Food and Fermentation Industries >

2022 , Vol. 48 >Issue 17: 21 - 28

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.030855

Fermentation process for preparation of low-molecular-weight β-1,3-glucan using glycerol as carbon source

  • LI Min ,
  • ZHU Li ,
  • ZHAN Xiaobei
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  • 1(Key Laboratory of Carbohydrate Chemistry and Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China)
    2(Wuxi Galaxy Biotech Co.Ltd., Wuxi 214125, China)

Received date: 2022-01-17

  Revised date: 2022-02-19

  Online published: 2022-10-01

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Abstract

Agrobacterium sp. WS-8E3T produced low-molecular-weight β-1,3-glucan was obtained by adaptive domestication of Agrobacterium sp. ATCC 31749. Before optimization, the yield of low-molecular-weight β-1,3-glucan by the bacterium used glycerol as a carbon source was 1.648 g/L. To obtain high-yield oligosaccharides with lower cost, the fermentation conditions were optimized. The optimal initial glycerol concentration was 20 g/L, after replenished 20 g/L glycerol when the residual glycerol content was about 10 g/L. The optimal initial ratio of compound nitrogen source was 4 g/L of yeast extract and 3 g/L of ammonium sulfate, with a liquid volume of 75 mL/500 mL and pH of 6.3. In 7 L fermenter, during the growth period of the bacteria, the optimal pH was 7.0, and the glycerol was 20 g/L. During the oligosaccharide-producing period, the pH was 6.3, and the glycerol was 30 g/L. The final yield of oligosaccharides reached 8.03 g/L, which was 386.7% higher than that of before optimization. The results of FT-IR analysis and monosaccharide composition was consistent with curdlan, and the degree of polymerization was 18-22. The single-strain fermentation strategy was used to significantly increase the yield of low molecular weight β-1,3-glucan for the first time, which provide a theoretical basis for further low-cost and large-scale fermentation production of low molecular weight β-1,3-glucan.

Key words： β-1,3-glucan; single-strain fermentation; glycerol; feeding fermentation; two-stage feeding

Cite this article

LI Min , ZHU Li , ZHAN Xiaobei . Fermentation process for preparation of low-molecular-weight β-1,3-glucan using glycerol as carbon source[J]. Food and Fermentation Industries, 2022 , 48(17) : 21 -28 . DOI: 10.13995/j.cnki.11-1802/ts.030855

