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Variations in caulerpenyne contents in Caulerpa taxifolia and Caulerpa racemosa.

Caulerpenyne (CYN) contents was measured in two Chlorophyceae algae, Caulerpa taxifolia and Caulerpa racemosa, between July 1999 and July 2000. Sampling was performed at three stations exhibiting increasing levels of competition with the seagrass Posidonia oceanica. Significant differences were observed as a function of the Caulerpa species, the season, and the level of competition. CYN concentrations were always greater in C. taxifolia, regardless of either season or level of competition (35-80 times greater, according to the season). For a given species, maximum concentrations were recorded in autumn (September/November) and minimum values occurred in spring (April/May). CYN contents decreased with increasing level of competition, whereas frond length increased over this same gradient. It would appear that when the algae are in competition with P oceanica, Caulerpa is more inclined to accelerate vegetative growth (competition for light) than to produce secondary metabolites.

Chlorophyta↗

Seaweeds along KwaZulu-Natal Coast of South Africa--4: Elemental uptake by edible seaweed Caulerpa racemosa (sea grapes) and the arsenic speciation.

The elemental uptake by edible seaweed Caulerpa racemosa (Sea grapes), a marine macroalgae (chlorophyta, green alga) grown richly along KwaZulu-Natal coastline. The total concentrations of seven elements, namely Mn, Fe, As, B, Ti, Zn and Hg in Caulerpa racemosa were monitored for a one-year cycle (June 2002 to May 2003) at four selected sampling sites spread over 150 km wide from North to South. The C. racemosa possess high arsenic, boron and titanium accumulating ability, but low iron uptake. A typical C. racemosa sample at Treasure Beach in the vicinity of Durban Metropolis in autumn contained Mn (5.2+/-ppm), Fe (0.21+/-0.01 ppm), As (8.5+/-0. ppm), B (1090+/-ppb), Ti (159+/-ppb), Zn (3.8+/-0.1 ppb), and Hg (189+/-ppb). The general trend found at all sites was high elemental concentrations in winter and a decrease in concentrations from winter to spring and summer. C. racemosa recorded highest mercury levels (>205 ppb) during the summer season at the Zinkwasi site. The arsenic speciation in four abundant seaweeds from the beaches of Indian Ocean in the KwaZulu-Natal coast is investigated. The speciation of arsenic in two rhodophyta seaweeds, Plocamium corallorhiza and Gelidium abbottiorum, and two chlorophyta seaweeds, Ulva lactuca and Caulerpa racemosa at four sampling sites during the summer of 2003, is elucidated. Caulerpa racemosa had highest total arsenic (in ppb) reaching (8850+/-200) at Zinkwasi, and the concentrations of the other arsenic species analysed are As (III) (194+/-10), As(V) (568+/-27), methylarsonic acid (494+/-22) and dimethylarsinic acid (373+/-12). In an indirect estimation, C. racemosa had 81+/-2% aresenosugars.

Arsenic↗

Preparation of tubulin from Caulerpa, a marine green alga, using casein as a protective agent against proteolytic degradation.

Bidirectional organelle movements taking place in the cytoplasm of the rhizomes of Caulerpa, a coenocytic marine green alga, have been indicated to be dependent on microtubules (Kuroda, K. & Manabe, E. (1983) Proc. Jpn. Acad. 59B, 131-134; Manabe, E. & Kuroda, K. (1984) Proc. Jpn. Acad. 60B, 118-121). However, when a crude extract of Caulerpa rhizomes was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and subjected to immunoblotting with monoclonal anti-tubulin antibody, no reacting band could be detected. This apparent absence of tubulin in the extract was found to be a result of the complete degradation of tubulin by potent intrinsic proteolytic activity. All of the commercially available protease inhibitors so far tested (p-chloromercuriphenylsulfonic acid, phenyl methylsulfonyl fluoride, 1-chloro-4-phenyl-3-tosylamido-2-butanone, 7-amino-1-chloro-3-tosylamido-2-heptanone, p-tosyl-L-arginine methyl ester, soybean trypsin inhibitor, antipain, chymostatin, leupeptin, and pepstatin) failed to inhibit the activity completely. But addition of casein at the concentration of 1% (weight per volume) to the solutions used for preparation was effective in protecting tubulin from proteolytic degradation, thus making it possible to prepare tubulin from the crude extract of Caulerpa. On SDS-PAGE, the Caulerpa alpha-tubulin thus prepared was a little smaller in molecular weight than that of rabbit brain.

