半导体激光对紫球藻生物学效应的影响

采用波长650nm,功率40mW,功率密度13W／cm^2的半导体激光,对紫球藻（Porphyridium cruentum)进行诱变,辐照时间分别为5min、10min、20min。通过对紫球藻每天细胞数、叶绿素a、第7d细胞干重和胞外多糖产量的测定可知：与对照组比较,3个处理对紫球藻生长及胞外多糖产生影响,5min、10min均有不同程度的促进作用,其中5min作用最明显,而20min则有抑制作用。     

紫球藻胞外多糖的分离纯化与性质分析

采用Q Sepharose Fast Flow凝胶,从紫球藻胞外多糖（ESPS）获得3个组分ESPS0,ESPS1.0和ESPS1.6,它们经过Sephadex G200凝胶过滤纯化后,紫外光谱、红外光谱、氨基酸分析、单糖组成分析表明,ESPS0中未测出蛋白,ESPS1.0和ESPS1.6的蛋白含量分别为7.8%和7.6%,三者硫酸基含量分别为16.3%,11.6%和8.3%,同时分别在201 nT,207 nm和199 nm处出现特征吸收峰,而在280 nm和260 nm波长处无吸收峰.ESPS0、ESPS1.0中单糖的连接方式为β-糖苷键,而ESPS16中主要为α-糖苷键.ESPS1.0、ESPS1.6都含有十七种氨基酸.三种组分都是由D-木糖、D-葡萄糖、D-甘露糖、D-艾杜糖以不同的比例所组成.     

硒化紫球藻胞外多糖组成与结构的初步分析

通过在ASW培养基中加入适量的亚硒酸制备硒化紫球藻胞外多糖,经分离纯化、纯度鉴定后,利用下列手段对其进行分析:通过紫外可见光谱扫描、红外光谱扫描了解其结构信息;通过高效液相色谱对其单糖组分进行分析;通过硫酸-咔唑法测定其糖醛酸含量;通过硫酸比浊法测定其硫酸根含量,等。Se-PSP和PSP一样,分离后分别得到两种成分,紫外光谱也和PSP相似,不含有蛋白质和核酸;红外光谱显示Se-PSP中Se可能取代了C-H上的H和SO42-中的S;HPLC显示其单糖组分种类相似,含量稍有差别;另外,PSP和Se-PSP所含的糖醛酸含量没有统计学差异,Se-PSP所含SO42-比PSP少。     

紫球藻及其胞外多糖对小鼠免疫功能的调节作用

探讨紫球藻及其胞外多糖对免疫低下小鼠免疫功能的调节作用,用环磷酰胺(CY)建立昆明种小鼠免疫功能低下的实验模型。小鼠随机均分6组,其中2组每天分别给予紫球藻藻粉1200、600 mg/kg灌胃,2组给予紫球藻胞外多糖300、150 mg/kg灌胃,正常对照组及CY组用等容积生理盐水灌胃,连续14 d。实验结果表明紫球藻及其胞外多糖均可显著提高免疫抑制小鼠的脾指数、胸腺指数、碳廓清能力、单核细胞吞噬功能,并可对抗环磷酰胺引起的外周血白细胞数目下降,因此紫球藻藻粉及其胞外多糖对小鼠免疫功能具有一定的正向调节作用。毒性实验表明,小鼠腹腔注射300、150 mg/kg紫球藻胞外多糖溶液和分别给予紫球藻藻粉1200、600 mg/kg灌胃,小鼠未出现死亡,也未有明显毒性反应出现的一些指标变化,说明紫球藻藻粉及胞外多糖的安全性较高。     

倍频Nd：YAG激光对紫球藻生长与胞外多糖产量的影响

紫球藻经Nd:YAG激光（波长1060nm)照射,以不同时间设置处理剂量,比较紫球藻细胞活力,生长速率,生物量,胞外多糖产量等方面的变化。高剂量处理组（Y-20、Y-30和Y-40）致死率为17-44%,继代培养后生长迅速,K值分别比对照组提高18%、23.4%和15.3%,各处理组培养10天后收获的生物量与对照组无显著差异,但胞外多糖产量均有较大幅度提高,增幅达50-150%。此外,YAG激光照射对藻细胞叶绿素含量也有影响,YAG激光有望成为紫球藻优良藻株选育的较佳诱变剂。     

超声波降解紫球藻胞外多糖研究

通过测定溶液粘度,判断超声波降解紫球藻(Porphyridium cruentumi)胞外多糖的效果。运用均匀设计对超声波降解紫球藻胞外多糖的影响因素(处理振幅、时间、脉冲)进行优化,获得超声波处理的最佳条件:振幅39%、处理时间245s和脉冲9.5s。在最佳条件下,胞外多糖的粘度为2.98mm²/s,与预测值一致。采用DPS软件对实验结果进行二次多项式分析与拟合,并对模型和回归系数进行显著性检验,建立了以胞外多糖粘度为目标的回归方程式。     

