Sodium Hydroxide → Acetate

Temperature

10 °C

Into a 14 L fermenter was placed 4.60 L of the 1.10M filtered fermentation broth described in Example 18, then 5.40 L of distilled water was added to produce 10.0 L of an aqueous solution containing 0.51M ammonium lactate, and the mixture cooled to 10° C. To the fermenter was then added 500 g (wet weight) of Pichia pastoris transformant strain GS115-MSP10 (2.52 IU/mL glycolate oxidase and 6,900 IU/mL catalase) which had been permeabilized by treatment with 0.1% benzalkonium chloride ("BARQUAT" MB-50), and the mixture adjusted to pH 7.5 with 50% NaOH and cooled to 10° C. The fermenter was purged with oxygen, then the mixture was stirred at 250 rpm and at 10° C. under 25 psig of oxygen while sparging the mixture with oxygen at 4.5 slpm. 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 0.10M isobutyric acid as internal standard. After 20 hours, the HPLC yields of pyruvate and acetate were 93.0% and 5.8%, respectively, and 1.5% lactate remained. The recovered activities of permeabilized-cell glycolate oxidase and catalase were 260% and 62% of their initial values, respectively.

1. temperaturecooled to 10° C
2. customThe fermenter was purged with oxygen
3. customwhile sparging the mixture with oxygen at 4.5 slpm
4. filtrationfiltering the aliquot
5. filtrationFilter Unit

ORD record

ord-3df7960d02fe4aec8377d2e76632a0d3

Source dataset

uspto-grants-1996_07

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05538875