Extraction of β-glucan from <i>Saccharomyces cerevisiae</i>: Comparison of different extraction methods and <i>in vivo</i> assessment of immunomodulatory effect in mice

PENGKUMSRI, Noppawat; SIVAMARUTHI, Bhagavathi Sundaram; SIRILUN, Sasithorn; PEERAJAN, Sartjin; KESIKA, Periyanaina; CHAIYASUT, Khontaros; CHAIYASUT, Chaiyavat

doi:10.1590/1678-457X.10716

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Research Paper • Food Sci. Technol (Campinas) 37 (1) • Jan-Mar 2017 • https://doi.org/10.1590/1678-457X.10716 copy

Extraction of β-glucan from Saccharomyces cerevisiae: Comparison of different extraction methods and in vivo assessment of immunomodulatory effect in mice

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Beta-glucan (BG) is a conserved cell wall components of bacteria, fungi, and yeast. BG is an immunomodulator and stimulates the host immune system. This study was performed to screen Saccharomyces cerevisiae strain with high BG, extraction of BG using different chemical extraction methods, composition analysis of BG, and evaluation of the immunomodulatory effect of high-quality BG using mice model. Ten yeast strains were screened for high BG content using total glucan extraction kit and were subjected to FT-IR analysis. The kit based extraction revealed that HII31 showed a high content of total glucan and BG. HII31 cells were subjected to four different acid/base extractions, which indicated that combination of a strong base (NaOH) and weak acid (CH₃COOH) extraction recovered high BG and a high ratio of polysaccharide, protein, and lipid. Further, the immunomodulatory effect of the selected BG was evaluated using mice, which suggested that low dose of HII31-BG induces the expression of selected pro-inflammatory (IL-17, IFN-γ) and anti-inflammatory cytokine (IL-10) significantly, whereas relatively high dose was required to alter the IL-6 and TGF-β expression. Overall, the present study revealed that BG extracted from HII31 cells alters the expression of studied cytokines, which can be used as a potent immunomodulator in pharmaceutical products.

Keywords:
beta-glucan; Saccharomyces cerevisiae; immunomodulation; FTIR; cytokines

S. No.	S. cerevisiae strains	Yield (g/L)	Total-glucan (%, w/w)	α-glucan (%, w/w)	β-glucan (%, w/w)
1	HII31	3.53 ± 0.26	49.21 ± 3.96a	7.52 ± 0.93	41.69 ± 2.83^a^* * Significant value (p < 0.05);
2	TISTR5003	3.30 ± 0.28	44.52 ± 3.73	8.21 ± 0.96	36.31 ± 2.57
3	TISTR5024	3.41 ± 0.26	46.34 ± 3.82	7.86 ± 0.94	38.48 ± 2.67
4	TISTR5051	3.66 ± 0.30b	47.45 ± 3.87	8.03 ± 0.95	39.42 ± 2.72
5	TISTR5059	3.52 ± 0.28	48.73 ± 3.94b	7.49 ± 0.92	41.24 ± 2.83b
6	TISTR5191	3.23 ± 0.25	46.42 ± 3.82	7.84 ± 0.94	38.58 ± 2.68
7	TISTR5197	3.63 ± 0.22c	45.91 ± 3.80	7.84 ± 0.90	38.07 ± 2.65
8	TISTR5278	3.38 ± 0.27	47.92 ± 3.90	7.85 ± 0.89	40.07 ± 2.76
9	TISTR5328	3.11 ± 0.20	46.25 ± 3.81	7.72 ± 0.95	38.53 ± 2.68
10	TISTR5623	3.72 ± 0.31a	47.93 ± 3.91c	7.33 ± 0.92	40.60 ± 2.78c

S. No.	S. cerevisiae strains	Band area (%)			Range ratio (1:2:3)
S. No.	S. cerevisiae strains	Range 1 925-1,190 cm^-1 (Polysaccharide) (1)	Range 2 1,500-1,700 cm^-1 (Protein) (2)	Range 3 2,800-3,000 cm^-1 (Lipid) (3)	Range ratio (1:2:3)
1	HII31	13.21	9.44	6.29	2.1:1.5:1.0
2	TISTR5003	10.96	9.14	6.09	1.8:1.5:1.0
3	TISTR5024	9.03	9.04	6.02	1.5:1.5:1.0
4	TISTR5051	10.40	9.75	6.50	1.6:1.5:1.0
5	TISTR5059	13.68	10.26	6.84	2.0:1.5:1.0
6	TISTR5191	11.49	10.82	6.76	1.7:1.6:1.0
7	TISTR5197	10.64	9.98	6.65	1.6:1.5:1.0
8	TISTR5278	10.84	8.43	6.02	1.8:1.4:1.0
9	TISTR5328	11.75	7.84	6.53	1.8:1.2:1.0
10	TISTR5623	11.95	10.62	6.64	1.8:1.6:1.0

S. No	S. cerevisiae strains	Band area (%)			Range ratio (1:2:3)
S. No	S. cerevisiae strains	β-1,3-glucan (1)	β-1,4-glucan (2)	β-1,6-glucan (3)	Range ratio (1:2:3)
1	HII31	48.70	15.22	27.40	3.2:1.0:1.8
2	TISTR5003	47.32	18.20	30.94	2.6:1.0:1.7
3	TISTR5024	33.06	12.24	19.59	2.7:1.0:1.6
4	TISTR5051	34.61	12.82	16.67	2.7:1.0:1.3
5	TISTR5059	52.47	17.49	31.48	3.0:1.0:1.8
6	TISTR5191	31.13	14.15	16.98	2.2:1.0:1.2
7	TISTR5197	48.64	15.69	23.54	3.1:1.0:1.5
8	TISTR5278	45.09	15.03	22.55	3.0:1.0:1.5
9	TISTR5328	38.95	13.43	30.89	2.9:1.0:2.3
10	TISTR5623	38.82	16.17	24.26	2.4:1.0:1.5

S. No.	Fractions	Band area (%)			Range ratio (1:2:3)	Content (%) of polysaccharide
S. No.	Fractions	Range 1 925-1,190 cm^-1 (Polysaccharide)	Range 2 1,500-1,700 cm^-1 (Protein)	Range 3 2,800-3,000 cm^-1 (Lipid)	Range ratio (1:2:3)	Content (%) of polysaccharide
1	E1	18.98	4.52	4.97	4.2:1.0:1.1	66.67
2	E2	18.36	4.08	4.49	4.5:1.0:1.1	68.18
3	E3	16.18	4.76	5.24	3.4:1.0:1.1	61.82
4	E4	15.40	5.50	4.43	2.8:1.0:1.0	58.33

S. No.	Strains	Band area (%)		β-glucan ratio (β-1,3:β-1,6)
S. No.	Strains	β-1,3-glucan	β-1,6-glucan	β-glucan ratio (β-1,3:β-1,6)
1	E1	45.85	6.44	7.12
2	E2	78.19	6.22	12.57
3	E3	32.70	6.54	5.00
4	E4	15.59	7.12	2.19

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[1] *Corresponding author: chaiyavat@gmail.com