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Purification, Chemical Characterization, and Bioactivity of an Extracellular Polysaccharide Produced by the Marine Sponge Endogenous Fungus Alternaria sp. SP-32

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Abstract

Marine sponges are ancient and simple multicellular filter-feeding invertebrates attached to solid substrates in benthic habitats and host a variety of fungi both inside and on their surface because of its unique ingestion and digest system. Investigation on marine sponge-associated fungi mainly focused on the small molecular metabolites, yet little attention had been paid to the extracellular polysaccharides. In this study, a homogeneous extracellular polysaccharide AS2-1 was obtained from the fermented broth of the marine sponge endogenous fungus Alternaria sp. SP-32 using ethanol precipitation, anion-exchange, and size-exclusion chromatography. Results of chemical and spectroscopic analyses showed that AS2-1 was composed of mannose, glucose, and galactose with a molar ratio of 1.00:0.67:0.35, and its molecular weight was 27.4 kDa. AS2-1 consists of a mannan core and a galactoglucan chain. The mannan core is composed of (1→6)-α-Manp substituted at C-2 by (1→2)-α-Manp with different degrees of polymerization. The galactoglucan chain consists of (1→6)-α-Glcp residues with (1→6)-β-Galf residues attached to the last glucopyranose residue at C-6. (1→6)-β-Galf residues have additional branches at C-2 consisting of disaccharide units of (1→2)-β-Galf and (1→2)-α-Glcp residues. The glucopyranose residue of the galactoglucan chain is linked to the mannan core. AS2-1 possessed a high antioxidant activity as evaluated by scavenging of 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radicals in vitro. AS2-1 was also evaluated for cytotoxic activity on Hela, HL-60, and K562 cell lines by the MTT and SRB methods. The investigation demonstrated that AS2-1 was a novel extracellular polysaccharide with different characterization from extracellular polysaccharides produced by other marine microorganisms.

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References

  • Abdelmohsen UR, Pimentel-Elardo SM, Hanora A, Radwan M, Abou-El-Ela SH, Ahmed S, Hentschel U (2010) Isolation, phylogenetic analysis and anti-infective activity screening of marine sponge-associated Actinomycetes. Mar Drugs 8:399–412

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ahrazem O, Prieto A, Leal JA, Giménez-Abián MI, Jiménez-Barbero J, Bernabé M (2007) Fungal cell wall polysaccharides isolated from Discula destructiva spp. Carbohydr Res 342:1138–1143

    Article  CAS  PubMed  Google Scholar 

  • Al-Nahas MO, Darwish MM, Ali AE, Amin MA (2011) Characterization of an exopolysaccharide-producing marine bacterium, isolate Pseudoalteromonas sp. AM. Afr J Microbiol Res 5:3823–3831

    Article  CAS  Google Scholar 

  • Aruoma OI (1998) Free radicals, oxidative stress, and antioxidants in human health and disease. J Am Oil Chem Soc 75:199–212

    Article  CAS  Google Scholar 

  • Baker PW, Kennedy J, Dobson A, Marchesi JR (2009) Phylogenetic diversity and antimicrobial activities of fungi associated with Haliclona simulans isolated from Irish coastal waters. Mar Biotechnol 11:540–547

    Article  CAS  PubMed  Google Scholar 

  • Bitter T, Muir HM (1962) A modified uronic acid carbazole reaction. Anal Biochem 4:330–334

    Article  CAS  PubMed  Google Scholar 

  • Braca A, De Tommasi N, Di Bari L, Pizza C, Politi M, Morelli I (2001) Antioxidant principles from Bauhinia terapotensis. J Nat Prod 64:892–895

    Article  CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Chen Y, Mao WJ, Yang YP, Teng XC, Zhu WM, Qi XH, Chen YL, Zhao CQ, Hou YJ, Wang CY, Li N (2012) Structure and antioxidant activity of an extracellular polysaccharide from coral-associated fungus, Aspergillus versicolor LCJ-5-4. Carbohydr Polym 87:218–226

    Article  CAS  Google Scholar 

  • Croft MT, Lawrence AD, Raux-Deery E, Martin J, Warren MJ, Smith AG (2005) Algae acquire vitamin B12 through a symbiotic relationship with bacteria. Nature 383:90–93

