Esterase PHE21 from Pseudomonas oryzihabitans HUP022

Enzyme Description

Species

Extremophile

EC Number

3.1.1.1 ↗

Sequence

Length: 321 amino acids

MPDVFARLTPEVAGLLAQLAAHPQPPLDSLTPAQARAAYVQLNRALGLPPAPVKRVEALSAPGPLGPIPLRLYRPYAEDDRPRPLVIYLHGGGWVIGDLDSHDSLCRQIAIGTGYAVLAVHYALAPEHPAPASPDDVLAALHWLAEAGSAFHLDLDRIAVAGDSAGGGLAAMAALYLREGPLRLKAQVLIYPGVDNTPEAWNHPSRIENAQVPPLTRPMMDYFSGMYLQDVDTRDPRVSPLYAASHADLPPALIFGAECDALRDDARLYAQALIGAGNAVEYHELPGMIHGFIEMLGVLPSVRWTLARMNAFLREHLQQAA

Yilong Wang et al. (2016) — Functional Characterization of a Robust Marine Microbial Esterase and Its Utilization in the Stereo-Selective Preparation of Ethyl (S)-3-Hydroxybutyrate

Applied Biochemistry and Biotechnology · doi:10.1007/s12010-016-2161-1 ↗ · Stability - Incubation Activity + Stability - Conversion Specificity - Enantioselectivity

51 measurements ▼

Database ID

UniProt: A0A1B1PFW8 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (51 measurements)

51 measurements — page 3 of 3

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	n-Hexane	10%	100.0	95.21	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	1-Butanol	10%	100.0	108.81	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	1-Decanol	10%	100.0	94.4	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Isopropanol	10%	100.0	78.73	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100.0	82.81	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Chloroform	10%	100.0	112.89	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Tetrahydrofuran (THF)	10%	100.0	110.87	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Toluene	10%	100.0	106.43	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	1-Pentanol	10%	100.0	144.98	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	n-Decane	10%	100.0	87.99	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Methanol	10%	100.0	102.64	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid , p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Cyclohexanone	10%	51.0	28.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Dimethylformamide (DMF)	10%	51.0	26.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Isooctane	10%	51.0	34.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	n-Hexane	10%	51.0	28.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Tetrahydrofuran (THF)	10%	51.0	34.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Chloroform	10%	51.0	37.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Acetonitrile	10%	51.0	46.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Methanol	10%	51.0	34.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Toluene	10%	51.0	23.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Dimethyl Sulfoxide (DMSO)	10%	51.0	36.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	Cyclohexane	10%	51.0	36.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	1-Pentanol	10%	51.0	28.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Activity + Stability - Conversion	Conversion after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC	1-Butanol	10%	51.0	32.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Dimethylformamide (DMF)	10%	69.0	13.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Toluene	10%	9.63	12.75	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Dimethyl Sulfoxide (DMSO)	10%	9.63	10.27	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	1-Butanol	10%	69.0	32.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	1-Pentanol	10%	69.0	26.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Cyclohexanone	10%	69.0	5.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Cyclohexane	10%	69.0	16.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Dimethyl Sulfoxide (DMSO)	10%	69.0	42.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Methanol	10%	9.63	91.01	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Acetonitrile	10%	9.63	7.8	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Chloroform	10%	9.63	2.8	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Tetrahydrofuran (THF)	10%	9.63	3.04	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	n-Hexane	10%	9.63	6.17	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Isooctane	10%	9.63	11.43	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Dimethylformamide (DMF)	10%	9.63	2.41	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Cyclohexane	10%	9.63	2.08	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	Cyclohexanone	10%	9.63	1.35	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	1-Pentanol	10%	9.63	10.47	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric ratio (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ln((1-C)(1-ees))/ln((1+C)(1+ees))	1-Butanol	10%	9.63	6.96	enantiomeric ratio	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in enantiomeric ratio) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	n-Hexane	10%	69.0	26.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Tetrahydrofuran (THF)	10%	69.0	22.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Chloroform	10%	69.0	22.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Acetonitrile	10%	69.0	54.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Methanol	10%	69.0	50.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Toluene	10%	69.0	24.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Specificity - Enantioselectivity	Enantiomeric excess (toward s form) after incubation in the presence of organic solvent (2 hours at 40°C) measured by GC and computed as ([S]-[R])/([S]+[R])	Isooctane	10%	69.0	40.0	%	50 mM phosphate buffer	8	40°C	50 mM Ethyl 3-hydroxybutyrate	3-Hydroxybutyric Acid, Ethanol	—	—	Classical aqueous control (in %) \| Uncertainty about organic solvent concentration
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Ethanol	10%	100.0	84.59	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	n-Hexane	10%	100.0	95.21	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	1-Butanol	10%	100.0	108.81	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	1-Decanol	10%	100.0	94.4	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Isopropanol	10%	100.0	78.73	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100.0	82.81	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Chloroform	10%	100.0	112.89	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Tetrahydrofuran (THF)	10%	100.0	110.87	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Toluene	10%	100.0	106.43	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	1-Pentanol	10%	100.0	144.98	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	n-Decane	10%	100.0	87.99	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control
Stability - Incubation	Activity after incubation (12 hours at 4°C), measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in aqueous phase	Methanol	10%	100.0	102.64	%	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer, 1% acetonitrile, 4% Ethanol	Incubation: 7.5, Assay: 7.5	Incubation: 4°C, Assay: 35°C	0.1 mM p-Nitrophenyl butyrate	Butyric Acid, p-Nitrophenol	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Uncertainty about whether assay in aqueous phase or in the presence of organic solvent, uncertainty about control

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Visualization : Stability — Incubation

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

Visualization : Activity + Stability — Conversion

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

Visualization : Specificity — Enantioselectivity

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

Structure

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Esterase PHE21 from Pseudomonas oryzihabitans HUP022

Enzyme Description

Sequence

Experimental Data (51 measurements)

Visualization : Stability — Incubation

Visualization : Activity + Stability — Conversion

Visualization : Specificity — Enantioselectivity

Structure

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