Influence of temperature on growth of Scenedesmus obliquus in diluted olive mill wastewater as culture medium

Scenedesmus obliquus can help to reduce the environmental impact of industrial olive mill wastewater from olive oil extraction in the three‐phase system. This work examines the effect of temperature changes (288–308 K) on algal growth, culture medium, and biochemical composition of S. obliquus. The maximum specific growth rate of 0.024 h^?1 occurred at an optimal temperature of 302.7 K. The apparent activation energies of cell growth and cell death were determined as 61.8 and 142.8 kJ/mol, respectively. At the end of culture the percentages of pigments, proteins, and carbohydrates were greater at the two ends of the temperature range studied, as it was also observed for the maximum elimination of biochemical oxygen demand (BOD₅). The mono‐ and polyunsaturated fatty acid content of the biomass was greater at the lowest temperature used (288 K).     

Functional characteristics of green alga Scenedesmus obliquus (Chlorophyceae): 276–6 wild type and its two photosystems deficient mutants cultured under photoautotrophic,mixotrophic and heterotrophic conditions

Functional features of Scenedesmus obliquus: wild type 276–6 strain (WT) and its two mutants reported as photosystem I‐deficient (mutant 56.80) and photosystem II‐deficient (mutant 57.80) were characterized. Algae were cultured aseptically under continuous light or in darkness on mineral bold basal medium (BBM), yeast extract‐enriched BBM and yeast extract to evaluate the physiology of algal cells under photoautotrophic, mixotrophic and heterotrophic conditions. Growth, superoxide dismutase activity and photosynthetic parameters, including polyphasic fluorescence rise during the first seconds of chlorophyll a illumination (OJIP), were analyzed to find relationships between the photosynthetic/respiratory activity of the cells, occurrence of oxidative stress and trophic conditions applied to PSs‐deficient algae. Despite the highest superoxide dismutase activity, indicating the presence of oxidative stress, mixotrophic conditions appeared to be optimal for S. obliquus WT and mutant strains kept in non‐aerated cultures. OJIP analysis indicated that in mutant 56.80 part of photosystem (PS) I was functional and in mutant 57.80 residual PS II activity was found.     

斜生栅藻中虾青素的积累过程及其光合活性变化

分析了斜生栅藻（Scenedesmus obliquus）在光温（30℃,180 μmol/m2·s）胁迫条件下积累虾青素的过程,观察了该过程中细胞形态及细胞光合生理的变化。胁迫条件下,细胞在48h内生成并积累了包括海胆酮、角黄素、金盏花黄素和金盏花红素在内的多种次生类胡萝卜素,并合成了虾青素及其酯。该过程中,细胞形态由两端尖细变得不规则、膨大,原来由4、8个细胞组成的定形群体变为游离的单个细胞或2个细胞组成的群体。藻细胞光合速率在24h内先下降后上升,而后又呈现下降趋势,从34.29 μmol O2/mg Chla/h迅速下降为5.21 μmol O2/mg Chla/h;呼吸速率在前24h内升高至60.37 μmol O2/mg Chla/h,而后缓慢下降到38.40 μmol O2/mg Chla/h;光合系统Ⅱ的活性随着胁迫时间的延续而逐步下降,较初始值降低了63.9％。结果表明,斜生栅藻细胞在高光照条件下可以合成虾青素,并通过调节光合速率、呼吸速率以及光合系统Ⅱ的效率来应对胁迫。     

角突网纹溞培养滤液对铜绿微囊藻和栅藻形态及生长率的影响

在藻类中,随着环境条件的变化而产生相应的单细胞和群体之间的形态转化是一种广泛存在的现象,称为表型可塑性。例如,栅藻属成员就是这些藻类的代表,在野外具有典型的群体特征,但在室内纯培养条件下栅藻常较少形成群体而是保持以单细胞形态为主。     

水芹水浸提液对斜生栅藻的化感效应研究

采用栅藻在不同浓度的水芹水浸提液中纯培养的方法,研究了水芹水浸提液对斜生栅藻细胞数、叶绿素含量及藻细胞超微结构的影响。结果显示,10 g·L^-1水芹水浸提液对斜生栅藻的生长和叶绿素含量的增加具有明显的促进效应;20 g·L^-1水芹水浸提液对藻细胞数和叶绿素含量的增加持续到第3 d,3 d后出现显著的抑制效应;高浓度（30~50 g·L^-1）水芹水浸提液对藻细胞数和叶绿素含量的明显抑制在第2 d始现,随时间延长而加剧,并具有浓度效应;40 g·L^-1水芹水浸提液处理后,斜生栅藻细胞壁断裂甚至消失,细胞中叶绿体片层肿胀甚至解体,核膜破裂,核质外渗。水芹水浸提液对斜生栅藻具有化感效应,低浓度促进生长,高浓度抑制。     

