Green Alga Codium Research Articles

Content Analysis, Cytotoxic, and Anti-metastasis Potential of Bioactive Polysaccharides from Green Alga Codium intricatum Okamura

Background: Codium intricatum, locally known as Pukpuklo, is a seasonal and edible green alga found in Ilocos Norte, Philippines. In this study, the biochemical content, cytotoxic and inhibitory potential against Matrix Metalloproteinase-1 (MMP-1) production of the polysaccharide-protein fractions from C. intricatum is first reported. MMPs are novel targets for therapeutic intervention with the potential to inhibit tumor growth, metastasis and invasion either on their own or in conjunction with cytotoxic treatments. Methods: Water-soluble Crude Polysaccharide (CP) and its fractions (CFs) from C. intricatum were isolated using hot water and ion-exchange chromatography and analyzed using different analytical techniques. Cytotoxicity against MCF-7 breast cancer cells and Human normal Dermal Fibroblasts (HnDFs) was determined by MTT assay. The MMP-1 inhibitory potential was tested in UVB exposed Human normal Dermal Fibroblast Cells. Results: CP and CFs afforded carbohydrates (2.07-16.1%), sulfates (1.81%- 9.9%), protein (0.05-2.7%), ash (<77.2%), lipids (<1.0%) and uronic acid (0.15- 4.49%). FT- IR and NMR spectra of CP and CFs exhibited absorption peaks comparable to sulfated galactans. Both CP and CFs significantly inhibited the growth of breast cancer (MCF-7) in a concentration-dependent manner (p<0.05), induced proliferation of HnDFs, inhibited the production of Matrix metalloproteinase-1 (MMP-1) in UV-B induced HnDFs (p<0.05). Conclusion: Results are highly suggestive that polysaccharide fractions from C. intricatum are bioactive molecules with cytotoxic and anti-metastasis potential.

Siphonaxanthin, a carotenoid from green algae Codium cylindricum, protects Ob/Ob mice fed on a high-fat diet against lipotoxicity by ameliorating somatic stresses and restoring anti-oxidative capacity

Oxidative stress is implicated in the pathogenesis of many diseases including obesity, non-alcoholic fatty liver disease, and diabetes mellitus. Previously, we reported that siphonaxanthin, a carotenoid from green algae, elicited a potent inhibitory effect on hepatic de novo lipogenesis, and an anti-obesity effect in both 3T3L1 cells and KKAy mice. Thus, we hypothesized that consumption of siphonaxanthin could improve metabolic disorders including hepatic steatosis and systemic adiposity, as well as ameliorate somatic stress under obese conditions. Both the hepatocyte cell line HepG2 and a mouse model of severe obesity, produced by feeding Ob/Ob mice on a high-fat diet (HFD), were used to test this hypothesis. In obese mice, siphonaxanthin intake did not improve liver steatosis or systemic adiposity. However, intake did lower plasma glucose and alanine aminotransferase (ALT) levels and diminished hepatic lipid peroxidation products and antioxidant gene expression, which increased significantly in control group obese mice. Renal protein carbonyl content decreased significantly in the siphonaxanthin group, which might also indicate an ameliorated oxidative stress. Siphonaxanthin restored gene expression related to antioxidant signaling, lipid β-oxidation, and endoplasmic-reticulum-associated protein degradation in the kidney, which decreased significantly in obese mice. Liver and kidney responded to obesity-induced somatic stress in a divergent pattern. In addition, we confirmed that siphonaxanthin potently induced Nrf2-regulated antioxidant signaling in HepG2 cells. In conclusion, our results indicated that siphonaxanthin might protect obesity-leading somatic stress through restoration of Nrf2-regulated antioxidant signaling, and might be a promising nutritional supplement.

