Structure and Metabolically Oriented Efficacy of Fucoidan from Brown Alga Sargassum muticum in the Model of Colony Formation of Melanoma and Breast Cancer Cells.

This work reports the detailed structure of fucoidan from Sargassum miticum (2SmF2) and its ability to potentiate the inhibitory effect of glycolysis inhibitor 2-deoxy-d-glucose (2-DG). 2SmF2 was shown to be sulfated and acetylated galactofucan containing a main chain of alternating residues of 1,3- and 1,4-linked α-l-fucopyranose, fucose fragments with monotonous 1,3- and 1,4-type linkages (DP up to 3), α-d-Gal-(1→3)-α-L-Fuc disaccharides, and 1,3,4- and 1,2,4-linked fucose branching points. The sulfate groups were found at positions 2 and 4 of fucose and galactose residues. 2SmF2 (up to 800 µg/mL) and 2-DG (up to 8 mM) were not cytotoxic against MDA-MB-231 and SK-MEL-28 as determined by MTS assay. In the soft agar-based model of cancer cell colony formation, fucoidan exhibited weak inhibitory activity at the concentration of 400 µg/mL. However, in combination with low non-cytotoxic concentrations of 2-DG (0.5 or 2 mM), 2SmF2 could effectively inhibit the colony formation of SK-MEL-28 and MDA-MB-231 cells and decreased the number of colonies by more than 50% compared to control at the concentration of 200 µg/mL. Our findings reveal the metabolically oriented effect of fucoidan in combination with a glycolysis inhibitor that may be beneficial for a therapy for aggressive cancers.

Rapid mass spectrometric analysis of a novel fucoidan, extracted from the brown alga Coccophora langsdorfii.

The novel highly sulfated (35%) fucoidan fraction Cf2 , which contained, along with fucose, galactose and traces of xylose and uronic acids was purified from the brown alga Coccophora langsdorfii. Its structural features were predominantly determined (in comparison with fragments of known structure) by a rapid mass spectrometric investigation of the low-molecular-weight fragments, obtained by "mild" (5 mg/mL) and "exhaustive" (maximal concentration) autohydrolysis. Tandem matrix-assisted laser desorption/ionization mass spectra (MALDI-TOF/TOFMS) of fucooligosaccharides with even degree of polymerization (DP), obtained by "mild" autohydrolysis, were the same as that observed for fucoidan from Fucus evanescens, which have a backbone of alternating (1 → 3)- and (1 → 4) linked sulfated at C-2 and sometimes at C-4 of 3-linked α -L-Fucp residues. Fragmentation patterns of oligosaccharides with odd DP indicated sulfation at C-2 and at C-4 of (1 → 3) linked α -L-Fucp residues on the reducing terminus. Minor sulfation at C-3 was also suggested. The "exhaustive" autohydrolysis allowed us to observe the "mixed" oligosaccharides, built up of fucose/xylose and fucose/galactose. Xylose residues were found to occupy both the reducing and nonreducing termini of FucXyl disaccharides. Nonreducing galactose residues as part of GalFuc disaccharides were found to be linked, possibly, by 2-type of linkage to fucose residues and were found to be sulfated, most likely, at position C-2.

Structural characteristics and anticancer activity of fucoidan from the brown alga Sargassum mcclurei.

Three different fucoidan fractions were isolated and purified from the brown alga, Sargassum mcclurei. The SmF1 and SmF2 fucoidans are sulfated heteropolysaccharides that contain fucose, galactose, mannose, xylose and glucose. The SmF3 fucoidan is highly sulfated (35%) galactofucan, and the main chain of the polysaccharide contains a →3)-α-L-Fucp(2,4SO3⁻)-(1→3)-α-L-Fucp(2,4SO3⁻)-(1→ motif with 1,4-linked 3-sulfated α-L-Fucp inserts and 6-linked galactose on reducing end. Possible branching points include the 1,2,6- or 1,3,6-linked galactose and/or 1,3,4-linked fucose residues that could be glycosylated with terminal ß-D-Galp residues or chains of alternating sulfated 1,3-linked α-L-Fucp and 1,4-linked ß-D-Galp residues, which have been identified in galactofucans for the first time. Both α-L-Fucp and ß-D-Galp residues are sulfated at C-2 and/or C-4 (and some C-6 of ß-D-Galp) and potentially the C-3 of terminal ß-D-Galp, 1,4-linked ß-D-Galp and 1,4-linked α-L-Fucp residues. All fucoidans fractions were less cytotoxic and displayed colony formation inhibition in colon cancer DLD-1 cells. Therefore, these fucoidan fractions are potential antitumor agents.

