Application | Comment | Organism |
---|---|---|
additional information | both (R,S)-(-)-ephedrine and (S,S)-(+)-pseudoephedrine are constituents of various over-the-counter (OTC) drugs and are also used as decongestants and stimulants. Arthrobacter sp. TS-15 and its isolated ephedrine-oxidizing enzymes have potential for use in decontamination and synthetic applications | Arthrobacter sp. TS-15 |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Arthrobacter sp. TS-15 |
recombinant expression of His-tagged enzyme in Escherichia coli strain T7 SHuffle(DE3) | Arthrobacter sp. TS-15 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.26 | - |
(+)-(1S,2S)-pseudoephedrine | pH 7.8, 25°C | Arthrobacter sp. TS-15 | |
0.263 | - |
(S,S)-(+)-pseudoephedrine | pH 7.8, 25°C | Arthrobacter sp. TS-15 | |
0.33 | - |
(S,R)-(+)-ephedrine | pH 7.8, 25°C | Arthrobacter sp. TS-15 | |
3.4 | - |
NAD+ | pH 7.8, 25°C | Arthrobacter sp. TS-15 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
30000 | - |
- |
Arthrobacter sp. TS-15 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(+)-(1S,2S)-pseudoephedrine + NAD+ | Arthrobacter sp. TS-15 | - |
(S)-2-(methylamino)-1-phenylpropan-1-one + NADH + H+ | - |
? | |
(S,R)-(+)-ephedrine + NAD+ | Arthrobacter sp. TS-15 | - |
(R)-2-methylimino-1-phenylpropan-1-ol + NADH + H+ | - |
? | |
(S,S)-(+)-pseudoephedrine + NAD+ | Arthrobacter sp. TS-15 | - |
(S)-2-(methylamino)-1-phenylpropan-1-one + NADH + H+ | - |
? | |
1-phenylpropan-1,2-dione + NAD+ | Arthrobacter sp. TS-15 | - |
phenylacetylcarbinol + NADH + H+ | - |
? | |
1-phenylpropan-1,2-dione + NAD+ | Arthrobacter sp. TS-15 | PseDH overcomes the stability of the conjugated molecule PPD by reducing it to alpha-hydroxyketone (S)-PAC. Phenylacetylcarbinol (PAC) cannot be oxidized by these dehydrogenases and possibly undergoes a cleavage reaction to yield benzaldehyde and acetaldehyde | phenylacetylcarbinol + NADH + H+ | - |
? | |
additional information | Arthrobacter sp. TS-15 | PseDH is strictly stereospecific for the other diastereomers, (S,S)-(+)-pseudoephedrine and (S,R)-(+)-ephedrine. Conversion of all ephedrine isomers required the addition of NAD+ as an oxidant. Without a surplus oxidant or in the presence of NADP+, cell lysates do not significantly convert (pseudo)ephedrine | ? | - |
- |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arthrobacter sp. TS-15 | A0A545BBS8 | - |
- |
Arthrobacter sp. TS-15 | A0A545BBS8 | DSM 32400, isolated from soil | - |
Purification (Comment) | Organism |
---|---|
- |
Arthrobacter sp. TS-15 |
recombinant His-tagged enzyme from Escherichia coli strain T7 SHuffle(DE3) by nickel affinity chromatography | Arthrobacter sp. TS-15 |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | the Gram-positive soil bacterium Arthrobacter sp. strain TS-15 (DSM 32400) is capable of metabolizing all isomers of ephedrine as a sole source of carbon and energy. (S,S)-(+)-pseudoephedrine is consumed within the shortest time period (half-life of approx. 6.3 h) and is accompanied by the fastest culture growth (generation time of approximately 4.4 h) | Arthrobacter sp. TS-15 | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
4.7 | - |
substrate: (+)-(1S,2S)-pseudoephedrine, pH 7.8, 25°C | Arthrobacter sp. TS-15 |
4.7 | - |
purified native enzyme, substrate (-)-ephedrine, pH 7.8, 25°C | Arthrobacter sp. TS-15 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(+)-(1S,2S)-pseudoephedrine + NAD+ | - |
Arthrobacter sp. TS-15 | (S)-2-(methylamino)-1-phenylpropan-1-one + NADH + H+ | - |
? | |
