钝顶螺旋藻突变株FBL细胞超微结构

利用透射电镜技术观察钝顶螺旋藻出发株和突变株FBL的细胞超微结构。观察结果表明L出发株和突变株均为多细胞丝状体,细胞间横隔膜清晰,细胞壁均由四层结构组成,细胞质膜内陷形成类囊体,类囊体由双层膜堆积而成,膜上附着藻胆体,类囊体与细胞壁呈垂直方向排列,细胞质内包含有充气液泡等细胞器。与出发株相比,突变株细胞壁表面较光滑,四层结构电子密度较深;类囊体膜增多、变发达;羧化体数量增多;横隔膜收缢明显。     

富硒螺旋藻中硒别藻蓝蛋白的纯化及其特性

从富硒螺旋藻(Se richSpirulina platensis,Se-SP)中分离纯化高纯度的含硒别藻蓝蛋白(Se-containingallophycocyanin,Se-APC)并观察其生化特性。羟基磷灰石和DEAE-52柱层析方法结合制备电泳技术纯化Se-APC;光谱扫描、Native-PAGE、SDS-PAGE和IEF方法鉴定Se-APC生化特性;2,3-DAN荧光光度法检测蛋白质中Se含量。结果发现3种高纯度Se-APC的光谱特征分别与APCI、APCII、APCIII吻合;电泳鉴定它们可能都是(αβ)3,α、β亚基分别为18.3和15.7 kDa,其pI值分别为:4.76、4.85和5.02;3种Se-APC中Se含量分别为316、273和408μg/g,Se-APC经0.5mol/L NaSCN解聚和β-巯基乙醇变性处理后,蛋白质中Se含量依次减低并趋于稳定。结果提示Se-SP中APC可结合Se,APC中Se含量与其分子聚态有关,亚基中含Se量稳定,可能是以共价键方式结合,Se-APC生物活性及硒在蛋白质中的结合位点值得深入研究。     

Cadmium biosorption on Spirulina platensis biomass

Dry biomass of Spirulina platensis re-hydrated for 48 h was employed as a biosorbent in tests of cadmium(II) removal from water. Various concentrations of biomass (from 1 to 4 g l⁻¹) and metal (from 100 to 800 mg l⁻¹) were tested. Low biomass levels (X_o  2 g l⁻¹) ensured metal removal up to 98% only at Cd₀= 100 and 200 mg l⁻¹, while X_o  2.0 g l⁻¹ were needed at Cd₀ = 400 mg l⁻¹ to achieve satisfactory results. Whereas X_o = 4.0 g l⁻¹ was effective to remove up to Cd₀ = 500 mg l⁻¹, a further increase in metal concentration (Cd₀ = 600 and 800 mg l⁻¹) led to progressive worsening of the system performance. At a given biomass levels, the kinetics of the process was better at low Cd²⁺ concentrations, while, raising the adsorbent level from 1.0 to 2.0 g l⁻¹ and then to 4.0 g l⁻¹, the rate constant of biosorption increased by about one order of magnitude in both cases and the adsorption capacity of the system progressively decreased from 357 to 149 mg g⁻¹.     

Production and regeneration of spheroplasts from the cyanobacterium Spirulina platensis

Abstract The development of a micro-method for the production and regeneration of spheroplasts starting from S. platensis trichomes is presented. The influence of the growth stage along with different treatments and conditions on the efficiency of spheroplast formation and regeneration are analyzed.     

钝顶螺旋藻(Spirulina platensis)光系统Ⅰ的分离表征及稳定性

采用蔗糖密度梯度离心法对钝顶螺旋藻光合膜蛋白(PSI)进行分离纯化,并对其光谱学性质、热稳定性及光合放氧活性进行分析表征。结果发现,采用蔗糖密度精度离心法,可以成功分离出4条色素蛋白质复合体条带,其中最下层条带为完整的PSI三聚体,其每毫克叶绿素a光合放氧活性达到420μmol/h。当温度达到50℃左右时,分离得到的PSI在溶液开始变性失活。     

Separation and nanoencapsulation of antitumor polypeptide from Spirulina platensis

