鱿鱼肝脏中大分子核酸的组成及分离纯化条件研究

A study on composing and separating conditions of large molecular nucleic acid from squid liver

目的 转化鱼类脏器废弃物为高值生物大分子 ,利用活性载体处理鱿鱼肝脏 ,获得有效成分核酸。方法 在分析鱿鱼肝脏中核酸类物质含量的基础上 ,研究活性载体处理鱿鱼肝脏的最适浓度、最适培养时间、最适温度、最适光照条件及氧含量等作用条件 ,得到核酸分离纯化的有效方案。进一步 ,通过离子交换色谱法研究了核酸精化的分离条件。结果 从鱿鱼肝脏中分离核酸的最适条件 :10 %的活性载体与鱿鱼肝脏废弃物混合 ,37℃、培养 36h、暗光、无氧。色谱分析条件为 :固定相为DEAE 纤维素 ,色谱柱 :8cm× 1cm ,流速 :1ml/min ,2 6 0nm紫外检测 ,浓度梯度NaCl (pH 7 4Tris Cl缓冲液 )溶液洗脱。 结论 产物经透析、真空冻干 ,可获得纯度达 87%～ 90 %的核酸纯品 ,可用于医药及食品工业。

Objective Nucleic acid in squid liver was extracted and separated to produce raw material used for medical industry and food additive.Methods Nucleic acid in squid liver was cultured with active carriers.The optimal conditions of carriers,including their concentration,culture time,culture temperature,with or without illumination,and oxygen content in the culture.Results An efficient separation condition of nucleic acid was found out as:the liver-homogenate culture made by blending 10% active carrier and squid liver is the best medium.The best incubation condition is 37 ℃,36 hours,dark and anaerobic.Nucleic acid was eluted with Tris-Cl (pH 7.4) containing 0.6 mol/L NaCl and selected with ion-exchange chromatography.Conclusion After dialyzed and lyophilized,87%～90% pure product was obtainable.