Aminopeptidase (gene LUCA-DmpA) sequence construced by ancestral sequence reconstruction

Enzyme Description

Species

Na (ancestral sequence reconstruction)

Extremophile

No t applicable (ancestral sequence reconstruction)

EC Number

3.4.11.- ↗

Sequence

Length: 359 amino acids

MKRARLRDLGITIGRLPTGPYNAITDVPGVRVGHTTIIEDDPHVVNTGVTTILPQDGEVWEHHVFAGYFRFNGSGEMTGSHWLEESGLLSSPVIITNSFGVGACYDALVKYAAEQDPTAPFTLPVIAETFDGWLSDIGAMAVTPEHVREALENARSGPVAEGNVGGGTGMITMGFKAGTGTSSRVVEVEGEGYTVGALVQSNFGGARFLTINGVPVGRLIPADQVPVPWEEPPREDGSIIVIIATDAPLLPHQCKRLARRATLGVARTGGWGSNYSGDIFLAFSTGNRLPRQPEEPVYGLKMLPNEEMDPLFQGAVEATEEAILNAICMAETMKGRKGREVKALPLDRLKEILKRPGRR

Fan Liu et al. (2022) — Preparation and Characterization of an Ancient Aminopeptidase Obtained from Ancestral Sequence Reconstruction for L-Carnosine Synthesis

Molecules · doi:10.3390/molecules27196620 ↗ · Stability - Incubation

8 measurements ▼

Database ID

—

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (C2530)

Experimental Data (8 measurements)

8 measurements

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 (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	n-Hexane	50%	100	124	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine , Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Toluene	50%	100	104	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine , Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	tert-Butanol	50%	100	108	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine , Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Isopropanol	50%	100	88	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine , Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Ethanol	50%	100	94	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine , Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Acetonitrile	50%	100	80	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine , Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Methanol	50%	100	89	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine , Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Dimethyl Sulfoxide (DMSO)	50%	100	70	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine , Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase

Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	n-Hexane	50%	100	124	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine, Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Toluene	50%	100	104	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine, Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	tert-Butanol	50%	100	108	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine, Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Isopropanol	50%	100	88	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine, Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Ethanol	50%	100	94	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine, Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Acetonitrile	50%	100	80	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine, Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Methanol	50%	100	89	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine, Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase
Stability - Incubation	Activity after incubation (30 minutes at room temperature), measured by HPLC (L-carnosine product formation measurement) in aqueous phase	Dimethyl Sulfoxide (DMSO)	50%	100	70	%	Incubation: ?, Assay: 20 mM phosphate buffer	Incubation: ?, Assay: 9	Incubation: Room temperature, Assay: 45°C	100 mM L-Histidine, 50 mM Methyl β-alanate	L-Carnosine, Methanol	—	—	Non-incubated control (in %), assay in aqueous phase \| High uncertainty about whether assay really was in the presence of organic solvent or in aqueous phase

Visualization : Stability — Incubation

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

Structure

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