Animals↗

Are the Mediterranean ascoglossan molluscs Oxynoe olivacea and Lobiger serradifalci suitable agents for a biological control against the invading tropical alga Caulerpa taxifolia?

The Mediterranean ascoglossan Oxynoe olivacea and Lobiger serradifalci, which are scarce in meadows of their usual food the alga Caulerpa prolifera, have become adapted to feeding on the invading tropical alga Caulerpa taxifolia. Grazing rates are low (O. olivacea destroys only a 5-cm C. taxifolia frond in 3-7 d). The grazing rates are a function of temperature. Despite a high spawning frequency (0.25 egg-mass.day-1) and a large number of eggs released per spawning event (O. olivacea: from 2,545 to 29,128 eggs.spawning-1; L. serradifalci: from 12,112 to 48,448 eggs.spawning-1), the recruitment on Caulerpa meadows is usually low due to the hazardous pelagic development of the larvae. The possible use of these molluscs as agents of biological control against C. taxifolia appears to be possible only through an artificial enhancement of their populations after cultivation of the veligers and release of juveniles during the winter season. This means of control is discussed and compared with the possible use of tropical ascoglossan species.

Animals↗

Comparison of the wound-activated transformation of caulerpenyne by invasive and noninvasive Caulerpa species of the Mediterranean.

The invasive green alga, Caulerpa taxifolia, that has spread rapidly after its introduction into the Mediterranean and the North American Pacific, reacts to wounding by transforming its major metabolite caulerpenyne. This wound-activated reaction involves the transformation of the bis-enol acetate moiety of 1, releasing reactive 1,4-dialdehydes. The ability to perform this transformation is found also in both the noninvasive Mediterranean C. prolifera and the invasive C. racemosa. Trapping experiments, as well as transformation of the model substrate geranyl acetate, suggest that all three investigated Caulerpa spp. rely on esterases that act upon wounding of the algae by subsequently removing the three acetate residues of caulerpenyne. The resulting reactive 1,4-dialdehyde oxytoxin 2 can be identified by liquid chromatography-mass spectrometry and is unstable in the wounded tissue. Caulerpenyne transformation occurs rapidly, and severe tissue damage caused degradation of more than 50% of the stored caulerpenyne within 1 min in all three algae. Prevention of the enzymatic reaction before extraction, by shock freezing the tissue with liquid nitrogen, was used for the determination of the caulerpenyne content in intact algae. It gives about twofold higher values compared to an established methanol extraction protocol. The speed and mechanism of the wound-activated transformation, as well as the caulerpenyne content in intact tissue of invasive and noninvasive Caulerpa spp., are comparable. Thus, this enzymatic transformation, despite being fast and efficient, is likely not the key for the success of the investigated invasive species.

Adaptation, Physiological↗

Removal of basic yellow dye from aqueous solution by sorption on green alga Caulerpa scalpelliformis.

Dynamic batch experiments were carried out for the biosorption of basic yellow dye on to the green macroalgae Caulerpa scalpelliformis. The factors affecting the sorption process such as the initial concentration of the dye and pH of the solution, the adsorbent dosage and the time of contact were studied. The sorption kinetics followed pseudo-second order kinetic model. The Caulerpa species exhibited a maximum uptake of 27 mg of dye per gram of seaweed. The Boyd's plot confirmed the external mass transfer as the rate-limiting step. The average effective diffusion coefficient was found to be 2.47 x 10(-4)cm(2)/s. Sorption equilibrium studies demonstrated that the biosorption followed Freundlich isotherm model, which implies a heterogeneous sorption phenomenon. Various thermodynamic parameters such as enthalpy of sorption DeltaH degrees , free energy change DeltaG degrees and entropy DeltaS degrees were estimated. The negative value of DeltaH degrees and negative values of DeltaG degrees show the sorption process is exothermic and spontaneous. The negative value of entropy DeltaS degrees shows the decreased randomness at the solid-liquid interface during the sorption of dyes onto green seaweed.

Adsorption↗

Cell growth inhibitory effects of caulerpenyne, a sesquiterpenoid from the marine algae Caulerpa taxifolia.

