反应详情

温度

5 °C

A 300-mL EZE-Seal stirred autoclave reactor (Autoclave Engineers) was charged with 100 mL of a solution containing glycolic acid (0.75M), ethylenediamine (0.86M, pH 9.0), propionic acid (0.075M, HPLC internal standard), and flavin mononucleotide (0.01 mM), and the solution cooled to 5° C. To the reactor was then added 26 g of frozen (-80° C.) Aspergillus nidulans FT17SYG/OL (29.9 IU glycolate oxidase and 177,000 IU catalase), and the cells were allowed to thaw at 5° C. The resulting mixture was stirred at 400 rpm and 5° C. under 70 psig (483 kPa) of oxygen, while bubbling oxygen through the mixture at 50 mL/min. The reaction was monitored by taking a 100 μL aliquot of the reaction mixture at regular intervals, mixing the aliquot with 300 μL of 0.1N sulfuric acid to quench the reaction, filtering the aliquot and analyzing by HPLC. After 23 hours, the yields of glyoxylic acid, oxalic acid, and formic acid were 95%, 0%, and 0%, respectively, with complete conversion of glycolic acid. The final activities of glycolate oxidase and catalase were 12% and 76% of their initial values.

1. additionTo the reactor was then added 26 g
2. temperatureof frozen (-80° C.)
3. customthaw at 5° C
4. customwhile bubbling oxygen through the mixture at 50 mL/min
5. additionmixing the aliquot with 300 μL of 0.1N sulfuric acid
6. customto quench
7. customthe reaction
8. filtrationfiltering the aliquot and analyzing by HPLC