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Biotechnological potential of immobilized algae for wastewater N, P and metal removal: a review.

This presentation comprises a review on the use of immobilized algae for wastewater nitrogen, phosphorus and metal removal purposes. Details of the use of immobilized algae, the techniques of immobilization and the effects of immobilization on cell function are included. Particularly relevant in their use for heavy metal removal from wastewaters; upon enriching the biomass in metal, can be recoverd, thereby providing economic advantages. The use of immobilized microalgae in these processes is very adequate and offers significant advantages in bioreactors. The future of this area of algal cell biotechnology is considered.

Bioreactors↗

Double-stranded RNA replicons associated with chloroplasts of a green alga, Bryopsis cinicola.

Double-stranded RNAs (dsRNAs) associated with chloroplasts and mitochondria have been found in the coenocytic green alga Bryopsis cinicola. In this study we report molecular properties of the four chloroplast-associated dsRNAs (BDRC1 to BDRC4). The longest dsRNA molecule (BDRC1) was sequenced entirely (1959 bp) and a single large ORF of 1722 bp was found within it. Database searches revealed similarities between the deduced amino acid sequence of this ORF and RNA-dependent RNA polymerase (RdRp) sequences from several RNA viruses. The most similar sequence in the database was the RdRp of beet cryptic virus 3. Phylogenetic analysis revealed that the RdRp-like sequence of BDRC1 can be placed in the Partitiviridae clade. To detect autonomous replication of these dsRNAs, RdRp assays were carried out with actinomycin D, which is an inhibitor of DNA-dependent RNA synthesis. Incorporation of [alpha-32P]UTP was detected specifically in the chloroplast and mitochondrial dsRNAs, indicating that both the chloroplast dsRNAs (BDRCs) and the mitochondrial dsRNA (BDRM) of B. cinicola are RNA replicons. The green alga B. cinicola harbors different dsRNA replicons in its chloroplasts and mitochondria.

Amino Acid Sequence↗

The respiratory inhibitor antimycin A specifically binds Fe(III) ions and mediates utilization of iron by the halotolerant alga Dunaliella salina (Chlorophyta).

It is demonstrated that Antimycin A (AA), a respiratory inhibitor produced by Streptomyces bacteria, forms lipophylic complexes with Fe(III) ions. Spectroscopic titration indicates that Fe(III) ions interact with 2AA molecules. At growth-limiting Fe concentrations, AA mediates Fe uptake and promotes growth and chlorophyll synthesis better than other Fe chelators in the halotolerant alga Dunaliella salina. It is proposed that AA enhances Fe bioavailability in hypersaline solutions by formation of lipophylic Fe-AA complexes which are taken-up and utilized by the algae. The results suggest that the respiratory inhibitor AA can affect Fe metabolism in microorganisms.

Antimycin A↗

Hydrogen metabolism of green algae: discovery and early research - a tribute to Hans Gaffron and his coworkers.

The detection of hydrogen metabolism in green algae more than 60 years ago by Hans Gaffron dispelled the widely accepted dogma at that time that this feature was unique to prokaryotic organisms. Research on this unexpected aspect of algal physiology has continued until today because of its evolutionary implications and possible practical significance. This minireview focuses on the work of Gaffron and his collaborators, whose experiments provided most of the information about the mechanism of hydrogen metabolism in algae during the 35 years following its discovery. It is shown that the emergence of our present mechanistic concepts was closely linked to the changing perception of the process of photosynthetic water oxidation. Whereas the mechanism of 'photoreduction,' i.e., the photoassimilation of carbon dioxide with hydrogen as the electron donor, was well understood already by Gaffron's group as being a reaction mediated by Photosystem I only, a clear concept of the mechanism of light-dependent hydrogen production has been more difficult to establish. Gaffron and his collaborators provided ample evidence, however, that 'photohydrogen' evolution can be fueled by reducing equivalents derived from a photolysis of water as well as by an oxidation of internal and external organic molecules. The presently prevailing view embraces this concept of multiple pathways, but the relative contribution of each of them, and the regulatory mechanisms determining it, remain a matter of debate.

Journal Article↗

Trails of green alga hydrogen research - from hans gaffron to new frontiers.

