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PubMed2005

Antioxidant and antiproliferative activities of Spirulina and Chlorella water extracts.

Liver fibrosis is a chronic liver disease that will further develop to cirrhosis if severe damage continues to form. A potential treatment for liver fibrosis is to inhibit activated hepatic stellate cell (HSC) proliferation and, subsequently, to induce HSC apoptosis. It has been reported that antioxidants are able to inhibit the proliferation of HSCs. In this study, the aqueous extract of spirulina was chosen as the source of antioxidant to investigate the inhibitory effect on the proliferation of HSC. The growth inhibitory effects of aqueous spirulina and chlorella extract on human liver cancer cells, HepG2, were also studied and compared in pairs. Results indicated that the total phenol content of spirulina was almost five times greater than that of chlorella (6.86 +/- 0.58 vs 1.44 +/- 0.04 mg tannic acid equivalent/g of algae powder, respectively). The antioxidant activity of spirulina determined by the ABTS*+ method was higher than chlorella (EC50: 72.44 +/- 0.24 micromol of trolox equivalent/g of spirulina extract vs 56.09 +/- 1.99 micromol of trolox equivalent/g of chlorella extract). Results of DPPH* assay also showed a similar trend as the ABTS*+ assay (EC50: 19.39 +/- 0.65 micromol of ascorbic acid equivalent/g of spirulina extract vs 14.04 +/- 1.06 micromol of ascorbic acid equivalent/g of chlorella extract). The aqueous extracts of these two algae both showed antiproliferative effects on HSC and HepG2, but spirulina was a stronger inhibitor than chlorella. Annexin-V staining showed that aqueous extract of spirulina induced apoptosis of HSC after 12 h of treatment. In addition, the aqueous extract of spirulina triggered a cell cycle arrest of HSC at the G2/M phase.

Animals↗
PubMed2002

Phylogenetic relationships and taxonomic position of Chlorella-like isolates from low pH environments (pH < 3.0).

BACKGROUND: Little is known about phytoplankton communities inhabiting low pH environments such as volcanic and geothermal sites or acidic waters. Only specialised organisms are able to tolerate such extreme conditions. There is, thus, low species diversity. We have characterised the previously isolated acid tolerant Chlorella-like microalgae Viridiella fridericiana and Chlorella protothecoides var. acidicola by microscopical and biomolecular methods in order to assess their phylogenetic relationships. RESULTS: Both isolates belong to the trebouxiophycean lineage of chlorophytes. 18S and ITS1 sequence data clearly confirm that Viridiella fridericiana constitutes a new genus apart from the morphologically similar and likewise acid tolerant microalga Chlorella saccharophila. Chlorella protothecoides var. acidicola on the other hand is not a variety of Chlorella protothecoides but falls within a heterogeneous cluster consisting of Nannochloris, "Chlorella" spec. Yanaqocha, and Koliella, and is most closely related to algae which were also isolated from extreme environments. CONCLUSIONS: The distribution of acid tolerant strains in the 18S rRNA tree shows that acquisition of acid tolerance was unlikely a monophyletic event in green microalgae. We propose that different strains have independently adapted to extreme environments. Some of them have spread worldwide and were able to colonise other extreme habitats. Considering the problems of successfully isolating acid tolerant strains, acidic soils could represent an unsuspected source of biological diversity with high potential for biotechnological utilisations.

Adaptation, Physiological↗
PubMed1995

Detection of hydroxyl radical in intact cells of Chlorella vulgaris.

Using ESR with 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin-trapping reagent, we measured the levels of free radical species generated from living cells of Chlorella vulgaris var. vulgails (IAM C-534). To investigate the production of free radicals in the living Chlorella vulgaris cells, the influence of DMPO toward the intact cells of the Chlorella vulgaris using the O2 evolution rate was first studied as a guide. Since the O2 evolution rate was not changed by DMPO, it was judged that DMPO has no toxicity toward the intact cells of Chlorella vulgaris. Only hydroxyl radicals (.OH) were detected as the DMPO-OH adduct in the suspension of intact cells of Chlorella vulgaris irradiated with visible light. Moreover, since production of .OH was inhibited by some hydroxyl radical scavengers such as KI and ethanol, production of .OH was proved to be due to hydroxyl radicals. It was also clear that the intensity of .OH increased with increasing irradiation intensity of visible light. Therefore, it was suggested that .OH might be one of the photoinhibition factors of the intact Chlorella vulgaris cells in severe light conditions.

