Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 4.2.3.129 extracted from

  • Lopez-Gallego, F.; Wawrzyn, G.; Schmidt-Dannert, C.
    Selectivity of fungal sesquiterpene synthases: Role of the active sites H-1alpha loop in catalysis (2010), Appl. Environ. Microbiol., 76, 7723-7733.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.2.3.129

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain JM109 Coprinopsis cinerea

Protein Variants

Protein Variants Comment Organism
H235P site-directed mutagenesis, no production of sativene, the mutation converts Cop4 into a much more selective enzyme that produces (-)-germacrene D as the major cyclization product with 50% of total sesquiterpenes products. The mutant makes beta-ylangene, which is a diastereomer of beta-copaene and not synthesized by wild-type Cop4 Coprinopsis cinerea
K233I site-directed mutagenesis, mutation of K233, interacting with the second Asp92 in the DDXXD motif of Cop4, does not significantly change the overall product promiscuity of Cop4, though beta-cubebene 4 does become the major product and püroduction of sativene is reduced by 50% compared to the wild-type enzyme Coprinopsis cinerea
additional information directed mutations of the H-alpha1 loop have a marked effect on the product profile Cop4, loop mutations in Cop4 also implicate specific residues responsible for the pH sensitivity of the enzyme. H-alpha1 loop swap between Cop4 and Cop6 shifts Cop4 to a germacrene D synthase Coprinopsis cinerea
N238L site-directed mutagenesis, the mutant shows a altered product profile compared to the wild-type enzyme with only slight reduction in beta-cubebene synthesis, sativene levels are similar to the wild-type. The mutant does no longer show production of cubebol and has reduced (-)-germacrene D synthesis activity compared to the wild-type enzyme, synthesis of beta-cubebene, beta-copaene, delta-cadinene, and alpha-cubebene Coprinopsis cinerea
N239L site-directed mutagenesis, no production of sativene, the mutation converts Cop4 into a much more selective enzyme that produces (-)-germacrene D as the major cyclization product with 50% of total sesquiterpenes products. The mutant makes beta-ylangene, which is a diastereomer of beta-copaene and not synthesized by wild-type Cop4 Coprinopsis cinerea
T236L site-directed mutagenesis, the mutant shows a altered product profile compared to the wild-type enzyme with an increase in sativene synthesis. The mutant does no longer show production of cubebol and (-)-germacrene D compared to the wild-type enzyme Coprinopsis cinerea

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.011
-
 (2E,6E)-farnesyl diphosphate wild-type enzyme, and mutants H235P and N239L, pH 8.0, 30° Coprinopsis cinerea
0.048
-
 (2E,6E)-farnesyl diphosphate Cop6 loop graft mutant Cop6L4, pH 8.0, 30° Coprinopsis cinerea
0.0707
-
 (2E,6E)-farnesyl diphosphate mutant K233I, pH 8.0, 30° Coprinopsis cinerea

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(2E,6E)-farnesyl diphosphate Coprinopsis cinerea
-
 (+)-sativene + diphosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Coprinopsis cinerea A8NU13
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2E,6E)-farnesyl diphosphate
-
 Coprinopsis cinerea (+)-sativene + diphosphate
-
 ?

Subunits

Subunits Comment Organism
More structure comparison, homology structural modeling of Cop enzymes, overview. Cop4 has a large active site cavity that undergoes substantial conformational change in the model upon ligand binding Coprinopsis cinerea

Synonyms

Synonyms Comment Organism
Cop4
-
 Coprinopsis cinerea

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25 30 assay at Coprinopsis cinerea

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 about Coprinopsis cinerea

General Information

General Information Comment Organism
evolution the enzyme belongs to the sesquiterpene synthases Coprinopsis cinerea
additional information directed mutations of the H-alpha1 loop have a marked effect on the product profile Cop4, loop mutations in Cop4 also implicate specific residues responsible for the pH sensitivity of the enzyme. In vivo analysis of sesquiterpene product profiles of H-alpha1 loop mutants, overview. Mutation of K233, presumed to interact with the second Asp92 in the DDXXD motif of Cop4, does not significantly change the overall product promiscuity of Cop4, though beta-cubebene, with 27% of total sesquiterpene products, does become the major product Coprinopsis cinerea
physiological function the enzyme belongs to the sesquiterpene synthases that are responsible for the cyclization of farnesyl diphosphate into a myriad of structurally diverse compounds with various biological activities Coprinopsis cinerea