This paper summarizes aspects of the history of photosynthetic hydrogen research, from the pioneering discovery of Hans Gaffron over 60 years ago to the potential exploitation of green algae in commercial H(2)-production. The trail started as a mere scientific curiosity, but promises to be a most important discovery, one that leads photosynthesis research to important commercial applications. Progress achieved in the field of photosynthetic hydrogen production by green algae includes elucidation of the mechanism, the ability to modify photosynthesis by physiological means and to produce bulk amounts of H(2) gas, and cloning of the [Fe]-hydrogenase genes in several green algal species.

Journal Article↗

Consistent fractionation of 13C in nature and in the laboratory: growth-rate effects in some haptophyte algae.

The carbon isotopic fractionation accompanying formation of biomass by alkenone-producing algae in natural marine environments varies systematically with the concentration of dissolved phosphate. Specifically, if the fractionation is expressed by epsilon p approximately delta e - delta p, where delta e and delta p are the delta 13C values for dissolved CO2 and for algal biomass (determined by isotopic analysis of C37 alkadienones), respectively, and if Ce is the concentration of dissolved CO2, micromole kg-1, then b = 38 + 160*[PO4], where [PO4] is the concentration of dissolved phosphate, microM, and b = (25 - epsilon p)Ce. The correlation found between b and [PO4] is due to effects linking nutrient levels to growth rates and cellular carbon budgets for alkenone-containing algae, most likely by trace-metal limitations on algal growth. The relationship reported here is characteristic of 39 samples (r2 = 0.95) from the Santa Monica Basin (six different times during the annual cycle), the equatorial Pacific (boreal spring and fall cruises as well as during an iron-enrichment experiment), and the Peru upwelling zone. Points representative of samples from the Sargasso Sea ([PO4] < or = 0.1 microM) fall above the b = f[PO4] line. Analysis of correlations expected between mu (growth rate), epsilon p, and Ce shows that, for our entire data set, most variations in epsilon p result from variations in mu rather than Ce. Accordingly, before concentrations of dissolved CO2 can be estimated from isotopic fractionations, some means of accounting for variations in growth rate must be found, perhaps by drawing on relationships between [PO4] and Cd/Ca ratios in shells of planktonic foraminifera.

Algorithms↗

Origin of the algae.

Eukaryotic algae are traditionally separated into three broad divisions: the rhodophytes, the chromophytes and the chlorophytes. The evolutionary relationships between these groups, their links with other eukaryotes and with other photosynthetic groups, such as euglenophytes and cryptophytes, have been the subject of much debate and speculation. Here we analyse partial sequences of the large (28S) cytoplasmic ribosomal RNA from ten new species of protists belonging to various groups of unicellular algae. By combining them with the homologous sequences from 14 other unicellular and multicellular eukaryotes, we show that rhodophytes, chromophytes and chlorophytes emerge as three distinct groups late among eukaryotes, that is, close to the metazoa-metaphytes radiation. This implies a relatively late occurrence of eukaryotic photosynthetic symbiosis. We also provide details of intra- and inter-phyla relationships.

Base Sequence↗

Cryptophyte algae are robbed of their organelles by the marine ciliate Mesodinium rubrum.

Mesodinium rubrum (Lohmann 1908) Jankowski 1976 (= Myrionecta rubra) is a common photosynthetic marine planktonic ciliate which can form coastal red-tides. It may represent a 'species complex' and since Darwin's voyage on the Beagle, it has been of great cytological, physiological and evolutionary interest. It is considered to be functionally a phytoplankter because it was thought to have lost the capacity to feed and possesses a highly modified algal endosymbiont. Whether M. rubrum is the result of a permanent endosymbiosis or a transient association between a ciliate and an alga is controversial. We conducted 'feeding' experiments to determine how exposure to a cryptophyte alga affects M. rubrum. Here we show that although M. rubrum lacks a cytostome (oral cavity), it ingests cryptophytes and steals their organelles, and may not maintain a permanent endosymbiont. M. rubrum does not fall into recognized cellular or functional categories, but may be a chimaera partially supported by organelle robbery.

Animals↗

A new route for synthesis of dimethylsulphoniopropionate in marine algae.