Caulerpa taxifolia (Vahl) C. Agardh (Ulvophyceae, Caulerpales) is an algae of tropical origin that was accidentally introduced into the Mediterranean in 1984. Caulerpenyne (Cau) is the major metabolite present in Caulerpa taxifolia. This metabolite has previously been shown to be cytotoxic against cell lines in culture as in KB cells and fibroblasts from hamsters. Cau along with 6 other drugs representative of the major classes of anticancer products was tested against 8 cancer cell lines of human origin. Cau demonstrated growth-inhibitory effects in all cases with some variability between cell lines; this inter-cell variability was, however, less marked than that observed with the anticancer drug tested. Cells of colorectal cancer origin were the most sensitive to the presence of Cau with IC50 values of 6.1 and 7.7 microM. Increasing the duration of contact between Cau and the cells from 75 min to 29.5 hr did not improve the cytotoxic efficacy of this compound. When Cau was pre-incubated in the culture medium for from 7 to 83 min before being exposed to CAL 27 cells (head and neck cancer origin), there was a constant loss of cytostatic action of Cau as a function of Cau pre-incubation time. As the bovine serum albumin concentration increased in the culture medium, the concentration-response curves showed a constant shift towards the right, indicating a loss of cytostatic activity of Cau. In the presence of Cau, cells in culture clearly exhibited an early and marked shift into S phase followed by a blockade into the premitotic G2 M phase. Possible targets for CAU remain to be identified. Cau needs to be tested on tumor bearing animals to confirm this promising antiproliferative activity.

Animals↗

Biosorption of Cu2+, Cd2+, Pb2+, and Zn2+ using dried marine green macroalga Caulerpa lentillifera.

The sorption of Cu2+, Cd2+, Pb2+, and Zn2+ by a dried green macroalga Caulerpa lentillifera was investigated. The removal efficiency increased with pH. The analysis with FT-IR indicated that possible functional groups involved in metal sorption by this alga were O-H bending, N-H bending, N-H stretching, C-N stretching, C-O, SO stretching, and S-O stretching. The sorption of all metal ions rapidly reached equilibrium within 20min. The sorption kinetics of these metals were governed by external mass transfer and intraparticle diffusion processes. The sorption isotherm followed the Langmuir isotherm where the maximum sorption capacities was Pb2+>Cu2+>Cd2+>Zn2+.

Adsorption↗

High performance biosorbent (Caulerpa lentillifera) for basic dye removal.

The sorptions of three basic dyes, Astrazon((R)) Blue FGRL (AB), Astrazon((R)) Red GTLN (AR), and methylene blue (MB) onto green macroalga Caulerpa lentillifera were investigated. The results were compared to the sorption performance of a commercial activated carbon (CARBON). The results revealed that the alga exhibited greater sorption capacities than activated carbon for the three basic dyes investigated in this work. The sorption process for all mixture systems (ALGA/AB, ALGA/AR, ALGA/MB, CARBON/AB, CARBON/AR, and CARBON/MB) obeyed the pseudo-second order kinetic model. C. lentillifera could more rapidly sequester AR when compared with activated carbon, but was more slowly in the sorption of AB. For the sorption of MB, both ALGA and CARBON seemed to have the same sorption rate. The sorption processes were initially controlled by both film and pore-diffusion, and only were limited by pore diffusion in the later stage. The isotherms followed Langmuir model which suggested that the sorption was monolayer coverage.

Absorption↗

Two novel aromatic valerenane-type sesquiterpenes from the Chinese green alga Caulerpa taxifolia.

Caulerpal A (2) and B (3), two novel sesquiterpenes possessing an uncommon aromatic valerenane-type carbon skeleton, along with one known metabolite, caulerpin (4), have been isolated from the Chinese green alga Caulerpa taxifolia (Vahl) C. Agardh. Their structures and relative stereochemistry were elucidated on the basis of extensive spectroscopic analysis. Compounds 2-4 were evaluated for their inhibitory activity against hPTP1B and the result showed that only compound 4 had a strong PTP1B inhibitory activity with an IC50 value of 3.77 microM.

Caulerpa↗

Removal of basic dye (Astrazon Blue FGRL) using macroalga Caulerpa lentillifera.