Chlorella↗
PubMed1996

Evaluation of active oxygen effect on photosynthesis of Chlorella vulgaris.

The relationship between O2 and an active oxygen scavenging system in Chlorella vulgaris var.vulgaris (IAM C-534) was investigated. When Chlorella vulgaris was exposed to 2% O2, only traces of active oxygen scavenging enzymes were found. When the Chlorella vulgaris was treated with 20% or 50% O2, it was shown that the level of enzyme activity increased as the O2 concentration increased. An increase in enzyme activity was not found in any specific enzyme but in all of the enzymes, but the level of glutathione and ascorbate remained the same in all the cases. In addition, the photosynthetic efficiency also decreased as the concentration of O2 was increased. These results suggest that an O2 enriched environment can lead to an increase in the production of active oxygen species such as O2.- and H2O2 and to a decrease in the photosynthetic efficiency in Chlorella vulgaris. The hydroxyl radical (.OH) was detected directly in the Chlorella vulgaris suspension with a spin trapping reagent. It was also clear that the increase in the .OH intensity as the visible light intensity increased was unrelated to the O2 concentration. It was suggested that the conditions for producting .OH and the other active oxygen species were different, and that two types of oxygen stress should exist in the Chlorella vulgaris.

Antioxidants↗
PubMed2000

Algal-lytic activities encoded by Chlorella virus CVK2.

Using a halo assay with E. coli lysates expressing Chlorella virus CVK2 genes on a cosmid contig, two different algal-lytic activities against Chlorella strain NC64A cells were found to be encoded on the CVK2 genome. The gene for vAL-1, one of the two activities, encoded a 349-aa ORF, which was homologous to PBCV-1 A215L and CVN1 CL-2. The vAL-1 gene was expressed at relatively early stages of the virus life cycle; transcripts and translation products appeared at 60 and 90 min postinfection, respectively. The vAL-1 protein was not incorporated into the viral particles but remained in the cell lysate, suggesting a role in the digestion of the cell wall before viral release at the final stage of infection. Cell wall materials isolated from Chlorella strain NC64A cells were digested by vAL-1 and degradation products were detected on TLC. In addition to Chlorella strain NC64A, vAL-1 lysed cells of four C. vulgaris strains as well as Chlorella sp. SAG-241-80.

Amino Acid Sequence↗
PubMed1976

The cryopreservation of Chlorella. 1. Interactions of rate of cooling, protective additive and warming rate.

The cryoprotective additives glycerol and dimethylsulphoxide were found to be toxic to Chlorella cells at concentrations greater then 2.5% w/v. Polyvinylpyrrolidone, was not damaging up to a concentration of 15%w/v. Chlorella 211/7a had a recovery rate greater than 95% at all rates of cooling studied. With Chlorella 211/8h the survival was lower than 0.1% at all rates examined. The addition of dimethylsulphoxide (5% w/v) to Chlorella 211/8h increased the recovery, particularly at the faster rates of cooling; with polyvinylpyrrolidone (10% w/v) there was an optimum range of cooling rate. Cells of Chlorella 211/7a from the exponential phase of growth were found to be damaged both by a temperature reduction from 25 degrees C to 0 degrees C (thermal shock) and by freezing and thawing. In contrast cells from the stationary phase of growth were resistant to these stresses.

Cell Division↗
PubMed2006

Reliability of microbial surrogate Chlorella for determining Cryptosporidium oocysts removal.