Pharmacological prospection and structural characterization of two purified sulfated and pyruvylated homogalactans from green algae Codium isthmocladum

Two sulfated polysaccharides (SPs), F2 and F3, isolated from Codium isthmocladum were found to contain galactose, sulfate, and pyruvate. The apparent molecular weights of F2 and F3 were determined to be 62 and 61 kDa, respectively. NMR spectroscopy combined with chemical analysis showed that F2 and F3 have the same structural features. However, F3 showed higher sulfate/sugar ratio (1/2.6) than F2 (1/4). F2 and F3 are essentially (1 → 3)-β-D-galactans with some branching at C6. Pyruvylation occurs at O3 and O4, forming 3,4-O-(1-carboxyethylidene)-β-D-Galp residues; some of these pyruvylated residues contain sulfate groups at C6. Some non-branching residues contain sulfate at C4. None of the SPs exhibited antioxidant activity. MTT results indicated that 1 mg/mL of both SPs about 40% of PANC-1 cell viability. At 10 μg/mL, F2 and F3 had 1.7-fold longer clotting times compared to that of Clexane® at the same concentration. The higher sulfate content of F3 is not a determining factor for pharmacological activities of galactans, considering that both F2 and F3 exerted the effects.

Isolation and characterization of three pairs of indolediketopiperazine enantiomers containing infrequent N-methoxy substitution from the marine algal-derived endophytic fungus Acrostalagmus luteoalbus TK-43

Three pairs of new N-methoxy-containing indolediketopiperazine enantiomers, acrozines A–C (1–3), were isolated from the culture extract of Acrostalagmus luteoalbus TK-43, an endophytic fungus obtained from the marine green alga Codium fragile. The optical resolution of compounds 1–3 by chiral HPLC successfully afforded individual enantiomers (+)-1/(–)-1, (+)-2/(–)-2, and (+)-3/(–)-3, respectively. The structures of all these compounds were established on the basis of detailed interpretation of their NMR and mass spectroscopic data. X-ray crystallographic analysis confirmed the structures of compounds 1–3, while the absolute configurations were determined by TDDFT-ECD calculations. All these compounds containing a N-methoxy group which is uncommon in indolediketopiperazines. The enantiomers, (+)-2/(–)-2, showed different antimicrobial activities against several plant-pathogenic fungi, while (+)-1 displayed better inhibitory activity against acetylcholinesterase than that of (–)-1.

First record of the invasive green alga Codium fragile ssp. fragile (Chlorophyta, Bryopsidales) in Abruzzi waters, central Adriatic Sea

The authors report for the first time on the invasive green alga Codium fragile ssp. fragile (Suringar) Hariot from the Adriatic coast of the Abruzzi Region of Italy (central Adriatic Sea). Several thalli have been found stranded on the beach of Silvi Marina since November 2015. This species, native to East Asia, has widely invaded several locations of the Mediterranean where it has been probably introduced through boating or shellfish aquaculture.

Anti-Obesity Properties of the Dietary Green Alga, Codium cylindricum, in High-Fat Diet-Induced Obese Mice.

Marine carotenoids have been reported to prevent and alleviate lifestyle-related diseases including diabetes and obesity. We previously reported that siphonaxanthin, a green algal carotenoid, effectively suppresses adipogenesis in 3T3-L1 cells and prevents lipid accumulation in mesenteric adipose tissue of KK-Ay mice. Thus, we expect that dietary siphonaxanthin-rich marine green algae may effectively prevent obesity. Here, we assessed the effect of dietary siphonaxanthin-rich green algae (Codium cylindricum) on the development of diet-induced obesity in C57BL/6J mice. The mice were fed a low-fat diet (LF; 7% fat, w/w), a high-fat diet (HF; 35% fat, w/w), or a high-fat diet supplemented with 1% or 5% green algal powder (1GA or 5GA) for 78 d. Our results showed that weights of body and perirenal white adipose tissue (WAT) in the 5GA group were significantly lower than that in the HF group. The mesenteric and total WAT, as well as plasma and hepatic cholesterol concentrations tended to be lower in both the 1GA and 5GA groups compared to the HF group. Dietary green algal powder reduced the expression of lipogenesis-related genes and enhanced the expression of energy expenditure-related genes in the mesenteric WAT. Siphonaxanthin accumulated in the mesenteric WAT may contribute to the prevention of adiposity in mesenteric WAT. Furthermore, the reduction in the weight of WAT was attributed to the inhibitory effect on fat absorption of dietary fiber contained in the green algae. Overall, these results indicated that siphonaxanthin-rich green algae may be beneficial for the prevention of obesity and regulation of lipid metabolism.