Spirulina platensis is a multicellular edible blue‐green alga with abundant proteins (～60%). No report is available on the antitumor polypeptides from the whole proteins of S. platensis. In this study, for the first time, an antitumor polypeptide Y2 from trypsin digest of S. platensis proteins was obtained by using freeze‐thawing plus ultrasonication extraction, hydrolysis with four enzymes (trypsin, alcalase, papain, and pepsin), and gel filtration chromatography. The results showed that the degree of hydrolysis can be ordered as: trypsin (38.5%) > alcalase (31.2%) > papain (27.8%) > pepsin (7.1%). For MCF‐7 and HepG2 cells, at 250 µg/mL, the maximum inhibitory rate of Y2 was 97%, while standard drug 5‐FU was 55 and 97%, respectively. Furthermore, the nanoencapsulation of Y2 with chitosan (CS) was also investigated. After nanoencapsulation, the maximum encapsulation efficiency and polypeptides contents are 49 and 15%, respectively; and the antitumor activity is basically not lost. These data demonstrated the potential of nanopolypeptides (Y2‐CS) in food and pharmaceutical applications. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1230–1238, 2013     

螺旋藻多糖抗单纯疱疹病毒作用的实验研究

在体外进行了钝顶螺旋藻多糖(polysaccharides fromSpirulina platensis,PSP)抗单纯疱疹病毒活性的研究。以不同剂量的PSP分别作用于HSV-1及HSV-2病毒复制周期的各个环节,以病毒半数感染量(TCID50),细胞病变效应(CPE),蚀斑形成单位(PFU),MTT染色细胞保护率(MTT法)作为评价指标,判断PSP的抗病毒效果;FQ-PCR检测PSP抗病毒作用的时效关系。结果表明PSP对Vero细胞毒性极低(TC50为1750μg/mL),对HSV-1及HSV-2均无直接灭活作用,可阻滞HSV-1及HSV-2病毒吸附和抑制感染细胞内病毒的复制,但不影响病毒的释放;FQ-PCR结果显示随着PSP浓度及作用时间的增加,PSP对HSV-1病毒DNA的抑制作用明显增强,具有良好的剂量和时效关系。提示PSP抗HSV-1及HSV-2病毒作用的机制与抑制病毒吸附和感染细胞内病毒的生物合成有关。     

钝顶螺旋藻FtsZ在大肠杆菌中的表达和定位

FtsZ是与真核微管蛋白类似的原核骨架蛋白,能在细胞分裂位点聚合组装成环状结构而调控细胞分裂过程。为了研究钝顶螺旋藻（Spirulina platensis）FtsZ蛋白的功能,构建了钝顶螺旋藻FtsZ与绿色荧光蛋白GFP融合表达的质粒,并在大肠杆菌中进行了表达和定位研究,结果发现,表达融合蛋白GFP-FtsZ的大肠杆菌细胞由短杆状变为长丝状,且菌丝体长度与融合蛋白的表达量呈正比。在荧光显微镜下观察到融合蛋白GFP-FtsZ在长丝状体细菌中呈有规律的点状分布,这说明FtsZ蛋白功能高度保守,钝顶螺旋藻FtsZ蛋白能识别大肠杆菌分裂位点并装配成环状结构调控大肠杆菌细胞分裂,FtsZ蛋白的过量表达能抑制大肠杆菌正常的细胞分裂而导致长丝状体细胞的形成。     

螺旋藻多糖抗柯萨奇B3病毒的体外实验研究

在体外进行了钝顶螺旋藻多糖(polysaccharides from Spirulina platensis,PSP)抗柯萨奇病毒B3(Coxsackie virus B3,CVB3)活性的研究.以不同剂量的PSP作用于病毒复制周期的各个环节,以病毒半数感染量(TCID50),细胞病变效应(CPE),蚀斑形成单位(PFU),MTT染色细胞保护率(MTT法)作为评价指标,判断PSP的抗病毒效果.结果表明PSP对Vero细胞毒性极低(TC50为1750μg/mL),对CVB3无直接灭活作用,PSP可干扰病毒向宿主细胞的吸附,其抗病毒生物合成的活性最高(IC50为5.26,TI为237.6),是阳性对照药病毒唑的3.95倍,但不影响病毒的释放.提示PSP抗病毒靶位可能在于抑制病毒吸附和感染细胞内病毒的生物合成.     

Mutants of Spirulina platensis resistant to valine inhibition

Abstract Growth of the cyanobacterium Spirulina platensis is inhibited by low concentrations of valine. Spontaneous valine-resistant mutants were isolated from S. platensis and a preliminary characterization of three of them indicates that one (strains DR2) is defective in valine uptake and two (strains DR5 and DR9) carry alterations in a valine-mediated mechanism of synthesis of acetohydroxy acid synthase, the first common enzyme of the pathway.     