The green alga, Chlorella, abundant in water sources and similar in size to the Cryptosporidium oocysts, may be a suitable candidate to evaluate the removal capacities of the waterworks for Cryptosporidium oocysts. The zeta potentials of Cryptosporidium oocysts in different water samples were more negative than Chlorella. Although the hydrophobicities of Chlorella were similar to Cryptosporidium oocysts, the experimental collision efficiencies for Cryptosporidium oocysts were higher than for Chlorella. It is indicated that Cryptosporidium oocysts would consume more coagulants during coagulation, with higher removal rate in the filtration process than Chlorella.

Animals↗
PubMed2005

Chronic toxicity of uranium to a tropical green alga (Chlorella sp.) in natural waters and the influence of dissolved organic carbon.

The chronic toxicity (72-h cell division rate) of uranium (U) to the unicellular alga, Chlorella sp., was assessed in natural Magela Creek water (NMCW) to provide data for the derivation of a site-specific water quality trigger value for U in Magela Creek, NT, Australia. In addition, the data were compared to those for Chlorella sp. when tested for U toxicity using synthetic Magela Creek water (SMCW), which simulates the inorganic composition of Magela Creek water and contains no organic component. Based on one rangefinder and four definitive toxicity tests, concentrations causing a 50% inhibition of algal growth after 72 h exposure (72 h IC50s) ranged between 137 and 238 microg/LU, no-observed-effect concentrations (NOECs) from 72 to 157 microg/LU and lowest-observed-effect concentrations (LOECs) from 120 to 187 microg/LU. Based on these data, Chlorella sp. was the second most sensitive organism to U of five local species that have been assessed using NMCW. The U toxicity data for Chlorella sp. were incorporated with existing data for the four other species to derive a site-specific guideline value for Magela Creek that is protective of 99% of species of 6 microg/L. The toxicity of U to Chlorella sp. in NMCW was approximately two to four times lower than in SMCW. Based on geochemical speciation modelling, this difference corresponded to a four-fold decrease in the proportion of free uranyl ion (UO2(2+)) in NMCW compared to SMCW, most likely due to the presence of dissolved organic carbon (DOC) in NMCW. Relatively, large variability in U toxicity across the tests conducted in NMCW was found to be inversely related to DOC concentration (r2 = 0.996, n = 4, P = 0.002). Speciation modelling indicated that the increase in DOC was associated with an increase in the proportion of U complexed with DOC (r2 = 0.986, n = 4, P < 0.001) and a decrease in the proportion of the UO2(2+) (r2 = 0.989, n = 4, P = 0.006). When the proportion of UO2(2+) was regressed against U toxicity, a very strong, positive relationship was observed (r2 = 1, n = 4, P < 0.001). The results indicate that the bioavailability and toxicity of U is highly influenced by dissolved organic matter and that the relationship should be further quantified.

Carbon↗
PubMed2005

Preventing dyslipidemia by Chlorella pyrenoidosa in rats and hamsters after chronic high fat diet treatment.

The effects of Chlorella pyrenoidosa on serum lipid profiles, after concomitant long-term treatment of high-fat diet (HFD) in rats and hamsters was studied. Wistar rats and Syrian hamsters were fed with or without various concentrations of Chlorella pyrenoidosa contained high-fat diet (CHFD) for 2, 4 and 8 weeks prior to assay of serum lipids. Fasting triglycerides, total cholesterol, and LDL cholesterol as well as HDL cholesterol levels in high-fat diet treated rats and hamster were determined. Results showed that triglycerides, total cholesterol and LDL cholesterol levels in HFD treated rats and hamsters were increased from the normal rodent diet (NRD) treated controls after 2, 4, and 8-week treatments. However, the presence of Chlorella pyrenoidosa in high-fat diets significantly decreased the levels of triglycerides, total cholesterol and LDL cholesterol with comparison to HFD group in rats and hamsters. The total cholesterol/HDL ratios, an indication of occurrence of coronary heart disease, were decreased in all CHFD treated grouped rats and hamsters which suggests administration of Chlorella pyrenoidosa could lower the occurring risk of heart diseases. In conclusion, Chlorella pyrenoidosa has the ability to prevent dyslipidemia in chronic high-fat fed animals and could be potential in use to prevent intestinal absorption of redundant lipid from our daily intake and subsequently to prevent hyperlipidemia as well as atherosclerosis.