    Article  Google Scholar 

  • Ding X, Tang J, Cao M, Guo CX, Zhang X, Zhong J, Zhang SQ, Feng S, Yang ZR, Zhao J (2010) Structure elucidation and antioxidant activity of a novel polysaccharide isolated from Tricholoma matsutake. Int J Biol Macromol 47:271–275

    Article  CAS  PubMed  Google Scholar 

  • Ding B, Yin Y, Zhang FL, Li ZY (2011) Recovery and phylogenetic diversity of culturable fungi associated with marine sponges Clathrina luteoculcitella and Holoxea sp. in the South China Sea. Mar Biotechnol 13:713–721

    Article  CAS  PubMed  Google Scholar 

  • Dobretsov S, Qian PY (2004) The role of epibiotic bacteria from the surface of the soft coral Dendronephthya sp. in the inhibition of larval settlement. J Exp Mar Biol Ecol 299:35–50

    Article  Google Scholar 

  • Dubois C, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356

    Article  CAS  Google Scholar 

  • Guo SD, Mao WJ, Li YL, Tian JH, Xu J (2013) Structural elucidation of the exopolysaccharide produced by fungus Fusarium oxysporum Y24-2. Carbohydr Res 365:9–13

    Article  CAS  PubMed  Google Scholar 

  • Guo SD, Mao WJ, Yan MX, Zhao CQ, Li N, Shan JM, Lin C, Liu X, Guo T, Guo TT, Wang SY (2014) Galactomannan with novel structure produced by the coral endophytic fungus Aspergillus ochraceus. Carbohydr Polym 105:325–333

    Article  CAS  PubMed  Google Scholar 

  • Hakomori SJ (1964) A rapid permethylation of glycolipid and polysaccharide catalyzed by methylsulfinyl carbanion in dimethyl sulfoxide. J Biochem 55:205–208

    CAS  PubMed  Google Scholar 

  • Hiort J, Maksimenka K, Reichert M, Perović-Ottstadt S, Lin WH, Wray V, Steube K, Schaumann K, Weber H, Proksch P, Ebel R, Müller WEG, Bringmann G (2004) New natural products from the sponge-derived fungus Aspergillus niger. J Nat Prod 67:1532–1543

    Article  CAS  PubMed  Google Scholar 

  • Kiho T, Ookubo K, Usui S, Ukai S, Hirano K (1999) Structural features and hypoglycemic activity of a polysaccharide (CS-F10) from the cultured mycelium of Cordyceps sinensis. Biol Pharm Bull 22:966–970

    Article  CAS  PubMed  Google Scholar 

  • Kimura Y, Sumiyoshi M, Suzuki T, Sakanaka M (2006) Antitumor and antimetastatic activity of a novel water-soluble low molecular weight beta-1,3-d-glucan (branch beta-1,6) isolated from Aureobasidium pullulans 1A1 strain black yeast. Anticancer Res 26:4131–4141

    CAS  PubMed  Google Scholar 

  • Kobayashi J, Tsuda M (2004) Bioactive products from Okinawan marine micro- and macroorganisms. Phytochem Rev 3:267–274

    Article  CAS  Google Scholar 

  • Leal JA, Giménez-Abián MI, Canales Á, Jiménez-Barbero J, Bernabé M, Prieto A (2009) Cell wall polysaccharides isolated from the fungus Neotestudina rosatii, one of the etiologic agents of mycetoma in man. Glycoconj J 26:1047–1054

    Article  CAS  PubMed  Google Scholar 

  • Li ZY (2009) Advances in marine microbial symbionts in the China Sea and related pharmaceutical metabolites. Mar Drugs 7:113–129

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li HY, Mao WJ, Chen Y, Ren SM, Qi XH, Chen YL, Zhao CQ, Li N, Wang CY, Lin C, Yan MX, Shan JM (2012) Sequence analysis of the sulfated rhamno-oligosaccharides derived from a sulfated rhamnan. Carbohydr Polym 90:1299–1304

    Article  CAS  PubMed  Google Scholar 

  • Li W, Xia XD, Tang WZ, Ji J, Rui X, Chen XH, Jiang M, Zhou JZ (2015) Structural characterization and anticancer activity of cell-bound exopolysaccharide from Lactobacillus helveticus MB2-1. J Agric Food Chem 63:3454–3463

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Sun ZL, Zhang MS, Meng XM, Xia XK, Yuan WP, Xue F, Liu CH (2012) Antioxidant and antihyperlipidemic activities of polysaccharides from sea cucumber Apostichopus japonicus. Carbohydr Polym 90:1664–1670

