Glutaraldehyde → Penicillin G

Participants

Reaction Steps

Escherichia coli ATCC 9637 is used for the production of penicillin acylase. The nutrient solution consists of 2% of corn steep liquor plus 1% of ammonium phenylacetate, and the pH value is 6.5. 200 ml of nutrient solution are inoculated with 1 ml of an Escherichia coli suspension in a 1 liter conical flask. The fermentation proceeds at 31° C. for 20 hours on a rotary shaking machine at 290 rpm. 20 ml of 4% strength tannin solution are first added dropwise to 500 ml of cell suspension, and the suspension is then flocculated completely with the aid of 100 ml of BETZ/glutaraldehyde mixture (60 g/liter of BETZ, 2.2% v/v of glutaraldehyde, pH 9.0). When flocculation is complete, a 50% excess of BETZ/glutaraldehyde are immediately added. Half of the batch is immediately replaced with solid (I) to a final concentration of 0.1M, in addition to the excess of BETZ/glutaraldehyde, and the other half is worked up without (I). After the cell mass has been separated off, washed and extruded, it is dried overnight at 40° to 45° C. and then ground and sieved. Without (I), splitting of penicillin G in 6-aminopenicillanic acid and phenylacetic acid gave a specific activity of 2.17 pen G units per gram of dry catalyst, corresponding to 8.36 NIPAB units/g.

Provenance & Evidence