Purification and in vitro anti-proliferative effect of novel neutral polysaccharides from Lentinus edodes

A (1 -> 6)-beta-D-glucan (WPLE-N-1) and two mannogalactoglucans (WPLE-N-2 and WPLE-N-3) were isolated from the basidiocarps of Lentinus edodes by hot water extraction, ethanol precipitation, anion exchange chromatography, and further purified by gel-permeation chromatography (GPC). Their structural features were investigated by high performance liquid chromatography (HPLC), high performance gel-permeation chromatography (HPGPC), methylation analysis, periodate oxidation-Smith degradation, Fourier transform-infrared (FT-IR) and NMR spectroscopy, including two-dimensional (2D) NMR. HPLC analysis revealed that WPLE-N-1 was mainly composed of glucose (92%) with small amount of galactose (3.9%) and mannose (4.1%), WPLE-N-2 and WPLE-N-3 contained mannose-galactose-glucose in the molar ratio of 10:27:63 and 5:12:83, respectively. GPC and HPGPC showed that WPLE-N-1, WPLE-N-2 and WPLE-N-3 are homogeneous fractions and their molecular weights were estimated to be 757.5 kDa, 20.9 kDa and 4.7 kDa, respectively. Chemical and spectroscopic studies indicated that WPLE-N-1 consisted of (1 -> 6)-beta-D-glucopyranosyl residues; while WPLE-N-2 and WPLE-N-3 were found to contain (1 -> 6)-, (1 -> 4)- and (1 -> 3)-alpha-D-glucopyranosyl residues, (1 -> 6)-alpha-D-galactopyranosyl residues, (1 -> 3,6)- and (1 -> 2,4)-alpha-D-mannopyranosyl residues and terminal residues of D-glucopyranosyl residues. On a preliminary bioactivity test, these three polysaccharides exhibited antitumor activity against Sarcoma S-180, Carcinoma HCT-116 and HT-29 in vitro. This finding suggests that mannogalactoglucan should be explored as potential antitumor agents and utilized as tumor cell growth inhibitors for food and pharmaceutical industries. (C) 2012 Elsevier B.V. All rights reserved.