Allobodo chlorophagus n. gen. n. sp., a Kinetoplastid that Infiltrates and Feeds on the Invasive Alga Codium fragile

A novel biflagellate protist that consumed chloroplasts inside material of the invasive marine green alga Codium fragile was reported from the U.S. east coast in 2003. We observed a similar association in C. fragile from five sites in Nova Scotia, Canada during 2013 and 2014. After incubating Codium fragments for 2-3 days, some utricles and filaments contained numerous chloroplast-consuming cells. Transmission electron microscopy (TEM) confirmed that these were kinetoplastids with a pankinetoplast, large electron-dense droplets in the cytoplasm and a connective between the paraxonemal rod bases, but no conspicuous para-cytopharyngeal rod, all consistent with U.S. material observed in 2003. The ITS1-5.8S rRNA-ITS2 sequences from 13 Nova Scotia isolates were identical. SSU rRNA gene phylogenies placed the Codium-associated kinetoplastid in neobodonid clade '1E'. Clade 1E likely contains no previously described species, and branches outside all other major neobodonid groups, either as their sister or as a separate lineage, depending on rooting. These results indicate that the kinetoplastid represents a single species that merits a new genus (and family), and we describe it as Allobodo chlorophagus n. gen., n. sp. The lack of evidence for food sources other than Codium is consistent with a parasitic association, but other possibilities exist (e.g. necrotrophy).

Spatio-temporal dynamics of Codium populations along the rocky shores of N and NW Spain

The green alga Codium fragile ssp. fragile (hereafter C. fragile) has long been assumed to outcompete and displace its native congeners via cryptic invasion. We analysed the population dynamics of the exotic C. fragile and native congeners and their relative abundance on intertidal shores in N-NW Spain. Our results did not support the existence of current competitive displacement by the exotic species. The presence of C. fragile was clearly seasonal, while the native C. tomentosum was more persistent throughout the year, due to a higher frequency of frond sprouting from perennial basal fragments. However, our results also indicated an increase in the proportion of C. fragile relative to native species towards inner areas of the Bay of Biscay, which was correlated with environmental gradients. The greater tolerance of C. fragile to environmental stress and its opportunistic abilities may favour establishment and spread of the species under a warming scenario.

Algal biomass: A sustainable, economical and renewable approach for microbial production of pectinolytic enzymes using submerged and solid state fermentation techniques

In the past decade, algal waste has been used as useful natural resource for production of enormous range of products that have wide economical and commercial importance. Pectinases are group of enzymes that have wide commercial applications. Hence, current study was designed to utilize algal biomass for the production of pectinases using submerged (SmF) and solid state fermentation (SSF) techniques. Different algal sources including brown (Dictyopteris polypodioides, Sargassum wightii and Dictyopteris divaricata) and green algae (Ulva lactuca and Codium tomentosum) were used and U. lactuca was found to be the most suitable substrate. Several bacterial and fungal strains were screened and among them Bacillus licheniformis KIBGE-IB4 was selected based on maximum pectinase production. SmF and SSF were studied utilizing U. lactuca as a substrate and results revealed that enzyme production was favoured by SmF (2457 ± 3.31 U mg−1) as compared to SSF (1432 ± 1.46 U mg−1). Parametric optimization of pectinase production indicated that B. licheniformis KIBGE-IB4 requires 10.0 g L–1 U. lactuca as a biomass in the medium with a pH 7.0 when incubated at 37 °C for 24 hours. Likewise, production of pectinase using algal resource was also compared with that of the conventional agricultural biomass and it was observed that when U. lactuca was used, the selected bacterial isolate produced a higher yield of enzyme than sugarcane bagasse and rice husk. Hence, it is anticipated that algal biomass can be efficiently utilized as an environmental friendly bioresource for the production of industrially important hydrolytic enzymes.

