A new arabinomannan from the cell wall of the chlorococcal algae Chlorella vulgaris

Abstract

A new arabinomannan has been isolated from the cell wall of the green algae Chlorella vulgaris by extraction with 0.1M NaOH, dialysis and SEC fractionation. Mw was about 8000u. Terminal, 2- and 5-O-linked arabinofuranosyl residues, and 2,6-O-linked mannopyranosyl residues were detected as the main constituents beside some minor mannose components by methylation analysis. Electrospray ionisation mass spectrometry (ESI–MS) with collision induced dissociation (CID) up to MS3 experiments of oligosaccharides obtained by partial methanolysis or partial acid hydrolysis indicated the presence of Man→Man- and Ara→Ara- as well as Ara→Man- and also Man→Ara-sequences. MS2 experiments gave evidence of a (1→6)-linked mannan, probably also including some (1→2)-linked mannosyl and 5-linked arabinofuranosyl residues. Mannooligosaccharides up to DP5 with mainly (1→6)-, but at higher DP also (1→2)-linkages were obtained by acid hydrolysis, and arabino oligomers up to DP4 could be detected after mild methanolysis. In accordance with results from methylation analysis and ESI–MS/CID t-β-Araf, 2-α-Araf, 5-α-Araf and 2,6-α-Manp were identified from homo- and heteronuclear 1D- and 2D NMR experiments in a molar ratio of ∼2:2:1:2. A highly branched structure is suggested with a 6- and 2-O-linked mannan main chain, comprising 5-Araf residues. Araf-β-[(1→2)-Araf-α-(1→]n side chains with an average chain length of 2 are linked to the mannan main chain. The configuration of Ara is d. The new polysaccharide shows structural similarities with the (lipo)arabinomannans of Mycobacterium tuberculosis.