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

BRENDA support

Literature summary extracted from

  • Stickforth, P.; Sandmann, G.
    Structural and kinetics properties of a mutated phytoene desaturase from Rubrivivax gelatinosus with modified product specificity (2011), Arch. Biochem. Biophys., 505, 118-122.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.5.6 crtI gene, construction of different mutation libraries used for screening for mutants which synthesize almost exclusively either neurosporene or lycopene, cloning n Escherichia coli strain DH5alpha Rubrivivax gelatinosus
1.3.99.28 gene crtI, cloning and expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha Rubrivivax gelatinosus
1.3.99.31 expression in Escherichia coli Rubrivivax gelatinosus

Protein Variants

EC Number Protein Variants Comment Organism
1.3.5.6 D355G naturally occuring mutation Rubrivivax gelatinosus
1.3.5.6 D53G naturally occuring mutation Rubrivivax gelatinosus
1.3.5.6 L153P naturally occuring mutation, which influences the secondary structure of the enzyme Rubrivivax gelatinosus
1.3.5.6 L208F naturally occuring mutation, which influences the secondary structure of the enzyme Rubrivivax gelatinosus
1.3.5.6 L208P naturally occuring mutation, which influences the secondary structure of the enzyme Rubrivivax gelatinosus
1.3.5.6 L278P naturally occuring mutation, which influences the secondary structure of the enzyme Rubrivivax gelatinosus
1.3.5.6 L424P naturally occuring mutation, which influences the secondary structure of the enzyme Rubrivivax gelatinosus
1.3.5.6 additional information in mutant EP21, zeta-carotene and phytofluene, which are both early intermediates of the desaturation pathway and which are absent in the wild-type, are the main carotene products Rubrivivax gelatinosus
1.3.5.6 P134L naturally occuring mutation, which influences the secondary structure of the enzyme Rubrivivax gelatinosus
1.3.5.6 T256M naturally occuring mutation Rubrivivax gelatinosus
1.3.5.6 V395A naturally occuring mutation Rubrivivax gelatinosus
1.3.5.6 Y44C naturally occuring mutation Rubrivivax gelatinosus
1.3.99.28 L153P/L278P site-directed mutagenesis, the mutant shows altered secondary structure and substrate desaturation level with increased lycopene production compared to the wild-type enzyme Rubrivivax gelatinosus
1.3.99.28 L208F site-directed mutagenesis, the mutant shows altered secondary structure and substrate desaturation level with increased lycopene production compared to the wild-type enzyme Rubrivivax gelatinosus
1.3.99.28 L208P site-directed mutagenesis, the mutant shows altered secondary structure compared to the wild-type enzyme Rubrivivax gelatinosus
1.3.99.28 T256M/D355G/L424P site-directed mutagenesis, the mutant shows altered secondary structure and substrate desaturation level with increased lycopene production compared to the wild-type enzyme Rubrivivax gelatinosus
1.3.99.28 Y44C/D53G/P134L/V395A site-directed mutagenesis, the mutant shows altered secondary structure and substrate desaturation level with increased lycopene production compared to the wild-type enzyme Rubrivivax gelatinosus
1.3.99.31 L153P/L278P increased formation of neurosporene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg), the ratio of the mutant enzyme is 12:88 Rubrivivax gelatinosus
1.3.99.31 L208F increased formation of lycopene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 26:74 after complementation of CrtI in pPEU(DH5a/pACCrtEBEU/pPEUCrtIRg), the ratio of the mutant enzyme is 89:11 Rubrivivax gelatinosus
1.3.99.31 L208P increased formation of neurosporene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg), the ratio of the mutant enzyme is 3:97 Rubrivivax gelatinosus