倍频Nd∶YAG激光对紫球藻生长与胞外多糖产量的影响

紫球藻经Nd:YAG激光(波长1060 nm)照射,以不同时间设置处理剂量,比较紫球藻细胞活力、生长速率、生物量、胞外多糖产量等方面的变化.高剂量处理组(Y-20、Y-30和Y-40)致死率为17～44%,继代培养后生长迅速,K值分别比对照组提高18%、23.4%和15.3%.各处理组培养10天后收获的生物量与对照组无显著差别,但胞外多糖产量均有较大幅度提高,增幅达50～150%.此外,YAG激光照射对藻细胞叶绿素含量也有影响.YAG激光有望成为紫球藻优良藻株选育的较佳诱变剂.     

紫球藻及其多糖抗菌性能初探

本文报道紫球藻提取液及其多糖溶液抗病原细菌、真菌性能的研究结果。采用管碟法分别测定紫球藻培养分泌物、藻体破碎后的水提物和醇提物、胞外多糖和胞内多糖水溶液等的抗菌能力。结果表明,各样品均有一定抗菌能力,紫球藻水提物和胞外分泌物抗菌性能较强。经比较分析,多糖是抗菌作用的主要组分。紫球藻提取物及其多糖对革兰氏阳性细菌(特别是对金黄色葡萄球菌和八叠球菌)的抑菌作用较为明显,具有一定开发价值。     

紫球藻的培养与利用（综述）

紫球藻为单细胞红藻,其细胞富含红藻多糖、多不饱和脂肪酸(主要为AA和EPA）及藻胆蛋白等高价值生物活性物质,在医药和精细化工领域有广泛的应用前景。本文综述紫球藻的生物学特性、生物活性物质、培养与培养特性等方面的研究概况。     

紫球藻胞外多糖抗呼吸道合胞病毒(RSV)活性研究

采用体外细胞培养的方法,在Hela细胞系上检测了来自紫球藻的胞外多糖及其组分的抗呼吸道病毒(RSV)活性。发现紫球藻胞外多糖对呼吸道合胞病毒具有强烈的抑制活性,同时对宿主细胞的抑制作用很小。分离组分中的强带电性组分ESPSⅥ活性最高,其TI值达3125,为阳性对照药病毒唑的40余倍。     

Increased production of extracellular polysaccharide by Porphyridium cruentum immobilized in foam sheets

Large‐size plate bioreactors were used to compare the production of extracellular polysaccharide by the red microalga Porphyridium cruentum when grown in suspension and in a foam sheet. A well‐defined illuminated area and unidirectional light propagation allowed us to generate information that is better quantified when expressed in terms of illuminated area. This is essential for meaningful comparison of data, especially considering that for a well‐designed and managed bioreactor, the culture production rates are believed to be light limited. At the same level of illumination, the culture immobilized in foam showed double the production rate of extracellular polysaccharide compared with the culture in suspension. The saturation level of biomass density per unit of illuminated area was eight times higher for the immobilized culture compared with the culture in suspension. Despite the increased biomass density for the immobilized culture, an increase in the light level above the optimum found for the culture in suspension reduced the extracellular polysaccharide production, suggesting that the photoinhibition light level was surpassed.     

紫球藻藻红蛋白的分离纯化及光谱学特性

采用4℃反复浸提、离心、硫酸铵沉淀、DEAE—Sepharose Fast Flow离子交换柱层析,从紫球藻（Porphyridium cruentum Naegeli）冻干粉中分离纯化藻红蛋白,分离纯度达到4．85,总收率51．9％;经羟基磷灰石柱层析纯化,藻红蛋白纯度达到5．10,总收率34．0％,聚丙烯酰胺凝胶电泳显示1条带。所分离纯化的藻红蛋白含有3个亚基（α、β、γ）,在可见光区545nm和560nm处有2个吸收峰,在498nm处有1个吸收肩峰。实验结果说明所分离纯化的藻红蛋白纯度符合要求。     

Comparison of artificial light photobioreactors and other production systems using Porphyridium cruentum

A new type of preparative photobioreactor for high quality production of microalgae is developed for hatchery-nursery of marine animals, as well as for fine chemicals extraction. Of modular conception, two artificial light photobioreactors in plastic and stainless steel are designed so as to provide strictly controlled conditions in an attempt to increase quality and diminish cost prices. They are assessed for production of Porphyridum cruentum and compared to conventional transparent tanks and solar photobioreactors. The concentration and productivity obtained are ten-fold higher than with hatchery tanks, which leads to a significant drop in cost price of biomass. Comparison is also made with a 10 m² solar photobioreactor operated in the south of France, for which biomass cost price is half that of 1.5 m² artificial light photobioreactor. Extrapolations erasing size discrepancy show that the cost price of the two technologies are not very different. This revised version was published online in June 2006 with corrections to the Cover Date.     

