生物转化法产D-阿洛糖的分离纯化

Separation and purification of D-allose produced by biotransformation

摘要: 利用重组菌E.coli BL21/p ET-22b-tl-rpib,将原料D-阿洛酮糖生物转化为具有抑癌效果的稀有糖D-阿洛糖。经液相色谱和串联质谱联用技术（LC-MS/MS）鉴定,反应混合液中25%的D-阿洛酮糖转化成了D-阿洛糖。等电聚焦法测定了重组蛋白的等电点为6.3,并成功运用到产物混合液的去蛋白沉淀中。经过预处理的混合糖液,通过DTFCa2+型离子交换树脂实现了分离纯化。最佳分离条件为:柱温60℃,进样量4m L,流速1m L/min。

Abstract: The anticancer rare sugar D-allose was converted from D-psicose by E. coli BL21/ p ET-22b-tl-rpib.The LC-MS/MS results showed that 25% of D-psicose was converted to D-allose in the reaction product mixture. The p I of the recombinant protein was 6.3 determined by isoelectrofocusing and was successfully applied in isoelectric precipitation. The methods of separation and purification of D-allose was established according to DTF-Ca2 +cation exchange chromatography with a flow rate of 1m L/min, column temperature at60℃ and the loading volume of 4m L.