    Article  CAS  PubMed  Google Scholar 

  • Maina NH, Tenkanen M, Maaheimo H, Juvonen R, Virkki L (2008) NMR spectroscopic analysis of exopolysaccharides produced by Leuconostoc citreum and Weissella confusa. Carbohydr Res 343:1446–1455

    Article  CAS  PubMed  Google Scholar 

  • Mao WJ, Li HY, Zhang HJ, Qi XH, Sun HH, Chen Y, Guo SD (2009) Chemical characteristic and anticoagulant activity of the sulfated polysaccharide isolated from Monostroma latissimum (Chlorophyta). Int J Biol Macromol 44:70–74

    Article  CAS  PubMed  Google Scholar 

  • Martinichen-Herrero JC, Carbonero ER, Sassaki GL, Gorin PAJ, Iacomini M (2005) Anticoagulant and antithrombotic activities of a chemically sulfated galactoglucomannan obtained from the lichen Cladonia ibitipocae. Int J Biol Macromol 35:97–102

    Article  CAS  PubMed  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  CAS  PubMed  Google Scholar 

  • Olson JB, Kellogg CA (2010) Microbial ecology of corals, sponges, and algae in mesophotic coral environments. FEMS Microbiol Ecol 73:17–30

    Article  CAS  PubMed  Google Scholar 

  • Paul WB, Jonathan K, Alan WD, Julian RM (2009) Phylogenetic diversity and antimicrobial activities of fungi associated with Haliclona simulans isolated from Irish coastal waters. Mar Biotechnol 11:540–547

    Article  Google Scholar 

  • Prieto A, Leal JA, Giménez-Abián MI, Canales Á, Jiménez-Barbero J, Bernabé M (2007) Isolation and structural determination of a unique polysaccharide containing mannofuranose from the cell wall of the fungus Acrospermum compressum. Glycoconj J 24:421–428

    Article  CAS  PubMed  Google Scholar 

  • Qi HM, Zhang QB, Zhao TT, Chen R, Zhang H, Niu XZ, Li ZE (2005) Antioxidant activity of different sulfate content derivatives of polysaccharide extracted from Ulva pertusa (Chlorophyta) in vitro. Int J Biol Macromol 37:195–199

    Article  CAS  PubMed  Google Scholar 

  • Qi XH, Mao WJ, Gao Y, Chen Y, Chen YL, Zhao CQ, Li N, Wang CY, Yan MX, Lin C, Shan JM (2012) Chemical characteristic of an anticoagulant-active sulfated polysaccharide from Enteromorpha clathrata. Carbohydr Polym 90:1804–1810

    Article  CAS  PubMed  Google Scholar 

  • Raghukumar C (2008) Marine fungal biotechnology: an ecological perspective. Fungal Divers 31:19–35

    Google Scholar 

  • Rateb ME, Ebel R (2011) Secondary metabolites of fungi from marine habitats. Nat Prod Rep 28:290–344

    Article  CAS  PubMed  Google Scholar 

  • Saleem M, Ali MS, Hussain S, Jabbar A, Ashraf M, Lee YS (2007) Marine natural products of fungal origin. Nat Prod Rep 24:1142–1152

    Article  CAS  PubMed  Google Scholar 

  • Sassaki GL, Iacomini M, Gorin PAJ (2005) Methylation-GC-MS analysis of arabinofuranose- and galactofuranose-containing structures: rapid synthesis of partially O-methylated alditol acetate standards. An Acad Bras Cienc 77:223–234

  • Selbmann L, Onofri S, Fenice M, Federici F, Petruccioli M (2002) Production and structural characterization of the exopolysaccharide of the Antarctic fungus Phoma herbarum CCFEE 5080. Res Microbiol 153:585–592

    Article  CAS  PubMed  Google Scholar 

  • Shimada K, Fujikawa K, Yahara K, Nakamura T (1992) Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem 40:945–948

    Article  CAS  Google Scholar 

  • Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistic D, Warren JT, Bokesch H, Kenney S, Boyd MR (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82:1107–1112

    Article  CAS  PubMed  Google Scholar 

  • Smirnoff N, Cumbes QJ (1989) Hydroxyl radical scavenging activity of compatible solutes. Phytochemistry 28:1057–1060