Animals↗
PubMed2006

Sequence and annotation of the 369-kb NY-2A and the 345-kb AR158 viruses that infect Chlorella NC64A.

Viruses NY-2A and AR158, members of the family Phycodnaviridae, genus Chlorovirus, infect the fresh water, unicellular, eukaryotic, chlorella-like green alga, Chlorella NC64A. The 368,683-bp genome of NY-2A and the 344,690-bp genome of AR158 are the two largest chlorella virus genomes sequenced to date; NY-2A contains 404 putative protein-encoding and 7 tRNA-encoding genes and AR158 contains 360 putative protein-encoding and 6 tRNA-encoding genes. The protein-encoding genes are almost evenly distributed on both strands, and intergenic space is minimal. Two of the NY-2A genes encode inteins, the large subunit of ribonucleotide reductase and a superfamily II helicase. These are the first inteins to be detected in the chlorella viruses. Approximately 40% of the viral gene products resemble entries in the public databases, including some that are unexpected for a virus. These include GDP-d-mannose dehydratase, fucose synthase, aspartate transcarbamylase, Ca(++) transporting ATPase and ubiquitin. Comparison of NY-2A and AR158 protein-encoding genes with the prototype chlorella virus PBCV-1 indicates that 85% of the genes are present in all three viruses.

Amino Acid Sequence↗
PubMed2006

Sequence and annotation of the 314-kb MT325 and the 321-kb FR483 viruses that infect Chlorella Pbi.

Viruses MT325 and FR483, members of the family Phycodnaviridae, genus Chlorovirus, infect the fresh water, unicellular, eukaryotic, chlorella-like green alga, Chlorella Pbi. The 314,335-bp genome of MT325 and the 321,240-bp genome of FR483 are the first viruses that infect Chlorella Pbi to have their genomes sequenced and annotated. Furthermore, these genomes are the two smallest chlorella virus genomes sequenced to date, MT325 has 331 putative protein-encoding and 10 tRNA-encoding genes and FR483 has 335 putative protein-encoding and 9 tRNA-encoding genes. The protein-encoding genes are almost evenly distributed on both strands, and intergenic space is minimal. Approximately 40% of the viral gene products resemble entries in public databases, including some that are the first of their kind to be detected in a virus. For example, these unique gene products include an aquaglyceroporin in MT325, a potassium ion transporter protein and an alkyl sulfatase in FR483, and a dTDP-glucose pyrophosphorylase in both viruses. Comparison of MT325 and FR483 protein-encoding genes with the prototype chlorella virus PBCV-1 indicates that approximately 82% of the genes are present in all three viruses.

Aquaglyceroporins↗
PubMed2006

Chlorella viruses.

Chlorella viruses or chloroviruses are large, icosahedral, plaque-forming, double-stranded-DNA-containing viruses that replicate in certain strains of the unicellular green alga Chlorella. DNA sequence analysis of the 330-kbp genome of Paramecium bursaria chlorella virus 1 (PBCV-1), the prototype of this virus family (Phycodnaviridae), predict approximately 366 protein-encoding genes and 11 tRNA genes. The predicted gene products of approximately 50% of these genes resemble proteins of known function, including many that are completely unexpected for a virus. In addition, the chlorella viruses have several features and encode many gene products that distinguish them from most viruses. These products include: (1) multiple DNA methyltransferases and DNA site-specific endonucleases, (2) the enzymes required to glycosylate their proteins and synthesize polysaccharides such as hyaluronan and chitin, (3) a virus-encoded K(+) channel (called Kcv) located in the internal membrane of the virions, (4) a SET domain containing protein (referred to as vSET) that dimethylates Lys27 in histone 3, and (5) PBCV-1 has three types of introns; a self-splicing intron, a spliceosomal processed intron, and a small tRNA intron. Accumulating evidence indicates that the chlorella viruses have a very long evolutionary history. This review mainly deals with research on the virion structure, genome rearrangements, gene expression, cell wall degradation, polysaccharide synthesis, and evolution of PBCV-1 as well as other related viruses.

