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Information on EC 4.2.3.118 - 2-methylisoborneol synthase and Organism(s) Streptomyces coelicolor and UniProt Accession Q9F1Y6

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EC Tree
     4 Lyases
         4.2 Carbon-oxygen lyases
             4.2.3 Acting on phosphates
                4.2.3.118 2-methylisoborneol synthase
IUBMB Comments
The product, 2-methylisoborneol, is a characteristc odiferous compound with a musty smell produced by soil microorganisms.
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This record set is specific for:
Streptomyces coelicolor
UNIPROT: Q9F1Y6
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Word Map
The taxonomic range for the selected organisms is: Streptomyces coelicolor
The enzyme appears in selected viruses and cellular organisms
Synonyms
monoterpene cyclase, mib synthase, 2-methylisoborneol synthase, sco7700, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2-MIB cyclase
-
-
-
-
MIB synthase
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-
-
-
MIBS
-
-
-
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sco7700
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-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(E)-2-methylgeranyl diphosphate + H2O = 2-methylisoborneol + diphosphate
show the reaction diagram
catalytic mechanism, overview
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
(E)-2-methylgeranyl-diphosphate diphosphate-lyase (cyclizing, 2-methylisoborneol-forming)
The product, 2-methylisoborneol, is a characteristc odiferous compound with a musty smell produced by soil microorganisms.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(E)-2-methylgeranyl diphosphate + H2O
2-methylisoborneol + diphosphate
show the reaction diagram
2-fluorolinalyl diphosphate + H2O
(1R)-(+)-camphor + HF + diphosphate
show the reaction diagram
-
single cyclic product, obtained via the (4R)-alpha-2-fluoroterpinyl carbocation intermediate. The initially generated 2-fluoroisoborneol cyclization product is unstable and undergoes elimination of hydrogen fluoride to yield (1R)-(+)-camphor
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?
2-methyllinalyl diphosphate + H2O
2-methylisoborneol + diphosphate
show the reaction diagram
natural intermediate of the cyclization reaction
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-
?
2-methylneryl diphosphate + H2O
2-methyllimonene
show the reaction diagram
-
-
-
?
geranyl diphosphate + H2O
limonene + diphosphate
show the reaction diagram
-
products are 6% alpha-pinene, 23% beta-pinene , 32% limonene, 29% gamma-terpinene, and 120% delta-terpinene
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?
additional information
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(E)-2-methylgeranyl diphosphate + H2O
2-methylisoborneol + diphosphate
show the reaction diagram
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
three Mg2+ ions are presumed to be required for the cyclization reaction
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.026 - 0.095
(E)-2-methylgeranyl diphosphate
0.018
2-methylneryl diphosphate
pH 6.7, 30°C
0.0028
geranyl diphosphate
pH 7.0, 30°C
additional information
additional information
Michaelis-Menten steady-state kinetics
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.039 - 0.105
(E)-2-methylgeranyl diphosphate
0.0046
2-methylneryl diphosphate
pH 6.7, 30°C
0.0003
geranyl diphosphate
pH 7.0, 30°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.105
(E)-2-methylgeranyl diphosphate
pH 6.7, 30°C
0.256
2-methylneryl diphosphate
pH 6.7, 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
2-methylisoborneol (2-MIB) has a muddy smell with a low odor and flavor threshold and is a contaminant of drinking water, wine and fish. Moreover it is related to the musty-earthy aroma in camembert and brie
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
47000
x * 47000, calculated
49937
x * 49937, MALDI-TOF
80000 - 90000
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 49937, MALDI-TOF
multimer
x * 47000, calculated
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
in complex with substrate analogues geranyl-S-thiolodiphosphate and 2-fluorogeranyl diphosphate at 1.80 and 1.95 A resolution, respectively. MIBS adopts the tertiary structure of a class I terpenoid cyclase. The dimeric quaternary structure is different, the active sites in the MIBS dimer are oriented in parallel fashion and the dimer interface in MIBS is formed by helices D1, D2
in unliganded state and in complex with two Mg2+ ions and 2-fluoroneryl diphosphate at 1.85 and 2.00 A resolution, respectively. Cocrystallization with the fluorinated analogue of the tertiary allylic diphosphate intermediate, 2-fluorolinalyl diphosphate yields the crystal structure of the complex with the primary allylic diphosphate, 2-fluoroneryl diphosphate. The crystalline enzyme active site remains partially open, presumably due to the binding of only two Mg2+ ions
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
usage of Escherichia coli strain MG1655 and derivatives for heterologous production of different C11-terpene synthases together with the GPP methyltransferase and the mevalonate pathway enzymes, 35 different C11-terpenes can be produced, overview
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged enzyme by nickel affinity chromatography
recombinant protein
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
gene sco7700, recombinant overexpression of His-tagged enzyme
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wang, C.M.; Cane, D.E.
Biochemistry and molecular genetics of the biosynthesis of the earthy odorant methylisoborneol in Streptomyces coelicolor
J. Am. Chem. Soc.
130
8908-8909
2008
Streptomyces coelicolor (Q9F1Y6), Streptomyces coelicolor
Manually annotated by BRENDA team
Koeksal, M.; Chou, W.K.; Cane, D.E.; Christianson, D.W.
Structure of 2-methylisoborneol synthase from Streptomyces coelicolor and implications for the cyclization of a noncanonical C-methylated monoterpenoid substrate
Biochemistry
51
3011-3020
2012
Streptomyces coelicolor (Q9F1Y6), Streptomyces coelicolor
Manually annotated by BRENDA team
Koeksal, M.; Chou, W.K.; Cane, D.E.; Christianson, D.W.
Unexpected reactivity of 2-fluorolinalyl diphosphate in the active site of crystalline 2-methylisoborneol synthase
Biochemistry
52
5247-5255
2013
Streptomyces coelicolor (Q9F1Y6), Streptomyces coelicolor A3(2) (Q9F1Y6), Streptomyces coelicolor A3(2)
Manually annotated by BRENDA team
Chou, W.K.; Gould, C.A.; Cane, D.E.
Incubation of 2-methylisoborneol synthase with the intermediate analog 2-methylneryl diphosphate
J. Antibiot.
70
625-631
2017
Streptomyces coelicolor (Q9F1Y6), Streptomyces coelicolor ATCC BAA-471 / A3(2) / M145 (Q9F1Y6)
Manually annotated by BRENDA team
Kschowak, M.J.; Wortmann, H.; Dickschat, J.S.; Schrader, J.; Buchhaupt, M.
Heterologous expression of 2-methylisoborneol / 2 methylenebornane biosynthesis genes in Escherichia coli yields novel C11-terpenes
PLoS ONE
13
e0196082
2018
Streptomyces coelicolor (Q9F1Y6), Streptomyces coelicolor ATCC BAA-471 / A3(2) / M145 (Q9F1Y6), Streptomyces griseus subsp. griseus (Q9F1V8)
Manually annotated by BRENDA team