Structure and chemopreventive activity of fucoidans from the brown alga Alaria angusta.

Six fucoidan fractions were isolated from the brown alga Alaria angusta. Structures of enzymatic hydrolysis products of the fraction 1AaF2 (Fuc:Gal ~ 1:1; 33 % of sulfates) by fucanase from Wenyingzhuangia fucanilytica were studied by chemical and instrumental (NMR spectroscopy and mass-spectrometry) methods. It was shown that 1AaF2 consisted of two structurally different fucoidans: a sulfated 1,3;1,4-α-L-fucan and an enzyme-resistant sulfated and acetylated complex fucogalactan (Fuc:Gal ~ 1:2; 19 % of sulfates) 1AaF2_HMP containing extended 1,3-linked fucose and 1,3/1,4-linked galactose fragments (up to 5 residues). The fractions 1AaF2 and 1AaF2_HMP were a non-cytotoxic, possessed dose-dependent chemopreventive effect on EGF-induced neoplastic cell transformation of mouse normal epidermal JB6 Cl41 cells and inhibited the colony formation of human melanoma SK-MEL-28 cells.

Determination of the structure and in vitro anticancer activity of fucan from Saccharina dentigera and its derivatives.

The fucoidan SdeF was isolated from brown alga Saccharina dentigera. The structure of the obtained polysaccharide was studied by chemical methods, NMR spectroscopy of the fully and partially desulfated derivatives, and mass spectrometry of the fucoidan fragments, labeled with 18O. The SdeF was shown to be sulfated (40%) 1,3-linked α-L-fucan, with branches at C2. The sulfate groups were found at positions C2 and C4. Derivatives SdeFDS and SdeFPL were obtained by solvolytic desulfation and autohydrolysis of SdeF, respectively. According to 13C NMR data, SdeFDS is 1,3-linked α-L-fucan, while SdeFPL is 4-sulfated 1,3-linked α-L-fucan. Native fucoidan SdeF was shown to be a non-toxic anticancer substance in the model of human malignant melanoma RPMI-7951, colorectal adenocarcinoma HCT-116, and small intestine adenocarcinoma HuTu 80 cells. The partial desulfation of SdeF at C2 and/or the reduction of its Mw, from 229 to 28 kDa, decreased the anticancer activity; complete removal of the sulfated groups and/or Mw reduction to 4.7 kDa further reduced the effect of this polysaccharide.

The structure of fucoidan from Sargassum oligocystum and radiosensitizing activity of galactofucans from some algae of genus Sargassum.

The aim of this study was to establish the fine structure of fucoidan from Sargassum oligocystum and to study the radiosensitizing effect of fucoidans from three algae of genus Sargassum (S. oligocystum, S. duplicatum, and S. feldmannii) with different structures. The fucoidan SoF2 from S. oligocystum was sulfated (32%) galactofucan (Fuc:Gal = 2:1), with a Mw of 183 kDa (Mw/Mn = 2.0). Its supposed structure was found to be predominantly 1,3-linked fucose as the main chain, with branching points at C2 and C4. The branches could be single galactose and/or fucose short chains with terminal galactose residues. Sulfate groups were found at positions C3, C2, and/or C4 of fucose residues and at C2 and/or C4 of galactose residues. The radiosensitizing effect of galactofucans from S. oligocystum, S. duplicatum, and S. feldmannii against human melanoma SK-MEL-28, colon HT-29, and breast MDA-MB-231 cancer cells was investigated. The influence of all investigated polysaccharides treatments with/without X-ray radiation on colony formation of human melanoma cells SK-MEL-28 was weak. Fucoidan from S. feldmannii has been shown to be the most promising radiosensitizing compound against human colon HT-29 and breast MDA-MB-231 cancer cells.

Structural characteristics of carrageenans of red alga Mastocarpus pacificus from sea of Japan.

