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

Mar Biotechnol (NY). 2016 Jun;18(3):301-13. doi: 10.1007/s10126-016-9696-6. Epub 2016 May 6.

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.

Keywords: Alternaria sp. SP-32; Bioactivity; Chemical characterization; Extracellular polysaccharide; Marine sponge endogenous fungus.

MeSH terms

  • Alternaria / chemistry*
  • Alternaria / metabolism
  • Animals
  • Antioxidants / chemistry*
  • Antioxidants / isolation & purification
  • Antioxidants / pharmacology
  • Biphenyl Compounds / antagonists & inhibitors
  • Carbohydrate Sequence
  • Cell Survival / drug effects
  • Fermentation
  • Fungal Polysaccharides / chemistry*
  • Fungal Polysaccharides / isolation & purification
  • Fungal Polysaccharides / pharmacology
  • Galactans / chemistry
  • Galactose / chemistry
  • Glucans / chemistry
  • Glucose / chemistry
  • HL-60 Cells
  • HeLa Cells
  • Humans
  • Hydroxyl Radical / antagonists & inhibitors
  • K562 Cells
  • Mannans / chemistry
  • Mannose / chemistry
  • Molecular Weight
  • Picrates / antagonists & inhibitors
  • Polysaccharides, Bacterial / chemistry
  • Porifera / microbiology*
  • Seawater / microbiology
  • Solid Phase Extraction / methods
  • Symbiosis / physiology

Substances

  • Antioxidants
  • Biphenyl Compounds
  • Fungal Polysaccharides
  • Galactans
  • Glucans
  • Mannans
  • Picrates
  • Polysaccharides, Bacterial
  • Hydroxyl Radical
  • galactoglucan
  • 1,1-diphenyl-2-picrylhydrazyl
  • Glucose
  • Mannose
  • Galactose