(+)-(1S,2S)-pseudoephedrine + NAD+ | the enzyme is strictly stereospecific for the diastereomers (S,S)-(+)-pseudoephedrine and (S,R)-(+)-ephedrine | Arthrobacter sp. TS-15 | (S)-2-(methylamino)-1-phenylpropan-1-one + NADH + H+ | - |
? | |
(S,R)-(+)-ephedrine + NAD+ | the enzyme is strictly stereospecific for the diastereomers (S,S)-(+)-pseudoephedrine and (S,R)-(+)-ephedrine | Arthrobacter sp. TS-15 | ? + NADH + H+ | - |
? | |
(S,R)-(+)-ephedrine + NAD+ | - |
Arthrobacter sp. TS-15 | (R)-2-methylimino-1-phenylpropan-1-ol + NADH + H+ | - |
? | |
(S,S)-(+)-pseudoephedrine + NAD+ | - |
Arthrobacter sp. TS-15 | (S)-2-(methylamino)-1-phenylpropan-1-one + NADH + H+ | - |
? | |
1-phenylpropan-1,2-dione + NAD+ | - |
Arthrobacter sp. TS-15 | phenylacetylcarbinol + NADH + H+ | - |
? | |
1-phenylpropan-1,2-dione + NAD+ | PseDH overcomes the stability of the conjugated molecule PPD by reducing it to alpha-hydroxyketone (S)-PAC. Phenylacetylcarbinol (PAC) cannot be oxidized by these dehydrogenases and possibly undergoes a cleavage reaction to yield benzaldehyde and acetaldehyde | Arthrobacter sp. TS-15 | phenylacetylcarbinol + NADH + H+ | - |
? | |
additional information | no activity on (S,N)-(+)-(pseudo)ephedrine or (R,N)-(-)-(pseudo)ephedrine | Arthrobacter sp. TS-15 | ? | - |
- |
|
additional information | PseDH is strictly stereospecific for the other diastereomers, (S,S)-(+)-pseudoephedrine and (S,R)-(+)-ephedrine. Conversion of all ephedrine isomers required the addition of NAD+ as an oxidant. Without a surplus oxidant or in the presence of NADP+, cell lysates do not significantly convert (pseudo)ephedrine | Arthrobacter sp. TS-15 | ? | - |
- |
|
additional information | pseudoephedrine dehydrogenase (PseDH) selectively converts (S,S)-(+)-pseudoephedrine and (S,R)-(+)-ephedrine to (S)- and (R)-methcathinone, respectively | Arthrobacter sp. TS-15 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
PseDH | - |
Arthrobacter sp. TS-15 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Arthrobacter sp. TS-15 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.45 | - |
(S,R)-(+)-ephedrine | pH 7.8, 25°C | Arthrobacter sp. TS-15 | |
3.77 | - |
(+)-(1S,2S)-pseudoephedrine | pH 7.8, 25°C | Arthrobacter sp. TS-15 | |
3.77 | - |
(S,S)-(+)-pseudoephedrine | pH 7.8, 25°C | Arthrobacter sp. TS-15 | |
9.22 | - |
NAD+ | pH 7.8, 25°C | Arthrobacter sp. TS-15 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
assay at | Arthrobacter sp. TS-15 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no activity with NADP+ | Arthrobacter sp. TS-15 | |
NAD+ | - |
Arthrobacter sp. TS-15 | |
NAD+ | the enzyme is strictly NAD+-dependent | Arthrobacter sp. TS-15 |
Organism | Comment | Expression |
---|---|---|
Arthrobacter sp. TS-15 | (S,S)-(+)-pseudoephedrine induces the enzyme | up |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the superfamily of short-chain dehydrogenases | Arthrobacter sp. TS-15 |
metabolism | the enzyme is involved in the ephedrine metabolism, overview | Arthrobacter sp. TS-15 |
physiological function | Arthrobacter sp. TS-15 is capable of metabolizing all isomers of ephedrine as a sole source of carbon and energy. Enantioselective oxidation of (-)-(R,N)-(pseudo)ephedrine and (+)-(S,N)-(pseudo)ephedrine catalyzed by PseDH and EDH, respectively, overview | Arthrobacter sp. TS-15 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.71 | - |
NAD+ | pH 7.8, 25°C | Arthrobacter sp. TS-15 | |
4.39 | - |
(S,R)-(+)-ephedrine | pH 7.8, 25°C | Arthrobacter sp. TS-15 | |
14.34 | - |
(S,S)-(+)-pseudoephedrine | pH 7.8, 25°C | Arthrobacter sp. TS-15 |