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J. Biochem, 1984, Vol. 96, No. 4 1005-1011
© 1984 Japanese Biochemical Society

research-article

Structure of the Cell Wall Proteogalactomannan from Neurospora crassa

I. Purification of the Proteoheteroglycan and Characterization of Alkali-Labile Oligosaccharides

Tasuku NAKAJIMA, Masahiro YOSHIDA, Nozomi HIURA and Kazuo MATSUDA

Department of Agricultural Chemistry, Tohoku University Tsutsumidori-Amamiyamachi, Sendai, Miyagi 980

Proteoheteroglycan (PHG) was prepared from Neurospora crassa cells by extraction with hot water followed by cetyltrimethylammoniumbromide fractionation. The polymer was purified by DEAE-celluIose chromatography followed by gel filtrations. The PHG was fractionated into five subfractions containing carbohydrate (65–88%), protein (19–36%), and a trace amount of phosphate (0.3–1.9%). The sugar compositions of the fractions were similar to each other (D-mannose, 47–60%, D-galactose, 35–50%, glucose, 2–5%) while the fractions showed significant heterogeneity in molecular size.

Mild alkali treatment of the PHG in the presence of sodium borohydride yielded three kinds of reduced oligosaccharides. Structural studies using a methylation-GC-MS method, and proton and carbon NMR indicated that the tetrasaccharide fragment is ß-D-Galf(l-5)-beta;-D-Galf(l-2)-{alpha}-D-Manp(l-2)mannitol, the trisaccharide is ß-D-Galf(l-2)-{alpha}-D-Manp(l-2)mannitol, and the disaccharide is {alpha}-D-Manp(l-2)mannitol.


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