红胞藻藻红蛋白的分离纯化及稳定性研究  

Isolation,Purification and Stability of Phycoerythrin from Rhodomonas sp.

作　　者：董硕 蒲洋 聂岩 薛欣然 袁新雨 李文军[3] 唐志红[1] DONG Shuo;PU Yang;NIE Yan;XUE Xinran;YUAN Xinyu;LI Wenjun;TANG Zhihong(College of Life Science,Yantai University,Yantai 264005,China;School of Agriculture,Ludong University,Yantai 264000,China;Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences,Yantai 264003,China)

机构地区：[1]烟台大学生命科学学院,山东烟台264005 [2]鲁东大学农学院,山东烟台264000 [3]中国科学院烟台海岸带研究所,山东烟台264003

出　　处：《食品科技》2023年第4期230-236,共7页Food Science and Technology

基　　金：山东省自然科学基金联合基金项目(ZR2021LLZ003);烟台大学研究生科研创新基金项目(GGIFYTU2224)。

摘　　要：文章旨在从红胞藻中分离纯化得到试剂级的藻红蛋白,并对其相关性质进行研究。以红胞藻为原料,采用反复冻融、硫酸铵盐析和疏水层析的方法分离纯化藻红蛋白,并对其稳定性进行研究。结果表明,红胞藻经过4次反复冻融后藻红蛋白溶出率最高,两步硫酸铵分级沉淀可使藻红蛋白纯度(A_(545)/A_(280))提高到2.45,进一步采用Butyl-S-QZT 6FF疏水层析纯化后藻红蛋白纯度(A_(545)/A_(280))达到试剂级,为6.63,回收率为48.9%。纯化的藻红蛋白在545 nm处有最大特征吸收峰,荧光激发峰位于541 nm和553 nm处,发射峰位于587.5 nm处,二级结构主要以α-螺旋的形式存在。藻红蛋白含有3个亚基,分子质量分别为9、10 ku和20 ku,在30~50℃、pH6~8条件下能保持相对稳定。Cu^(2+)对藻红蛋白的光谱学性质有一定的影响,在Ca^(2+)、Mn^(2+)、Zn^(2+)、Fe^(2+)、Fe^(3+)等离子存在时,藻红蛋白光谱学性质没有发生明显变化。结论:采用硫酸铵分级沉淀结合一步疏水层析获得了试剂级的藻红蛋白,得率高,工艺简便。研究结果为红胞藻藻红蛋白的大规模生产提供了参考。The purpose of this paper is to isolate and purify the reagent grade phycoerythrin from Rhodomonas sp.,and investigate its related properties.In this study,the phycoerythrin was separated and purified from Rhodomonas sp.by repeated freezing and thawing,ammonium sulfate salting out and hydrophobic chromatography,and its stability was studied.The results showed that the dissolution rate of phycoerythrin from Rhodomonas sp.was the highest after four repeated freeze-thaw cycles.The purity of phycoerythrin(A_(545)/A_(280))could be increased to 2.45 by two-step ammonium sulfate fractional precipitation.After further using Butyl-S-QzT 6FF hydrophobic chromatography,the purity of phycoerythrin(A_(545)/A_(280)=6.63)reached reagent level,and the recovery rate was 48.9%.The purified phycoerythrin has the largest characteristic absorption peak at 545 nm,the fluorescence excitation peak at 541 nm and 553 nm,and the emission peak at 587.5 nm.The secondary structure is mainly composed of α-helix.Phycoerythrin contains three subunits,with molecular weights of 9 ku,10 ku and 20 ku respectively.It can remain relatively stable at 30~50℃and pH 6~8.Cu^(2+)has a certain effect on the spectral properties of phycoerythrin.In the presence of Ca^(2+),Mn^(2+),Zn^(2+),Fe^(2+),Fe^(3+),the spectral properties of phycoerythrin have not changed significantly.Conclusion:Reagent grade phycoerythrin was obtained by ammonium sulfate fractionation precipitation and one-step hydrophobic chromatography,with high yield and simple process.This study provides a basis for the large-scale production of phycoerythrin from Rhodomonas sp.

关 键 词：红胞藻 藻红蛋白 分离纯化 稳定性 

分 类 号：TS201.2[轻工技术与工程—食品科学]