摘要
目的:获得高活力5′-磷酸二酯酶液,提高核酸RNA酶解效率。方法:采用超滤和盐析技术对从麦芽根浸提液中纯化5′-磷酸二酯酶工艺进行研究,采用单因子试验法优化酶解工艺条件。结果:浸提液依次经过5万Da超滤膜浓缩、40%饱和度硫酸铵盐析、5万Da超滤膜脱盐后,酶活力可达1 500U/ml;第1次超滤膜透过液可作为浸提液循环使用,酶活力是水浸提的1.15倍;第2次超滤膜透过液浓缩5倍后,可回收56.46%硫酸铵,浓缩母液可按1∶2比例循环使用;在底物浓度5.8%、酶用量8%、反应时间2h条件下,RNA酶解率可达95%。结论:初步建立了适合工业化规模的核苷酸生产新工艺。
Objective： Obtained the high activity 5＇ -phosphodiesterase liquid, and raised the enzymatie hydrolysis yield of RNA. Method： The purification teehnology and enzyme hydrolysis condition were studied by uhrafihration, salting out technique and individual faetor experiment. Result： The best teehnology was eoneentrating by 50 000 Da uhrafihration membrane, salting out with 40% saturation （NH4）2SO4 ,and desalination by 50 000 .Da uhrafihration membrane in turn, so that the liquid of 1500U/ml activity could be obtained; the filtrate of first uhrafil/ration eouId be reused as new extraet liquid, and its enzyme activity was 1.15 times than the water extract; the filtrate of second uhrafihration was concentrated to 1/5 volume, then 56.46% （NH4）2SO4 could be harvest, and the mother liquor could be reused in the ratio of 1：2; unde, the condition of 5.8% RNA,8% enzyme eoneention, and 2 hours reaction time, the 95% enzymatic hydrolysis yield of RNA eould be reaehed. Conclusion： The technology of this paper could be used in nueleotide manufacture on industry scale.
出处
《生物技术》
CAS
CSCD
北大核心
2010年第1期62-65,共4页
Biotechnology
基金
农业部淡水鱼类遗传育种和养殖生物学重点开放实验室开放基金项目("新型安全高效水产饲料添加剂--酵母核苷酸的研究开发
"BZ2009)
农业部农业科技成果转化资金项目("高效规范化淡水养殖关键技术示范
"2008GB2C100110)资助
关键词
麦芽根
5'-磷酸二酯酶
核苷酸
超滤
盐析
barley rootlets
5＇ - phosphodiesterase
5 ＇ - nueleotides
uhrafiltration
salting out
作者简介
张一平（1967-），男，工学士，副研究员，研究方向：核苷与核苷酸的研究开发。
通讯作者：Email：heyj@firc．cn。