Biosynthesis of chlorophyll b in pigment mutant C-2A' of Scenedesmus obliquus

It is demonstrated that chlorophyll b does not only derive from chlorophyll a , but is also formed separately from an in vivo-occurring chlorophyllide b . The branching point for the latter synthesis is at the level of chlorophyllide, since no protochlorophyllide b was detectable. We have indications that the enzyme oxidizing chlorophyll a to chlorophyll b accepts also non-phytylated 17,18 dihydroporphyrins and is not restricted to chlorophylls. Preparations of chlorophyllide a and chlorophyll a could both be transferred with the same enzyme fraction to chlorophyllide b and chlorophyll b , respectively. Preliminary experiments show this enzyme to be membrane bound and light independent. An updated scheme for chlorophyll b biosynthesis is presented.     

Enantioselective toxicity of metolachlor to Scenedesmus obliquus in the presence of cyclodextrins

Cyclodextrins (CDs) possess a variety of chiral centers and are capable of recognizing enantiomeric molecules through the formation of inclusion complexes. Two types of CDs, α-cyclodextrin (α-CD) and β-cyclodextrin (β-CD), were selected to evaluate the effects of the enantioselective ecotoxicity of racemic metolachlor (Rac-metolachlor) and its S-enantiomer (S-metolachlor) on the freshwater algae Scenedesmus obliquus (S. obliquus) by acute toxicity test. The results showed that the aquatic toxicity of S-metolachlor was higher than Rac-metolachlor and that CDs enhanced the toxicity of metolachlor enantioselectively by increasing the aquatic toxicity of Rac-metolachlor rather than that of S-metolachlor to S. obliquus. The equilibrium constant for Rac-metolachlor-CD complexes was higher than that of S-metolachlor-CDs, which was responsible for the greater aquatic toxicity shift effect of Rac-metolachlor. Thermodynamic studies of CD complexes showed that inclusion for all of the complexes was primarily a spontaneous, enthalpy-driven process. These results will help to understand the preliminary mechanism of shifting aquatic toxicity of metolachlor by CDs and the CDs mediated environmental processes of metolachlor, to correctly apply CDs to chiral pesticides formulation and environmental remediation of chiral contaminants.     

壶状臂尾轮虫(Brachionus urceus)培养滤液对斜生栅藻(Scenedesmus obliquus)形态和多聚糖含量的影响

分离自太湖的壶状臂尾轮虫(Brachionus urceus (Linnaeus))按1000 ind·L-1的密度置于斜生栅藻(Scenedesmus obliquus)中培养24h后,用孔径0.10 靘的微孔滤膜抽滤,得到壶状臂尾轮虫培养滤液.将壶状臂尾轮虫培养滤液以20%的比例添加到纯培养的斜生栅藻中,进行为期7d的试验.结果表明壶状臂尾轮虫培养滤液能显著地影响斜生栅藻的形态,处理组中诱发性多细胞群体增加,导致处理组中斜生栅藻种群平均每个个体的细胞数量显著高于对照组.斜生栅藻在壶状臂尾轮虫诱发下形成群聚体可以解释为一种诱发性的反牧食防御,同时也进一步证实了栅藻对浮游动物牧食的响应是一种广泛存在的现象.壶状臂尾轮虫培养滤液处理后的斜生栅藻在生长率上与对照组没有显著性差异,说明诱发性防御群聚体形成的代价没有体现在对生长的影响上.在壶状臂尾轮虫培养滤液诱发下,较多栅藻细胞聚积在一起形成群聚体后,栅藻溶解性胞外多聚糖和固着性胞外多聚糖均有显著提高,而胞内多聚糖略有下降.但从总多聚糖含量看,处理组明显高于对照组,这也说明轮虫培养滤液刺激了栅藻多聚糖合成的额外增加,且被分泌到胞外,充当细胞之间的粘合物,促使细胞容易聚合形成群聚体,有效防御象轮虫这些小型浮游动物的进一步牧食,保持种群得以延续.     

