Carotenoid 9,10-dioxygenase from Vitis vinifera (grape)

Enzyme Description

Species
Vitis vinifera (grape) β†—
Extremophile
No
EC Number
1.14.99.n4 β†—

Sequence

Length: 739 amino acids
MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRGAGKTAPEGIIPKVLKQHAAELGHDLKEKGLKDLDRLQWLGKQLKQLPGVLPALADGAGPKGQPGMTEGSLAVYQAGDTLVVSTQTALAMAEKEEQGGAGVVVVDPKPSKGFTSKAVDWLEKLIVKLMHDSSQPLHYLSGNFAPVRDETPPCKNLPVIGYLPECLNGEFVRVGPNPKFSPVAGYHWFDGDGMIHGLHIKDGKATYVSRYVRTSRLKQEEYFGGAKFMRIGDLKGLFGLLMVNMQMLRAKLKILDVSYGTGTGNTALVFHHGKLLALSEADKPYVLKVLEDGDLQTLGMLDYDKRLTHSFTAHPKVDPFTGEMFSFGYSHTPPYITYRVISKDGFMHEPVPITISDPIMMHDFAITENYAIFMDLPLYFRPKEMVKEKKLIFTFDATKKARFGVLPRYAKNELHIKWFELPNCFIFHNANAWEEEDEVVLITCRLENPDLDLVGGDVKEKLENFGNELYEMRFNMKTGIASQRKLSASSVDFPRVNESYTGRKQRYVYGTILDSIAKVTGIIKFDLHAEPDTGKSKLEVGGNVQGIFDLGVGRFGSEAVFVPREPGITSEEDDGYLIFFVHDEKTGKSYVNVIDAKTMSPDPIAIVELPNRVPYGFHAFFVTEEQLKEQAKL
Sandrine Mathieu et al. (2007) β€” Production of a recombinant carotenoid cleavage dioxygenase from grape and enzyme assay in water-miscible organic solvents
Biotechnology Letters  Β· doi:10.1007/s10529-007-9315-8 β†—  Β· Activity - Classical
12 measurements ▼
Database ID
UniProt: Q3T4H1 β†—
Sequence Annotation
Inferred - previous publication for protein sequence, with N-terminal GST-tag
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (12 measurements)

12 measurements
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Extraction method Data type Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent 1,4-Dioxane 10 % v/v 0.044 β€” 0.14 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent 1,4-Dioxane 15 % v/v 0.044 β€” 0.26 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent 1,4-Dioxane 20 % v/v 0.044 β€” 0.13 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10 % v/v 0.044 β€” 0.93 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20 % v/v 0.044 β€” 0.23 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Methanol 10 % v/v 0.044 β€” 0.36 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Methanol 20 % v/v 0.044 β€” 1.09 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Methanol 30 % v/v 0.044 β€” 0.54 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Acetone 5 % v/v 0.044 β€” 0.53 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Acetone 10 % v/v 0.044 β€” 1.16 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Acetone 15 % v/v 0.044 β€” 0.73 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)
No, presence GST-tag Activity - Classical Activity measured by absorbance spectrophotometry (lutein absorbance decrease measurement, 450 nm), in the presence of organic solvent Acetone 20 % v/v 0.044 β€” 0.25 nmol/(min * mg of protein) Direct Continuous 70% Bis-Tris (v/v), 1 mM DTT, 5 Β΅M FeSO4, 0.11% Triton X-100 6.5 30Β°C 35 Β΅M Lutein 3-hydrox-beta-ionine , C14-dialdehyde β€” 500 rpm Classical aqueous control (in nmol/(min * mg of protein)), measured after 1 hour incubation | Presence of an N-terminal GST-fusion (uncleaved)

Mutations in this dataset (1)

No, presence GST-tag

Visualization : Activity β€” Classical

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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