Pyruvate decarboxylase from Acetobacter pasteurianus

Enzyme Description

Species

Extremophile

EC Number

4.1.1.1 ↗

Sequence

Length: 558 amino acids

MTYTVGMYLAERLVQIGLKHHFAVAGDYNLVLLDQLLLNKDMKQIYCCNELNCGFSAEGYARSNGAAAAVVTFSVGAISAMNALGGAYAENLPVILISGAPNSNDQGTGHILHHTIGKTDYSYQLEMARQVTCAAESITDAHSAPAKIDHVIRTALRERKPAYLDIACNIASEPCVRPGPVSSLLSEPEIDHTSLKAAVDATVALLEKSASPVMLLGSKLRAANALAATETLADKLQCAVTIMAAAKGFFPEDHAGFRGLYWGEVSNPGVQELVETSDALLCIAPVFNDYSTVGWSAWPKGPNVILAEPDRVTVDGRAYDGFTLRAFLQALAEKAPARPASAQKSSVPTCSLTATSDEAGLTNDEIVRHINALLTSNTTLVAETGDSWFNAMRMTLPRGARVELEMQWGHIGWSVPSAFGNAMGSQDRQHVVMVGDGSFQLTAQEVAQMVRYELPVIIFLINNRGYVIEIAIHDGPYNYIKNWDYAGLMEVFNAGEGHGLGLKATTPKELTEAIARAKANTRGPTLIECQIDRTDCTDMLVQWGRKVASTNARKTTLA

Samuel Sutiono et al. (2019) — To beat the heat–engineering of the most thermostable pyruvate decarboxylase to date

RSC Advances · doi:10.1039/c9ra06251c ↗ · Stability - Incubation

59 measurements ▼

Database ID

UniProt: A0A4Y3TS39 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (59 measurements)

59 measurements — page 2 of 3

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Extraction method	Data type	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
WT	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	8 % v/v	100	—	3	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
WT	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	7 % v/v	100	—	3	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
WT	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	6 % v/v	100	—	22	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
WT	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	5 % v/v	100	—	70	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
WT	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	4 % v/v	100	—	86	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
WT	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	3 % v/v	100	—	95	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
WT	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	2 % v/v	100	—	94	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
WT	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	0.5 % v/v	100	—	99	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	15 % v/v	100	—	107	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	1-Butanol	9 % v/v	100	2	6	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 60°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	10 % v/v	100	—	105	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	5 % v/v	100	—	102	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	18 % v/v	100	—	109	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	20 % v/v	100	74	109	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	25 % v/v	100	—	108	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	30 % v/v	100	—	103	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	35 % v/v	100	—	89	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	40 % v/v	100	—	52	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	45 % v/v	100	—	2	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans
L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V	Stability - Incubation	Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase	Ethanol	50 % v/v	100	—	0	%	Digital	Continuous	Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer	Incubation: 6.5, Assay: 6.5	Incubation: 50°C, Assay: 25°C	25 mM Pyruvate	Acetaldehyde , Carbon Dioxide (CO2)	Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

9 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Non-incubated control (in %), assay in aqueous phase | Clear about assay in aqueous conditions/Explicit protein sequence given corresponds to UniProt A0A4Y3TS39 which is annotated as for a protein from Acetobacter peroxydans

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

0.5 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

1 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

2 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

3 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

4 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

5 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

6 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

7 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

8 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

1 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

4 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

8 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

12 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

16 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

20 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

24 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

28 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

32 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

9 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

8 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

7 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

6 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

5 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

4 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

3 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

2 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

0.5 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

15 % v/v

100

—

107

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

9 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

10 % v/v

100

—

105

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

5 % v/v

100

—

102

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

18 % v/v

100

—

109

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

20 % v/v

100

109

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

25 % v/v

100

—

108

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

30 % v/v

100

—

103

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

35 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

40 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

45 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

Ethanol

50 % v/v

100

—

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

0.5 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

8 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

7 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

6 % v/v

100

102

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

5 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

4 % v/v

100

104

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

3 % v/v

100

103

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

2 % v/v

100

102

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

1 % v/v

100

101

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

0.5 % v/v

100

102

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 50°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

9 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 50°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

1 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

2 % v/v

100

102

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

3 % v/v

100

101

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

4 % v/v

100

102

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

5 % v/v

100

101

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

6 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

7 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Stability - Incubation

Activity after incubation (1 hour at 60°C), measured by absorbance spectrophotometry (change in pyruvate absorbance measurement, 315 nm) in aqueous phase

1-Butanol

8 % v/v

100

Digital

Continuous

Incubation: 50 mM phosphate buffer, Assay: 50 mM phosphate buffer

Incubation: 6.5, Assay: 6.5

Incubation: 60°C, Assay: 25°C

25 mM Pyruvate

Acetaldehyde, Carbon Dioxide (CO2)

Incubation: 0.1 mM TDP, 5 mM MgSO₄, Assay: 0.1 mM TDP, 5 mM MgSO₄

—

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Mutations in this dataset (2)

WT L84I/A100S/S103T/Q106Y/K118T/S122N/A128V/R129K/Q130H/T139V/D140S/H142E/S143E/L441V

Visualization : Stability — Incubation

One bar per measurement. Colour = solvent, shade = solvent volume. — — — Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Structure

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Pyruvate decarboxylase from Acetobacter pasteurianus

Enzyme Description

Sequence

Experimental Data (59 measurements)

Visualization : Stability — Incubation

Mutation Effect

Structure

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