The macroalga Caulerpa lentillifera was found to have adsorption capacity for a basic dye, Astrazon Blue FGRL. For the whole range of concentrations employed in this work (20-1280 mgl(-1)), the adsorption reached equilibrium within the first hour. The kinetic data corresponded well with the pseudo second-order kinetic model where the rate constants decreased as initial dye concentrations increased. At low dye concentrations (20-80 mgl(-1)), an increase in the adsorbent dosage resulted in a higher removal percentage of the dye, but a lower amount of dye adsorbed per unit mass (q). The adsorption isotherm followed both the Langmuir and Freundlich models within the temperature range employed in this work (18-70 degrees C). The highest maximum adsorption capacity (q(m)) was obtained at 50 degrees C. The enthalpy of adsorption was estimated at 14.87 kJmol(-1) suggesting a chemical adsorption mechanism.

Adsorption↗

First assessment of the Caulerpa racemosa (Caulerpales, Chlorophyta) invasion along the French Mediterranean coast.

The introduced green alga Caulerpa racemosa var. cylindracea has been rapidly spreading in the Mediterranean Sea since 1990. It was first observed in France in 1997 (Marseilles). In early 2004, the stretch of the French Mediterranean coastline and the surface area affected by the invasion were estimated at about 83 km and 4014 ha, respectively. The depth range of colonized areas was usually 10-35 m depth. Shallow (0-10 m) and deep (down to 40 m) dense meadows were rarely observed. In contrast to the dead matte of Posidonia oceanica, which constituted the most widely colonized substratum, dense P. oceanica meadows and fine sand with large ripple-marks were not invaded. Few rocky areas were colonized and coarse sand bottoms were usually colonized below 20 m depth. All the colonized areas were exposed to human activities and more than 40% were fishing areas. Mild climate, suitable substrata, presence of vectors of dispersal and absence of efficient biological control make the French Mediterranean coast particularly vulnerable to the further spread of the alga.

Caulerpa↗

In vitro anti-herpetic activity of sulfated polysaccharide fractions from Caulerpa racemosa.

A sulfated polysaccharide fraction was isolated from the hot water extract of the green alga Caulerpa racemosa and designated HWE. This polymer, which contained galactose, glucose, arabinose and xylose as the major component sugars, had [alpha](D)(30) + 46.2 degrees in water and contained 9% sulfate hemiester groups. Sugar linkage analysis indicates that HWE was branched and mainly contained 1,3- and 1,3,6-linked galactose, 1,3,4-linked arabinose, 1,4-linked glucose and terminal- and 1,4-linked xylose residues. Sulfation was deduced from infrared spectroscopy and methylation analysis to occur on O-6 of galactose and O-3 of arabinose. The native polysaccharide could be fractionated by size exclusion chromatography into two overlapping fractions and the major fraction has a hydrodynamic volume similar to that of 70 kDa dextran. HWE was a selective inhibitor of reference strains and TK(-) acyclovir-resistant strains of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in Vero cells, with antiviral effective concentration 50% (EC(50)) values in the range of 2.2-4.2 microg/ml and lacking cytotoxic effects. Furthermore, HWE did not exhibit anticoagulant properties at concentrations near the EC(50).

Animals↗

Structure of beta-1,3-xylooligosaccharides generated from Caulerpa racemosa var. laete-virens beta-1,3-xylan by the action of beta-1,3-xylanase.

Recently we reported the molecular cloning and characterization of a novel beta-1,3-xylanase from the marine bacterium Vibrio sp. AX-4 [Kiyohara et al. (2005) Biochem. J. 388, 949-957]. We report here the structural analysis of oligosaccharides generated from beta-1,3-xylan of a siphonous green alga, Caulerpa racemosa var. laete-virens, by the action of beta-1,3-xylanase. The enzyme degraded the polysaccharide producing oligosaccharides with different R(f)s on TLC (EX2-EX5). Sugar component, linkage, and MALDI-TOF-MS analyses revealed that EX2 and EX3 were Xyl-1,3-Xyl and Xyl-1,3-Xyl-1,3-Xyl, respectively. On the other hand, EX4 was a mixture of Glc-1,3-Xyl-1,3-Xyl, Xyl-1,4-Xyl-1,3-Xyl and Xyl-1,3-Xyl-1,4-Xyl, while EX5 was a mixture of tetra-saccharides containing 3-substitued Glc in addition to the same components of EX4. Branching was not likely present in EXOs prepared from the polysaccharide by the enzyme. These results strongly suggest that the C. racemosa beta-1,3-xylan is a linear heteropolysaccharide containing 1,3-Glc and 1,4-Xyl both of which are thought to be located within a beta-1,3-Xyl chain and linked via covalent bonds. This report indicates the usefulness of the enzyme for the structural analysis of beta-1,3-xylan.