The sulfated polysaccharide from sterile alga Mastocarpus pacificus was investigated. Partial reductive hydrolysis and NMR spectroscopy showed that the extracted polysaccharides were only carrageenans. According to FT-IR- and NMR spectroscopy this polysaccharide was a hybrid kappa/iota-carrageenan with a predominance of kappa-type units. According to MALDI-TOFMS, oligosaccharide fragments obtained by mild acid hydrolysis had a polymerization degree of 1-9, while chains built up of galactose residues were up to 3. Tandem ESI mass spectrometry together with innovative 18O-labelling method showed that the polymer chain of the carrageenan included kappa-carrabiose, kappa-carratetraose, iota-carrabiose, hybrid kappa/iota oligosaccharide units and contained minor insertions of mu-carrageenan (the precursor of kappa-carrageenan). Parallel artificial membrane permeability assay shown that the studied carrageenan inhibited bile salts permeation through an artificial membrane imitating the gastrointestinal barrier by 50 % on average compared to negative control independent of incubation time. However, its action was less pronounced than the hindering ability of cholestyramine.

Composition of polysaccharides and radiosensitizing activity of native and sulfated laminarans from the Tаuуа basicrassa Kloczc. et Krupn.

Polysaccharide fractions of alginate, laminarans and fucoidans were obtained from the brown alga Tauya basicrassa. Yields of alginate and laminarans were large (19.7 % and 5.62 %, respectively), whereas the content of fucoidans (0.52 %) was not significant. Alginate and laminarans had typical structures for those substances. Fucoidans were low- and medium-sulfated heterogeneous polysaccharides. The fucoidan fraction 1TbF1 was sulfated fucogalactan containing a backbone from 1,6-linked residues of ß-d-galactopyranose with branches at C3 and C4, terminal fucose and galactose residues and fragments from 1,3-; 1,4-; and 1,2-fucose residues. Sulfate groups were found at positions 2 and 4 of fucose, and positions 2, 3 and 4 of galactose residues. Laminaran 2TbL was subjected to a sulfation to obtain the derivative 2TbLS with partial sulfation (46 %) at C2, C4 and C6. It was shown that 2TbL and 2TbLS inhibited colony formation of sensitize-tested colon cancer cells HT-29 and HCT-116 to X-ray radiation.

Triterpene glycosides from Antarctic sea cucumbers. 2. Structure of Achlioniceosides A(1), A(2), and A(3) from the sea cucumber Achlionice violaecuspidata (=Rhipidothuria racowitzai).

Three new triterpene glycosides, achlioniceosides A(1) (1), A(2) (2), and A(3) (3), have been isolated from the Antarctic sea cucumber Achlionice violaecuspidata. The glycoside structures were elucidated using extensive NMR spectroscopic analysis including one-dimensional (1)H and (13)C spectra, (1)H-(1)H-COSY, HMBC, HMQC, and NOESY and mass spectrometry. Gycosides 1-3 are disulfated pentaosides that are branched at the first xylose residue. The sulfates are attached to C-6 of the glucose residues. Glycosides 1-3 are the first triterpene glycosides isolated from a sea cucumber belonging to the order Elasipodida.

Comparison of structure and in vitro anticancer activity of native and modified fucoidans from Sargassum feldmannii and S. duplicatum.

Fucoidans are valuable biologically active polysaccharides of brown algae. The aim of this study was to investigate the structure of fucoidan from Sargassum feldmannii and the anticancer effects of native and modified polysaccharides from S. feldmannii and S. duplicatum. The structure of sulfated (25.3%) galactofucan SfF2 (Fuc/Gal = 72/28 mol%) from S. feldmannii was investigated by NMR spectroscopy of desulfated derivative and mass spectrometry of fucoidan fragments labelled with 18O. SfF2 was shown to contain the main chain from 1,3-linked α-l-fucopyranose and ß-d-galactopyranose residues with fucose branches at C4 and C6 of galactose residues and C2 of fucose residues. The following fragments were also identified in SfF2: Fuc-(1,4)-Fuc, Gal-(1,3)-Gal, and Gal-(1,4)-Gal. The sulfate groups occupied positions C2, C3, and C4 of fucose residues and C2, C3, C4, and C6 of galactose residues. The galactofucans from S. feldmannii, S. duplicatum, and their derivatives exhibited no cytotoxicity in vitro. The native and deacetylated fucoidans (200 µg/mL) inhibited colony formation of human colon cancer cells (DLD-1, HT-29, and HCT-116). Both desulfated fucoidans possessed weak anticancer activity.

Fucoidans from brown algae Laminaria longipes and Saccharina cichorioides: Structural characteristics, anticancer and radiosensitizing activity in vitro.