The 3-dimethylsulphoniopropionate (DMSP) produced by marine algae is the main biogenic precursor of atmospheric dimethylsulphide (DMS). This biogenic DMS, formed by bacterial and algal degradation of DMSP, contributes about 1.5 x 10(13) g of sulphur to the atmosphere annually, and plays a major part in the global sulphur cycle, in cloud formation and potentially in climate regulation. Although DMSP biosynthesis has been partially elucidated in a higher plant, nothing is known about how algae make DMSP except that the whole molecule is derived from methionine. Here we use in vivo isotope labelling to demonstrate that DMSP synthesis in the green macroalga Enteromorpha intestinalis proceeds by a route entirely distinct from that in higher plants. From methionine, the steps are transamination, reduction and S-methylation to give the novel sulphonium compound 4-dimethylsulphonio-2-hydroxybutyrate (DMSHB), which is oxidatively decarboxylated to DMSP. The key intermediate DMSHB was also identified in three diverse phytoplankton species, indicating that the same pathway operates in other algal classes that are important sources of DMS. The fact that a transamination initiates this pathway could help explain how algal DMSP (and thereby DMS) production is enhanced by nitrogen deficiency.

Chlorophyta↗

A chromosome-level assembly of the alpine snow alga Chloromonas typhlos.

Chloromonas typhlos is a cosmopolitan alpine snow alga distributed across continents, and its blooming accelerates snow melting by decreasing the amount of snow albedo. To elucidate the genetic traits underlying the adaptation of C. typhlos to the alpine habitat, we combined PacBio sequencing and Hi-C to generate a high-quality chromosome-level genome assembly (contig N50: 1.29&#x2009;Mb; scaffold N50: 7.23&#x2009;Mb) with 31 chromosomes and a genome size of 200.86&#x2009;Mb. Repetitive elements constituted 11.05% of the genome, and 16,133 protein-coding genes were predicted, of which 82% were functionally annotated. This study provides a set of omics resources both for snow algae and the genus Chloromonas.

Snow↗

Quality assurance in measuring the elemental composition of the alga Fucus vesiculosus.

Algae are increasingly used for the purpose of environmental biomonitoring, for instance in the long-term program of the Federal Environmental Specimen Bank of Germany. Therefore, for the studies presented here, freeze-dried samples of Fucus vesiculosus collected from the North Sea shores were analyzed for a broad range of elements by instrumental neutron activation analysis (INAA), while inductively coupled plasma atomic emission spectrometry (ICP-AES) and mass spectrometry (ICP-MS) were carried out on digests. The entire analytical procedures, including field sampling, cleaning of material, sample handling, determination of blanks and instrumental parameters, are described. Certified reference materials analyzed in parallel with real samples and intermethod comparisons were used for assuring the accuracy of the analytical data. Reproducibility of INAA measurements was between 4 and 6% depending on the element considered. Possible sources of uncertainty and variation of the contaminant origins are discussed. Boundary conditions for the performance of algae sampling within environmental monitoring programs and the application of this marine bioindicator for the purpose of controlling time-dependent and local differences in element patterns are presented.

Cryopreservation↗

Assessing metal sorption on the marine alga Pilayella littoralis.

Increasing interest in the development of biological materials for metal sorption led us to investigate the brown marine alga, Pilayella littoralis, as a biological sorbent. This work focuses on the harvest, preparation and evaluation of P. littoralis from Nahant beaches for use as a metal biosorbent. This biomass was used in batch tests with synthetic solutions. Its metal uptake properties, including metal binding capacity, the pH dependence of metal uptake and the kinetics of metal sorption, were investigated. Most metal sorption occurred within the first 5 min of exposure and the metals were optimally bound to the algae at pH 5.5. The algal binding capacities for Al(III), Cd(II), Co(II), Cr(VI), Cu(II), Fe(III), Ni(II) and Zn(II), were 2,000, 430, 560, 90, 850, 700, 390 and 450 micromol g(-1) of dried biomass, respectively. Metals were desorbed with 0.12 mol l(-1) HCl and determined by inductively coupled plasma atomic emission spectrometry (ICP-AES).