The effect of arsenate and arsenite on photosynthesis in Scenedesmus obliquus. A Potentiometric study in a closed CO2-system

Abstract

A Potentiometric titration method was used to study the adverse effect of arsenate (As(V)) and arsenite (As(III)) on inorganic carbon uptake in suspensions of the green alga Scenedesmus obliquus. The measurements were performed in a closed CO₂-system with diluted synthetic seawater (1‰ salinity) as ionic medium. Usually, the algal chlorophyll concentration was 0.4 mg dm^?3, while the arsenate- and arsenite-concentrations were varied within the limits 0.1 to 200 μmol dm^?3. In some experiments arsenate toxicity was studied in the presence of 1 to 100 μmol dm^?3 of phosphate (P(V)).

With concentrations of arsenate or arsenite less than 0.1 μmol dm^?3 no toxic effects were observed. However, at As-concentrations of 200 μmol dm^?3, the algal carbon uptake was reduced by 41% with arsenate and 29% with arsenite, i.e., arsenate is more toxic to Scenedesmus obliquus than arsenite. The toxicity of arsenate was negligible in the presence of a ten fold excess of phosphate. This is probably due to chemical similarities between arsenate and phosphate causing competition between the ions for the binding sites.

The importance of taking the speciation as well as the buffer capacity of the algal system into account, when calculating the carbon uptake, is also discussed.     

Evidence for the Presence of Chlorophyllide b in the Green Alga Scenedesmus obliquus in vivo

Chlorophyllide b could be extracted from the wild type of Scenedesmus obliquus and its pigment mutant C-2A'. Its identity was proved by absorption and fluorescence spectroscopy and by a positive hydroxylamine test. Chlorophyllide b could be transformed into pheophorbide b and methylpheophorbide b. The formation of chlorophyllide b from chlorophyll b by dephytylation with chlorophyllase could be ruled out. The stimulation of chlorophyllide b biosynthesis with o-phenanthroline, as described in the literature, could not be confirmed under physiological conditions.     

Enantioselective toxicological response of the green alga Scenedesmus obliquus to isocarbophos

Chiral compounds usually behave enantioselectively in phyto‐biochemical processes. Isocarbophos (ICP) is a chiral pesticide that is widely used. To evaluate the toxicological response of ICP and its enantiomers to Scenedesmus obliquus, algal growth, total chlorophyll, total soluble protein, and the superoxide anion radicals (O₂^?‐) were investigated. The microalgae were treated with ICP and its enantiomers at 0.01–10 mg/l for 96 h. The growth of S. obliquus was stimulated at low levels of ICP and its enantiomers (0.01–1 mg/l), but all were inhibited at high concentrations (10 mg/l). The total soluble protein content and total chlorophyll content of the tested green alga S. obliquus gradually increased, depending on the growth of algal cells in the medium. Meanwhile, the content of O₂^?‐ was decreased. Interestingly, the cell number and content of the chlorophylls and protein decreased with increasing levels of concentration, whereas O₂^?‐ increased. Our results indicated that enantioselectivity was observed in the dose–response of ICP and its enantiomers in S. obliquus. The high O₂^?‐ level might lead to the death of S. obliquus. The stimulation of growth suggests a regulatory mechanism that is related to the capability of the algae to adapt to the O₂^?‐. Chirality 24:481–485, 2012. © 2012 Wiley Periodicals, Inc.     

Occurrence of protochlorophyll and its phototransformation to chlorophyll in mutant C-2A' of Scenedesmus obliquus

The pigment mutant C-2A' of the unicellular green alga Scenedesmus obliquus accumulates protochlorophyllide and small amounts of protochlorophyll in darkness. Protochlororphyll was isolated and characterized by thin layer chromatography and absorption and fluorescence spectroscopy. The protochlorophyll was photoreduced by light to chlorophyll both in vitro and in vivo.     

Ultrahigh-cell-density heterotrophic cultivation of the unicellular green microalga Scenedesmus acuminatus and application of the cells to photoautotrophic culture enhance biomass and lipid production

Although production of biodiesels from microalgae is proved to be technically feasible, a commercially viable system has yet to emerge. High-cell-density fermentation of microalgae can be coupled with photoautotrophic cultivation to produce oils. In this study, by optimizing culturing conditions and employing a sophisticated substrate feed control strategy, ultrahigh-cell-density of 286 and 283.5 g/L was achieved for the unicellular alga Scenedesmus acuminatus grown in 7.5-L bench-scale and 1,000-L pilot-scale fermenters, respectively. The outdoor scale-up experiments indicated that heterotrophically grown S. acuminatus cells are more productive in terms of both biomass and lipid accumulation when they are inoculated in photobioreactors for lipid production as compared to the cells originally grown under photoautotrophic conditions. Technoeconomic analysis based on the pilot-scale data indicated that the cost of heterotrophic cultivation of microalgae for biomass production is comparable with that of the open-pond system and much lower than that of tubular PBR, if the biomass yield was higher than 200 g/L. This study demonstrated the economic viability of heterotrophic cultivation on large-scale microalgal inocula production, but ultrahigh-productivity fermentation is a prerequisite. Moreover, the advantages of the combined heterotrophic and photoautotrophic cultivation of microalgae for biofuels production were also verified in the pilot-scale.     