Caulerpa↗

Antiviral properties of a crude extract from a green alga Caulerpa taxifolia (Vahl) C. Agardh.

A crude extract of Caulerpa taxifolia was tested for its antiviral activity. The chloroform-methanol residue showed an interesting inhibitor effect in vitro toward the feline immunodeficiency virus (FIV), a valid model for studying the acquired immunodeficiency syndrome. This extract reduced the virus-induced syncytia in the cultured cells, the viral reverse transcriptase activity and the viral capsid protein P24 expression.

Animals↗

Effect of caulerpenyne, a toxin extracted from Caulerpa taxifolia on mechanisms regulating intracellular pH in sea urchin eggs and sea bream hepatocytes.

The proliferation of the green marine alga Caulerpa taxifolia in the Mediterranean led us to investigate the toxic effects on marine organisms of caulerpenyne (Cyn), the major secondary metabolite synthesized by the alga. This study was performed on sea urchin eggs (Paracentrotus lividus) and isolated hepatocytes from the sea bream (Sparus aurata), in which accumulation of the toxins by metabolic processes may be of significance. Cyn provoked an acidification of seawater containing both unfertilized and fertilized eggs, as revealed by a titrable efflux of protons. The pHi in unfertilized eggs continuously increased in the presence of Cyn, whereas there was a biphasic response in both fertilized eggs and isolated hepatocytes, with a decrease of the pHi followed by recovery to the initial value. Cyn inhibited the accumulation of 14C-methylamine in acidic granules present in the cortical zone of sea urchin eggs. Dicyclocarbodiimide (DCCD), a well-known H(+)-ATPase inhibitor, provoked a similar inhibition. Both molecules increased pH in the acidic compartments of isolated bream hepatocytes. These results suggest that Cyn inhibits intracellular sequestration of protons and thus liberates protons into the cell cytoplasm from which they leak toward the extracellular medium.

Animals↗

Caulerpenyne from Caulerpa taxifolia has an antiproliferative activity on tumor cell line SK-N-SH and modifies the microtubule network.

Caulerpenyne, the major secondary metabolite synthesized by the green marine alga Caulerpa taxifolia, is cytotoxic against several cell lines. To identify possible targets of this toxin, we investigated the effect of caulerpenyne on the neuroblastoma SK-N-SH cell line. Caulerpenyne induced an inhibition of SK-N-SH cell proliferation with an IC50 of 10 +/- 2 microM after 2 hr of incubation. We observed no blockage in G2/M phase and an increase in cell death. On immunofluorescence microscopy, caulerpenyne affected the microtubule network in SK-N-SH cell line; we observed a loss of neurites and a compaction of the microtubule network at the cell periphery. In vitro, after 35 min of incubation, caulerpenyne inhibited the polymerization of pig brain purified tubulin or microtubule proteins, with an IC50 of 21 +/- 2 microM and 51 +/- 6 microM respectively. Analysis by electron microscopy indicated that caulerpenyne induced aggregation of tubulin, which may be responsible for inhibition of microtubule polymerization and bundling of residual microtubules.

Antineoplastic Agents↗

Using genetic techniques to investigate the sources of the invasive alga Caulerpa taxifolia in three new locations in Australia.

The invasive green alga Caulerpa taxifolia has gained a high profile due to 'outbreaks' in the Mediterranean and California. During the year 2000 three new discrete locations colonised by abundant C. taxifolia were discovered in New South Wales (NSW), Australia. Sequencing of the internal transcribed spacer (ITS) region of the ribosomal DNA was used to explore the source(s) of these new records, which is an important prerequisite for subsequent environmental management responses. Our results indicate that the NSW C. taxifolia originated from several sources and, hence, through different invasion events. For two of the new records (Port Hacking, Careel Bay) it can be excluded that they are derived from the so-called "aquarium strain" of C. taxifolia, closely related to the invasive Mediterranean populations. Port Hacking is likely to have originated from tropical native populations. However, samples from Lake Conjola cannot be sufficiently distinguished with the applied technique from native C. taxifolia in Moreton Bay and the Mediterranean/"aquarium strain".

Base Sequence↗