Absorption↗

Photosynthetic electron transport in a cell-free preparation from the thermophilic blue-green alga Phormidium laminosum.

1. A cell-free preparation of membrane fragments was prepared from the thermophilic blue-green alga Phormidium laminosum by lysozyme treatment of the cells followed by osmotic shock to lyse the spheroplasts. The membrane fragments showed high rates of photosynthetic electron transport and O2 evolution (180-250 mumol of O2/h per mg of chlorophyll a with 2,6-dimethyl-1,4-benzoquinone as electron acceptor). O2-evolution activity was stable provided that cations (e.g. 10mM-Mg2+ or 100mM-Na+) or glycerol (25%, v/v) were present in the suspending medium. 2. The components of the electron-transport chain in P. laminosum were similar to those of other blue-green algae: the cells contained Pigment P700, plastocyanin, soluble high-potential cytochrome c-553, soluble low-potential cytochrome c-54 and membrane-bound cytochromes f, b-563 and b-559 (both low- and high-potential forms). The amounts and midpoint potentials of the membrane-bound cytochromes were similar to those in higher-plant chloroplasts. 3. Although O2 evolution in P. laminosum spheroplasts was resistant to high temperatures, thermal stability was not retained in the cell-free preparation. However, in contrast with higher plants, O2 evolution in P. laminosum membrane fragments was remarkably resistant to the non-ionic detergent Triton X-100.

Cell Membrane↗

The physical state of osmoregulatory solutes in unicellular algae. A natural-abundance carbon-13 nuclear-magnetic-resonance relaxation study.

Natural-abundance 13C n.m.r. spin-lattice relaxation-time measurements have been carried out on intact cells of the unicellular blue--green alga Synechococcus sp. and the unicellular green alga Dunaliella salina, with the aim of characterizing the environments of the organic osmoregulatory solutes in these salt-tolerant organisms. In Synechococcus sp., all of the major organic osmoregulatory solute, 2-O-alpha-D-glucopyranosylglycerol, is visible in spectra of intact cells. Its rotational motion in the cell is slower by a factor of approx. 2.4 than in aqueous solution, but the molecule is still freely mobile and therefore able to contribute to the osmotic balance. In D. salina, only about 60% of the osmoregulatory solute glycerol is visible in spectra of intact cells. The rotational mobility of this observable fraction is approximately half that found in aqueous solution, but the data also indicate that there is a significant concentration of some paramagnetic species in D. salina which contributes to the overall spin-lattice relaxation of the glycerol carbon atoms. The non-observable fraction, which must correspond to glycerol molecules that have very broad 13C resonances and that are in slow exchange with bulk glycerol, has not been properly characterized as yet, but may represent glycerol in the chloroplast. The implications of these findings in relation to the physical state of the cytoplasm and the mechanism of osmoregulation in these cells are discussed.

Chlorophyta↗

Intermediates of tocopherol biosynthesis in the unicellular alga Scenedesmus obliquus. The presence of three isomeric methylphytylbenzoquinones.

Three isomers of methylphytylbenzoquinone have been isolated from lipids of the unicellular alga Scenedesmus obliquus, the most abundant being 2-methyl-6-phytylbenzoquinone (65% of the total). The 2-methyl-3-phytyl and 2-methyl-5-phytyl isomers amounted to 8 and 27% respectively. Previously problems have been encountered in the separation of the 3-phytyl and the 6-phytyl isomers, but in the present study it was found that they separated readily as quinols. Phytyl plastoquinone was also found and the relevance of these compounds to the biosynthesis of alpha-tocopherol is discussed. As well as phylloquinone, a hydroxyphylloquinone was detected, and studies indicated that it is the 5' carbon atom to which the hydroxy group is attached. Such a compound has been found by workers using other unicellular algae.

Chlorophyta↗

The exceptional photofermentative hydrogen metabolism of the green alga Chlamydomonas reinhardtii.

