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

BRENDA support

Literature summary extracted from

  • Hirte, M.; Meese, N.; Mertz, M.; Fuchs, M.; Brueck, T.B.
    Insights into the bifunctional aphidicolan-16-beta-ol synthase through rapid biomolecular modeling approaches (2018), Front. Chem., 6, 101 .
    View publication on PubMedView publication on EuropePMC

Protein Variants

EC Number Protein Variants Comment Organism
4.2.3.B35 D661A site-directed mutagenesis, the mutation results in a premature termination of the cyclization reaction cascade en-route from syn-copalyl diphosphate to aphidicolan-16-beta-ol, the mutant generates the diterpene macrocycle syn-copalol and a minor, non-hydroxylated labdane related diterpene Neocamarosporium betae
4.2.3.B35 Y658L site-directed mutagenesis, the mutation results in a premature termination of the cyclization reaction cascade en-route from syn-copalyl diphosphate to aphidicolan-16-beta-ol, the mutant generates the diterpene macrocycle syn-copalol and a minor, non-hydroxylated labdane related diterpene Neocamarosporium betae
4.2.3.42 C831G site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Neocamarosporium betae
4.2.3.42 C831T site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Neocamarosporium betae
4.2.3.42 D661A site-directed mutagenesis, the mutation results in a premature termination of the cyclization reaction cascade en-route from syn-copalyl diphosphate to aphidicolan-16-beta-ol, the mutant generates the diterpene macrocycle syn-copalol and a minor, non-hydroxylated labdane related diterpene Neocamarosporium betae
4.2.3.42 F629L site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Neocamarosporium betae
4.2.3.42 F789L site-directed mutagenesis, inactive mutant Neocamarosporium betae
4.2.3.42 I626A site-directed mutagenesis, inactive mutant Neocamarosporium betae
4.2.3.42 T920G site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Neocamarosporium betae
4.2.3.42 Y658F site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Neocamarosporium betae
4.2.3.42 Y658L site-directed mutagenesis, the mutation results in a premature termination of the cyclization reaction cascade en-route from syn-copalyl diphosphate to aphidicolan-16-beta-ol, the mutant generates the diterpene macrocycle syn-copalol and a minor, non-hydroxylated labdane related diterpene Neocamarosporium betae
4.2.3.42 Y923F site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Neocamarosporium betae
4.2.3.42 Y923L site-directed mutagenesis, inactive mutant Neocamarosporium betae

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.3.B35 Mg2+ required Neocamarosporium betae
4.2.3.42 Mg2+ required Neocamarosporium betae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.3.42 9alpha-copalyl diphosphate + H2O Neocamarosporium betae
-
 aphidicolan-16beta-ol + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.3.B35 Neocamarosporium betae
-
 i.e. Phoma betae
-
4.2.3.42 Neocamarosporium betae Q96WT2 i.e. Phoma betae
-

Reaction

EC Number Reaction Comment Organism Reaction ID
4.2.3.42 9alpha-copalyl diphosphate + H2O = aphidicolan-16beta-ol + diphosphate the reaction includes a Wagner-Meerwein rearrangement and proceeds via several cation intermediates stages, modeling, overview Neocamarosporium betae
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.3.B35 syn-copalyl diphosphate + H2O reaction of aphidicolan-16beta-ol synthase (EC 4.2.3.42) enzyme mutants Y658L and D661A, no activity with wild-type aphidicolan-16beta-ol synthase Neocamarosporium betae syn-copalol + diphosphate
-
 ?
4.2.3.42 9alpha-copalyl diphosphate + H2O
-
 Neocamarosporium betae aphidicolan-16beta-ol + diphosphate
-
 ?
4.2.3.42 9alpha-copalyl diphosphate + H2O syn-CDP Neocamarosporium betae aphidicolan-16beta-ol + diphosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.3.42 ACS
-
 Neocamarosporium betae

General Information

EC Number General Information Comment Organism
4.2.3.B35 evolution enzyme ACS belongs to the diterpene synthase family Neocamarosporium betae
4.2.3.B35 malfunction aphidicolan-16beta-ol synthase (EC 4.2.3.42) enzyme mutants Y658L and D661A show a premature termination of the cyclization reaction cascade en-route from syn-copalyl diphosphate to aphidicolan-16-beta-ol, the mutants generates the diterpene macrocycle syn-copalol and a minor, non-hydroxylated labdane related diterpene. The structural changes D661A and Y658L still allow syn-copalyl diphosphate binding in the active site with subsequent hydrolyses of the diphosphate group. The syn-copalyl carbocation is then quenched either by water (release of syn-copalol) or an amino acid side chain (release of non-hydroxylated diterpene). No other substitution is found that stops cyclization at the proposed transitional states nor changes in the byproduct formation of the active mutants. The ACS cyclization occurs in a spatially restricted area and the diphosphate group remains in the active site Neocamarosporium betae
4.2.3.B35 additional information modeling of the protein-ligand complex structure of fungal, bifunctional aphidicolan-16-beta-ol synthase, molecular dynamic studies, only the ACS alpha-domain sequence is used for homology prediction. The catalytically relevant ACS amino acid network includes residues I626, T657, Y658, A786, F789, and Y923. The initial conversion from the universal diterpene precursor GGDP to syn-CDP occurs in class II active site, located between the ACS beta- and gamma-domain. The subsequent syn-CDP cyclization to aphidicolan-16beta-ol is then conducted in the class I active site that is positioned in the middle of an alpha-helical bundle forming the ACS alpha-domain Neocamarosporium betae
4.2.3.42 evolution enzyme ACS belongs to the diterpene synthase family Neocamarosporium betae
4.2.3.42 malfunction aphidicolan-16beta-ol synthase (EC 4.2.3.42) enzyme mutants Y658L and D661A show a premature termination of the cyclization reaction cascade en-route from syn-copalyl diphosphate to aphidicolan-16-beta-ol, the mutants generates the diterpene macrocycle syn-copalol and a minor, non-hydroxylated labdane related diterpene. The structural changes D661A and Y658L still allow syn-CDP binding in the active site with subsequent hydrolyses of the diphosphate group. The syn-copalyl carbocation is then quenched either by water (release of syn-copalol) or an amino acid side chain (release of non-hydroxylated diterpene). No other substitution is found that stops cyclization at the proposed transitional states nor changes in the byproduct formation of the active mutants. The ACS cyclization occurs in a spatially restricted area and the diphosphate group remains in the active site Neocamarosporium betae
4.2.3.42 additional information modeling of the protein-ligand complex structure of fungal, bifunctional aphidicolan-16-beta-ol synthase, molecular dynamic studies, only the ACS alpha-domain sequence is used for homology prediction. The catalytically relevant ACS amino acid network includes residues I626, T657, Y658, A786, F789, and Y923. The initial conversion from the universal diterpene precursor GGDP to syn-CDP occurs in class II active site, located between the ACS beta- and gamma-domain. The subsequent syn-CDP cyclization to aphidicolan-16beta-ol is then conducted in the class I active site that is positioned in the middle of an alpha-helical bundle forming the ACS alpha-domain Neocamarosporium betae
4.2.3.42 physiological function the bifunctional enzyme aphidicolan-16beta-ol synthase (ACS) catalyzes the complex conversion of GGDP via syn-copalyl diphosphate to the tetracyclic aphidicolan-16beta-ol which is the core structure of the cytostatic compound aphidicolin, cyclization reaction cascade en-route from syn-copalyl diphosphate to aphidicolan-16-beta-ol, overview Neocamarosporium betae