    Article  CAS  Google Scholar 

  • Sun HH, Mao WJ, Chen Y, Guo SD, Li HY, Qi XH, Chen YL, Xu J (2009) Isolation, chemical characteristics and antioxidant properties of the polysaccharides from marine fungus Penicillium sp. F23-2. Carbohydr Polym 78:117–124

    Article  CAS  Google Scholar 

  • Sun HH, Mao WJ, Jiao JY, Xu JC, Li HY, Chen Y, Qi XH, Chen YL, Xu J, Zhao CQ, Hou YJ, Yang YP (2011) Structural characterization of the extracellular polysaccharides produced by the marine fungus Epicoccum nigrum JJY-40 and their antioxidant activities. Mar Biotechnol 13:1048–1055

    Article  CAS  PubMed  Google Scholar 

  • Svensson MV, Zhang X, Huttunen E (2011) Structural studies of the capsular polysaccharide produced by Leuconostoc mesenteroides ssp. cremoris PIA2. Biomacromolecules 12:2496–2501

    Article  CAS  PubMed  Google Scholar 

  • Therho TT, Hartiala K (1971) Method for determination of the sulfate content of glycosamino glycans. Anal Biochem 41:471–476

    Article  Google Scholar 

  • Thomas TRA, Kavlekar DP, LokaBharathi PA (2010) Marine drugs from sponge-microbe association—a review. Mar Drugs 8:1417–1468

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tsiapali E, Whaley S, Kalbfleisch J, Ensley HE, Browder IW, Williams DL (2001) Glucans exhibit weak antioxidant activity, but stimulate macrophage free radical activity. Free Radical Biol Med 30:393–402

    Article  CAS  Google Scholar 

  • Umezawa H, Okami Y, Kurasawa S, Ohnuki T, Ishizuka M, Takeuchi T, Shiio T, Yugari Y (1983) Marinactan, antitumor polysaccharide produced by marine bacteria. J Antibiot 5:471–477

    Article  Google Scholar 

  • Wang K, Li W, Rui X, Chen X, Jiang M, Dong M (2014) Characterization of a novel exopolysaccharide with antitumor activity from Lactobacillus plantarum 70810. Int J Biol Macromol 63:133–139

    Article  CAS  PubMed  Google Scholar 

  • Yang XB, Gao XD, Han F, Xu BS, Song YC, Tan RX (2005) Purification, characterization and enzymatic degradation of YCP, a polysaccharide from marine filamentous fungus Phoma herbarum YS4108. Biochimie 87:747–754

    Article  CAS  PubMed  Google Scholar 

  • Zhao HZ, Wang J, Lv FX, Bie XM, Lu ZX (2015) Chemical characterization and antitumor activity of an exopolysaccharide from Pholiota Squarrosa Quel. AS 5.245. Food Sci Biotechnol 24:659–664

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (41476108, 41406142), NSFC-Shandong Joint Fund for Marine Science Research Centers (U1406402-1), the Science and Technology Development Program of Shandong Province, China (2014GHY115015), and the Natural Science Foundation of Zhejiang Province, China (LQ14H300001).

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Authors and Affiliations

  1. Key Laboratory of Marine Drugs, Ministry of Education, Institute of Marine Drugs and Foods, Ocean University of China, 5 Yushan Road, Qingdao, 266003, People’s Republic of China

    Yin Chen, Wen-Jun Mao, Meng-Xia Yan, Xue Liu, Shu-Yao Wang, Zheng Xia, Bo Xiao, Su-Jian Cao, Bao-Qin Yang & Jie Li

  2. College of Food and Pharmacy, Zhejiang Ocean University, 18 Haiyuan Road, Zhoushan, 316000, People’s Republic of China

    Yin Chen

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  1. Yin Chen
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Correspondence to Wen-Jun Mao.

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Supplemental Fig. 1

Total ion gas chromatogram of AS2-1 on GC-MS and mass spectra. (A) Total ion gas chromatogram on GC-MS of AS2-1. (B) mass spectra of the peaks a, b, c, d, e, f. (C) mass spectra of the peaks g, h, i, j, k. (TIF 1524 kb)

(TIF 2764 kb)

(TIF 2808 kb)

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Chen, Y., Mao, WJ., Yan, MX. et al. Purification, Chemical Characterization, and Bioactivity of an Extracellular Polysaccharide Produced by the Marine Sponge Endogenous Fungus Alternaria sp. SP-32. Mar Biotechnol 18, 301–313 (2016). https://doi.org/10.1007/s10126-016-9696-6

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