1.3.99.31 additional information two different mutation libraries for the crtI gene are constructed to screen for modified enzymes which synthesize almost exclusively either neurosporene or lycopene. The resulting mutants carry between one and four amino acid exchanges and at least one of them affects the secondary protein structure by shortening or extending one of the helices. A prominent amino acid which is exchanged in the neurosporene or lycopene-forming desaturase is leucine 208. Enzyme kinetic studies are carried out with the L208 modified desaturase and the specificities for phytoene and neurosporene as substrates determined. Higher and lower values correlate well with the higher or lower potential for the synthesis of lycopene from neurosporene. TopPred analysis of the mutations of L208 indicate that the location is in a highly hydrophobic membrane-integrated region which is a good candidate for the substrate-binding site of the desaturase Rubrivivax gelatinosus
1.3.99.31 T256M/D355GL424P increased formation of neurosporene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg), the ratio of the mutant enzyme is 0:100 Rubrivivax gelatinosus
1.3.99.31 Y44C/D53G/P134L/V395A increased formation of neurosporene compared to wild-type enzyme. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg), the ratio of the mutant enzyme is 3:97 Rubrivivax gelatinosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.5.6 0.0048
-
 phytoene pH 8.0, 28°C, recombinant mutant L208F Rubrivivax gelatinosus
1.3.5.6 0.0148
-
 phytoene pH 8.0, 28°C, recombinant wild-type enzyme Rubrivivax gelatinosus
1.3.5.6 0.0171
-
 phytoene pH 8.0, 28°C, recombinant mutant L208P Rubrivivax gelatinosus
1.3.5.6 0.0331
-
 7,9,9'-tricis-neurosporene pH 8.0, 28°C, recombinant wild-type enzyme Rubrivivax gelatinosus
1.3.5.6 0.0417
-
 7,9,9'-tricis-neurosporene pH 8.0, 28°C, recombinant mutant L208P Rubrivivax gelatinosus
1.3.5.6 0.0658
-
 7,9,9'-tricis-neurosporene pH 8.0, 28°C, recombinant mutant L208F Rubrivivax gelatinosus
1.3.99.28 0.0048
-
 15-cis-phytoene recombinant mutant L208F, pH 8.0, 28°C Rubrivivax gelatinosus
1.3.99.28 0.0148
-
 15-cis-phytoene recombinant wild-type enzyme, pH 8.0, 28°C Rubrivivax gelatinosus
1.3.99.28 0.0171
-
 15-cis-phytoene recombinant mutant L208P, pH 8.0, 28°C Rubrivivax gelatinosus
1.3.99.28 0.0331
-
 all-trans-neurosporene recombinant wild-type enzyme, pH 8.0, 28°C Rubrivivax gelatinosus
1.3.99.28 0.0417
-
 all-trans-neurosporene recombinant mutant L208P, pH 8.0, 28°C Rubrivivax gelatinosus
1.3.99.28 0.0658
-
 all-trans-neurosporene recombinant mutant L208F, pH 8.0, 28°C Rubrivivax gelatinosus
1.3.99.31 4.8
-
 15-cis-phytoene pH 8, 28°C, mutant enzyme L208F (complementation of CrtI in pPEU (DH5a/pACCrtEBEU/pPEUCrtIRg)) Rubrivivax gelatinosus
1.3.99.31 14.8
-
 15-cis-phytoene pH 8, 28°C, wild-type enzyme Rubrivivax gelatinosus
1.3.99.31 17.1
-
 15-cis-phytoene pH 8, 28°C, mutant enzyme L208P (complementation of CrtI in pUC8 (DH5a/pACCrtEBEU/pUCCrtIRg)) Rubrivivax gelatinosus
1.3.99.31 33.1
-
 all-trans-neurosporene pH 8, 28°C, wild-type enzyme Rubrivivax gelatinosus
1.3.99.31 41.7
-
 all-trans-neurosporene pH 8, 28°C, mutant enzyme (complementation of CrtI in pUC8 (DH5a/pACCrtEBEU/pUCCrtIRg)) Rubrivivax gelatinosus
1.3.99.31 65.8
-
 all-trans-neurosporene pH 8, 28°C, mutant enzyme L208F (complementation of CrtI in pPEU (DH5a/pACCrtEBEU/pPEUCrtIRg)) Rubrivivax gelatinosus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.3.5.6 membrane transmembrane enzyme, model, overview Rubrivivax gelatinosus 16020
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.5.6 7,9,9'-tricis-neurosporene + a quinone Rubrivivax gelatinosus
-
 7,9,7',9'-tetracis-lycopene + a quinol
-
 ?
1.3.5.6 7,9,9'-tricis-neurosporene + a quinone Rubrivivax gelatinosus CGMCC 6086
-
 7,9,7',9'-tetracis-lycopene + a quinol
-
 ?
1.3.5.6 phytoene + quinone Rubrivivax gelatinosus
-
