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PubMedDate not supplied

[Structural changes of chloroplasts and galactolipid contents in Chlorella cells during clinorotation].

Research of chloroplast ultrastructure in Chlorella cells grown during long-term period under clinorotation has been carried out. Different changes of the chloroplast structure, concerning both the amount of starch grains and stroma electron density as well as membrane system have been revealed. Occurrence of more significant bends of the thylakoids compared to the control and more loose arrangement of the thylakoids in a bunch were noted. The most significant changes were observed in a membrane system, in particular, appearance of the non-uniform expansions of the inter- and intrathylakoid spaces in Chlorella chloroplasts. Taking into account the role of galactolipids as important components of the photosynthetic membranes, the content of general galactolipids, monogalactosyldiacylglycerol and digalactosyldiacylglicerol in Chlorella cells was determined. It was assumed that statistically significant increase of the galactolipid content, especially of MDG, can probably be one of the reasons of membrane system reorganizations in Chlorella cells under altered gravity.

Chlorella↗
PubMed1977

[Regulation of glutamine metabolism in Chlorella pyrenoidosa. Regulation of glutamine synthetase activity by adenylic system components].

A decrease of glutamine synthetase (E. C. 6.3.1.2.) activity was observed under the assimilation of ammonium nitrogen in Chlorella. At the same time a decrease of ATP content in Chlorella cells took place. The ATP content was 7-fold decreased, while ADP and AMP contents were 4-fold and 3-fold increased respectively, after 15 min. of Chlorella incubation on "ammonium" medium. Further incubation for 45 min, resulted in gradual increase of ATP content and in decrease of ADP and AMP contents. The value of energy charge in ammonium assimilating Chlorella cells sharply decreased for first 15 min. of incubation and then it normalized gradually. The experiments with glutamine synthetase preparation, isolated from ammonium assimilating cells, have shown that ADP and AMP are strong inhibitors of the enzyme in the presence of Mg2+, and only ADP produces the inhibitory effect in the presence of Mn2+. No enzyme reactivation was observed after the transfer of ammonium assimilating cells into nitrogen-free medium or nitrate medium, the enzyme activity increasing at the expense of enzyme protein synthesis denovo.

Adenine Nucleotides↗
PubMedDate not supplied

[Study of the compatibility of certain higher plants and chlorella used as a bioregenerative human life support system].

The phototrophic component of a bioregenerative life support system should be of a multispecies structure. It should incorporate not only higher plants but also different algae. The paper discusses the studies concerning mutual effects of Chlorella and higher plants cultivated together in a closed atmosphere. It can be inferred from the studies that gaseous products of Chlorella did not exert a significant effect on the carbon dioxide consumption by wheat and radish plants or on their biomass increment. In turn, gaseous products of higher plants did not influence Chlorella growth. It can, therefore, be concluded that Chlorella and the above higher plants, when cultivated in a common atmosphere, do not inhibit each other and can be regarded as biologically compatible constituents of the photoautotrophic component of future bioregenerative life support systems.

Carbon Dioxide↗
PubMed1981

[Effect of pyridazinone herbicides on the photosynthetic electron transport chain of chloroplasts and Chlorella].

In order to establish the site of pyridazinone herbicides action on the photosynthetic electron transport chain, their effect on the photochemical activity of chloroplasts and Chlorella was studied. It was shown that these compounds similar to diuron inhibit the delta F of chloroplasts but enhance the delta F and cause the disappearance of slow transient processes in Chlorella and change the light-off time course of delta F both in Chlorella and in the chloroplasts. The inhibiting effect is observed at herbicide concentration of 5 x 10(-6) M and is maximal at 10(-4) M. However, in contrast to diuron the herbicides enhance the msec afterglow in Chlorella cells; besides, even at concentration as high as 10(-4) M they only partly block photosynthetic oxygen evolution and the light-induced change of pH. Pyridazinone herbicides retard the delay of light-off delta F at-196 degrees C more efficiently than diuron. It is suggested that the herbicides under study inhibit the photosynthetic electron transport chain, however less efficiently than diuron; the inhibiting effect is decreased in the following order: SAN 9785, SAN 6706, SAN 9789. The herbicides affect mainly the acceptor part of the photosystem 2, retarding the electron transport from the intermediary acceptor to plastoquinone. In addition these herbicides may also have other sites of action in the region of photosystem 2.

