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), aminomethylphosphonic acid (0.375M), 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 0.75 g of a Pichia pastoris transformant GS115-MSP10 (13.2 IU glycolate oxidase and 21,200 IU catalase) which had been permeabilized by treatment with 0.1% "TRITON" X-100/1 freeze-thaw. The pH of the resulting mixture was re-adjusted to 8.3 with 50% NaOH, 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 16 hours, the HPLC yields of glyoxylate, formate, and oxalate were 30.5%, 59.2%, and 10.7%, respectively, and 0.8% glycolate remained.

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-ffed8b0e73d94c87b3d5c9f60ea5eefb

Source dataset

uspto-grants-1996_07

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05541094