyl-CoA + NADPH + H+ + S-adenosyl-L-methionine
4-hydroxy-6-[(1E,3E,5E)-1-methylhepta-1,3,5-trien-1-yl]-2H-pyran-2-one + 4-hydroxy-6-[(1E,3E,5E,7E)-3-methylnona-1,3,5,7-tetraen-1-yl]-2H-pyran-2-one + CoA + CO2 + NADP+ + S-adenosyl-L-homocysteine + H2O
show the reaction diagram
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in absence of accessory protein LovC, formation of truncated pyrones 4-hydroxy-6-[(1E,3E,5E)-1-methylhepta-1,3,5-trien-1-yl]-2H-pyran-2-one + 4-hydroxy-6-[(1E,3E,5E,7E)-3-methylnona-1,3,5,7-tetraen-1-yl]-2H-pyran-2-one. Chemical synthesis of these truncated pyrones for analysis of enzyme reaction
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?
acetyl-CoA + malonyl-CoA + NADPH + H+ + S-adenosyl-L-methionine
?
show the reaction diagram
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in the absence of the accessory protein LovC, LovB forms conjugated pyrones as truncated polyketide synthase products
-
-
?
acetyl-CoA + malonyl-CoA + NADPH + H+ + S-adenosyl-L-methionine
dihydromonacolin L + CoA + CO2 + NADP+ + S-adenosyl-L-homocysteine + H2O
show the reaction diagram
additional information
?
-
-
the methyltransferase domain displays methylation activity toward different beta-ketoacyl groups, it is exceptionally selective toward its naturally programmed beta-keto-dienyltetraketide substrate with respect to both chain length and functionalization. The ketoreductase domain displays broader substrate specificity toward different beta-ketoacyl groups
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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
(4E,6E)-2-methyl-3-oxoocta-4,6-dienoyl-S-ACP + NAPDH + H+
(4E,6E)-2-methyl-3-hydroxyocta-4,6-dienoyl-S-ACP + NAPDH
show the reaction diagram
-
reaction of ketoreductase subunit, natural substrate. ACP = acyl-carrier protein
-
-
?
(4E,6E)-3-oxoocta-4,6-dienoyl-S-ACP + S-adenosyl-L-methionine
(4E,6E)-2-methyl-3-oxoocta-4,6-dienoyl-S-ACP + S-adenosyl-L-homocysteine
show the reaction diagram
-
reaction of methyltransferase subunit, natural substrate. ACP = acyl-carrier protein
-
-
?
2-methyl-3-oxohexanoyl-N-acetylcysteamine + NAPDH + H+
3-hydroxy-2-methyl-hexanoyl-N-acetylcysteamine + NAPDH
show the reaction diagram
-
reaction of ketoreductase subunit, natural substrate
-
-
?
3-oxohexanoyl-N-acetylcysteamine + NAPDH + H+
3-hydroxyhexanoyl-N-acetylcysteamine + NAPDH
show the reaction diagram
-
reaction of ketoreductase subunit, natural substrate
-
-
?
acetyl-CoA + 8 malonyl-CoA + 11 NADPH + S-adenosyl-L-methionine + 11 H+
dihydromonacolin L + 9 CoA + 8 CO2 + 11 NADP+ + S-adenosyl-L-homocysteine + 6 H2O
show the reaction diagram
acetyl-CoA + malonyl-CoA + NADPH + H+ + S-adenosyl-L-methionine
?
show the reaction diagram
-
in the absence of the accessory protein LovC, LovB forms conjugated pyrones as truncated polyketide synthase products
-
-
?
acetyl-CoA + malonyl-CoA + NADPH + H+ + S-adenosyl-L-methionine
dihydromonacolin L + CoA + CO2 + NADP+ + S-adenosyl-L-homocysteine + H2O
show the reaction diagram
additional information
?