Chlorella↗
PubMed1997

Stable Transformation of Chlorella: Rescue of Nitrate Reductase-Deficient Mutants with the Nitrate Reductase Gene

Unicellular green algae, like Chlorella, offer a potentially useful system for the expression of heterologous proteins. However, the development of Chlorella as a bioreactor has been delayed owing to the lack of a stable transformation technique. Here we report on the use of micro-projectile bombardment to introduce the nitrate reductase (NR) gene from Chlorella vulgaris into NR-deficient Chlorella sorokiniana mutants, resulting in stable transformants. The stable transformants were able to grow on nitrate medium after repeated passages between selective and nonselective medium and exhibited inducible nitrate reductase activity comparable to that of wild-type cells. Southern analysis suggests homologous recombination occurs with insertion of the wild type gene into the mutated gene and that the genes of the two Chlorellaspecies used are very similar. Specific RNase protection assays, selecting for a poorly conserved region of the gene, identified the presence of the C. vulgaris NR transcript only in the transformed C. sorkiniana mutant and not in the mutant.

Journal Article↗
PubMed1989

Survival, growth and reproduction of Daphnia carinata (Crustacea: Cladocera) exposed to chronic cadmium stress at different food (Chlorella) levels.

In nature, organisms have to respond to a diversity of factors acting simultaneously. The present investigation was conducted to study whether changes in food (Chlorella) levels could modify the chronic toxicity of cadmium on the various life-history parameters, such as survivorship, longevity, life expectancy, fecundity, age at first reproduction, R(0), T, r and growth rates of the cladoceran Daphnia carinata. The study indicated that at low food levels (0.5 x 10(6) cells ml(-1) Chlorella), cadmium concentrations in the range of 27-162 microg litre(-1) reduced these life-history parameters by 50% (EC(50)). At medium food levels (1.5 x 10(6) ml(-1) Chlorella) the EC(50) of cadmium was in the range of 51-127 microg litre(-1). At high food levels (4.5 x 10(6) cells ml(-1) Chlorella), the toxic effect of cadmium was greatly reduced. The decreases in survival, growth and reproduction of D. carinata at high cadmium-low food levels affected the fitness parameter 'r'. The study emphasises the need to include reproductive parameters other than mere survival in toxicity bioassays. The study also stresses the need to incorporate in laboratory tests other relevant factors that might modify pollutant toxicity.

Journal Article↗
PubMed1999

Chlorella viruses as a source of novel enzymes.

A special advantage has been conferred upon Chlorella cells as tools in biotechnology when viruses (Phycodnaviridae) infecting Chlorella cells were discovered and isolated. The viruses are large icosahedral particles (150-200 nm in diameter), containing a giant, 330-380 kbp long, linear dsDNA genome. Recently, the nucleotide sequence of the 330,740-bp genome of PBCV-1, the prototype virus of Phycodnaviridae, was determined, and up to 702 open reading frames (ORFs) were identified along the genome. The possible genes present include those encoding a variety of enzymes involved in the modification of DNA, RNA, protein and polysaccharides as well as those involved in the metabolism of sugars, amino acids, lipids, nucleotides and nucleosides. Many of these genes are actually expressed during viral infection, with functional enzymes detected in the host cytoplasm or incorporated into the virion. The successful utilization of these viral enzymes as various DNA restriction and modification enzymes (Cvi enzymes) that are now commercially available is well documented. Also noteworthy are virion-associated chitinase and chitosanase activities that have potentially important applications in the recycling of natural resources. The virions of Chlorella viruses contain more than 50 different structural proteins, ranging in size from 10 to 200 kDa. Some of these proteins may be replaced with useful foreign proteins using recombinant DNA technology. The proteins of interest can be recovered easily from the viral particles, and collected by centrifugation after complete lysis of the host Chlorella cells.

Journal Article↗
PubMed1990

Functional expression of the Chlorella hexose transporter in Schizosaccharomyces pombe.

