Spectral Analyses During the Isolation Course of Spirulina Platensis Phycobilisomes Using Low-Speed Centrifugation
FAN Shou-jin1,ZHANG Xi-ying2*,ZHANG Yu-zhong2
1. School of Life Sciences, Shandong Normal University, Ji’nan 250014, China 2. State Key Laboratory of Microbial Technology, Shandong University, Ji’nan 250100, China
Abstract:The phycobilisomes were isolated from Spirulina platensis using low-speed centrifugation. The crude phycobilisomes solution extracted by Triton X-100 was centrifugated (13 000 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.
Key words:Spirulina platensis;Phycobilisomes;Absorption and fluorescence spectra;Chromatography
樊守金1,张熙颖2*,张玉忠2. 钝顶螺旋藻藻胆体低速离心制备过程的光谱表征[J]. 光谱学与光谱分析, 2008, 28(09): 2119-2121.
FAN Shou-jin1,ZHANG Xi-ying2*,ZHANG Yu-zhong2. Spectral Analyses During the Isolation Course of Spirulina Platensis Phycobilisomes Using Low-Speed Centrifugation. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2008, 28(09): 2119-2121.
[1] ZHANG Xi-ying, LIU Lu-ning, CHE Xiu-lan, et al(张熙颖,刘鲁宁,陈秀兰,等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2004, 24(10): 1224. [2] MacColl R. Journal of Structural Biology, 1998, 124(2): 311. [3] Liu L N, Chen X L, Zhang Y Z, et al.Biochimica et Biophysica Acta.Bioenergetics, 2005, 1708(2): 133. [4] YAN Shi-gan, CHEN Xiu-lan, ZHANG Xi-ying, et al(颜世敢, 陈秀兰, 张熙颖, 等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2008, 28(5): 1115. [5] Cheng Y, Prussin C. Journal of Allergy and Clinical Immunology, 2004, 113(2): 209. [6] Fujihira M. In Honda K(Editor). Photochemical Processes in Organized Molecular Systems. Elsevier Science Publishers, Amsterdam, The Netherlands, 1991. 463. [7] Chen C, Zhang Y Z, Chen X L, et al. Acta Biochimica et Biophysica Sinica, 2003, 35(10): 952. [8] Gantt E, Lipschultz C A, Grabowski J, et al. Plant Physiology, 1979, 63( 6): 615. [9] Zhang Y Z, Zhou B C, Tseng C K. Marine Sciences, 1997, 21(6): 39. [10] Rosinski J, Hainfeld J F, Rigbi M, et al. Annals of Botany, 1981, 47(1): 1. [11] Siegelman H W, Kycia J H. Plant Physiology, 1982,70(3): 887. [12] ZHOU Zhi-gang, YIN Chang-song(周志刚,尹长松). Journal of Shanghai Fisheries University(上海水产大学学报), 2002, 11(3): 208.