High-affinity siderophore-mediated iron-transport system in the green alga Scenedesmus incrassatulus

Under iron-deficient conditions a high-affinity siderophore-mediated iron-transport system is induced in the green alga Scenedesmus incrassatulus R-83. Algal siderophores have a strong avidity for ferric versus ferrous iron, quickly oxidate Fe^II and efficiently solubilize Fe^III hydroxides. The entire ferrated molecule is translocated across the membrane by the specific transport system. The iron-uptake rate in Fe-deficient cells is higher at higher pH adjusted with bicarbonate in the medium, suggesting the presence of an inducible membrane-bound translocator. The iron-reduction step is not involved in uptake of ferrated siderophores. The total absorbed iron from siderophores is high and does not strongly depend on the nutritional status of cells, showing that the critical step for iron uptake is siderophore secretion rather than the membrane-bound iron-transport system.Abbreviations DFOB desferrioxamine B - EDDHA ethylenediamine di (o-hydroxyphenyl) acetic acid - BPDS bathophenanthrolinedisulphonate This work was supported by grant No. B-69 from the National Fund for Scientific Investigations at the Ministery of Education and Science in Bulgaria.     

Glutamate dehydrogenase of the unicellular green alga Scenedesmus acutus

The coenzyme-non-specific glutamate dehydrogenase (EC 1.4.1.3) from Scenedesmus acutus in inhibited by p-hydroxymercuribenzoate only in the deamination reaction. From this result and from its stability in the presence of urea it is concluded that this enzyme exhibits and equilibrium between three conformations: aminating and deaminating conformations induced by NADH-2-oxoglutarate and NAD⁺-glutamate, respectively, and the “native” conformation in the absence of substrates.     

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     

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.     

Volatile compounds of the green alga, Capsosiphon fulvescens

Essential oils extracted by static vacuum simultaneous distillation–extraction (V-SDE) and conventional SDE from a green alga, Capsosiphon fulvescens, were analyzed by gas chromatography (GC) and GC-mass spectrometry. The essential oil extracted with V-SDE and SDE has totals of 151 and 140 compounds, respectively. A combined total of 208 compounds were identified and 81 volatiles were common in both extracts. These included 8 acids, 28 alcohols, 34 aldehydes, 11 esters, 25 ketones, 19 aliphatic hydrocarbons, 43 branched hydrocarbons, 6 unsaturated hydrocarbons, 19 cyclic hydrocarbons, and 15 miscellaneous. The major volatile compounds of the oil extracted with V-SDE were (E)-β-ionone, octane, (E,E)-2,4-heptadienal, hexadecanoic acid, and β-cyclocitral, while those extracted with SDE were hexadecanoic acid, (Z,Z)-1,5-octadien-3-ol, tetradecanoic acid, (E,E)-2,4-heptadienal, and benzaldehyde. The characteristics of the flavor of the green alga might be contributed by the presence of a large number of aldehydes and ketones. Many of the compounds extracted with SDE might originate from thermal degradation and/or thermal interactions among the constituents in the alga during steam distillation.     

The effect of small-scale fluid motion on the green alga Scenedesmus quadricauda

The effect of shear flow on the green alga Scenedesmus quadricauda grown in Bristol's medium wastested. The shear flow was generated using a Couettetype rotating cylinder apparatus. Growth of Scenedesmus quadricauda, measured in terms ofchlorophyll a concentration, was inhibited underdifferent fluid motions. Inhibition was mostpronounced at high Reynolds number (Re) and thecorresponding mean rate of energy dissipation(). Algal growth was maximum during thestagnant fluid flow experiment. The flocs comprised of dead and living cells of algae formed as a resultof shear flow. Cell morphometry did not changeconsistently under different flow conditions but celldestruction was evident. A two step model isproposed, relating algal growth as a function of Re,and . The attenuation factor, _F for growth limiting conditions underdifferent fluid motions, was defined as the ratio of the algal growth rate constant to the maximum algalgrowth constant under stagnant fluid flowconditions.     