Schizosaccharomyces pombe cells were transformed with an S. pombe expression vector containing a full-length cDNA of the Chlorella hexose transporter. The transformed cells accumulated 3-O-methylglucose up to 10-fold, whereas wild-type S. pombe and control transformants could only equilibrate this sugar analogue. In a pH-jump experiment, in which extracellular pH was lowered by 1.9 units, the accumulation ratio was increased in transformed cells but not in control cells. This result indicates that the gene product, Chlorella H+/glucose-symporter protein, and a pH gradient suffice for active sugar uptake. Km values for glucose, 6-deoxyglucose, and 3-O-methylglucose of 1.5 x 10(-5) M, 2.7 x 10(-4) M, and 1.0 x 10(-3) M, respectively, were identical in Chlorella and in S. pombe cells transformed with Chlorella cDNA and approximately 100-fold lower than those of the endogenous transport system of S. pombe.

Journal Article↗
PubMed1999

Footprinting of Chlorella virus DNA ligase bound at a nick in duplex DNA.

The 298-amino acid ATP-dependent DNA ligase of Chlorella virus PBCV-1 is the smallest eukaryotic DNA ligase known. The enzyme has intrinsic specificity for binding to nicked duplex DNA. To delineate the ligase-DNA interface, we have footprinted the enzyme binding site on DNA and the DNA binding site on ligase. The size of the exonuclease III footprint of ligase bound a single nick in duplex DNA is 19-21 nucleotides. The footprint is asymmetric, extending 8-9 nucleotides on the 3'-OH side of the nick and 11-12 nucleotides on the 5'-phosphate side. The 5'-phosphate moiety is essential for the binding of Chlorella virus ligase to nicked DNA. Here we show that the 3'-OH moiety is not required for nick recognition. The Chlorella virus ligase binds to a nicked ligand containing 2',3'-dideoxy and 5'-phosphate termini, but cannot catalyze adenylation of the 5'-end. Hence, the 3'-OH is important for step 2 chemistry even though it is not itself chemically transformed during DNA-adenylate formation. A 2'-OH cannot substitute for the essential 3'-OH in adenylation at a nick or even in strand closure at a preadenylated nick. The protein side of the ligase-DNA interface was probed by limited proteolysis of ligase with trypsin and chymotrypsin in the presence and absence of nicked DNA. Protease accessible sites are clustered within a short segment from amino acids 210-225 located distal to conserved motif V. The ligase is protected from proteolysis by nicked DNA. Protease cleavage of the native enzyme prior to DNA addition results in loss of DNA binding. These results suggest a bipartite domain structure in which the interdomain segment either comprises part of the DNA binding site or undergoes a conformational change upon DNA binding. The domain structure of Chlorella virus ligase inferred from the solution experiments is consistent with the structure of T7 DNA ligase determined by x-ray crystallography.

Amino Acid Sequence↗
PubMed1995

Photosystem II Excitation Pressure and Development of Resistance to Photoinhibition (I. Light-Harvesting Complex II Abundance and Zeaxanthin Content in Chlorella vulgaris).

The basis of the increased resistance to photoinhibition upon growth at low temperature was investigated. Photosystem II (PSII) excitation pressure was estimated in vivo as 1 - qp (photochemical quenching). We established that Chlorella vulgaris exposed to either 5[deg]C/150 [mu]mol m-2 s-1 or 27[deg]C/2200 [mu]mol m-2 s-1 experienced a high PSII excitation pressure of 0.70 to 0.75. In contrast, Chlorella exposed to either 27[deg]C/150 [mu]mol m-2 s-1 or 5[deg]C/20 [mu]mol m-2 s-1 experienced a low PSII excitation pressure of 0.10 to 0.20. Chlorella grown under either regime at high PSII excitation pressure exhibited: (a) 3-fold higher light-saturated rates of O2 evolution; (b) the complete conversion of PSII[alpha] centers to PSII[beta] centers; (c) a 3-fold lower epoxidation state of the xanthophyll cycle intermediates; (d) a 2.4-fold higher ratio of chlorophyll a/b; and (e) a lower abundance of light-harvesting polypeptides than Chlorella grown at either regime at low PSII excitation pressure. In addition, cells grown at 5[deg]C/150 [mu]mol m-2 s-1 exhibited resistance to photoinhibition comparable to that of cells grown at 27[deg]C/2200 [mu]mol m-2 s-1 and were 3- to 4-fold more resistant to photoinhibition than cells grown at either regime at low excitation pressure. We conclude that increased resistance to photoinhibition upon growth at low temperature reflects photosynthetic adjustment to high excitation pressure, which results in an increased capacity for nonradiative dissipation of excess light through zeaxanthin coupled with a lower probability of light absorption due to reduced chlorophyll per cell and decreased abundance of light-harvesting polypeptides.

