Fan Shou-jin, Zhang Xi-ying, Zhang Yu-zhong
School of Life Sciences, Shandong Normal University, Ji'nan 250014, China.
Guang Pu Xue Yu Guang Pu Fen Xi. 2008 Sep;28(9):2119-21.
The phycobilisomes were isolated from Spirulina platensis using low-speed centrifugation. The crude phycobilisomes solution extracted by Triton X-100 was centrifugated (13000 rpm) four times. The centrifugated phycobilisomes solution was spectrally analyzed using absorption spectrum each time. The absorption spectrum showed that the ultraviolet absorption maximum of the phycobilisomes solution was still at 263 nm, and also exhibited the characteristic chlorophyllous absorption in the rang of 400-450 nm after the fourth centrifugation. This indicated that there existed small quantities of Triton X-100 and chlorophyll in the centrifugated phycobilisomes solution. But the ultraviolet absorption maximum was red-shifted to 277 nm and the chlorophyllous absorption was not observed in the absorption spectrum of the phycobilisomes solution obtained by high concentration salt precipitation, which meant that the method of high concentration salt precipitation could effectively remove Triton X-100 and chlorophyll from the phycobilisomes solution. The precipitated phycobilisomes of Spirulina platensis were further purified by using Sepharose CL-6B column chromatography. The fluorescence emission maximum of the purified phycobilisomes at room temperature was at 680 nm, which indicated that the purified phycobilisomes were intact.
采用低速离心法从钝顶螺旋藻中分离藻胆体。用Triton X - 100提取的藻胆体粗溶液进行4次离心(13000转/分钟)。每次对离心后的藻胆体溶液进行吸收光谱分析。吸收光谱表明,藻胆体溶液的紫外吸收最大值仍在263 nm处,且在第4次离心后,在400 - 450 nm范围内也呈现出叶绿素特征吸收。这表明离心后的藻胆体溶液中存在少量Triton X - 100和叶绿素。但在高浓度盐沉淀法得到的藻胆体溶液的吸收光谱中,紫外吸收最大值红移至277 nm,且未观察到叶绿素吸收,这意味着高浓度盐沉淀法可有效去除藻胆体溶液中的Triton X - 100和叶绿素。采用Sepharose CL - 6B柱色谱法对钝顶螺旋藻沉淀的藻胆体进行进一步纯化。纯化后的藻胆体在室温下的荧光发射最大值为680 nm,表明纯化后的藻胆体是完整的。