Benzalkonium Chloride → Acetate

Participants

Reaction Conditions

Reaction Steps

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 (1.06M), 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 (5.54 IU/mL glycolate oxidase and 14,400 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, filtering 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 5 hours, the HPLC yields of pyruvate and acetate were 95.7% and 2.5%, respectively, and 1.9% lactate remained. The remaining permeabilized cell activity of glycolate oxidase and catalase were 116% and 107% of their initial values, respectively.

Provenance & Evidence