Accumulation of uranium at low concentration by the green alga Scenedesmus obliquus 34

Accumulation of UO ₂ ^{2 +} by Scenedesmus obliquus 34 was rapid and energy-independent and the biosorption of UO ₂ ^{2 +} could be described by the Freundlich adsorption isotherm below the maximum adsorption capacity (75 mg g-¹ dry wt). The optimum pH for uranium uptake was between 5.0_8.5.0.1_2.0 M NaCl enhanced uranyl, while Cu²⁺, Ni²⁺, Zn²⁺, Cd²⁺ and Mn²⁺ competed slightly with uranyl. Pretreatment had an unexpected effect on biosorption. After being killed by 0.1 M HCl, S. Obliquus 34 showed 45% of the uptake capacity of the control in which fresh cells were suspended directly in uranyl solution, while the pretreatment of cells by 0.1 M NaOH, 2.0 M NaCl, ethanol or heating decreased uptake slightly. Fresh S. obliquus 34 at 1.2_2.4 mg dry wt mL-1 was able to decrease U from 5.0 to 0.05 mg L-1 after 4_6 equilibrium stages with batch adsorption. Deposited U could be desorbed by pH 4.0 buffer. It is suggested that U was captured by effective groups or by capillary action in the cell wall in the form of [UO2OH]+. This revised version was published online in August 2006 with corrections to the Cover Date.     

萼花臂尾轮虫培养滤液对铜绿微囊藻、斜生栅藻和小球藻群体形成及生长的影响

为探索浮游动物和藻类之间可能存在的信息传递,研究了萼花臂尾轮虫培养滤液对铜绿微囊藻、斜生栅藻和小球藻的生长及群体形成的影响.把萼花臂尾轮虫按1000个·L^-1的初始密度置于小球藻中培养24h后,用孔径0.15μm的微孔滤膜抽滤,得到轮虫培养滤液,此滤液中含有轮虫在生活过程中释放的一些信息化学物质.将轮虫培养滤液以20%的比例分别加入纯培养的铜绿微囊藻、斜生栅藻和小球藻中,进行为期7d的试验.结果表明,萼花臂尾轮虫培养滤液能显著地促进斜生栅藻的群体形成,而对铜绿微囊藻和小球藻在群体形成方面没有显著作用.另外,该滤液能显著提高小球藻种群的增长,对铜绿微囊藻和斜生栅藻的生长无明显影响.3种藻类对萼花臂尾轮虫的潜在牧食采取了不同的生态策略:斜生栅藻形成群体,增大摄食阻力,从而降低被摄食的风险;小球藻通过提高增长率来抵消被取食的损失;铜绿微囊藻是通过其它方式来降低被牧食(例如毒素).这些方式分别是这些藻类维持种群规模的反牧食防御策略之一.     

Monovinyl and divinyl protochlorophyll in different stages of esterification isolated from mutant C-2A'of the unicellular green alga Scenedesmus obliquus

The mutant C-2A'of the unicellular green alga Scenedesmus obliquus accumulates the chlorophyll-precursors protocbloropbyllide and the already eslerified protochlorophyll when grown heterotrophically in the dark. Two derivatives of protochlorophyll. monovinyl protochlorophyll (MV-PChl) and divinyl protochlorophyll (DV-PChl), were isolated from dark-grown cells of mutani C-2A'and characterized by absorption and fluorescence spectroscopy. Their molecular masses were determined by plasma desorption mass spectrometry. Both MV- and DV-PChl were mainly esterified with geranylgeraniol. However, some esterification with the more saturated alcohols dihydrogeranylgeraniol. tetrahydrogeranylgeraniol and phytol could also be detected.     

