反应详情

温度

60 °C

50 kg of wet cell paste obtained by fermenting E. aerogenes were resuspended in approximately 200 liters of 30 mM phosphate buffer at pH 7 and mixed with 800 liters of phosphate buffer in which had been dissolved at elevated temperature 5.4 kg of adenine (final concentration 40 mM) and 8.9 kg of Ara-U (final concentration 40 mM). The mixture was maintained at 60° C., with agitation, for 20 hours, diluted to approximately 3000 liters with hot H20 and subjected to diafiltration on a membrane, collecting approximately 5000 liters of ultrafiltrate. The bioconversion yield determined by RP-HPLC was approximately 55%. The residue containing the cell paste is used for a subsequent reaction. The ultrafiltrate was concentrated to approximately 1000 liters and cooled to collect the precipitate constituted by Ara-A contaminated with non-reacted adenine (approximately 30 g of adenine per 100 g of Ara-A). 5 kg of Ara-A (total yield approximately 46%) with a degree of purity higher than 99.5% were finally obtained after crystallisation with H2O.

1. custom50 kg of wet cell paste obtained
2. additiondiluted to approximately 3000 liters with hot H20
3. customcollecting approximately 5000 liters of ultrafiltrate
4. customThe bioconversion yield
5. additionThe residue containing the cell paste
6. customis used for a subsequent reaction
7. concentrationThe ultrafiltrate was concentrated to approximately 1000 liters
8. temperaturecooled
9. customto collect the precipitate