Isolation, physico-chemical characteristics and acute toxicity determination of water-soluble polysaccharide from basidial raw material Ganoderma lucidum

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Abstract

As a result of the study, branched polysaccharides were isolated from the basidiomycete raw materials of Ganoderma lucidum. It has been established that the isolated fractions contain branched polysaccharides in the form of complexes with melanin. After purification of polysaccharides by ion-exchange chromatography, two fractions were obtained from basidial raw materials: neutral polysaccharides GW-1 with a yield of 25.71% and anionic polysaccharides GW-2, the yield of which was 5.26%, respectively. The physicochemical properties of the obtained samples were studied by IR and UV spectroscopy. The degree of purity of the obtained fractions of branched polysaccharides was established. Using gas chromatography, one-dimensional (13C NMR, 1H NMR) and two-dimensional (COSY, TOCSY, HSQC, HMBC, NOESY) NMR spectroscopy, the composition and molecular structure of the obtained polysaccharide samples were determined. The results showed that the isolated and purified polysaccharides are branched glucans with 1,4,6- and 1,3,6-bonds between glucopyranose units. Pharmacotoxicological studies were carried out on white outbred mice and it was found that the resulting polysaccharides belong to class V, practically non-toxic compounds (LD50 ≥ 2000 mg/kg). Isolated polysaccharides (GW) are promising biologically active components, on the basis of which it is possible to create drugs with hepatoprotective and antitumor activity.

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About the authors

S. B. Khaytmetova

Institute of Bioorganic Chemistry named A.S. Sadykov AS RUz

Author for correspondence.
Email: xsb75@mail.ru
Uzbekistan, ul. Mirzo Ulugbek 83, Tashkent, 100125

A. S. Turaev

Institute of Bioorganic Chemistry named A.S. Sadykov AS RUz

Email: xsb75@mail.ru
Uzbekistan, ul. Mirzo Ulugbek 83, Tashkent, 100125

G. A. Khalilova

Institute of Bioorganic Chemistry named A.S. Sadykov AS RUz

Email: xsb75@mail.ru
Uzbekistan, ul. Mirzo Ulugbek 83, Tashkent, 100125

B. I. Muhitdinov

Institute of Bioorganic Chemistry named A.S. Sadykov AS RUz

Email: xsb75@mail.ru
Uzbekistan, ul. Mirzo Ulugbek 83, Tashkent, 100125

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Supplementary files

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2. Fig. 1. Structure of β-glucans of basidiomycetes.

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3. Fig. 2. UV spectrum of water-soluble fractions of polysaccharides isolated from basidial raw materials of Ganoderma lucidum.

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4. Fig. 3. Chromatogram of the separation of polysaccharides GW-1 and GW-2 on an ion exchange column with DEAE-52-cellulose (eluent 0–1 M NaCl gradient, elution rate 1 ml/min).

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5. Fig. 4. IR spectra of samples of crude (GW) and purified (GW-1 and GW2) polysaccharide fractions from Ganoderma lucidum.

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6. Fig. 5. Permethylated alditol acetals: 1,5-di-O-acetyl-2,3,4,6-tetra-O-methyl-glucitol (I); 1,3,5-tri-O-acetyl-2,4,6-tri-O-methyl-glucitol (II); 1,4,5-tri-O-acetyl-2,3,6-tri-O-methyl-glucitol (III); 1,5,6-tri-O-acetyl-2,3,4-tri-O-methylglucitol (IV); 1,4,5,6-tetra-O-acetyl-2,3-di-O-methylglucitol (V); 1,3,5,6-tetra-O-acetyl-2,4-di-O-methyl-glucitol (VI).

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7. Fig. 6. Spectra of 13C-NMR (a), 1H-NMR (b), HSQC (c) and HMBC (d) of the GW-1 polysaccharide.

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8. Fig. 7. Binding order, monosaccharide sequence and possible polysaccharide structures of GW-1.

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