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Mechanochemical phosphorylation and solubilisation of β-D-glucan from yeast Saccharomyces cerevisiae and its biological activities.

Shi F, Shi J, Li Y - PLoS ONE (2014)

Bottom Line: The energy transferred was 3.03-11.98 KJ/g.Three GP products with different degree of substitution (DS) and degree of polymerisation (DP) were able to upregulate the functional events mediated by activated murine macrophage RAW264.7 cells, among which GP-2 with a DS of 1.24 and DP of 30.5 exerted the highest immunostimulating activity.Our results indicate that mechanochemical processing is an efficient method for preparing water-soluble and biologically active GP with high DS.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China; Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China; Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, China.

ABSTRACT
To obtain a water-soluble β-D-glucan derivative cleanly and conveniently, a highly efficient mechanochemical method, planetary ball milling, was used to phosphorylate β-D-glucan isolated from yeast Saccharomyces cerevisiae in solid state. Soluble β-D-glucan phosphate (GP) with a high degree of substitution (0.77-2.09) and an apparent PEAK molecular weight of 6.6-10.0 kDa was produced when β-D-glucan was co-milled with sodium hexametaphosphate at 139.5-186.0 rad/s for 12-20 min. The energy transferred was 3.03-11.98 KJ/g. The phosphorylation of GPs was demonstrated by Fourier transform infrared spectroscopy and 13C and 31P Nuclear magnetic resonance spectroscopy. Three GP products with different degree of substitution (DS) and degree of polymerisation (DP) were able to upregulate the functional events mediated by activated murine macrophage RAW264.7 cells, among which GP-2 with a DS of 1.24 and DP of 30.5 exerted the highest immunostimulating activity. Our results indicate that mechanochemical processing is an efficient method for preparing water-soluble and biologically active GP with high DS.

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31P NMR spectra of insoluble β-D-glucan particles NG, β-D-glucan phosphate GP-2 prepared by planetary ball milling, and sodium hexametaphosphate (NaPO3)6 milled alone.
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pone-0103494-g004: 31P NMR spectra of insoluble β-D-glucan particles NG, β-D-glucan phosphate GP-2 prepared by planetary ball milling, and sodium hexametaphosphate (NaPO3)6 milled alone.

Mentions: Further, 31P NMR spectral analysis of the initial insoluble β-D-glucan particles NG in DMSO-d6 and the soluble derivative GP-2 in D2O was undertaken to confirm the presence of the phosphate group in GP product. No signal was observed in NG, whereas a strong phosphorus signal was observed at 2.69 ppm in GP-2 (Fig. 4), similar with previously reported β-D-glucan phosphate [5], [42] as well as phosphated (1→3)-α-D-glucan [43], confirming that phosphate groups were bound to the molecular chain. In addition, two small signals were observed at –7.44 and –18.75 ppm, suggesting that other types of phosphate group were present in the phosphated β-D-glucan derivative, perhaps being minor portions of polysaccharide diphosphates and oligophosphates, as reported in starch phosphates [30]. This was further demonstrated by a weak signal at –9.87 ppm and a strong signal at –21.66 ppm observed in 31P NMR spectrum of sodium hexametaphosphate milled alone [milled (NaPO3)6] (Fig. 4). Therefore, in view of the 13C and 31P NMR results, phosphate groups have been chemically bound to β-D-glucan.


Mechanochemical phosphorylation and solubilisation of β-D-glucan from yeast Saccharomyces cerevisiae and its biological activities.

Shi F, Shi J, Li Y - PLoS ONE (2014)

31P NMR spectra of insoluble β-D-glucan particles NG, β-D-glucan phosphate GP-2 prepared by planetary ball milling, and sodium hexametaphosphate (NaPO3)6 milled alone.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4116193&req=5

pone-0103494-g004: 31P NMR spectra of insoluble β-D-glucan particles NG, β-D-glucan phosphate GP-2 prepared by planetary ball milling, and sodium hexametaphosphate (NaPO3)6 milled alone.
Mentions: Further, 31P NMR spectral analysis of the initial insoluble β-D-glucan particles NG in DMSO-d6 and the soluble derivative GP-2 in D2O was undertaken to confirm the presence of the phosphate group in GP product. No signal was observed in NG, whereas a strong phosphorus signal was observed at 2.69 ppm in GP-2 (Fig. 4), similar with previously reported β-D-glucan phosphate [5], [42] as well as phosphated (1→3)-α-D-glucan [43], confirming that phosphate groups were bound to the molecular chain. In addition, two small signals were observed at –7.44 and –18.75 ppm, suggesting that other types of phosphate group were present in the phosphated β-D-glucan derivative, perhaps being minor portions of polysaccharide diphosphates and oligophosphates, as reported in starch phosphates [30]. This was further demonstrated by a weak signal at –9.87 ppm and a strong signal at –21.66 ppm observed in 31P NMR spectrum of sodium hexametaphosphate milled alone [milled (NaPO3)6] (Fig. 4). Therefore, in view of the 13C and 31P NMR results, phosphate groups have been chemically bound to β-D-glucan.

Bottom Line: The energy transferred was 3.03-11.98 KJ/g.Three GP products with different degree of substitution (DS) and degree of polymerisation (DP) were able to upregulate the functional events mediated by activated murine macrophage RAW264.7 cells, among which GP-2 with a DS of 1.24 and DP of 30.5 exerted the highest immunostimulating activity.Our results indicate that mechanochemical processing is an efficient method for preparing water-soluble and biologically active GP with high DS.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China; Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China; Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, China.

ABSTRACT
To obtain a water-soluble β-D-glucan derivative cleanly and conveniently, a highly efficient mechanochemical method, planetary ball milling, was used to phosphorylate β-D-glucan isolated from yeast Saccharomyces cerevisiae in solid state. Soluble β-D-glucan phosphate (GP) with a high degree of substitution (0.77-2.09) and an apparent PEAK molecular weight of 6.6-10.0 kDa was produced when β-D-glucan was co-milled with sodium hexametaphosphate at 139.5-186.0 rad/s for 12-20 min. The energy transferred was 3.03-11.98 KJ/g. The phosphorylation of GPs was demonstrated by Fourier transform infrared spectroscopy and 13C and 31P Nuclear magnetic resonance spectroscopy. Three GP products with different degree of substitution (DS) and degree of polymerisation (DP) were able to upregulate the functional events mediated by activated murine macrophage RAW264.7 cells, among which GP-2 with a DS of 1.24 and DP of 30.5 exerted the highest immunostimulating activity. Our results indicate that mechanochemical processing is an efficient method for preparing water-soluble and biologically active GP with high DS.

Show MeSH