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。
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.
出处
《食品工业科技》
CAS
CSCD
北大核心
2014年第22期304-307,316,共5页
Science and Technology of Food Industry
基金
国家自然科学基金项目(21276001
31171705)
绍兴市科技计划项目(2013A23002)
关键词
D-阿洛糖
生物转化
等电点沉淀
纯化
D-allose
biotransformation
isoelectric precipitation
purification