Bioaccumulation and toxicity of selenium compounds in the green alga Scenedesmus quadricauda

Background  

Selenium is a trace element performing important biological functions in many organisms including humans. It usually affects organisms in a strictly dosage-dependent manner being essential at low and toxic at higher concentrations. The impact of selenium on mammalian and land plant cells has been quite extensively studied. Information about algal cells is rare despite of the fact that they could produce selenium enriched biomass for biotechnology purposes.     

Possible toxic effects on Daphnia resulting from the green alga Scenedesmus obliquus

We cultured individuals of two Daphnia species and their hybrid on two different algae, Scenedesmus obliquus and Chlamydomonas globosa, in different concentrations. Our results suggest that culture conditions of S. obliquus can be such that the algal cells become toxic to Daphnia     

Heavy metals removal by the microalga Scenedesmus incrassatulus in continuous cultures

The microalga Scenedesmus incrassatulus was grown under continuous regime in the presence of chromium(VI), cadmium(II) and copper(II), as single metal species and as mixtures of two or three metals, in a laboratory scale system. We used an artificial wastewater with low free ion activities (as determined by MINEQL+) due to the presence of EDTA (a strong chelating agent) but with total concentrations not suitable for acceptable environments. Chromium(VI) and cadmium(II) had positive interaction that increased the removal percentages of both these metals; we could not, however, detect any interaction with copper(II). S. incrassatulus was able to remove all the tested metals to some extent (25-78%), but bivalent metals were not removed as efficiently as reported in batch cultures, probably due to the high pH values there recorded. Chromium(VI) was more efficiently removed in continuous cultures than in batch culture, because the uptake of chromate could be favored by actively growing algae.     

The effect of substances from different zooplankton species and fish on the induction of defensive morphology in the green alga Scenedesmus obliquus

The common green alga Scenedesmus obliquus may respond morphologicallyto the presence of natural enemies. Exposure to water-bornecues from the herbivorous zooplankton Ceriodaphnia reticulata,Daphnia galeata x hyalina, Daphnia magna and Daphnia pulicariastimulated the formation of protective colonies in S. obliquus.This response seemed strongly related to the amount of algaegrazed upon, because a highly significant correlation betweenthe amount of S. obliquus consumed and the induced colony formationwas found. However, when exposed to medium that had been inhabitedby the carnivorous zooplankton Bythotrephes longimanus and Leptodorakindtii, no colony formation occurred. A similar result wasobtained in two different experiments when S. obliquus was exposedto filtrate from cultures of the freshwater fish ide (Leuciscusidus) and perch (Perca fluviatilis). These results support thehypothesis that S. obliquus responds to a herbivorous zooplanktonchemical cue, rather than to a more general animal excretoryproduct. No support was obtained for the hypothesis that cuesfrom the enemy of their enemy could serve as signals to thealgae: despite the presence of filtrate from fish culture, filtratefrom Daphnia cultures induced the formation of colonies in S.obliquus. The biological activity seems to be linked to thealga–grazer interaction, which ensures a reliable cuethat evokes the morphological response of S. obliquus only whennecessary. The grazer-induced colony formation can be viewedas an adaptive reaction in habitats with variable grazing pressurefrom an assemblage of many different herbivores to pare downmortality through grazing.     

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.     

Physiological and structural effects of phosphorus starvation on the unicellular green alga Scenedesmus

