海生红藻多管藻(Polysiphonia urceolata)中藻胆体的制备  

Preparation of Phycobilisomes from Red Alga Polysiphonia urceolata

作　　者：侯翠英 台丹丹 赵明日[1] 孙力[1] HOU Cui-ying;TAI Dan-dan;ZHAO Ming-ri;SUN Li(School of Life Sciences,Yantai University,Yantai 264005,China)

机构地区：[1]烟台大学生命科学学院,山东烟台264005

出　　处：《烟台大学学报（自然科学与工程版）》2020年第2期152-162,共11页Journal of Yantai University(Natural Science and Engineering Edition)

基　　金：山东省自然科学基金资助项目(ZR2018LD009).

摘　　要：本研究以海生大型红藻多管藻(Polysiphonia urceolata)为材料,采用亚超速蔗糖密度梯度离心,从2种多管藻藻胆体提取液中分离制备藻胆体.吸收与荧光光谱分析表明:2种样品位于0.2 mol/L处的藻胆体组分,在498 nm R-PE特征吸收光激发时F 670~680/F 580~590都大于3,证明2种藻胆体组分均为完整藻胆体.与Trion X-100增溶后制备的藻胆体(0.2M-Triton-Band)相比,无Trion X-100增溶的藻胆体(0.2M-Band)表现出叶绿素a-蛋白复合物的光吸收和荧光光谱特征,表明0.2M-Band是带有类囊体膜Chl a-蛋白复合物的藻胆体.在蔗糖密度梯度超速离心中,0.2M-Band和0.2M-Triton-Band均被进一步分离成2个结构相对稳定的藻胆体组分:一个是位于0.8 mol/L梯度含量较高、密度较小的藻胆体组分,另一个是位于1.2 mol/L梯度含量较小、密度较大的藻胆体组分.本研究建立的分离制备完整藻胆体的亚超速蔗糖密度梯度离心技术方法,不仅有效提高了藻胆体的制备效率,而且可用于其他海生大型红藻藻胆体的制备.In this study,phycobilisomes(PBSs)are extracted from marine red macroalga Polysiphonia urceolata.The PBSs are isolated and prepared in sucrose density gradient using sub-ultracentrifugation.The absorption and fluorescence spectrum analysis of the prepared PBS fractions show that when the samples are excited by the typical light which R-phycoerythrin absorbs at 498 nm,the ratios of F 670-680 to F 580-590 are higher than 3 for the PBS fractions situated in 0.2 mol/L sucrose band from both of the extracted PBS solutions.This demonstrats that both PBS fractions obtained with(0.2M-Triton-Band)and without(0.2M-Band)Trion X-100 solubilizing are intact phycobilisomes.In contrast to the 0.2M-Triton-Band PBSs,the 0.2M-Band ones display the light absorption and fluorescence properties of chlorophyll a-protein complexes,demonstrating that the 0.2M-Band PBSs carry the Chl a-protein complexes of thylakoid membranes.Using sucrose density gradient ultracentrifugation,both of the 0.2M-Band and 0.2M-Triton-Band PBSs are further isolated as two relatively stable PBS fractions.One fraction with higher content and lower density is situated in 0.8 mol/L sucrose layer,TM-PBS 0.8L/NTM-PBS 0.8L.The other fraction with lower content and higher density is in 1.2 mol/L sucrose layer,TM-PBS 1.2L/NTM-PBS 1.2L.The method of the PBS isolation and preparation using sucrose density gradient sub-ultracentrifugation established in this work increases the efficiency of the PBS preparation.This method can also be used to prepare PBSs from other marine red macroalgae.

关 键 词：多管藻 藻胆体 密度梯度超速离心 吸收光谱 荧光光谱 

分 类 号：Q946[生物学—植物学]