Glycolic Acid → Glyoxylic Acid

Temperature

5 °C

A 300-mL EZE-Seal stirred autoclave reactor equipped with Dispersimax Impeller (Autoclave Engineers) was charged with 100 mL of a solution containing glycolic acid (0.750M), ethylenediamine (0.863M), isobutyric acid (0.100M, HPLC internal standard), and flavin mononucleotide (0.01 mM), at pH 9.25, and the solution cooled to 5° C. To the reactor was then added 2.0 g of Pichia pastoris transformant strain GS115-MSP10 (2763 IU glycolate oxidase and 494,000 IU catalase) which had been permeabilized by treatment with 0.1% Triton X-100/6 freeze-thaws, and the reactor purged with oxygen. The mixture was then stirred at 1000 rpm, which bubbled oxygen through the mixture via the action of the turbine impeller, and at 5° C. under 120 psig of oxygen. The reaction was monitored by taking a 0.4 mL aliquot of the reaction mixture at regular intervals, filtering the aliquot using a Millipore Ultrafree-MC 10,000 NMWL Filter Unit, and analyzing the filtrate by HPLC. After 0.75 h, the yields of glyoxylic acid, oxalic acid, and formic acid were 99.1%, 0.3%, and 0%, respectively, with 0.6% glycolic acid remaining. The recovered activities of permeabilized-cell glycolate oxidase and catalase were 104% and 105% of their initial values, respectively.

1. customA 300-mL EZE-Seal stirred autoclave reactor equipped with Dispersimax Impeller (Autoclave Engineers)
2. customthe reactor purged with oxygen
3. custombubbled oxygen through the mixture via the action of the turbine impeller
4. customat 5° C.
5. filtrationfiltering the aliquot
6. filtrationFilter Unit

ORD record

ord-a515a73e91574aa28e99890f71022fdf

Source dataset

uspto-grants-1998_11

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05834262