Sulfuric Acid → 4-Hydroxy-2-oxoglutaric acid

Participants

Reaction Steps

Pseudomonas putida ATCC 795 strain was inoculated into 9 ml of L medium and cultured for 16 hours at 30° C. with shaking. Whole amount of this culture was then inoculated into 300 ml of GMS medium in 2 liter volume Erlenmeyer flask and cultured for 20 hours at 30° C. with shaking. The resulting culture was centrifuged, the collected cells were resuspended in 60 ml of the reactant solution (b), and the suspension was stirred in 300-ml beaker while maintaining pH at 6.5 with 1N sulfuric acid. After 6 hours, 3 ml of 2M sodium pyruvate and 3 ml of 2M sodium glyoxylate were added. After 24 hours from the starting point of this reaction, the cells were removed from the suspension by centrifugation, and the amount of 4-hydroxy-2-ketoglutaric acid formed in the supernatant was determined by HPLC. The amount of L-4-hydroxy-2-ketoglutaric acid was determined to be 150 mM and D-form was not detected.

Provenance & Evidence