Journal Article↗
PubMed1966

Sulfolipid metabolism in chlorella.

When S-deficient cells of Chlorella cllipsoidea were incubated in radio-sulfate in light or in aerobic darkness for 1 hour, equal amounts of radioactivity were found in sulfolipid and glutathione but none was detected in sulfoquinovosyl glycerol which is one of the major S-compounds in this alga. No assimilation of radiosulfate was observed under anaerobic darkness.To elucidate the function of sulfolipid in algal cells uniformly (35)S-labeled Chlorella cells were transferred to S-deficient culture medium or unlabeled normal culture medium and the changes of radioactivity in sulfolipid and the related compounds were followed. A) On incubating (35)S-labeled algal cells in S-deficient medium under photosynthetic conditions, the amounts of radioactivity in sulfate, sulfoquinovosyl glycerol and sulfolipid decreased rapidly. B) When (35)S-labeled cells were cultured photoautotrophically in unlabeled medium, no decrease of radioactivity was observed in sulfoquinovosyl glycerol and sulfolipid. C) A decrease of (35)S-sulfolipid and an increase of (35)S-sulfoquinovosyl glycerol were observed when the uniformly (35)S-labeled algal cells were illuminated in CO(2)-free air.When S-deficient Chlorella cells were incubated in (35)S-sulfolipid under photosynthetic conditions, significant radioactivity was found in the insoluble fraction of the cells. A similar result was observed when normal Chlorella cells were incubated in (14)C-sulfolipid and CO(2)-free air.It is inferred from these observations that sulfolipid is a reservoir of sulfur and carbon compounds.In order to ascertain if the sulfolipid is involved in the mechanism of photosynthetic oxygen evolution, the rate of photosynthesis was measured during the incubation of (35)S-labeled cells in a S-deficient medium. Parallelism was not observed between the rate of photosynthetic activity and the decrease of sulfolipid.

Journal Article↗
PubMed1975

Acetyl coenzyme a-glutamate acetyltransferase and N-acetylornithine-glutamate acetyltransferase of chlorella.

The enzymic formation of acetylglutamate has been studied in Chlorella vulgaris extracts. Acetyl CoA and N(2)-acetyl-l-ornithine served as substrates for glutamate acetylation whereas acetylphosphate, N(5)-acetyl-l-ornithine, and N(2)-acetyl-2,4-diamino butyrate were ineffective. Acetyl CoA-glutamate transacetylase and acetylornithine-glutamate transacetylase activities have been purified over 180-fold with no indication of any separation of activities. The acetyl CoA activity was more labile than acetylornithine activity so that preparations having acetylornithine-glutamate transacetylase activity but no acetyl CoA-glutamate transacetylase activity were obtained. The two acetylating activities appear to be properties of one enzyme with one portion more easily denatured.Both acetylating activities had pH optima between 8 and 8.5. The Km value for glutamate was 3 mm for both activities. The Km values were 0.2 mm for acetylornithine and 3.2 mm for acetyl CoA. Arginine inhibited acetyl CoA-glutamate transacetylase (Ki = 0.94 mm) and acetylglutamate phosphokinase (Ki = 0.5 mm) but had no effect on acetylornithine-glutamate transacetylase. The lack of an inhibitory effect of proline on any of the three enzymic activities indicates that acetylglutamate is not a normal intermediate in proline biosynthesis. Growth of Chlorella with arginine as a nitrogen source had no effect on enzyme levels, showing that end-product repression is not a control factor in arginine biosynthesis in Chlorella. In Chlorella, arginine controls its own biosynthesis by inhibiting acetylglutamate phosphokinase and controls the level of acetylated intermediates by inhibiting acetyl CoA-glutamate transacetylase.

Journal Article↗
PubMed1989

Characterization of Nitrate Reductase Deficient Mutants of Chlorella sorokiniana.