Steroids from an Endophytic Eurotium rubrum Strain

A new steroid, 3β,5α-dihydroxy-10α-methyl-6β-acetoxy-ergosta-7,22-diene (1), and ten known steroids (2–11), were isolated from the cultures of Eurotium rubrum, an endophytic fungus isolated from the marine green alga Codium fragile. Structures of these compounds were unambiguously established by spectroscopic techniques and by comparison with literature values. These steroids exhibited low activity to inhibit acetylcholinesterase and no activity against plant pathogenic fungi Colletotrichum lagenarium and Fusarium oxysporium.

Two New Sulfonoglycolipids from the Green Alga Codium dwarkense

Two new sulfonoglycosides named Codioside E (1) and Codioside F (2) have been isolated from the MeOH extract of the marine green alga Codium darkense.collected:from the coastal areas of the Gulf of Oman. The structures of these secondary metabolites were elucidated by combined analysis of lD (¹H- and ¹³C-) and 2D (H-C correlations; HSQC and HMBC) NMR spectroscopic and MS data.

DNA barcodes and morphometric data indicate that Codium fragile (Bryopsidales, Chlorophyta) may consist of two species

:The siphonous green alga Codium fragile currently has 10 accepted subspecies, among which is the problematic invasive subsp. fragile. The subspecies are defined on the basis of five polymorphic nucleotides in the plastid rps3–rpl16 spacer region, and this subdivision into subspecies has not been investigated in detail with other DNA markers. Here, we compiled a data set of the green algal DNA barcoding marker tufA that includes 110 C. fragile specimens and an additional 80 specimens of eight related Codium species. Our tufA haplotype tree showed two distinct clades within C. fragile, and the three algorithmic species delimitation methods that were applied agreed that these are likely to represent separate species. The first clade consisted of the invasive subspecies fragile, whereas the second contained the remaining subspecies present among our sequences. Using a newly developed morphometric method to capture fine details of utricle morphology, we show that the two molecular clades also have subtle morphological differences. Despite all this new evidence in favour of two species being present in C. fragile, we refrain from making formal taxonomic changes, chiefly because our tufA data do not agree with previous rps3–rpl16 results. We propose four complementary approaches to resolve these conflicts.

Sequence analysis of the pyruvylated galactan sulfate-derived oligosaccharides by negative-ion electrospray tandem mass spectrometry

Five sulfated oligosaccharide fragments, F1–F5, were prepared from a pyruvylated galactan sulfate from the green alga Codium divaricatum, by partial depolymerization using mild acid hydrolysis and purification with gel-permeation chromatography. Negative-ion electrospray tandem mass spectrometry with collision-induced dissociation (ES-CID-MS/MS) is attempted for sequence determination of the sulfated oligosaccharides. The sequence of F1 with homogeneous disaccharide composition was first characterized to be Galp-(4SO4)-(1 → 3)-Galp by detailed nuclear magnetic resonance spectroscopic analyses. The fragmentation pattern of F1 in the product ion spectra was established on the basis of negative-ion ES-CID MS/MS, which was then applied to sequence analysis of other sulfated oligosaccharides. The sequences of F2 and F3 were deduced to be Galp-(4SO4)-(1 → 3)-Galp-(1 → 3)-Galp-(1 → 3)-Galp and 3,4-O-(1-carboxyethylidene)-Galp-(6SO4)-(1 → 3)-Galp, respectively. The sequences of major fragments in F4 and F5 were also deduced. The investigation demonstrated that negative-ion ES-CID-MS/MS was an efficient method for the sequence analysis of the pyruvylated galactan sulfate-derived oligosaccharides which revealed the patterns of substitution and glycosidic linkages. The pyruvylated galactan sulfate-derived oligosaccharides were novel sulfated oligosaccharides different from other algal polysaccharide-derived oligosaccharides.