The effects of phosphorus starvation on morphology and intracellular structure and on reactions related to the energy metabolism of the unicellular green alga Scenedesmus obtusiusculus (Chod.) were studied over a period of 96 h by employing transmission electron microscopy and various methods for measurement of physiological reactions. Increase in cell size and shape and in cell wall thickness are dominating features of phosphorus starvation. There is also an increase in number and size of starch granules and lipid globules and the internal structure of the cells appears successively disorganized. Shortage of phosphorus in the medium initially induces an increase of the adenylate pool whereas the energy charge value remains the same as for the controls. The photosynthetic and respiratory activities are high during incipient phosphorus starvation. After 24 h, as shortage of phosphorus becomes critical, the internal phosphorus reaches a low steady-state value, and this is also true for the adenylate energy charge. The total content of adenylates, however, peaks after 24 h of starvation and then decreases with increasing length of phosphorus starvation. Light-induced oxygen evolution appears not to be as much inhibited by a low phosphorus content in the cells as by the concomitant starch accumulation. The data indicate that the strategy for survival of the cells in a phosphorus-poor environment includes morphological and physiological changes that facilitate the transfer and adaption of the cells to environments with a more favourable supply of phosphorus, such as the often oxygen-poor but phosphorus-rich bottom zones.     

A metabolomics approach to assessing phytotoxic effects on the green alga Scenedesmus vacuolatus

The potential of metabolomics for toxicity analysis with synchronized algal populations during growth was explored in a proof of principle study. Low molecular weight compounds from hydrophilic and lipophilic extracts of algal populations of the unicellular green alga Scenedesmus vacuolatus were analyzed using gas chromatography-mass spectrometry (GC-MS) and subsequent multivariate analysis to identify time-related patterns. Algal metabolite responses were studied under control and exposure conditions for the photosystem II-inhibiting herbicide prometryn. To define the typical metabolic profile of control S. vacuolatus cultures seven time points over a growth period of 14 h were evaluated. The results show a clear time-related trend in metabolite levels and a distinct separation of exposed and reference algal populations. The results suggest an impairment of the energy metabolism associated with an activation of catabolic processes and a retardation of carbohydrate biosynthesis in treated algae. Metabolite results were compared to observation parameters, currently used in phytotoxicity assessment, showing that metabolites respond faster to exposure than algal growth. The potential of metabolomics for toxicity evaluation, especially to identify physiological markers and to detect effects at an early state of exposure, are discussed. Therefore, we suggest a metabolomics approach utilizing synchronous algal cultures to be a suitable future tool in ecotoxicology.     

Characterisation of two forms of phosphoribulokinase isolated from the green alga, Scenedesmus obliquus

Two forms of phosphoribulokinase from the alga, Scenedesmus obliquus, have been purified to homogeneity by DEAE-cellulose, Ultrogel AcA34 and hydroxyapatite chromatography. An active form of the enzyme is a dimer of identical 42,000-Mr subunits. A latent form of phosphoribulokinase, requiring incubation with dithiothreitol for activity, is of Mr 470,000 and apparent subunit composition X8Y4. The subunits X and Y are of Mr 39,000 and 42,000 respectively. The latent form of phosphoribulokinase is lost during DEAE-cellulose chromatography but this is prevented by NAD. Depolymerisation of the latent phosphoribulokinase to give the low-Mr form of the enzyme accompanied its activation by dithiothreitol. An algal protein with all the properties of thioredoxin stimulates activation of the latent phosphoribulokinase when incubated with low concentrations of dithiothreitol. The latent form of phosphoribulokinase predominates in the heterotrophically grown algae whilst under photoheterotrophic conditions equal amounts of both enzyme forms are present in algal extracts. This is consistent with the suggestion that light activation of phosphoribulokinase in vivo is also due to depolymerisation of the large-Mr latent form of the enzyme.     

Influence of selenium on toxicity of some heavy metals in the green alga Scenedesmus obliquus

The green alga Scenedesmus obliquus was incubated with various heavy metals (Cd2+, Zn2+, Mn2+, Ni2+) in presence/absence of selenium. S. obliquus exhibited higher rates of growth and some metabolic activities in cultures containing 0.1 mM Se than those containing only the heavy metals. The positive effect of Se was found in presence of Cd2+ while that in the case of presence of Ni2+ was less pronounced.     

