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 9.3, and the solution cooled to 5° C. To the reactor was then added 15.0 g of Hansenula polymorpha transformant strain GO1 (262 IU glycolate oxidase and 1.135×106IU catalase) which had been permeabilized by treatment with 0.1% Triton X-100/1 freeze-thaw, 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 250 psig of oxygen. The reaction was monitored by taking a 0.40 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 1.0 h, the yields of glyoxylic acid, oxalic acid, and formic acid were 96.9%, 0.3%, and 0%, respectively, with no remaining glycolic acid. The recovered activities of permeabilized-cell glycolate oxidase and catalase were 98% and 124% 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-ebad4b4b3af04080843e752592e4f0b2

Source dataset

uspto-grants-1998_11

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05834262