The sulfated α-l-fucans ScF and LlF were obtained from brown algae of the Laminariaceae family (Saccharina cichorioides and Laminaria longipes). According to spectroscopy NMR, the LlF fucan predominantly contained the →3)-α-l-Fucp-(2SO3-)-(1→4)-α-l-Fucp-(1→2)-α-l-Fucp-(4SO3-)-(1→ repeating units, with small amounts of disaccharide 1,4-linked fragments and 3-sulfated fucose residues. Mass spectrometric analysis revealed the presence of the following fragments in the fucan structure: α-l-Fucp-(2SO3-)-(1→4)-α-l-Fucp-(2SO3-)-(1→3)-α-l-Fucp-(4SO3-); α-l-Fucp-(2,4SO3-)-(1→3)-α-l-Fucp-(1→3)-α-l-Fucp-(4SO3-); α-l-Fucp-(2SO3-)-(1→2)-α-l-Fucp; α-l-Fucp-(2SO3-)-(1→2)-α-l-Fucp-(4SO3-); α-l-Fucp-(2SO3-)-(1→3)-α-l-Fucp; α-l-Fucp-(2,4SO3-)-(1→3)-α-l-Fucp; α-l-Fucp-(4SO3-)-(1→4)-α-l-Fucp; and α-l-Fucp-(4SO3-)-(1→4)-α-l-Fucp-(2SO3-). Both ScF and LlF fucoidans inhibited colony formation and growth of melanoma and colon cancer cells and sensitize-tested cancer cells to X-ray radiation to a comparable degree.

Triterpene glycosides from Antarctic sea cucumbers. 1. Structure of liouvillosides A1, A2, A3, B1, and B2 from the sea cucumber Staurocucumis liouvillei: new procedure for separation of highly polar glycoside fractions and taxonomic revision.

Five new triterpene glycosides, liouvillosides A1 (1), A2 (2), A3 (3), B1 (4), and B2 (5), have been isolated from the Antarctic sea cucumber Staurocucumis liouviellei along with the known liouvilloside A(6), isolated earlier from the same species, and hemoiedemosides A (7) and B (8), isolated earlier from the Patagonian sea cucumber Hemioedema spectabilis. The isolation was carried out using a new chromatographic procedure including application of ion-pair reversed-phase chromatography followed by chiral chromatography on a cyclodextrin ChiraDex column. The structures of the new glycosides were elucidated using extensive NMR spectroscopy (1H and 13C NMR spectrometry, DEPT, 1H-(1)H COSY, HMBC, HMQC, and NOESY), ESI-FTMS, and CID MS/MS, and chemical transformations. Glycosides 1-3 are disulfated tetraosides and glycosides 4 and 5 are trisulfated tetraosides. Glycosides 2 and 3 contain 3-O-methylquinovose, found for the first time as a natural monosaccharide in sea cucumber glycosides. On the basis of analyses of glycoside structures a taxonomic revision is proposed.

Catalytic properties and mode of action of endo-(1-->3)-beta-D-glucanase and beta-D-glucosidase from the marine mollusk Littorina kurila.

A complex of the enzymes from the liver of the marine mollusk Littorina kurila that hydrolyzes laminaran was investigated. Two (1-->3)-beta-d-glucanases (G-I and G-II) were isolated. The molecular mass of G-I as estimated by gel-permeation chromatography and SDS-PAGE analysis was 32 and 40kDa, respectively. The G-II molecular mass according to SDS-PAGE analysis was about 200kDa. The pH optimum for both G-I and G-II was pH 5.4. The G-I had narrow substrate specificity and hydrolyzed only the (1-->3)-beta-d-glucosidic bonds in the mixed (1-->3),(1-->6)- and (1-->3),(1-->4)-beta-d-glucans down to glucose and glucooligosaccharides. This enzyme acted with retention of the anomeric configuration and catalyzed a transglycosylation reaction. G-I was classified as the glucan endo-(1-->3)-beta-d-glucosidase (EC 3.2.1.39). G-II exhibited both exo-glucanase and beta-d-glucoside activities. This enzyme released from the laminaran glucose as a single product, but retained the anomeric center configuration and possessed transglycosylation activity. The hydrolysis rate of glucooligosaccharides by G-I decreased with an increase of the substrate's degree of polymerization. In addition to (1-->3)-beta-d-glucanase activity, the enzyme had the ability to hydrolyze p-nitrophenyl beta-d-glucoside and beta-d-glucobioses: laminaribiose, gentiobiose, and cellobiose, with the rate ratio of 50:12:1. G-II may correspond to beta-d-glucoside glucohydrolase (EC 3.2.1.21).

Tandem mass spectrometry of fucoidan-derived fragments, labeled with heavy-oxygen.