Base Sequence↗
PubMed2001

Dielectrophoretic manipulation of a single chlorella cell with dual-microdisk electrode.

Dielectrophoretic manipulation of a single chlorella cell was performed using a dual-microdisk electrode, which consists of two Pt-Rh ultrafine wires (ca. 1-microm radius) sealed in a glass capillary. An attractive or repulsive force was induced on the chlorella depending on the frequency of the ac voltage applied between the two disk electrodes. To avoid the direct contact of a chlorella with the metal, a dual electrode with retracted disks was fabricated and used for forming a micropattern of chlorellas at a solid substrate. The effect of both the frequency and ion concentration of the solutions on the dielectrophoretic force exerted on a chlorella cell was investigated in detail based on the theories of dielectrophoresis.

Chlorella↗
PubMed1991

Expression of a cDNA clone encoding the haem-binding domain of Chlorella nitrate reductase.

A partial cDNA clone coding for the haem-binding domain of NADH:nitrate reductase (EC 1.6.6.1) (NR) from the unicellular green alga Chlorella vulgaris has been isolated, sequenced and expressed. A 1.2 kb cDNA (pCVNR1) was isolated from a lambda gt11 expression library produced from polyadenylated RNA extracted from nitrate-grown Chlorella cells. pCVNR1 hybridized to a 3.5 kb mRNA transcript that was nitrate-inducible and absent from ammonium-grown cells. The entire sequence of pCVNR1 was obtained and found to have a single uninterrupted reading frame. The derived amino acid sequence of 318 amino acids has a 45-50% similarity to higher-plant NRs, including Arabidopsis thaliana, spinach (Spinacia oleracea) and tobacco (Nicotiana tabacum). A comparison with the putative domain structure of higher-plant nitrate reductases suggested that this sequence contains the complete haem-binding domain, approximately one-third of the Mo-pterin domain and no FAD-binding domain. A 32% sequence similarity is evident when comparing the Chlorella NR haem domain with that of calf cytochrome b5. Expression of pCVNR1 in a pET vector synthesized a 35 kDa protein that was antigenic to anti-(Chlorella NR) antibody. The spectral properties of this protein (reduced and oxidized) in the 400-600 nm region are identical with those of native Chlorella NR and indicate that haem is associated with the protein.

Amino Acid Sequence↗
PubMed2007

Optimization for high-density cultivation of heterotrophic Chlorella based on a hybrid neural network model.

AIMS: The purpose of this study was to develop a reliable hybrid neural network (HNN) model for heterotrophic growth of Chlorella, based on which optimization for fed-batch (FB) cultivation of Chlorella may be successfully realized. METHODS AND RESULTS: Deterministic kinetic model was preliminarily developed for the optimization of FB cultivation of Chlorella. The highest biomass concentration and the maximum productivity were obtained as: 104.9 g l(-1) dry cell weight and 0.613 g l(-1) h(-1), respectively. After several cultivations had been performed, an HNN model was developed. The efficiency of biomass production was further increased by the optimization using this model. The highest biomass concentration and the maximum productivity attained was: 116.2 g l(-1) dry cell weight and 1.020 g l(-1) h(-1), respectively. CONCLUSION: The HNN model agreed well with experimental results in different cultivations. Comparison between the HNN model and the deterministic model showed that the former had better generalization ability, which made it a reliable tool in modelling and optimization. SIGNIFICANCE AND IMPACT OF THE STUDY: The high cell density and productivity of biomass obtained in this study is of significance for the commercial cultivation of Chlorella. The simple and efficient optimization strategy proposed in this paper may be employed in heterotrophic mass culture of Chlorella as well as other similar organisms.

Biomass↗
PubMed2003

Unusual life style of giant chlorella viruses.