 7,9,9'-tricis-neurosporene + quinol
-
 ?
1.3.5.6 phytoene + quinone Rubrivivax gelatinosus CGMCC 6086
-
 7,9,9'-tricis-neurosporene + quinol
-
 ?
1.3.99.28 15-cis-phytoene + FAD Rubrivivax gelatinosus
-
 all-trans-phytofluene + FADH2
-
 ?
1.3.99.28 15-cis-phytoene + FAD Rubrivivax gelatinosus CGMCC 6086
-
 all-trans-phytofluene + FADH2
-
 ?
1.3.99.28 all-trans-neurosporene + FAD Rubrivivax gelatinosus
-
 lycopene + FADH2
-
 ?
1.3.99.28 all-trans-neurosporene + FAD Rubrivivax gelatinosus CGMCC 6086
-
 lycopene + FADH2
-
 ?
1.3.99.28 all-trans-phytofluene + FAD Rubrivivax gelatinosus
-
 all-trans-zeta-carotene + FADH2
-
 ?
1.3.99.28 all-trans-zeta-carotene + FAD Rubrivivax gelatinosus
-
 all-trans-neurosporene + FADH2
-
 ?
1.3.99.28 additional information Rubrivivax gelatinosus the phytoene desaturase CrtI from Rubrivivax gelatinosus catalyzes simultaneously a three- and four-step desaturation producing both neurosporene and lycopene, determination of kinetic with the L208 modified desaturase and the specificities for phytoene and neurosporene as substrates, overview ?
-
 ?
1.3.99.28 additional information Rubrivivax gelatinosus CGMCC 6086 the phytoene desaturase CrtI from Rubrivivax gelatinosus catalyzes simultaneously a three- and four-step desaturation producing both neurosporene and lycopene, determination of kinetic with the L208 modified desaturase and the specificities for phytoene and neurosporene as substrates, overview ?
-
 ?
1.3.99.31 15-cis-phytoene + 3 acceptor Rubrivivax gelatinosus the enzyme simultaneously catalyzes a three- and four-step desaturation of phytoene producing both neurosporene and lycopene. These carotenes are intermediates for the synthesis of spheroidene and spirilloxanthin, respectively all-trans-neurosporene + 3 reduced acceptor
-
 ?
1.3.99.31 15-cis-phytoene + 4 acceptor Rubrivivax gelatinosus the enzyme simultaneously catalyzes a three- and four-step desaturation of phytoene producing both neurosporene and lycopene. These carotenes are intermediates for the synthesis of spheroidene and spirilloxanthin, respectively all-trans-lycopene + 4 reduced acceptor
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.3.5.6 Rubrivivax gelatinosus
-
 gene crtI
-
1.3.99.28 Rubrivivax gelatinosus
-
-
-
1.3.99.28 Rubrivivax gelatinosus CGMCC 6086
-
-
-
1.3.99.31 Rubrivivax gelatinosus
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.5.6 7,9,9'-tricis-neurosporene + a quinone
-
 Rubrivivax gelatinosus 7,9,7',9'-tetracis-lycopene + a quinol
-
 ?
1.3.5.6 7,9,9'-tricis-neurosporene + a quinone
-
 Rubrivivax gelatinosus CGMCC 6086 7,9,7',9'-tetracis-lycopene + a quinol
-
 ?
1.3.5.6 phytoene + quinone
-
 Rubrivivax gelatinosus 7,9,9'-tricis-neurosporene + quinol
-
 ?
1.3.5.6 phytoene + quinone
-
 Rubrivivax gelatinosus CGMCC 6086 7,9,9'-tricis-neurosporene + quinol
-
 ?
1.3.99.28 15-cis-phytoene + FAD
-
 Rubrivivax gelatinosus all-trans-phytofluene + FADH2
-
 ?
1.3.99.28 15-cis-phytoene + FAD
-
 Rubrivivax gelatinosus CGMCC 6086 all-trans-phytofluene + FADH2
-
 ?
1.3.99.28 all-trans-neurosporene + FAD
-
 Rubrivivax gelatinosus lycopene + FADH2
-
 ?
1.3.99.28 all-trans-neurosporene + FAD
-
 Rubrivivax gelatinosus CGMCC 6086 lycopene + FADH2
-
 ?
1.3.99.28 all-trans-phytofluene + FAD
-
 Rubrivivax gelatinosus all-trans-zeta-carotene + FADH2
-
 ?
1.3.99.28 all-trans-zeta-carotene + FAD
-
 Rubrivivax gelatinosus all-trans-neurosporene + FADH2
-
 ?
1.3.99.28 additional information the phytoene desaturase CrtI from Rubrivivax gelatinosus catalyzes simultaneously a three- and four-step desaturation producing both neurosporene and lycopene, determination of kinetic with the L208 modified desaturase and the specificities for phytoene and neurosporene as substrates, overview Rubrivivax gelatinosus ?
-
 ?