A procedure for the partial depolymerization of sulfated fucans and selective labeling with 18O was developed. A tandem electrospray ionization mass spectrometry (ESI MS/MS) was applied for the direct analysis of mixtures of structurally-different oligosaccharides, derived from the fucoidans of known structure. The presence of label allowed unambiguous distinguishing between the fragment ions of 0,2X0-type at m/z 287 and 2,4A-type at m/z 285, since 18O at the reducing end gave +2 mass shifting. Thus, ESI MS/MS was able to detect (1,2)-type of linkage in disaccharides from the fucoidan of brown alga S. cichorioides for the first time. It was also discovered that 2,4A-type fragments in 4-linked disaccharides that were incorrectly assigned to 0,2X-type previously, suggested, probably, substitution at C-4 in mono- and disaccharide fragments, derived from the fucoidan of the brown alga F. evanescens.

A novel sulfated fucan from Vietnamese sea cucumber Stichopus variegatus: Isolation, structure and anticancer activity in vitro.

In the present study, three sulfated polysaccharides, two fractions of fucosylated chondroitin sulfates, and one sulfated fucan were isolated from the body wall of the Vietnamese sea cucumber Stichopus variegatus. The structure of the sulfated fucan fraction SvF3 from S. variegatus was investigated for the first time. According to NMR spectroscopy data, the sulfated fucan SvF3 contained 1,2- and 1,3-linked α-l-fucopyranose residues. Sulfate groups were found at the 2 and/or 4 positions. The structural analysis of fucoidan was assisted by tandem mass spectrometry; the recently-developed technique of autohydrolysis in heavy­oxygen water for the obtaining of selectively labeled fucoidan fragments was applied. The labeling (+2 Da mass shift at the reducing end) allowed us to assign MS/MS data unambiguously, and thus to confirm the NMR data and revealed minor sulfation at position 3. It was shown that the sulfated fucan SvF3 was not cytotoxic to human breast cancer T-47D and MDA-MB-231 cell lines, and it inhibited colony formation of those cells in vitro. SvF3 also possessed slight activity against migration of MDA-MB-231 cells in vitro.

Structural characteristics and anticancer activity in vitro of fucoidan from brown alga Padina boryana.

The sulfated and acetylated fucoidan fraction, containing fucose, galactose, mannose, glucose and uronic acid residues, was isolated from the brown alga Padina boryana. The structure of galactofucan part was studied after different modifications by NMR spectroscopy and mass spectrometry. It was shown that galactofucan contained the main chain of alternating 1,4-linked α-l-fucopyranose and 1,3-linked ß-d-Galactopyranose. Single fucose residues were found as branches at C4 of galactose residues. Also, fucoidan contained 1,3- or 1,4-linked Fuc-Fuc and Gal-Gal fragments. The sulfate groups occupied positions C2, C3 and C4 of both fucose and galactose residues, which was shown by tandem mass spectrometry of fragments, labeled with heavy-oxygen. The anticancer effect of native and modified fucoidan fractions was studied in vitro on the colorectal carcinoma cells DLD-1 and HCT-116. All fucoidans had no cytotoxicity under 400 µg/mL and inhibited colony formation of cancer cells at concentration of 200 µg/mL.

Structural features and anticancer activity in vitro of fucoidan derivatives from brown alga Saccharina cichorioides.

A fucoidan ScF from brown alga Saccharina cichorioides was extracted, purified and partially depolymerized by autohydrolysis at 37°C for 24, 48 and 72h. Supernatant (SN) and pellet (PL) fractions were obtained by ethanol precipitation of each sample. Unlike spectral data of ScF, NMR of PL derivatives clearly suggested the structure: 1,3-linked α-l-Fucp-4-OSO3- repeating unit. Molecular weights (MWs) of PL fractions were 30, 26 and 18kDa for 24, 48 and 72h of autohydrolyis, respectively. MALDI-TOFMS, size-exclusion HPLC and carbohydrate polyacrylamide-gel electrophoresis (C-PAGE) indicated a similarity of SN mixtures. They consisted mainly of a polysaccharide part (MW 6kDa, C-PAGE data) with a structure similar to PL components (NMR data) and monosaccharides α-l-Fucp-4-OSO3-, α-l-Fucp-2,4-di-OSO3-. PL fractions exhibited almost identical antiproliferative activity in vitro as native fucoidan, while an SN sample for 72h of autohydrolysis was slightly more active against colony formation of colorectal carcinoma cells HT-29.

Polysaccharides from brown algae Sargassum duplicatum: the structure and anticancer activity in vitro.

