Benzalkonium Chloride → Acetate

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 sodium lactate (0.50M), at pH 7.5, and the solution cooled to 5° C. To the reactor was then added 5.0 g (wet weight) of Pichia pastoris transformant strain GS115-MSP 10 (5.19 IU/mL glycolate oxidase and 10,800 IU/mL catalase ) which had been permeabilized by treatment with 0.1% benzalkonium chloride ("BARQUAT" MB-50), and the reactor purged with oxygen. The mixture was then stirred at 750 rpm, which bubbled oxygen through the mixture via the action of the turbine impeller, and at 5° C. under 70 psig of oxygen. The reaction was monitored by taking a 0.40 mL aliquot of the reaction mixture at regular intervals, faltering the aliquot using a Millipore Ultrafree-MC 10,000 NMWL Filter Unit, and analyzing the filtrate by HPLC using 0.20M isobutyric acid added to the sample as internal standard. After 2 hours, the HPLC yields of pyruvate and acetate were 99.3% and 0.5%, respectively, and 2.4% lactate remained. The remaining permeabilized cell activity of glycolate oxidase and catalase were 75% and 81% 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. filtrationFilter Unit
6. additionadded to the sample as internal standard

ORD record

ord-6f68805e780a4cc9b100cb522b22d274

Source dataset

uspto-grants-1996_07

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05538875