Diketone hydrolase from Anabaena sp. PCC 7120

Enzyme Description

Species
Anabaena sp. PCC 7120 β†—
Extremophile
No
EC Number
3.7.1.- β†—

Sequence

Length: 263 amino acids
MGSSHHHHHHMTLNQPEYFTKYENLHFHRDENGILEVRMHTNGSSLVFTGKTHREFPDAFYDISRDRDNRVVILTGSGDAWMAEIDFPSLGDVTNPREWDKTYWEGKKVLQNLLDIEVPVISAVNGAALLHSEYILTTDIILASENTVFQDMPHLNAGIVPGDGVHILWPLALGLYRGRYFLFTQEKLTAQQAYELNVVHEVLPQSKLMERAWEIARTLAKQPTLNLRYTRVALTQRLKRLVNEGIGYGLALEGITATDLRNN
Elina Siirola et al. (2011) β€” Tolerance of Ξ²-diketone hydrolases as representatives of the crotonase superfamily towards organic solvents
Biotechnology and Bioengineering  Β· doi:10.1002/bit.23275 β†—  Β· Activity - Classical Selectivity - Enantioselectivity
24 measurements ▼
Database ID
UniParc: UPI0000EDA45E β†—
Sequence Annotation
Inferred - from protein name
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (24 measurements)

24 measurements β€” page 1 of 2
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Extraction method Data type Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Activity - Classical Activity measured by HPLC in the presence of organic solvent 1,4-Dioxane 80 % v/v 100 7 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Petroleum Ether 80 % v/v 100 82 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Diisopropyl Ether 80 % v/v 100 75 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent tert-Butyl Methyl Ether (MTBE) 80 % v/v 100 64 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent n-Hexane 80 % v/v 100 43 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Diethyl Ether 80 % v/v 100 47 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Toluene 80 % v/v 100 50 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Ethyl Acetate 80 % v/v 100 14 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Tetrahydrofuran (THF) 80 % v/v 100 0 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Glycerol 80 % v/v 100 4 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Ethylene Glycol 80 % v/v 100 7 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Acetonitrile 80 % v/v 100 0 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Activity - Classical Activity measured by HPLC in the presence of organic solvent Acetone 80 % v/v 100 4 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Selectivity - Enantioselectivity Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC Acetone 80 % v/v 93 54 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Selectivity - Enantioselectivity Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC Petroleum Ether 80 % v/v 93 96 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Selectivity - Enantioselectivity Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC Diisopropyl Ether 80 % v/v 93 98 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Selectivity - Enantioselectivity Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC tert-Butyl Methyl Ether (MTBE) 80 % v/v 93 98 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Selectivity - Enantioselectivity Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC n-Hexane 80 % v/v 93 96 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Selectivity - Enantioselectivity Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC Diethyl Ether 80 % v/v 93 94 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
Selectivity - Enantioselectivity Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC Toluene 80 % v/v 93 99 % Direct Continuous 50 mM phosphate buffer 7.5 30Β°C 50 mM Bicyclo(2.2.2)octane-2,6-dione (S)-2-(3-ketocyclohexyl)acetic acid β€” 120 rpm Classical aqueous control (in %)
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Visualization : Activity β€” Classical

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

Visualization : Selectivity - Enantioselectivity

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

Structure

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