Codium fragile subsp. fragile (Suringar) Hariot in Tunisia: morphological data and status of knowledge

The Mediterranean Sea is currently facing dramatic changes and threats, including change in native species and accidental introductions. The introduced green alga Codium fragile subsp. fragile (Suringar) Hariot influences diversity and community structure in some parts of the world. This paper docum...

Dynamics of recruitment and establishment of the invasive seaweed Codium fragile within an eelgrass habitat.

Knowledge of the potential distribution (i.e. abundance and spatial extent) of an invasive species is important to estimating its potential impacts on recipient communities. Most previous studies have focused on the potential spatial extent of invasive species populations at regional scales, but little is known on how species successfully recruit and establish at more local scales. In this study, we examined how recruitment of the green alga Codium fragile ssp. fragile (hereafter Codium) can vary spatially and the environmental factors associated with Codium establishment in eelgrass (Zostera marina) beds. Standardized recruitment blocks (65 blocks in a 720 × 240 m2 grid) were used to monitor the number of Codium recruits, juveniles and adults over 2 years. Environmental factors (depth, relative water flow, light and temperature) and attributes of the surrounding macrophyte assemblage (eelgrass density, eelgrass length, Codium biomass) were also measured. Recruitment occurred on all blocks or nearby artificial structures (i.e. buoys) and mainly originated from button stages (i.e. female gametes or utricles). Contrary to other studies, the abundance of Codium (recruits, juveniles and adults) was best predicted by the density of the native canopy-forming species, Z. marina, which highlights a positive interaction between native and non-native canopy-forming species. Seasonal variation in recruitment was observed; it was lower during the summer. Recruitment did not show any distinct spatial pattern (e.g. gradient or patch), but the same spatial pattern of recruitment was observed every sampling date, suggesting that there are “hotspots” for recruitment. In general, the total number of Codium fronds observed on a block at the end of the experiment was positively correlated with the cumulative number of recruits. However, recruitment occurred on some blocks but recruits never grew, suggesting that some environmental factors limit Codium distribution and abundance in eelgrass beds. Overall, the assessment of Codium recruitment over 2 years showed that the colonization of suitable locations by Codium within seagrass beds may take several years and that some factors may not only limit, but also inhibit Codium expansion within eelgrass beds.

Modelling the spread of the invasive alga Codium fragile driven by long-distance dispersal of buoyant propagules

The secondary spread of an invasive species after initial establishment is a major factor in determining its distribution and impacts. Determining and understanding the factors driving this secondary spread is therefore of great importance to manage and predict invasions. In this study we constructed an individual-based model for the spread of the invasive green alga Codium fragile ssp. fragile along a straight coastline, in order to understand the factors governing spreading speed. Codium can spread locally through non-buoyant propagules, while long-distance dispersal depends on the wind-driven dispersal of buoyant fragments. Since fragment buoyancy is determined by light conditions, we first modelled the buoyancy of fragments, yielding a dispersal time dependent on light conditions. We then used this dispersal time, along with empirical wind speeds and directions to calculate a dispersal kernel for fragments. Finally, we incorporated this dispersal kernel into a population growth model including survival rate and fragmentation rate, to calculate a population spreading speed. We found that under current environmental conditions along the east coast of Canada, (the northernmost front of this invasion) further spread towards the northeast is possible but limited (only 4kmyr−1). However, a sensitivity analysis showed that environmental shifts associated with climate change, such as more variable winds and increased disturbances that cause fragmentation, have the potential to increase spreading speed and particularly northward spread.

New α-Glucosidase Inhibitory Triterpenic Acid from Marine Macro Green Alga Codium dwarkense Boergs.

