Purification and photodynamic bioactivity of phycoerythrin and phycocyanin from Porphyra yezoensis Ueda

Phycoerythrin and phycocyanin were purified from Porphyra yezoensis Ueda with their bioactivity determined in this study. Continuous precipitation with ammonium sulfate at different concentrations （10%, 20%, 40%and 50%） increased the purity （A564：A280） of phycoerythrin to 1.49, 3.92 fold of the raw extract （0.38） and the purity （A615：A280） of phycocyanin to 0.70, 3.33 fold of the raw extract （0.21）. Two more times of chromatography with hydroxylapatites finally made the purity of phycoerythrin and phy-cocyanin reach 5.50, 14.47 fold of the raw extract, and 5.10, 24.29 fold of the raw extract, respectviely. The yield of high purity phycoerythrin and phycocyanin were 0.21%and 0.09%of dried P. yezoensis blade, respectively. The photodynamic cytotoxic ex-periment showed that both phycoerythrin and phycocyanin inhibited the growth of liver tumor cells significantly. It was found that 250 mg L-1 purified phycoerythrin and phycocyanin inhibited the growth of hepatocellular carcinoma cells 24 h after laser-irradiation by 80%and 59%, respectively, and 100 mg L-1 purified phycoerythrin and phycocyanin induced the apoptosis of 31.54%and 32.54%of the cells, respectively, 8 h after photodynamic therapy. Oue findings demonstrated that P. yezoensis can serve as photosensitizer （phycoerythrin and phycocyanin） producer.     

隐藻藻蓝蛋白与类囊体膜的动态结合模型

隐藻藻胆蛋白在类囊体腔中的存在状态始终存在争议,为了深入了解这一问题,作者以含有藻蓝蛋白(PC645)的蓝隐藻(Chroomonas placoidea)活体细胞及充分洗涤过的类囊体膜为材料,测定其室温吸收光谱、荧光谱以及77K低温荧光光谱,并根据光谱数据分析了PC在类囊体腔中的存在状态。结果显示,蓝隐藻的类囊体膜上始终紧密结合着一定量的PC,并且很可能是以PC的β亚基与膜相接触,证实藻胆蛋白在类囊体腔中不是完全游离的,与膜的接触也并非完全无倾向性排布;PC既可将激发能传递给光系统(PS)II,但也可传递给PSI,相较于叶绿素(Chl)a和Chl c,在PSI长波荧光中PC的贡献更明显,而Chl c吸收的光能,更倾向于传递给PSII或者短波长Chl a;蓝隐藻细胞中表现出超比例的PC荧光发射,说明除了结合在膜上的PC外,有一部分PC是游离的,其存在对于隐藻捕光功能来说可能是有冗余的;游离的PC可与类囊体膜重新接触而恢复能量传递功能。根据实验结果以及前人的报道,提出了隐藻藻胆蛋白与类囊体膜的结合是动态的观点,并给出了相应的模型。     

Formation and functional significance of storage products in cyanobacteria

The most common storage products of cyanobacteria are polyphosphate as a phosphorus storage compound, cyanophycin or phycobilin protein pigment as nitrogen storage products, and glycogen as a storage product of both carbon and energy. Nutrient uptake kinetics are regulated by the storage pools, and the patterns of regulation have a feedback effect on the amount of accumulated nutrient in the cells. Besides having a storage function the nutrient storage products are likely to act as metabolic sinks during conditions of energy stress. Regulation of storage products is especially strict in light‐limited cultures. By increasing the rate of polysaccharide formation during growth with short photoperiods, cyanobacteria are able to sustain relatively high growth rates. This effect is enhanced by keeping respiratory losses very low.     

富营养化水体浮游植物遥感监测研究进展

随着湖泊流域经济的快速发展,蓝藻水华频繁暴发的现象越来越严重,水体富营养化已经成为国内外重大环境问题.浮游植物是水体的初级生产者,是衡量水体富营养化程度的主要指标之一,遥感技术则是探测水体浮游植物时空分布的重要手段.在收集整理近千篇国内外水体浮游植物遥感研究论文的基础上,从卫星数据源、研究内容及研究方法等角度,总结了遥感技术在富营养化湖泊浮游植物监测应用的历史进展、研究重点及发展趋势.研究表明,现有的富营养化水体浮游植物遥感研究,以湖泊蓝藻水华问题为切入点,研究视角由水体表层(藻华面积、色素浓度)转至水下三维(藻总量),研究方法从定性识别转向定量反演,研究内容从监测蓝藻水华推进到探测不同类群蓝藻,逐渐形成了以应用为导向,"MODIS/VIIRS大中型湖泊日常监测—GF/Sentinel2小型湖泊针对性监测—无人机应急监测"的浮游植物遥感综合监测体系.上述研究梳理了富营养化水体浮游植物遥感监测湖泊水环境学科的发展动向,以期为从事蓝藻水华生态灾害监测和预警人员提供重要的技术支撑和理论参考.     

