Ammonia → HCID 23615384, compound

Participants

Reaction Steps

6.3 g (0.1 mole) ammonium formate and 1.67 g (10 mmoles), 2-keto-3,3-dimethyl pentanoic acid sodium salt are suspended in 80 ml of water, the pH value is adjusted with ammonia to 8.2, so that the solids dissolve and the volume is adjusted to 100 ml. subsequently, 14.34 mg (0.02 mmole) NAD+3H2O cofactor as well as 800 U of leucine dehydrogenase (LeUDH) and 500 U of rec-formate dehydrogenase (rec-FDH) are added. The temperature is set to 32° C . During the reaction the system is gently stirred and the pH value is adjusted to 8.2 via a pH stat unit. The completion point of the reaction is sdemonstrated by determining the degree of conversion with HPLC. The reaction solution is adjusted to pH 9,5 with ammonia and subsequently clarified with 2% active charcoal. The almost colourless solution is concentrated with a rotary evaporator, the amino acid is crystallized, is separated via a filter funnel, washed three times with a little ethanol and dried overnight under vacuum at 50° C.

Provenance & Evidence