The marine ecosystem has been a key resource for secondary metabolites with promising biological roles. In the current study, bioassay-guided phytochemical investigations were carried out to assess the presence of enzyme inhibitory chemical constituents from the methanolic extract of marine green alga—Codium dwarkense. The bioactive fractions were further subjected to chromatographic separations, which resulted in the isolation of a new triterpenic acid; dwarkenoic acid (1) and the known sterols; androst-5-en-3β-ol (2), stigmasta-5,25-dien-3β,7α-diol (3), ergosta-5,25-dien-3β-ol (4), 7-hydroxystigmasta-4,25-dien-3-one-7-O-β-d-fucopyranoside (5), 7-hydroxystigmasta-4,25-dien-3-one (6), and stigmasta-5,25-dien-3β-ol (7). The structure elucidation of the new compound was carried out by combined mass spectrometry and 1D (1H and 13C) and 2D (HSQC, HMBC, COSY, and NOESY) NMR spectroscopic data. The sub-fractions and pure constituents were assayed for enzymatic inhibition of alpha-glucosidase. Compound 1 showed significant inhibition at all concentrations. Compounds 2, 3, 5, and 7 exhibited a dose-dependent response, whereas compounds 4–6 showed moderate inhibition. Utilizing such marine-derived biological resources could lead to drug discoveries related to anti-diabetics.

Ecology ofCodium fragileSubsp.FragilePopulations Within Salt Marsh Pannes in Southern Maine

Abstract The chloroplast haplotypes, ecology, growth, and reproduction of the introduced Japanese green alga Codium fragile were evaluated in four salt marsh pannes from Brave Boat Harbor, Kittery, Maine. All chloroplast haplotypes were identical and they matched the subsp. fragile delineated by Provan et al. Frond stature was seasonally variable and showed a pronounced increase in length from summer to early fall. Residual (i.e., overwintering) spring fronds were highly variable in stature compared to previous fall values, with some fronds only being ~ 0.5–1.0 cm. Frond stature and frequency patterns in the different pannes were related to their proximity to tidal channels, plus patterns of storm damage. Thus, one panne, which was closest to a tidal channel and had the lowest tidal level, exhibited the largest population frequency and smallest mean frond stature. The highest percent occurrence of gametangia occurred during late summer to early fall when the largest fronds were present. The reproductive p...

Structural characterization and anticoagulant activity of a sulfated polysaccharide from the green alga Codium divaricatum

A sulfated polysaccharide, designated CP2-1, was isolated from the green alga Codium divaricatum by water extraction and purified by anion-exchange and size-exclusion chromatography. CP2-1 is a galactan which is highly sulfated and substituted with pyruvic acid ketals. On the basis of chemical and spectroscopic analyses, the backbone of CP2-1 was mainly composed of (1→3)-β-d-galactopyranose residues, branched by single (1→)-β-d-galactopyranose units attached to the main chain at C-4 positions. The degree of branching was estimated to be about 12.2%. Sulfate groups were at C-4 of (1→3)-β-d-galactopyranose and C-6 of non-reducing terminal galactose residues. In addition, the ketals of pyruvic acid were found at 3,4- of non-reducing terminal galactose residues forming a five-membered ring. CP2-1 possessed a high anticoagulant activity as assessed by the activated partial thromboplastin time and thrombin time assays. The investigation demonstrated that CP2-1 was an anticoagulant-active sulfated polysaccharide distinguishing from other sulfated polysaccharides from marine green algae.

Coexistence of congeneric native and invasive species: The case of the green algae Codium spp. in northwestern Spain

We examined the patterns of distribution and abundance, and reproductive traits (presence of gametophytes and size at time of reproduction) in the invasive Codium fragile ssp. fragile and the native C. tomentosum and C. vermilara on intertidal habitats of NW Spain at two dates. All three species coexist in the locations and habitats studied, although abundances were low. We found a greater proportion of C. fragile ssp. fragile towards the east of the Cantabrian coast and on upper levels on the shore, where conditions are more stressful. The proportion of thalli bearing gametangia in C. fragile ssp. fragile was greater than in the native species in all habitats. The presence of gametangia was size-dependent for all species, with the invasive species maturing at a smaller size, which combined with the previous features, might confer competitive advantages to this species over the native species. We also demonstrated that molecular analyses are necessary for the correct identification of C. fragile subspecies.