The photosynthetic green alga Chlamydomonas reinhardtii is capable of performing a complex fermentative metabolism which is related to the mixed acid fermentation of bacteria such as Escherichia coli. The fermentative pattern includes the products formate, ethanol, acetate, glycerol, lactate, carbon dioxide and molecular hydrogen (H(2)). H(2) production is catalysed by an active [Fe]-hydrogenase (HydA) which is coupled with the photosynthetic electron-transport chain. The most important enzyme of the classic fermentation pathway is pyruvate formate-lyase, which is common in bacteria but seldom found in eukaryotes. An interaction between fermentation, photosynthesis and H(2) evolution allows the algae to overcome long periods of anaerobiosis. In the absence of sulphur, the cells establish a photofermentative metabolism and accumulate large amounts of H(2).

Animals↗

Differential regulation of the Fe-hydrogenase during anaerobic adaptation in the green alga Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii, a unicellular green alga, contains a hydrogenase enzyme, which is induced by anaerobic adaptation of the cells. Using the suppression subtractive hybridization (SSH) approach, the differential expression of genes under anaerobiosis was analyzed. A PCR fragment with similarity to the genes of bacterial Fe-hydrogenases was isolated and used to screen an anaerobic cDNA expression library of C. reinhardtii. The cDNA sequence of hydA contains a 1494-bp ORF encoding a protein with an apparent molecular mass of 53.1 kDa. The transcription of the hydrogenase gene is very rapidly induced during anaerobic adaptation of the cells. The deduced amino-acid sequence corresponds to two polypeptide sequences determined by sequence analysis of the isolated native protein. The Fe-hydrogenase contains a short transit peptide of 56 amino acids, which routes the hydrogenase to the chloroplast stroma. The isolated protein belongs to a new class of Fe-hydrogenases. All four cysteine residues and 12 other amino acids, which are strictly conserved in the active site (H-cluster) of Fe-hydrogenases, have been identified. The N-terminus of the C. reinhardtii protein is markedly truncated compared to other non-algal Fe-hydrogenases. Further conserved cysteines that coordinate additional Fe-S-cluster in other Fe-hydrogenases are missing. Ferredoxin PetF, the natural electron donor, links the hydrogenase from C. reinhardtii to the photosynthetic electron transport chain. The hydrogenase enables the survival of the green algae under anaerobic conditions by transferring the electrons from reducing equivalents to the enzyme.

Adaptation, Physiological↗

rbcL sequences reveal multiple cryptic introductions of the Japanese red alga Polysiphonia harveyi.

In Europe, the last 20 years have seen a spectacular increase in accidental introductions of marine species, but it has recently been suggested that both the actual number of invaders and their impacts have been seriously underestimated because of the prevalence of sibling species in marine habitats. The red alga Polysiphonia harveyi is regarded as an alien in the British Isles and Atlantic Europe, having appeared in various locations there during the past 170 years. Similar or conspecific populations are known from Atlantic North America and Japan. To choose between three competing hypotheses concerning the origin of P. harveyi in Europe, we employed rbcL sequence analysis in conjunction with karyological and interbreeding data for samples and isolates of P. harveyi and various congeners from the Pacific and North Atlantic Oceans. All cultured isolates of P. harveyi were completely interfertile, and there was no evidence of polyploidy or aneuploidy. Thus, this biological species is both morphologically and genetically variable: intraspecific rbcL divergences of up to 2.1% are high even for red algae. Seven rbcL haplotypes were identified. The four most divergent haplotypes were observed in Japanese samples from Hokkaido and south-central Honshu, which are linked by hypothetical 'missing' haplotypes that may be located in northern Honshu. These data are consistent with Japan being the centre of diversity and origin for P. harveyi. Two non-Japanese lineages were linked to Hokkaido and Honshu, respectively. A single haplotype was found in all North Atlantic and Mediterranean accessions, except for North Carolina, where the haplotype found was the same as that invading in New Zealand and California. The introduction of P. harveyi into New Zealand has gone unnoticed because P. strictissima is a morphologically indistinguishable native sibling species. The sequence divergence between them is 4-5%, greater than between some morphologically distinct red algal species. Two different types of cryptic invasions of P. harveyi have therefore occurred. In addition to its introduction as a cryptic sibling species in New Zealand, P. harveyi has been introduced at least twice into the North Atlantic from presumed different source populations. These two introductions are genetically and probably also physiologically divergent but completely interfertile.

Fertility↗