A new model of phycobilisome in Spirulina platensis

Phycobilisomes (PBS) were isolated from blue-green alga Spirulina platensis. Scanning tunneling microscope was used to investigate the three-dimensional structure of PBS deposited on freshly cleaved highly oriented py-rolytic graphite (HOPG) in ambient condition at room temperature. The results showed that the rods of PBS radiated from the core to different directions in the space other than arrayed in one plane, which was different from the typical hemi-discoidal model structure. The diameter of PBS was up to 70 nm, and the rod was approximately 50 nm in length. Similar results were observed in Langmuir-Blodgett (LB) film of PBS. The dissociated PBS could reaggregate into rod-like structures and easily form two-dimensional membrane while being absorbed on HOPG, however, no intact PBS was observed. The filling-space model structure of PBS in Spirulina platensis with STM from three-dimensional real space at nanometer scale was found, which showed that this new structural model of PBS surely exists in blue     

从一种富含藻胆蛋白的螺旋藻中大量提取和纯化藻蓝蛋白的研究

采用新鲜藻丝为原料和分段梯度盐析分离纯化钝顶螺旋藻Sp(NS)-90020的藻蓝蛋白（PC）,经羟基磷灰石一次层析,能使提取的PC的纯度大于普遍认可的标准,由于该工艺流程较为简单,适合藻蓝蛋白的大量生产,藻胆蛋白经Sephadex凝胶过滤后的可达电泳纯度标准,经SDS-PAE测得PC,APC的分子量分别约为38.33kD。     

钝顶螺旋藻（Spirulina platensis）藻胆体Langmuir-Blodgett膜

从钝顶螺旋藻中分离制备完整藻胆体 ,然后滴加于空气 水界面上 ,应用LB膜技术制备藻胆体LB膜。结果表明 ,藻胆体在空气 水界面上具有很好的成膜性能。将藻胆体LB单层膜转移到刚揭开的云母表面 ,喷一层金 ,然后用扫描隧道显微镜观察。结果表明 ,藻胆体在Langmuir Blodgett膜中的排列方式与其在体内类囊体膜表面的排列方式类似 ,一排排聚集在一起 ,然后排列成膜。藻胆体的“核”吸附在云母表面 ,而藻胆体的“杆”伸向外面。由于钝顶螺旋藻易于规模化培养 ,藻胆体容易批量制备 ,加之藻胆体具有的独特的光物理、光化学特性和良好的成膜性能 ,以及本身就是纳米量级的颗粒 (5 0 70nm) ,预示着藻胆体在纳米光电子器件中具有很好的应用前景。     

The Effect of Selenite Ions on Growth and Selenium Accumulation in Spirulina platensis

Selenium accumulation and the growth of cyanobacterium Spirulina platensis (Nordst.) Geitl. were studied in a culture with sodium selenite-supplemented nutritional medium. Selenite concentrations below 20 mg/l did not inhibit the growth of S. platensis. The addition of 30 mg/l of this salt somewhat decreased the growth rate during the linear growth phase, induced the earlier suspension transition to the steady-state phase, and substantially lowered the highest optical density of the suspension. However, even at 170 mg/l Na₂SeO₃, the culture still demonstrated a capacity for growth. The content of selenium in the cells depended directly on its concentration in the medium, up to the lethal level. At high selenium concentrations (100–170 mg/l), S. platensis reduced Se(IV) up to Se(0). The latter was secreted onto the cell surface and into the cultural medium. The high concentrations of Na₂SeO₃ acidified the cytoplasmic pH as was measured by ³¹P-NMR spectroscopy. At the same time, the content of protein on a dry weight basis decreased and that of carbohydrates and lipids somewhat increased, just as was observed in S. platensis cells under other stress factors. In the presence of 20 mg/l Na₂SeO₃, the selenium content in the biomass increased by 20000 times as compared to that in the control cells, whereas the biochemical composition of biomass did not change. In this case, the selenium was incorporated almost completely in the protein fraction. The selenium concentration in this fraction increased more significantly when the sulfur content was lowered in the medium.     