Paramecium bursaria chlorella virus (PBCV-1) is the prototype of a family of large, icosahedral, plaque-forming, dsDNA viruses that replicate in certain unicellular, eukaryotic chlorella-like green algae. Its 330-kb genome contains approximately 373 protein-encoding genes and 11 tRNA genes. The predicted gene products of approximately 50% of these genes resemble proteins of known function, including many that are unexpected for a virus, e.g., ornithine decarboxylase, hyaluronan synthase, GDP-D-mannose 4,6 dehydratase, and a potassium ion channel protein. In addition to their large genome size, the chlorella viruses have other features that distinguish them from most viruses. These features include: (a) The viruses encode multiple DNA methyltransferases and DNA site-specific endonucleases. (b) The viruses encode at least some, if not all, of the enzymes required to glycosylate their proteins. (c) PBCV-1 has at least three types of introns, a self-splicing intron in a transcription factor-like gene, a spliceosomal processed intron in its DNA polymerase gene, and a small intron in one of its tRNA genes. (d) Many chlorella virus-encoded proteins are either the smallest or among the smallest proteins of their class. (e) Accumulating evidence indicates that the chlorella viruses have a very long evolutionary history.

Chlorella↗
PubMedDate not supplied

Antioxidant activity of the microalga Chlorella vulgaris cultered on special conditions.

The chemical composition of Chlorella vulgaris indicates that it has a high nutritional value to a wide range of essential nutrients, such as vitamins, minerals and proteins. Moreover, it contains other compounds such as n-3 and n-6 polynsaturated fatty acids, provitamins and phenolic compounds. In addition, this alga can be produced in large-scale systems. The objective of the present study was to evaluate the antioxidant capacity of a Chlorella cultured on three differents temperatures (15 degrees C, 20 degrees C and 30 degrees C) in 3 Klux. Chlorella cultured samples were submitted to sequential extration using as solvents: ether, methanol and water. The antioxidant activity in the extracts was measured by b-carotene/linoleic acid system, at 50 degrees C and absorbances reading at 470 nm. One control with BHT, 100 ppm was used in this determination. The total phenolic compounds was determined with Folin-Ciocalteu reagent using the spectrophotometric measured at 780 nm with catechin as standard. The phenolic acid analysis were carried out using gas chromatograph equipped with a capillary column and flame ionization detector. Non conjugated and total phenolic acids were identified on the basis of the relative retention time of their derivatives compared with the standard phenolic acids. The methanolic extract from Chlorella cultured at 30 degrees C showed higher antioxidant activity (85%) quite similar of BHT (86%). By the Rancimat test (lipidic medium) two fractions from methanolic extracts showed too higher antioxidant activity with induction times > 37.50 h at 60 degrees C and 11.5 h at 100 degrees C. The total phenolic compounds were 24.95 mg in 1 g of dry alga matter from methanolic extract and five phenolic acids were identified. The phenolic compounds salicylic, trans cinnamic, synaptic, chlorogenic, chimic and caffeic acids found in the methanolic Chlorella extract may be responsible for its higher antioxidant activity.

Antioxidants↗
PubMed2004

Near-UV radiation promotes growth of Chlorella.

The wavelength of natural sunlight reaching the Earth's surface was detected to be above 300 nm using a multichannel spectrodetector and the ratio of UV to photosynthetically active radiation (PAR) of sunlight in Japan (137 degrees E/35 degrees 11' N, altitude; 50 m) in early summer was estimated to be 0.07:1 (i.e., 7%). On the basis of this UV/PAR ratio, Chlorella ellipsoidea (IAM-27) cells were cultured in flasks under various conditions of UV irradiation in growth chambers. The growth (cell division) of these cells without near-UV radiation was inferior to that with near-UV radiation. Growth at a UV/PAR ratio of 7% (natural conditions), determined tentatively using our detector in the present study, was maximal similar to those at 14% and 28%; whereas that at 0.7% was somewhat less and that at 70% was considerably less. Growth was linked with the activities of stress enzymes. NAD(P)H-dependent oxidase [NAD(P)-DH] and xanthine oxidase (XOD), extracted from Chlorella exposed to near-UV radiation demonstrated lower activities than those from Chlorella not exposed to near-UV radiation. On the other hand, superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APOD) extracted from Chlorella exposed to near-UV radiation have higher activities than those from unexposed Chlorella. Near-UV radiation clearly acted as an important factor for growth (cell division), at least at UV/PAR ratios of up to 0.28:1.

Cell Division↗