Application | Comment | Organism |
---|---|---|
synthesis | preparation of (S)-1-cyclopropyl-2-methoxyethanamine, a key chiral intermediate for the synthesis of a corticotropin releasing factor-1 (CRF-1) receptor antagonist, by a chemoenzymatic route using leucine dehydrogenase. Synthesis of (S)-1-cyclopropyl-2-methoxyethanamine starting from methylcyclopropyl ketone. Permanganate oxidation of the ketone gives cyclopropylglyoxylic acid, which is converted to (S)-cyclopropylglycine by reductive amination using leucine dehydrogenase from Thermoactinomyces intermedius, recombinantly expressed in Escherichia coli, with NADH cofactor recycling by formate dehydrogenase from Pichia pastoris | Thermoactinomyces intermedius |
Cloned (Comment) | Organism |
---|---|
recombinant expression in Escherichia coli strain SC16591 | Thermoactinomyces intermedius |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-leucine + H2O + NAD+ | Thermoactinomyces intermedius | - |
4-methyl-2-oxopentanoate + NH3 + NADH + H+ | - |
? | |
additional information | Thermoactinomyces intermedius | synthesis of (S)-1-cyclopropyl-2-methoxyethanamine starting from methylcyclopropyl ketone. Permanganate oxidation of the ketone gives cyclopropylglyoxylic acid, which is converted to (S)-cyclopropylglycine by reductive amination using leucine dehydrogenase from Thermoactinomyces intermedius with NADH cofactor recycling by formate dehydrogenase from Pichia pastoris | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermoactinomyces intermedius | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
comparing extract activities with 5 mM 2-oxoisovalerate (3282 U/mL) versus 5 mM cyclopropylglyoxylic acid (86 U/mL), leucine dehydrogenase is 38fold more active with the natural substrate | Thermoactinomyces intermedius |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-leucine + H2O + NAD+ | - |
Thermoactinomyces intermedius | 4-methyl-2-oxopentanoate + NH3 + NADH + H+ | - |
? | |
additional information | synthesis of (S)-1-cyclopropyl-2-methoxyethanamine starting from methylcyclopropyl ketone. Permanganate oxidation of the ketone gives cyclopropylglyoxylic acid, which is converted to (S)-cyclopropylglycine by reductive amination using leucine dehydrogenase from Thermoactinomyces intermedius with NADH cofactor recycling by formate dehydrogenase from Pichia pastoris | Thermoactinomyces intermedius | ? | - |
? | |
additional information | comparing extract activities with 5 mM 2-oxoisovalerate (3282 U/mL) versus 5 mM cyclopropylglyoxylic acid (86 U/mL), leucine dehydrogenase is 38fold more active with the natural substrate | Thermoactinomyces intermedius | ? | - |
? |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
with cyclopropylglyoxylic acid as substrate, recombinant enzyme | Thermoactinomyces intermedius |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7.5 | 10 | activity range, profile, overview | Thermoactinomyces intermedius |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Thermoactinomyces intermedius | |
NADH | - |
Thermoactinomyces intermedius |