-
-
the methyltransferase domain displays methylation activity toward different beta-ketoacyl groups, it is exceptionally selective toward its naturally programmed beta-keto-dienyltetraketide substrate with respect to both chain length and functionalization. The ketoreductase domain displays broader substrate specificity toward different beta-ketoacyl groups
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
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-
additional information
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enzyme utilizes an oxidized quinone as cofactor
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.17
(4E,6E)-3-oxoocta-4,6-dienoyl-N-acetylcysteamine
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pH not specified in the publication, temperature not specified in the publication
2
3-oxohexanoyl-N-acetylcysteamine
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pH not specified in the publication, temperature not specified in the publication
5.2
3-oxooctanoyl-N-acetylcysteamine
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pH not specified in the publication, temperature not specified in the publication
0.5
malonyl-CoA
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-
additional information
additional information
-
in absence of accessory protein LovC, enzyme forms truncated pyrones 4-hydroxy-6-[(1E,3E,5E)-1-methylhepta-1,3,5-trien-1-yl]-2H-pyran-2-one + 4-hydroxy-6-[(1E,3E,5E,7E)-3-methylnona-1,3,5,7-tetraen-1-yl]-2H-pyran-2-one. Chemical synthesis of these truncated pyrones for analysis of enzyme reaction
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.25
(4E,6E)-3-oxoocta-4,6-dienoyl-N-acetylcysteamine
Aspergillus terreus
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pH not specified in the publication, temperature not specified in the publication
0.085
2-methyl-3-oxohexanoyl-N-acetylcysteamine
Aspergillus terreus
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pH not specified in the publication, temperature not specified in the publication
0.57
3-oxohexanoyl-N-acetylcysteamine
Aspergillus terreus
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pH not specified in the publication, temperature not specified in the publication
0.32
3-oxooctanoyl-N-acetylcysteamine
Aspergillus terreus
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pH not specified in the publication, temperature not specified in the publication
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
18.93
(4E,6E)-3-oxoocta-4,6-dienoyl-N-acetylcysteamine
Aspergillus terreus
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pH not specified in the publication, temperature not specified in the publication
209008
0.31
3-oxohexanoyl-N-acetylcysteamine
Aspergillus terreus
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pH not specified in the publication, temperature not specified in the publication
209013
0.06
3-oxooctanoyl-N-acetylcysteamine
Aspergillus terreus
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pH not specified in the publication, temperature not specified in the publication
209011
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11000
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LovB ACP, determined by SDS-PAGE
52000
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LovB MAT, determined by SDS-PAGE
66000
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LovB ACP-CON, determined by SDS-PAGE
103000
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LovB KS-MAT, determined by SDS-PAGE
335000
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
inactivated by heating
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
homogeneitiy
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mutant BX102
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the single domains are purified using Ni-nitrilotriacetic agarose resin and anion-exchange chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Saccharomyces cerevisiae
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lovB and lovC protein
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minimal polyketide synthase domains of subunit LovB as standalone proteins; the entire lovB gene is inserted into pET21a+ to create pSMa33, the LovB KS-MAT didomain, MAT, ACP, ACP-CON and CON are each amplified from pSMa33 and cloned into pET28a+
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wild type and different lov mutants
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
S208A
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LovB MAT mutant
S209A
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LovB MAT mutant
S656A
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inactivation of malonyl-coenzyme A:acyl carrier protein acyltransferase domain in ketosynthase-malonyl-coenzyme A:acyl carrier protein acyltransferase for assay of ketosynthase activity. Ketosynthase displays no activity towards acetyl groups but recognizes malonyl groups in the absence of cerulenin; LovB KS-MAT mutant, MAT domain inactivated
additional information
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expression of minimal polyketide synthase domains of subunit LovB as standalone proteins. The didomain proteins ketosynthase-malonyl-coenzyme A:acyl carrier protein acyltransferase KS-MAT can transfer the acyl group to both the cognate LovB acyl carrier protein and heterologous acyl carrier proteins from bacterial type I and type II polyketide synthases. It transfers malonyl and acetyl groups with kcat/Km values of 0.62 per min and 0.032 per min, resp
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine