title: Purification and partial characterization of an acidic α-glucan-protein complex from the fruiting body of Pleurotus sajor-caju and its effect on macrophage activation.
abstract: The aim of this study was to purify an acidic α-glucan-protein complex from the fruiting bodies of Pleurotus sajor-caju by using the cell wall-degrading enzymes, xylanase and cellulase. The acidic glucan-protein complex was separated from a polysaccharide extract by using DEAE Toyopearl 650M anion-exchange and Sepharose CL-6B chromatography. Its homogeneity was ensured by high-performance size-exclusion chromatography and agarose gel electrophoresis. The acidic glucan-protein complex had a molecular weight of approximately 182 kDa. Fourier transform infrared spectroscopy of the acidic glucan-protein complex revealed an α-glycosidic bond and the typical characteristics of polysaccharides and proteins. The amino acid composition of the protein moiety was dominated by proline, glycine, glutamic acid and aspartic acid, indicating that the protein was highly flexible and had a negative charge. Atomic force microscopy proved that the acidic α-glucan-protein complex existed in a spherical conformation. The acidic α-glucan-protein complex stimulated the activation of macrophages, including the production of nitric oxide and tumor necrosis factor-α.
authors.affiliation: School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
authors.forename: Saranya
authors.initials: S
authors.lastname: Satitmanwiwat
authors.forename: Khanok
authors.initials: K
authors.lastname: Ratanakhanokchai
authors.forename: Natta
authors.initials: N
authors.lastname: Laohakunjit
authors.forename: Patthra
authors.initials: P
authors.lastname: Pason
authors.forename: Chakrit
authors.initials: C
authors.lastname: Tachaapaikoon
authors.forename: Khin Lay
authors.initials: KL
authors.lastname: Kyu
mesh_terms: D000818:Animals; D002460:Cell Line; D002480:Cellulase / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases / Q000378:metabolism; D048690:Fruiting Bodies, Fungal / Q000737:chemistry*; D005656:Fungal Proteins / Q000378:metabolism*; D005936:Glucans / Q000302:isolation & purification / Q000378:metabolism*; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D008264:Macrophages / Q000187:drug effects*; D051379:Mice; D008970:Molecular Weight; D020076:Pleurotus / Q000737:chemistry*
chemical_list: D005656:Fungal Proteins; D005936:Glucans; D006023:Glycoproteins; D002480:Cellulase; D043364:Endo-1,4-beta Xylanases