After x-ray irradiation, 13 mutants of Chlorella sorokiniana incapable of using NO(3) (-) as N source were isolated using a pinpoint method. Using immunoprecipitation and Western blot assays, no nitrate reductase was found in five strains while in eight mutants the enzyme was detected. The latter strains contained different patterns of nitrate reductase partial reactions. All isolates were of the nia-type as indicated by the inducibility of purine hydroxylase I and by complementation of nitrate reductase activity in the Neurospora crassa mutant Nit-1. A restoration of NADP-nitrate reductase in Nit-1 was also obtained with NH(4) (+)-grown cells indicating that Mo-cofactor is constitutive in Chlorella. Complementation experiments among the Chlorella mutants resulted in restoration of NADH-nitrate reductase activity. The characteristics of some of the Chlorella mutants are discussed in view of an improper orientation of Mo-cofactor in the residual nitrate reductase protein.

Journal Article↗
PubMed1989

Glycolate Metabolism in Low and High CO(2)-Grown Chlorella pyrenoidosa and Pavlova lutheri as Determined by O-Labeling.

Photorespiration in Chlorella pyrenoidosa Chick. was assayed by measuring (18)O-labeled intermediates of the glycolate pathway. Glycolate, glycine, serine, and excreted glycolate were isolated and analyzed on a gas chromatograph/mass spectrometer to determine isotopic enrichment. Rates of glycolate synthesis were determined from (18)O-labeling kinetics of the intermediates, pool sizes, derived rate equations, and nonlinear regression techniques. Glycolate synthesis was higher in high CO(2)-grown cells than in air-grown cells when both were assayed under the same O(2) and CO(2) concentrations. Synthesis of glycolate, for both types of cells, was stimulated by high O(2) levels and inhibited by high CO(2) levels. Glycolate synthesis in 1.5% CO(2)-grown Chlorella, when exposed to a 0.035% CO(2) atmosphere, increased from about 41 to 86 nanomoles per milligram chlorophyll per minute when the O(2) concentration was increased from 21% to 40%. Glycolate synthesis in air-grown cells increased from 2 to 6 nanomoles per milligram chlorophyll per minute under the same gas levels. Synthesis was undetectable when either the O(2) concentration was lowered to 2% or the CO(2) concentration was raised to 1.5%. Glycolate excretion was also sensitive to O(2) and CO(2) concentrations in 1.5% CO(2)-grown cells and the glycolate that was excreted was (18)O-labeled. Air-grown cells did not excrete glycolate under any experimental condition. Indirect evidence indicated that glycolate may be excreted as a lactone in Chlorella. Photorespiratory (18)O-labeling kinetics were determined for Pavlova lutheri, which unlike Chlorella and higher plants did not directly synthesize glycine and serine from glycolate. This alga did excrete a significant proportion of newly synthesized glycolate into the media.

Journal Article↗
PubMed1987

Chelating Properties of Extracellular Polysaccharides from Chlorella spp.

Chlorella stigmatophora LB 993 was grown in artificial seawater under controlled conditions. The production of cell wall polysaccharides attached to the cells and dissolved in the growth medium was monitored during algal growth. Preliminary characterization of the dissolved polysaccharides of C. stigmatophora and other Chlorella species is presented. The capacity of dissolved polysaccharides of C. stigmatophora to bind toxic heavy metals was also studied and compared with that of polysaccharides produced by other marine Chlorella species. The differences in metal-complexing capacity observed for dissolved polysaccharides obtained from various Chlorella species is attributable to differences in the composition of the polysaccharides, notably the uronic acids content.

Journal Article↗
PubMed1990

Preparation of allergen extracts from the green alga Chlorella. Studies of growth variation, batch variation, and partial purification.