Swirling flow implementation in a photobioreactor for batch and continuous cultures of Porphyridium cruentum

Light is the main limiting factor in photoautotrophic-intensive production of microorganisms, and improvement of its use is an important concern for photobioreactor design and operation. Swirling flows, which are known to improve mass and photon transfers, were applied to annular light chambers of a photobioreactor and studied by simulation and microalgal culture. Two hydrodynamic conditions were compared: axial flow generating poor radial mixing, and tangential flow generating three-dimensional swirling motion. Batch and continuous cultures of the Rhodophyte Porphyridium cruentum were performed in a 100-L, 1.5-m(2), fully controlled photobioreactor with eight light chambers. The inlet design of these chambers was modified to create the hydrodynamic conditions for comparison. Various intensities of swirling motion were used, characterized by the velocity factor (VF), defined as the ratio between annular chamber flow and inlet aperture sections. Experiments were performed within the range of photon flux densities (PFD) optimizing the yield of light energy transformation into living substance for the species and the temperature used. Culture kinetics with swirling flows generated by apertures of VF = 2, 4, and 9 were compared with pseudoaxial VF = 2 chosen as reference. Batch cultures with VF = 4 swirling flow showed no significant difference, whereas continuous cultures proved more discriminating. Although no significant difference was obtained for VF = 2, a 7% increase of steady-state productivity and a 26% decrease in time required to reach this steady state were obtained with VF = 4 swirling flow. This beneficial effect of swirling flow could have accounted for increased mixing. Conversely, VF = 9 swirling flow resulted in a 9% decrease of steady-state productivity and a 9% increase in the time required to reach this steady state, a negative effect that could have accounted for increased shear stress. CO(2) bioconversion yield at steady state showed a 34% increase for VF = 4. These results suggest that swirling motion makes microalgal cultures more efficient, provided that the resulting adverse effects remain acceptable. Experimental investigation was completed by a theoretical approach in which simulation of continuous cultures of P. cruentum was based on the hydrodynamic conditions achieved in the photobioreactor. Although the results obtained with pseudoaxial flow were correctly predicted, simulations with swirling flow showed a marked enhancement of productivity not observed experimentally. The influence of side effects induced by increased mixing (particularly hydrodynamic shear stress) was considered with respect to modeling assumptions. Comparison of experimental results with theoretical simulation provided a better understanding of the mixing effect, a key factor in improving the efficiency of such bioprocesses.     

紫球藻多糖化学结构解析

为明确紫球藻多糖的化学结构,本文采用化学分析和光谱分析方法对紫球藻多糖的一级糖链结构进行了分析。GC分析表明该多糖由木糖、葡萄糖和半乳糖组成,为一种杂多糖,其摩尔比为:2.96∶1.25∶3.06;红外光谱分析结果显示紫球藻多糖为硫酸化多糖,糖苷键类型为β构型;化学分析结果推断紫球藻多糖糖链连接方式以1→3为主,存在少量1→2,1→4,1→6键型,且半乳糖在支链或主链末端有较大量的存在,木糖和葡萄糖在主链或靠近主链区域有特定分布;NMR分析显示紫球藻多糖的硫酸酯基连在C-6上,且多糖的糖苷键为β型;GC-MS联机分析进一步确定紫球藻多糖为一种主要含有1→3糖苷键,并含有1→4,1→6糖苷键的杂多糖。综合上述分析,推断出紫球藻多糖的糖链主链的重复单元结构。     

平板式光生物反应器中紫球藻培养条件的优化

研究了平板式光生物反应器中紫球藻(Porphyridium cruentumNaegeli)的培养条件,运用均匀设计法对光照强度、通气速率、装液量、接种密度以及pH等影响紫球藻生长的因素进行优化,获得了在平板式光生物反应器中培养紫球藻的最佳条件:光照强度10 000 lx、通气速率350 L.h-1、装液量6 L、藻细胞接种密度1.1×106mL-1、pH9.0。在最佳条件下藻体的生物量产率和生物量产量分别达到0.431 g.L-1.d-1和3.240 g.L-1,最大生长速率达0.652 g.L-1.d-1,胞外多糖含量高达0.665 g.L-1。另外,在培养过程中隔天补充培养液有利于紫球藻生物量的增加和胞外多糖的产生。