Glycolic Acid → Formate

Temperature

5 °C

Into a 3 oz. Fischer-Porter glass aerosol reaction vessel was placed a magnetic stirring bar and 10 mL of an aqueous solution containing glycolic acid (0.500M), DEAMPA (0.525M), isobutyric acid (0.100M, HPLC internal standard), and flavin mononucleotide (0.01 mM) at pH 8.3 (adjusted with 50% NaOH), and the solution cooled to 5° C. To the vessel was then added 1.5 g of Hansenula polymorpha transformant G 01 (8.0 IU glycolate oxidase and 38,000 IU catalase) which had been permeabilized by treatment with 0.1% Triton X-1 00/1 freeze-thaw, then the reaction vessel was sealed and the reaction mixture was cooled to 5° C. The vessel was flushed with oxygen by pressurizing to 70 psig and venting to atmospheric pressure five times with stirring, then the vessel was pressurized to 70 psig of oxygen and the mixture stirred at 5° C. Aliquots (0.10 mL) were removed by syringe through a sampling port (without loss of pressure in the vessel) at regular intervals for analysis by HPLC to monitor the progress of the reaction. After 9 h, the HPLC yields of glyoxylate, formate, and oxalate were 98.4%, 0%, and 2.0%, respectively, and no glycolme remained. The remaining permeabilized cell activity of glycolate oxidase and catalase were 63% and 250%, respectively, of their initial values.

1. customFischer-Porter glass aerosol reaction vessel
2. customthe reaction vessel was sealed
3. temperaturethe reaction mixture was cooled to 5° C
4. customThe vessel was flushed with oxygen
5. stirringthe mixture stirred at 5° C
6. customAliquots (0.10 mL) were removed by syringe through a sampling port (without loss of pressure in the vessel) at regular intervals for analysis by HPLC
7. customthe progress of the reaction

ORD record

ord-f5035798c3a24d4f98e166c6071df894

Source dataset

uspto-grants-1996_09

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05559020