Allergen extracts were prepared from different raw materials of three strains of the unicellular green alga Chlorella (C. vulgaris, C. homosphaera and C. saccharophila). Growth variation, batch variation and composition of partially purified extracts were studied by analyzing total protein and carbohydrate content, protein and IgE-binding patterns and allergenic potency. Chlorella allergens were produced in the same proportions in early and late exponential growth phases but appeared partly degraded in the stationary phase. Uniform growth and extraction from Chlorella cells in the late exponential phase were demonstrated. Partially purified extracts from cells in late exponential phase contained 50% protein of the dry weight compared to 20% in crude extracts. The carbohydrate content was 20-30% of the dry weight of both crude and purified extracts. The allergenic potency of the purified extracts was 150-340% of the crude extracts, as assayed by RAST inhibition. The purified C. vulgaris extract showed a reduced IgE binding compared to crude extracts, while C. homosphaera and C. saccharophila extracts exhibited consistent IgE binding and are therefore suitable for in vivo and in vitro studies of allergenic properties of Chlorella.

Allergens↗
PubMed2002

Photosynthetic productivity of conical helical tubular photobioreactor incorporating Chlorella sorokiniana under field conditions.

The photosynthetic performance of a conical, helical tubular photobioreactor (HTP) incorporating Chlorella sorokiniana was investigated under conditions of high temperature and light intensity during midsummer in an outdoor environment. Although the culture medium temperature exceeded 40 degrees C for approximately 5 h each day, peaking at 47.5 degrees C under sunny conditions, a photosynthetic productivity of 30.0 g x m(-2) (installation area) x day(-1) and a photosynthetic efficiency of 8.66% [photosynthetically active radiation (PAR), 400-700 nm] were achieved. A maximum photosynthetic productivity of 33.2 g x m(-2) x day(-1) was achieved on a sunny day, when solar energy input was also maximal (11.5 MJ x m(-2) x day(-1) [PAR]). On the other hand, a maximum photosynthetic efficiency of 9.54% was obtained on a day that was rainy in the morning and cloudy in the afternoon, and there was relatively little solar energy input. The average daily photosynthetic efficiency over the two culture periods (August 4 to 7 and August 10 to 13, 1999) was 7.25%. Thus, a high level of photosynthetic performance was achieved in the conical HTP incorporating Chlorella sorokiniana despite the fact that culture medium temperature was not controlled. The use of Chlorella sorokiniana in the conical HTP should be a good choice to produce microalgal biomass during the summer under field conditions.

Biomass↗
PubMed2002

Combined effects of algal (Chlorella vulgaris) density and ammonia concentration on the population dynamics of Ceriodaphnia dubia and Moina macrocopa (Cladocera).

Ammonia is a natural variable in ponds and lakes. Although an important source of nitrogen for microalgae, at high concentrations ammonia can affect the density and diversity of cladocerans. Using the cladocerans Ceriodaphnia dubia, and Moina macrocopa, the effect of nominal concentrations of ammonium chloride under acute and chronic exposures at different levels of algal food was tested. Regardless of food level, C. dubia was more sensitive than M. macrocopa to ammonia. In the absence of food, the median lethal concentration of ammonia (LC(50) 24 h) for C. dubia was (112 mg L(-1)) less than half that of M. macrocopa (232 mg L(-1)). When algal food (0.5 x 10(6) and 1.5 x 10(6) cells ml(-1) of Chlorella) was used, the LC(50) values were much higher for both cladoceran species. Based on the population growth studies, it was found that in controls of M. macrocopa an increase in the availability of Chlorella from 0.5 to 1.5 x 10(6) cells mL(-1) led to an increase in the maximum density from 4.7 +/- 0.2 to 16.4 +/- 1.2 ind.mL(-1), while in C. dubia the peak population density decreased from 7.9 +/- 0.6 to 5.0 +/- l.0 ind.mL(-1). An increase in ammonia concentration (10 to 40 mg L(-1) for C. dubia and 20 to 120 mg L(-1) for M. macrocopa) resulted in a corresponding decrease in peak population densities of the tested cladocerans. The rate of population increase (r) values for M. macrocopa in the controls ranged from 0.21 +/- 0.001 and 0.25 +/- 0.02 at 0.5 and 1.5 x 10(6) cells mL(-1) of Chlorella, respectively. The corresponding values of C. dubia in controls were 0.21 +/- 0.004 and 0.18 +/- 0.01. At 0.5 x 10(6) cells mL(-1) of algal food, the r values became negative under 40 and 120 mg L(-1) of ammonia for C. dubia and M. macrocopa, respectively. The role of algal food in ammonia toxicity to cladocerans was discussed. (c) 2002 Elsevier Science (USA).

Ammonia↗