The laminaran SdL and fucoidan SdF were isolated from brown algae Sargassum duplicatum. SdL was 1,3;1,6-ß-d-glucan (1,3:1,6=6:1) with a main chain, represented by 1,3-linked glucose residues, due to NMR spectroscopy data. Single glucose residues could form branches at C6. Unusual structure of fucoidan SdF was studied by chemical and enzymatic methods, NMR spectroscopy of desulfated and deacetylated polysaccharide and mass spectrometry of fucoidan fragments labeled with 18O. Fucoidan was sulfated (31.7%) and acetylated galactofucan (Fuc:Gal∼1:1) with a main chain of 1,4-linked alternating α-l-fucose and ß-d-galactose residues. Side chains were represented by extensive (DP≥5) 1,3-linked 2,4-disulfated α-l-fucose residues with branching points at C2. Fucose residues in the main chain were sulfated at C2 and less at C3, while galactose residues were sulfated at C2, C3, and less at C4, C6. The fucoidan SdF was effective against colony formation of colon cancer cells in vitro.

Engineering of Chimeric Protein Based on E Protein Domain III of Tick- Borne Encephalitis Virus and OmpF Porin of Yersinia pseudotuberculosis.

BACKGROUND: Tick-borne encephalitis poses a serious public health threat in the endemic regions. The disease treatment is restricted to symptomatic therapy, so great expectations are in the development of the prophylactic and therapeutic vaccines. The domain III of E protein of the tickborne encephalitis virus is the main antigenic domain which includes virus-specific epitopes recognized by neutralizing antibodies. OBJECTIVES: The main objective of this study was to design, express, isolate and characterize the chimeric protein based on the fusion of domain III of E protein of the tick-borne encephalitis virus and bacterial porin OmpF from Yersinia pseudotuberculosis. METHODS: The chimeric gene was obtained by the PCR based fusion method from two fragments containing overlapping linker sequences. Resulting plasmids were transformed into BL21(DE3) pLysS electrocompetent cells for subsequent heterologous protein expression. All recombinant proteins were purified using immobilized metal affinity chromatography under denaturing conditions. The identity of the chimeric protein was confirmed by MALDI-TOF mass spectrometry and immunoblot analysis. The content of antibodies against the EIII protein was estimated in mice blood serum by ELISA. RESULTS: The bacterial partner protein was used for decreasing toxicity and increasing immunogenicity of antigen. The chimeric protein was successfully expressed by the Escherichia coli cells. The purified protein was recognized with immunoblots by anti-E protein of tick-borne encephalitis virus monoclonal antibodies. Furthermore, the protein was able to elicit antibody response against domain III of E protein in immunized mice. CONCLUSION: The newly obtained chimeric antigen could be valuable for the development of the preventing tick-borne encephalitis subunit vaccines.

The comparison of structure and anticancer activity in vitro of polysaccharides from brown algae Alaria marginata and A. angusta.

Laminaran and three fucoidan fractions were obtained from the brown alga Alaria marginata. Alaria angusta, studied earlier by us, has the same polysaccharide composition. Galactofucan AmF3 from A. marginata has a main chain of →3)-α-l-Fucp-(2,4-SO3(-))-(1→residues, similar to galactofucan from A. angusta. However, the structure of the branches in fucoidan AmF3 can differ from those in the fucoidan from A. angusta. The following fragments were identified in AmF3: HexA-(1→2)-Fuc, HexA-(1→2)-Gal, Gal-(1→4)-HexA, Fuc-(1→2)-Gal-6-SO3(-), Fuc-4-SO3(-)-(1→6)-Gal, Gal-(1→2)-Gal-2-SO3(-), Gal-4-SO3(-)-(1 →6)-Gal, Gal-4-SO3(-)-(1→3)-Fuc-(1→3)-Fuc, Fuc-4-SO3(-)-(1→6)-Gal-(1→4)-Gal, Gal-(1→4)-Gal-(1→3)-Fuc, Gal-2-SO3(-)-(1→4)-Gal-(1→4)-Gal, Gal-(1→4)-Gal-6-SO3(-)-(1→2)-Gal. Chains of galactose residues (DP up to 9) were found in AmF3 fucoidan. The laminarans, galactofucans and their derivatives from both algae exhibited no cytotoxicity in vitro. Polysaccharides from A. angusta were more effective against colony formation of HT-29 cells, while those from A. marginata had a greater effect on T-47D cells. Sulfated and desulfated fucoidans possessed weak antitumor activity using SK-MEL-28 cells.