Ship-of-opportunity based phycocyanin fluorescence monitoring of the filamentous cyanobacteria bloom dynamics in the Baltic Sea

Distribution of cyanobacteria cannot be evaluated using chlorophyll a (Chla) in vivo fluorescence, as most of their Chla is located in non-fluorescing photosystem I. Phycobilin fluorescence, in turn, is noted as a useful tool in the detection of cyanobacterial blooms. We applied phycocyanin (PC) fluorometer in the monitoring of the filamentous cyanobacterial bloom in the Baltic Sea. For the bloom forming filamentous cyanobacteria Aphanizomenon flos-aquae and Nodularia spumigena, PC fluorescence maximum was identified using the excitation–emission fluorescence matrix. Consequently, the optical setup of our instrument was noted to be appropriate for the detection of PC, and with minor or no interference from Chla and phycoerythrin fluorescence, respectively.During summer 2005, the instrument was installed on a ferryboat commuting between Helsinki (Finland) and Travemünde (Germany), and data were collected during 32 transects providing altogether 200 000 fluorescence records. PC in vivo fluorescence was compared with Chla in vivo fluorescence and turbidity measured simultaneously, and with Chla concentration and biomass of the bloom forming filamentous cyanobacteria determined from discrete water samples.PC fluorescence showed a linear relation to the biomass of the bloom forming filamentous cyanobacteria, and the other sources of PC fluorescence are considered minor in the open Baltic Sea. Estimated by PC fluorescence, cyanobacterial bloom initiated late June at the Northern Baltic Proper, rapidly extended to the central Baltic Proper and the Gulf of Finland, and peaked in the mid-July with values up to 10 mg l⁻¹ (fresh weight). In late July, bloom vanished in most areas.During single transects, or for the whole summer, the variability in Chla concentrations was explained more by PC fluorescence than by Chla fluorescence. Thus, filamentous cyanobacteria dominated the overall variability in phytoplankton biomass. Consequently, we show that during the cyanobacterial blooms, the estimation of Chla concentration using only Chla in vivo fluorescence is not applicable, but PC in vivo fluorescence is required as a predictor as well.     

钝顶螺旋藻藻蓝蛋白提取纯化新工艺

对钝顶螺旋藻（Spirulina platensis）中藻蓝蛋白的提取和纯化方法进行了改进。用磷酸盐缓冲液循环冻融联合超声波破碎法,50％硫酸铵沉淀获得藻蓝蛋白（phycocyanin,PC）,提取率达到13．1%。粗蛋白提取液再经过两次羟基磷灰石柱（HA）层析和sephacryl-HR-200凝胶层析对其进行纯化,纯度达到4．71％。纯化后的PC在12％十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）中得到两条带,分别是α和β两个亚基,分子质量分别为21．4ku和17．0ku。结果表明,通过上述分离纯化过程得到了较高提取率和电泳纯度的藻蓝蛋白。     

Temperature Influences on the Expression of GFP Promoted by the Upstream Sequence of cpcB from Arthrospiraplatensis

In order to investigate the regulation mechanism of the phycocyanin gene,a series of functional analyses of the upstream sequence of cpcB gene from Arthrospira platensis were conducted in E. coli with green fluorescent protein encoding gene(gfp) as the reporter. Results showed that the gfp gene could express at a high level under the promotion of the upstream sequence,suggesting the existence of some strong promoter elements in it. The expression of GFP was influenced by temperature. Higher temperature led to higher expression level. The bioinformatics analyses followed by mutation analyses on the secondary structure of translation ini-tiation region(TIR) revealed that RNA thermosensor might account for the temperature regulation.     

江蓠属藻胆蛋白的研究──Ⅱ.龙须菜藻胆蛋白光谱特性和荧光动力学研究

本文以野生型和突变型龙须菜为材料,测定了完整藻体的室温吸收光谱、77K低温荧光发射光谱和叶绿素诱导荧光动力学曲线.野生型(Wt)和黄色突变体(Ye100)的室温吸收光谱有435、498、540、566、618、640和680nm7个吸收峰;绿色突变体Gr180和Gr184则只有除540nm以外的6个吸收峰.3种突变体的藻红蛋白含量均比野生型少,两种绿色突变体的藻蓝蛋白含量相对有所增加,别藻蓝蛋白含量变化不明显.77K低温荧光发射发谱(激发光分别为436、540和580nm)的结果表明:在发射峰的波长上,4种材料没有明显的区别,但Wt中PE吸收的光能传递效率比较低.叶绿素诱导荧光动力学曲线显示Wt的光系统Ⅱ活性、光系统Ⅱ原初光能转化效率及光合碳代谢活性都高于突变体.     

藻蓝蛋白皮肤毒性实验研究

藻蓝蛋白是蓝藻中的一种主要活性成分,研究发现藻蓝蛋白有多种功能,可以清除氧自由基,且含有多种氨基酸能够抗衰老、抗过敏,为对藻蓝蛋白用于化妆品行业的可行性进行探究,本文采用新西兰白兔作为动物模型,通过光毒性、急性经皮毒性以及皮肤变态反应实验,验证其对皮肤的毒性作用。结果显示,藻蓝蛋白对白兔没有明显的毒性作用,也未引起皮肤过敏反应,为其在美容化妆方面的应用提供了参考。     

太湖水华蓝藻底泥中复苏和水柱中生长的比较

为了研究太湖底泥中蓝藻的复苏和水柱中的生长,作者自行设计了原位藻类复苏收集器和生长培养器并安放在太湖梅梁湾监测蓝藻复苏释放和生长.实验在藻类的复苏期(2005年3-6月)进行．结果表明在复苏期太湖蓝藻的复苏呈现波动性增加,4月达到最大量．水柱中蓝藻同步的比生长率和生长量也呈现波动性上升,并在5月初达到最大值．浮游动物的存在并没有对蓝藻的生长造成显著的影响．对底泥蓝藻释放和水柱生长进行比较,底泥释放的蓝藻只占藻类生长量的很小一部分(<2．5％)．所以,水柱中蓝藻的生长对其优势的确立和水华的形成具有重要的作用,今后的研究将主要集中在水柱中蓝藻的动态变化