溴化1-己基-3-甲基咪唑对斜生栅藻谷胱甘肽及其代谢酶的影响

研究了浓度为0、1、5、10、15、20 mg/L的新兴离子液体溴化1-己基-3-甲基咪唑([C6mim]Br)在24h、48h、72h和96h对斜生栅藻还原型谷胱甘肽（GSH）及其代谢酶-谷胱甘肽过氧化物酶（GPX）、谷胱甘肽转硫酶（GST）和谷胱甘肽还原酶（GR）的影响。结果表明：GSH含量在24h、48h和72h时,在最低处理浓度下不变,其他处理浓度下随胁迫浓度增加而降低,96h时则与对照无差异或较小;GPX和GST的活性在72h之前明显升高（最高浓度组的GST活性有波动）,96h时均降低至对照水平;GR活性在24h时,[C6mim]Br=1 mg/L时升高,之后降低,在48h增高至对照水平,72h时,[C6mim]Br≥10 mg/L的处理组高于对照水平,96h时,除最低处理组外,均降至对照水平以下。GR是GSH系统中的限速酶,GST则是该系统中活性和灵敏性最高的酶,可作为[C6mim]Br胁迫时的敏感的生物标志物。1 mg/L的[C6mim]Br可引起藻细胞的氧化胁迫,具有环境毒性。     

Effects of temperature and pH on growth and antioxidant content of the microalga Scenedesmus obliquus

Reactive forms of oxygen can damage DNA (among other molecules), thus triggering, e.g., atherogenesis and carcinogenesis. However, such dietary antioxidants as lutein and β-carotene can effectively inactivate them; these compounds were found to high levels in a novel strain (M2-1) of the microalga Scenedesmus obliquus. The independent and combined effects of pH and temperature on its rates of growth and production of antioxidants were experimentally assessed, via a full factorial experimental design; the effects of each parameter independently, and of their interactions were accordingly quantified by ANOVA. Our results indicated that temperature plays a more important role on the maximum specific growth rate than pH; in terms of antioxidant content, pH and, to a lesser extent, temperature also have relevant effects. Consequently, the highest rate of biomass specific growth (0.294 ± 0.013 day(-1)) and biomass productivity (0.837 ± 0.054 mg L(-1) day(-1)) were associated with relatively low pH (6) and relatively high temperature (30°C). Conversely, the antioxidant production rate increased with pH; hence, the highest productivity (0.638 mg L(-1) day(-1)) was attained at pH 8 and 30°C. At the best operating conditions for antioxidant content, the levels of lutein and β-carotene were 203.57 ± 1.41 and 18.20 ± 0.33 mg mL(-1), respectively; the maximum production of either one occurred at the early exponential phase.     

Photosynthetic Capacity and Quantum Requirement of Three Secondary Mutants of Scenedesmus obliquus with Deletions in Carotenoid Biosynthesis*

Photosynthetic capacity and quantum requirement of photosynthesis were evaluated in three secondary mutants of Scenedesmus obliquus possessing specific blocks in carotenoid biosynthesis. These were derived from the mutant, C-2 A', a strain which develops chlorophyll only in the light. All strains were capable of a normal and rapid production of chlorophyll and the development of a functional photosynthetic apparatus when exposed to light. LHC-II levels in the mutants lacking the β,ε-carotenoids (strains C-2 A'-34,1 and C-2 A'-67, 3b) were considerably reduced in fully greened cells but normal in the β,β-epoxycarotenoid deficient-only strain (C-2 A'-67,1) (Bishop, 1996). The maximum photosynthetic capacity and the quantum efficiency (at λ=682nm) of photosynthesis of cells of C-2A'-34,1 and C-2 A'-67,1 showed no significant changes when compared to the original parent strain, C-2 A'. However, the quantum requirement of the strain lacking both types of carotenoids, C-2 A'-67,3b, was slightly increased. It is of particular interest that both strains lacking the β,ε-carotenoids also show a preferential loss of the oligomeric form of the LHC and its associated chlorophylls. The potential requirement of the different carotenoids in maintenance of photosynthetic efficiency in Scenedesmus is discussed.     

Biotransformation of 1-naphthalenesulfonic acid by the green alga Scenedesmus obliquus

Under sulfate limitation, axenic batch cultures of the green alga Scenedesmus obliquus metabolized 1-naphthalenesulfonic acid and partially used the sulfonate as a source of sulfur. The main metabolite, 1-hydroxy-2-naphthalenesulfonic acid, which was not metabolized further in the algal culture, was formed by hydroxylation of the substrate in position 1 and by migration of the sulfonic acid group to position 2 of the naphthalene ring (NIH shift). A smaller amount of 1-naphthalenesulfonic acid was desulfonated. The resulting 1-naphthol was mostly transformed into 1-naphthyl β-d-glucopyranoside. Received: 27 March 1996 / Revision received: 18 October 1996 / Accepted: 30 October 1996