L-ALANINE DEHYDROGENASE FROM BACILLUS SUBTILIS (C005B-002530)

Application

L-Alanine dehydrogenase converts L-alanine to pyruvate and ammonium. L-Alanine dehydrogenase from Bacillus subtilis may be used to study enzyme inactivation and protection .

Biochem/physiol Actions

L-Alanine dehydrogenase is an A-stereospecific dehydrogenase that catalyzes the reversible deamination of L-alanine to pyruvate and ammonium. It is important for the generation of pyruvate during sporulation. L-Alanine dehydrogenase from Bacillus subtilis has a predominately ordered kinetic mechanism in which NAD binds before L-alanine. Subsequently, ammonia, pyruvate, and NADH are released in that specific order. Optimal pH for the amination reaction is 8.8-9.0, whereas it is 10-10.5 for the deamination reaction. The enzyme is inactivated by divalent metal ions and p-chloromercuribenzoate, mercuric ion being most effective. The inactivation may be reversed by L- or D-cysteine.

Unit Definition

One unit will convert 1.0 μmole of L-alanine to pyruvate and NH₃ per min at pH 10.0 at 25 °C.

Physical form

Solution in 50% glycerol containing 10 mM potassium phosphate buffer, pH 7.7

biological source: Bacillus subtilis. Quality Level: 200. form: buffered aqueous glycerol solution. specific activity: ~30 . units/mg protein (Lowry). foreign activity: LDH ~1% (using pyruvate as substrate). storage temp.: −. 20°C. Pictograms: GHS08. Signal Word: Danger. Hazard Statements: H334. Precautionary Statements: P261 - P284 - P501. Hazard Classifications: Resp. Sens. 1. Storage Class Code: 10 - Combustible liquids. WGK: WGK 3. Personal Protective Equipment: Eyeshields, Gloves, multi-purpose combination respirator cartridge (US).