罗丹明B标记树舌多糖的条件  

Research on the Conditions of Rhodamine B in Labeling Ganoderma applanatum Polysaccharides

作　　者：张中北[1] 杨颜铭[1] 王丽娜[1] 吴波[1] 王宇[1] 宋慧[1,2] 

机构地区：[1]吉林农业大学生命科学学院,长春130118 [2]吉林农业大学教育部食药用菌工程研究中心,长春130118

出　　处：《菌物研究》2016年第4期233-238,共6页Journal of Fungal Research

基　　金：吉林省科技发展计划项目(210-00115);世行贷款农产品质量安全项目(2011-Y18)

摘　　要：通过对树舌多糖进行荧光标记,解决多糖本身缺少易于检测发光基团的难题,为进一步研究多糖的生物学活性提供一种新型可靠的辅助手段。利用树舌多糖还原性末端与罗丹明B活泼的氨基发生反应,形成树舌多糖-罗丹明B复合物,再经流水透析和分子筛进一步纯化得到罗丹明B标记的树舌多糖。对树舌多糖与罗丹明B的反应时间、反应温度和p H进行了单因素考察。试验结果证明:罗丹明B可以有效的标记树舌多糖,在反应时间24 h、p H 5、温度25℃的条件下标记效果最好。在单因素试验的基础上对罗丹明B标记树舌多糖的条件进行响应面优化,最终得到罗丹明B标记树舌多糖的最优条件为:反应时间23.10 h、温度25.83℃、p H 5.23,预测标记后样品在552 nm波长处的最大OD值为0.135。This work was to solve the issue that polysaccharides lack detectable luminescent groups, and provid a novel and reliable approach to study the biological activity of polysaccharides via fluorescently labeling the Gauoderma applanatum polysaccharides. The G. applanatum polysac- charides labeled by Rhodamine B were prepared through the reaction between the reducing end of G. applanatum plolysaccharide and the active amino group of Rhodamine B. The G. applanatum polysaccharide-Rhodamine B conjugates were further purified by water dialysis and molecular sieves. The optimal conditions of reacting interval, pH, and temperature on Rhodamine B labeling were studied. The results indicated that Rhodamine B was able to label G. applanatum polysaccharides effectively under following conditions ： reacting interval of 24 h, pH 5, and the temperature of 25℃. By optimizing the labeling conditions .based on the results of single factor experiment, best conditions was obtained as followings： reacting interval of 23. 10 h, the temperature of 25.83℃, and pH 5.23. We estimated the maximum OD value of the labeled sample was 0.135 at a wavelength of 552 nm.

关 键 词：树舌多糖 罗丹明B 荧光标记 

分 类 号：Q539[生物学—生物化学]