1.3.99.28 additional information the phytoene desaturase CrtI from Rubrivivax gelatinosus catalyzes simultaneously a three- and four-step desaturation producing both neurosporene and lycopene, determination of kinetic with the L208 modified desaturase and the specificities for phytoene and neurosporene as substrates, overview Rubrivivax gelatinosus CGMCC 6086 ?
-
 ?
1.3.99.31 15-cis-phytoene + 3 acceptor the enzyme simultaneously catalyzes a three- and four-step desaturation of phytoene producing both neurosporene and lycopene. These carotenes are intermediates for the synthesis of spheroidene and spirilloxanthin, respectively Rubrivivax gelatinosus all-trans-neurosporene + 3 reduced acceptor
-
 ?
1.3.99.31 15-cis-phytoene + 3 acceptor the enzyme simultaneously catalyzes a three- and four-step desaturation of phytoene producing both neurosporene and lycopene. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg) and 26:74 after complementation of CrtI in pPEU(DH5a/pACCrtEBEU/pPEUCrtIRg). The affinity for neurosporene conversion is poorer than for phytoene conversion. This explains the formation of two desaturation products Rubrivivax gelatinosus all-trans-neurosporene + 3 reduced acceptor
-
 ?
1.3.99.31 15-cis-phytoene + 4 acceptor the enzyme simultaneously catalyzes a three- and four-step desaturation of phytoene producing both neurosporene and lycopene. These carotenes are intermediates for the synthesis of spheroidene and spirilloxanthin, respectively Rubrivivax gelatinosus all-trans-lycopene + 4 reduced acceptor
-
 ?
1.3.99.31 15-cis-phytoene + 4 acceptor the enzyme simultaneously catalyzes a three- and four-step desaturation of phytoene producing both neurosporene and lycopene. The ratio of lycopene to neurosporene of the wild-type enzyme under assay conditions is 88:12 after complementation of CrtI in pUC8(DH5a/pACCrtEBEU/pUCCrtIRg) and 26:74 after complementation of CrtI in pPEU(DH5a/pACCrtEBEU/pPEUCrtIRg). The affinity for neurosporene conversion is poorer than for phytoene conversion. This explains the formation of two desaturation products Rubrivivax gelatinosus all-trans-lycopene + 4 reduced acceptor
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.3.5.6 CrtI
-
 Rubrivivax gelatinosus
1.3.5.6 phytoene desaturase
-
 Rubrivivax gelatinosus
1.3.99.28 CrtI
-
 Rubrivivax gelatinosus
1.3.99.28 phytoene desaturase
-
 Rubrivivax gelatinosus
1.3.99.31 CrtI
-
 Rubrivivax gelatinosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.5.6 28
-
 assay at Rubrivivax gelatinosus
1.3.99.28 28
-
 assay at Rubrivivax gelatinosus
1.3.99.31 28
-
 assay at Rubrivivax gelatinosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.5.6 8
-
 assay at Rubrivivax gelatinosus
1.3.99.28 8
-
 assay at Rubrivivax gelatinosus
1.3.99.31 8
-
 assay at Rubrivivax gelatinosus

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.99.28 FAD
-
 Rubrivivax gelatinosus
1.3.99.31 FAD added to the reaction mixture Rubrivivax gelatinosus

General Information

EC Number General Information Comment Organism
1.3.5.6 metabolism phytoene desaturase CrtI from Rubrivivax gelatinosus catalyzes simultaneously a three- and four-step desaturation producing both neurosporene and lycopene. These carotenes are intermediates for the synthesis of spheroidene and spirilloxanthin, respectively Rubrivivax gelatinosus
1.3.5.6 additional information Leu208, located in a highly hydrophobic membrane-integrated region, is important in substrate binding Rubrivivax gelatinosus
1.3.99.28 additional information Leu208 is exchanged in the neurosporene or lycopene-forming desaturase Rubrivivax gelatinosus
1.3.99.31 physiological function the enzyme simultaneously catalyzes a three- and four-step desaturation of phytoene producing both neurosporene and lycopene. These carotenes are intermediates for the synthesis of spheroidene and spirilloxanthin, respectively Rubrivivax gelatinosus