Extraction of pigments and fatty acids from the green alga Scenedesmus obliquus (Chlorophyceae)

In this paper, the efficiency of pigment and fatty acid extraction from resistant algae using Scenedesmus obliquus as an example was examined. We found that adding quartz sand and solvent to freeze-dried algal material and subsequent extraction in an ultrasound bath for 90min at –4°C resulted in excellent extraction of these compounds. This extraction method was compared with a method regularly used for extraction of fatty acids and pigments, i.e. addition of solvents to algal material with subsequent incubation. Our extraction using the ultrasound and sand method was about twice as efficient as this method for both pigments and fatty acids. The ultrasound method is simple, extracts over 90% of the different substances in one step and conserves the relationships of pigments and fatty acids. In addition, no alteration- or breakdown products were observed with the new method. Thus, this method allows accurate quantitative extraction of both pigments and fatty acids from Scenedesmus obliquus and other algae. The method was also been found to be as effective for Cryptomonas erosa (Cryptophyceae), Cyclotella meneghiniana (Bacillariophyceae), Microcystis aeruginosa (Cyanophyceae), and Staurastrum paradoxum (Chlorophyceae, Desmidiaceae) and is thus applicable to a wide spectrum of algae.     

Purification and kinetic studies on a porphobilinogen deaminase from the unicellular green alga Scenedesmus obliquus

Porphobilinogen deaminase, the enzyme condensing four molecules of porphobilinogen, was isolated and purified from light grown Scenedesmus obliquus (wild type). The purification procedure included heat treatment, ammonium sulphate fractionation, gel filtration, high-resolution anion-exchange chromatography and hydrophobic interaction chromatography. The enzyme was purified 1368-fold, compared to the initial crude extract. Its final specific activity was 6812 units · (mg · protein)^?1 at pH 7.4 with a recovery of 44%. The relative molecular mass was 33000, as determined by Sephadex G-100 gel filtration, and 35900 by lithium dodecyl sulfate-polyacrylamide-gel electrophoresis, indicating that the enzyme is a monomer. Studies of initial reaction velocities showed a linear progress curve for hydroxymethylbilane formation and a hyperbolic dependence of the initial reaction rate on substrate concentration, consistent with a sequential displacement mechanism. Apparent kinetic constants (K _m and V _max) for the conversion of porphobilinogen to hydroxymethylbilane at 37 ° C, pH 7.4, were 79 μM and 176 pmol · min^?1, respectively. Variation of both V _max and K _max with pH indicated the presence of ionizable groups in the enzyme-substrate complex(es), showing a single ionization (pK 7.15) in V _max/K _m plots. A sharp pH-profile for V _max was interpreted as a positive cooperative proton dissociation. In spite of the two pathways existing for 5-aminolevulinate biosynthesis in Scenedesmus, currently there is no indication of the existence of two porphobilinogen deaminases or even of isoenzymes.     

Electron pathways involved in H(2)-metabolism in the green alga Scenedesmus obliquus

The green alga Scenedesmus obliquus is capable of both uptake and production of H(2) after anaerobic adaptation (photoreduction of CO(2) or photohydrogen production). The essential enzyme for H(2)-metabolism is a NiFe-hydrogenase with a [2Fe-2S]-ferredoxin as its natural redox partner. Western blot analysis showed that the hydrogenase is constitutively expressed. The K(m) values were 79.5 microM and 12.5 microM, determined with ferredoxin and H(2), respectively, as electron donor for the hydrogenase. In vitro, NADP(+) was reduced by H(2) in the presence of the hydrogenase, the ferredoxin and a ferredoxin-NADP reductase. From these results and considerations on the stoichiometry we propose that this light-independent electron transfer is part of the photoreduction of CO(2) in vivo. For ATP synthesis, necessary for the photoreduction of CO(2), light-dependent cyclic electron transfer around Photosystem (PS) I accompanies this 'dark reaction'. PS II fluorescence data suggest that (a) in S. obliquus H(2)-reduction might function as the anaerobic counterpart of the O(2)-dependent Mehler reaction, and (b) the presence of either a ferredoxin quinone-reductase or NAD(P)-dehydrogenase (complex I) in S. obliquus chloroplasts.