Creatine → 2-Acetoxypropionic Acid

HRID 1248789Notes

Reaction equation

HRID 1248789 reaction equation
[OH:1]S(O)(=O)=O.[O:6]=[C:7]([CH2:9]N(C(=N)N)C)[OH:8].[C:15]1([OH:25])[C:24]2[C:19](=CC=CC=2)C=CC=1>[OH-].[Na+]>[CH3:19][CH:24]([O:8][C:7]([CH3:9])=[O:6])[C:15]([OH:25])=[O:1]

Participants

5

Reactant 3

HCID 586CreatineHCID 1118Sulfuric AcidHCID 70051-Naphthol

Product 1

HCID 790412-Acetoxypropionic Acid

Reagents, catalysts & solvents 1

HCID 14798Sodium HydroxideSolvent2×

Conditions

Temperature
55 °C

Procedure

Tubes containing assay mix were temperature equilibrated at 30° prior to addition of permeabilized cells to start each reaction. Reactions were stopped by addition of 0.1 mL 6N H2SO4 or 0.1 mL 2.5N NaOH. The tubes were heated for 15 minutes at 55° and then 0.4 mL of 0.5% creatine was added. followed by 0.4 mL of 4% α-naphthol in 2.5 N NaOH, with vigorous mixing after each addition. The tubes were heated again at 55° C. for 5 minutes and centrifuged to remove precipitate. Optical density at 530 nm was then determined. This assay measures acetoin. Acetolactate formed by enzymatic reaction is converted to acetoin by acidification in reactions stopped with H2SO4. Thus the difference in OD530 readings of acid-stopped and base-stopped reactions is a measure of acetolactate formation over background acetoin formation.

Workup

  1. additionTubes containing assay
  2. additionmix
  3. customequilibrated at 30°
  4. additionto addition of permeabilized cells
  5. customeach reaction
  6. temperatureThe tubes were heated for 15 minutes at 55°
  7. additionwith vigorous mixing
  8. additionafter each addition
  9. customcentrifuged to remove precipitate

Sources & evidence

ORD record
ord-693181cd90184f12a972c3eb508349e3
Source dataset
uspto-grants-1986_12
Patent number
US04626505

Reaction notes

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