硒胁迫对钝顶螺旋藻抗氧化酶系统的影响

在钝顶螺旋藻的对数生长期进行硒胁迫处理,研究硒胁迫强度和硒胁迫时间对钝顶螺旋藻抗氧化酶系统与脂质过氧化作用的影响。设置4个胁迫实验组,分别从第5天到第10天分次加硒,累计加硒量均为1000 mg/L。结果表明,硒胁迫实验组中钝顶螺旋藻的抗氧化酶活性均高于对照组或与对照组相当,其中GPX的变化最大,实验组Ⅰ中的GPX活性为对照组的20.5倍;SOD的变化趋势与GPX相近,但对较低的硒胁迫的响应不敏感;POD、CAT和APX的活性均随硒胁迫强度的减小而上升;各硒胁迫实验组中只有实验组Ⅰ的MDA含量高于对照组;藻体对.OH的清除能力与藻体的MDA含量呈负相关,提示了硒对钝顶螺旋藻抗氧化系统的双重效应。     

Isolation and Photosynthesis of Spheroplasts in Spirulina platensis

Spirulina platensis is a nonheterocystic filamentous blue-green alga (cyanobacterium). Large quantity of highly qualified spheroplasts were obtained by improved isolation method. The spheroplast has a wrinkled and porous surface. Their diameter ranged from 3.8 btm to 4. 6 μm. The activity of photosynthetic oxygen evolution in the spheroplasts was about 40 % of the intact cell. The absorption spectra of the filaments and spheroplasts at room temperature revealed that they had the same pigments, Chla, PC, PEC and carotenoid. In spheroplasts the relative content of PC and carotenoid decreased, and that of PEC increased. It implicated that the light absorption of Spirulina platensis could be influenced by the cell wall. Some differences existed between the original cells and spheroplasts in the low temperature fluorescence emission spectra. F757 of spheroplasts excited by 436 nm was reduced obviously and that excited by 580 nm was disapeared. F728/F685 and F640/F685 enhanced, and F693/F685 was reduced. F728/F640 was lower than that of the original cells. These results indicated that removing the cell wall may inhibit the PS Ⅱ activity and influence the F695 from core antenna pigment system.     

Application of Arthrospira (Spirulina) platensis biomass for silver removal from aqueous solutions

The cyanobacterium Arthrospira (Spirulina) platensis was used to study the process of silver biosorption. Effects of various parameters such as contact time, dosage of biosorbent, initial pH, temperature, and initial concentration of Ag(I) were investigated for a batch adsorption system. The optimal biosorption conditions were determined as pH 5.0, biosorbent dosage of 0.4 g, and initial silver concentration of 30 mg/L. Equilibrium adsorption data were analyzed by the Langmuir and Freundlich models – however, the Freundlich model provided a better fit to the experimental data. The kinetic data fit the pseudo-second-order model well, with a correlation coefficient of 0.99. The analysis of thermodynamic parameters (ΔG°, ΔH° and ΔS°) revealed that the adsorption process of silver ion by spirulina biomass was exothermic and spontaneous (ΔG° < 0), and exothermic (ΔH° < 0) process. The biosorption capacity of biomass A. platensis serves as a basis for the development of green technology for environmental remediation.     

不同生境下的钝顶螺旋藻RAPD分析

本文对不同生境下的钝顶螺旋藻Spirulina (Arthrospira) platensis进行了RAPD分析。结果表明, 鄂尔多斯高原碱湖和Chad湖的钝顶螺旋藻基因组DNA扩增多态性片段同源性为 48.23%。2 个样品在分子遗传水平上存在着较大的差异, 这是由于各自生态环境明显不同和长期地理隔离造成的。     

不同生境下的钝顶螺旋藻RAPD分析

本文对不同生境下的钝顶螺旋藻Spirulina(Arthrospira)platensis进行了RAPD分析.结果表明,鄂尔多斯高原碱湖和Chad湖的钝顶螺旋藻基因组DNA扩增多态性片段同源性为48.23%.2个样品在分子遗传水平上存在着较大的差异,这是由于各自生态环境明显不同和长期地理隔离造成的.     

钝顶螺旋藻表面微观形貌的原子力显微镜研究

该文应用原子力显微镜(AFM)纳米级的分辨率对钝顶螺旋藻(Spirulina platensis)表面微观形貌进行了研究,获得了扫描范围为5.000×5.000μm、1.000×1.000μm和400.0×400.0nm三组清晰、稳定的图像,并对其进行了线性分析。结果表明:螺旋藻表面由紧密且无序堆积的突起结构组成,其高度小于20nm;突起结构高度从3nm~15nm不等,平均高度约为8~9nm。此法用于生物体表面,操作简单、快速、灵敏度好且样品无损伤,结果令人满意。