Preparation and Immunological Activity Evaluation of Water-soluble β-Glucan from Saccharomyces cerevisiae †

doi:10.7503/cjcu20190195

Abstract

Abstract:

The wasted beer yeast(Saccharomyces cerevisiae) was used as raw material to obtain insoluble β-glucan(SCBG) through acid and alkaline treatment. The water-soluble β-glucan(SCBGs) was prepared by oxidative degradation in the NaOH/urea aqueous solution at low temperature. Four purified components(SCBGs-0-1, SCBGs-1-1, SCBGs-1-2 and SCBGs-1-3) were further obtained by column chromatography. The monosaccharide compositions, molecular weights and chemical structures of four purified components were analyzed by high performance liquid chromatography(HPLC), high performance gel permeation chromatography-multiple angle laser light scattering(HPGPC-MALLS), nuclear magnetic resonance spectroscopy(NMR) and Congo-red test. Their immunological activities, including the effects of phagocytosis and the release of NO and TNF-α, were studied on macrophage RAW 264.7. The results showed that all purified component mainly consisted of glucose. They were kinds of β-1,3/1,6-glucans composed of β-1,3-glucan as backbone with branches at the site of C6 hydroxyl groups. The molecular weights were determined to be 198000, 960000, 270000 and 18700, respectively. All the components still retained their original supercoil structure except SCBGs-1-3. Four purified components could significantly promote the immunological activity of RAW 264.7.

Key words: Wasted beer yeast, Water-soluble β-glucan;, Supercoil structure, Immunological activity

CLC Number:

O629.1

TrendMD:

WANG Shanshan, CAI Chao, HAO Jiejie, LI Guoyun, WANG Xueliang, HU Minghua, FAN Luodi, YU Guangli. Preparation and Immunological Activity Evaluation of Water-soluble β-Glucan from Saccharomyces cerevisiae ^†[J]. Chem. J. Chinese Universities, 2019, 40(9): 1873.

Figures/Tables 8

Fig.1 Thesolubility of SCBG in H2O(A), 6%NaOH(B) and NaOH-Urea-H2O(mass ratio 6∶18∶76) with(C) or without refrigeration(D)

Fig.2 Separation of water soluble β-glucan from the wasted beer yeast by QFF(A) and Sephacryl S-300HR column chromatography(B and C)

Sample	Total sugar content(%)	Protein content(%)	Molecular weight	M_w/M_n
SCBGs-0-1	95.36	1.88	198000	1.05
SCBGs-1-1	92.73	3.50	960000	1.32
SCBGs-1-2	96.69	1.93	270000	1.11
SCBGs-1-3	85.32	8.95	18700	1.15

Fig.3 HPGPC distribution of SCBGs-0-1(A), SCBGs-1-1(B), SCBGs-1-2(C) and SCBGs-1-3(D)

Fig.4 Absorption spectra of Congo-red without(a) or with(b) SCBGs-0-1(A), SCBGs-1-1(B), SCBGs-1-2(C) and SCBGs-1-3(D)

Fig.5 1H NMR(A), 13C NMR(B) and 1H-13C HSQC(C) of SCBGs at 25 ℃ and 1H NMR of SCBGs-0-1(a), SCBGs-1-1(b), SCBGs-1-2(c) and SCBGs-1-3(d) at 50 ℃(D)

Purified β-glucans	Integral area of A/B(H1)	Integral area of B(H6)	Branching degree(%)
SCBGs-0-1	1	0.11	11
SCBGs-1-1	1	0.12	12
SCBGs-1-2	1	0.13	13
SCBGs-1-3	1	0.23	23

Fig.6 Effects of different doses of samples on the cell viability(A), pinocytotic uptake(B), NO release(C) and TNF-α release(D) of the macrophage RAW 264.7 Results are expressed as means±S.D., *P<0.05, ** P<0.01, *** P<0.001, # P<0.0001 compared with control, as determined by unpaired student’s t-test. a. Control; b. LPS; c. LNT; d. SCBGs-0-1; e. SCBGs-1-1; f. SCBGs-1-2; g. SCBGs-1-3.

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