Diketone hydrolase from Anabaena sp. PCC 7120

Enzyme Description

Species

Extremophile

EC Number

3.7.1.- ↗

Sequence

Length: 263 amino acids

MGSSHHHHHHMTLNQPEYFTKYENLHFHRDENGILEVRMHTNGSSLVFTGKTHREFPDAFYDISRDRDNRVVILTGSGDAWMAEIDFPSLGDVTNPREWDKTYWEGKKVLQNLLDIEVPVISAVNGAALLHSEYILTTDIILASENTVFQDMPHLNAGIVPGDGVHILWPLALGLYRGRYFLFTQEKLTAQQAYELNVVHEVLPQSKLMERAWEIARTLAKQPTLNLRYTRVALTQRLKRLVNEGIGYGLALEGITATDLRNN

Elina Siirola et al. (2011) — Tolerance of β-diketone hydrolases as representatives of the crotonase superfamily towards organic solvents

Biotechnology and Bioengineering · doi:10.1002/bit.23275 ↗ · Activity - Classical Specificity - Enantioselectivity

24 measurements ▼

Database ID

UniParc: UPI0000EDA45E ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (24 measurements)

24 measurements — page 1 of 2

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	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%	100	7	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Petroleum Ether	80%	100	82	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Diisopropyl Ether	80%	100	75	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	tert-Butyl Methyl Ether (MTBE)	80%	100	64	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	n-Hexane	80%	100	43	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Diethyl Ether	80%	100	47	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Toluene	80%	100	50	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Ethyl Acetate	80%	100	14	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Tetrahydrofuran (THF)	80%	100	0	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Glycerol	80%	100	4	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Ethylene Glycol	80%	100	7	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Acetonitrile	80%	100	0	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Acetone	80%	100	4	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Acetone	80%	93	4	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Petroleum Ether	80%	93	82	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Diisopropyl Ether	80%	93	75	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - 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%	93	64	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	n-Hexane	80%	93	43	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Diethyl Ether	80%	93	47	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Toluene	80%	93	50	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)

Activity - Classical	Activity measured by HPLC in the presence of organic solvent	1,4-Dioxane	80%	100	7	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Petroleum Ether	80%	100	82	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Diisopropyl Ether	80%	100	75	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	tert-Butyl Methyl Ether (MTBE)	80%	100	64	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	n-Hexane	80%	100	43	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Diethyl Ether	80%	100	47	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Toluene	80%	100	50	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Ethyl Acetate	80%	100	14	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Tetrahydrofuran (THF)	80%	100	0	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Glycerol	80%	100	4	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Ethylene Glycol	80%	100	7	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Acetonitrile	80%	100	0	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Activity - Classical	Activity measured by HPLC in the presence of organic solvent	Acetone	80%	100	4	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Acetone	80%	93	4	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Petroleum Ether	80%	93	82	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Diisopropyl Ether	80%	93	75	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - 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%	93	64	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	n-Hexane	80%	93	43	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Diethyl Ether	80%	93	47	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Toluene	80%	93	50	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Ethyl Acetate	80%	93	14	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Glycerol	80%	93	0	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Ethylene Glycol	80%	93	4	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)
Specificity - Enantioselectivity	Enantiomeric excess (toward the S form) after 3 hours of incubation in the presence of organic solvent, measured by HPLC	Acetonitrile	80%	93	7	%	50 mM phosphate buffer	7.5	30°C	50 mM Bicyclo(2.2.2)octane-2,6-dione	(S)-2-(3-ketocyclohexyl)acetic acid	—	120 rpm	Classical aqueous control (in %)

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Visualization : Activity — Classical

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

Visualization : Specificity — Enantioselectivity