反応式
[N:1]1[C:9]([NH2:10])=[C:8]2[C:4]([N:5]=[CH:6][NH:7]2)=[N:3][CH:2]=1.C1C(=O)NC(=O)N([C@@H:19]2[O:23][C@H:22]([CH2:24][OH:25])[C@@H:21]([OH:26])[C@@H:20]2[OH:27])C=1>P([O-])([O-])([O-])=O>[CH:2]1[N:3]=[C:4]2[N:5]([C@@H:19]3[O:23][C@H:22]([CH2:24][OH:25])[C@@H:21]([OH:26])[C@@H:20]3[OH:27])[CH:6]=[N:7][C:8]2=[C:9]([NH2:10])[N:1]=1参加物
反応条件
- 温度
- 60 °C
実験手順
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.
後処理
- custom50 kg of wet cell paste obtained
- additiondiluted to approximately 3000 liters with hot H20
- customcollecting approximately 5000 liters of ultrafiltrate
- customThe bioconversion yield
- additionThe residue containing the cell paste
- customis used for a subsequent reaction
- concentrationThe ultrafiltrate was concentrated to approximately 1000 liters
- temperaturecooled
- customto collect the precipitate
出典と根拠
- ORDレコード
- ord-92a4ac3f6eee4a5fa9b7b10aef984213
- 出典データセット
- uspto-grants-2010_08
- 特許番号
- US07781189B2
関連ノート
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