References

[1] WU L J, ZHAO J, ZHANG X N, et al. Antitumor effect of soluble β-glucan as an immune stimulant[J]. International Journal of Biological Macromolecules,2021, 179: 116-124.
[2] SYLLA B, LEGENTIL L, SARASWAT-OHRI S, et al. Oligo-β-(1→3)-glucans: Impact of thio-bridges on immunostimulating activities and the development of cancer stem cells[J]. Journal of Medicinal Chemistry, 2014, 57(20): 8 280-8 292.
[3] SUN N X, TONG L T, LIANG T T, et al. Effect of oat and tartary buckwheat-based food on cholesterol-lowering and gut microbiota in hypercholesterolemic hamsters[J]. Journal of Oleo Science, 2019, 68(3): 251-259.
[4] KHAN A A, GANI A, SHAH A, et al. Effect of γ-irradiation on structural, functional and antioxidant properties of β-glucan extracted from button mushroom (Agaricus bisporus)[J]. Innovative Food Science & Emerging Technologies, 2015, 31:123-130.
[5] METHACANON P, WEERAWATSOPHON U, TANJAK P, et al. Interleukin-8 stimulating activity of low molecular weight β-glucan depolymerized by γ-irradiation[J]. Carbohydrate Polymers, 2011, 86(2):574-580.
[6] FU Y B, CHENG L K, MENG Y Y, et al. Cellulosimicrobium cellulans strain E4-5 enzymatic hydrolysis of curdlan for production of (1→3)-linked β-D-glucan oligosaccharides[J]. Carbohydrate Polymers. 2015, 134: 740-744.
[7] GAO M J, YAN J J, ZHAO Y, et al. Expression of a thermostable β-1,3-glucanase from Trichoderma harzianum in Pichia pastoris and use in oligoglucosides hydrolysis[J]. Process Biochemistry, 2021, 107: 74-82.
[8] WANG D M, KIM D H, YOON J J, et al. Production of high-value β-1,3-glucooligosaccharides by microwave-assisted hydrothermal hydrolysis of curdlan[J]. Process Biochemistry,2017, 52: 233-237.
[9] ISHIMOTO Y, ISHIBASHI K I, YAMANAKA D, et al. Production of low-molecular weight soluble yeast β-glucan by an acid degradation method[J]. International Journal of Biological Macromolecules. 2018, 107: 2 269-2 278.
[10] KUMAGAI Y, OKUYAMA M, KIMURA A. Heat treatment of curdlan enhances the enzymatic production of biologically active β-(1,3)-glucan oligosaccharides[J]. Carbohydrate Polymers,2016, 146: 396-401.
[11] ZHU Q, WU S J. Water-soluble β-1, 3-glucan prepared by degradation of curdlan with hydrogen peroxide[J]. Food Chemistry, 2019, 283:302-304.
[12] 宗玉, 李晶, 朱莉, 等. 两步法硫酸水解热凝胶生产β-1, 3-葡寡糖[J]. 工业微生物, 2014, 44(1):22-27.
ZONG Y, LI J, ZHU L, et al. Preparation of β-1, 3-glucan oligosaccharides by two-step sulfuric acid hydrolysis of curdlan[J]. Industrial Microbiology, 2014, 44(1):22-27.
[13] GRANDPIERRE C, JANSSEN H G, LAROCHE C, et al. Enzymatic and chemical degradation of curdlan targeting the production of β-(1→3) oligoglucans[J]. Carbohydrate Polymers, 2008, 71(2): 277-286.
[14] LIANG Y, ZHU L, GAO M J, et al. Effective production of biologically active water-soluble β-1,3-glucan by a coupled system of Agrobacterium sp. and Trichoderma harzianum[J]. Preparative Biochemistry & Biotechnology, 2018, 48(5): 446-456.
[15] 李菲菲, 金树霞, 朱莉, 等. 土壤杆菌-毕赤酵母耦合培养直接生产热凝胶低聚糖[J]. 过程工程学报, 2019, 19 (4): 801-808.
LI F F, JIN S X, ZHU L, et al. Direct production of curdlan oligosaccharides by coupled fermentation system of Agrobacterium sp.-Pichia pastoris[J]. The Chinese Journal of Process Engineering, 2019, 19 (4): 801- 808.
[16] 王冰, 朱莉, 李茂玮, 等. 适应性驯化生产低分子质量β-葡聚糖及其抗氧化活性研究[J]. 食品与发酵工业, 2021, 47(17):27-33.
WANG B, ZHU L, LI M W, et al. Improvement of low-mass β-glucan yield by adaptive laboratory evolution and its antioxidant activity[J]. Food and Fermentation Industries, 2021, 47(17):27-33.
[17] LIU Y M, GU Q Y, OFOSU F K, et al. Production, structural characterization and gel forming property of a new exopolysaccharide produced by Agrobacterium HX1126 using glycerol or D-mannitol as substrate[J]. Carbohydrate Polymers, 2016, 136:917-922.
[18] 崔连杰, 李科, 李震宇, 等. 寡糖提取分离与质谱结构解析研究进展[J]. 药学学报, 2020, 55(5):843-853.
CUI L J, LI K, LI Z Y, et al. Progress in oligosaccharide extraction, separation and structural analysis by mass spectrometry[J]. Acta Pharmaceutica Sinica, 2020, 55(5):843-853.
[19] LIU Y M, WANG Z H, ZHAO J, et al. Effects of glycerol concentrations on the bioproduction of PGHX by Agrobacterium HX1126[J]. Preparative Biochemistry & Biotechnology, 2019, 49(6):584-589.
[20] MORANO K A, GRANT C M, MOYE-ROWLEY W S. The response to heat shock and oxidative stress in Saccharomyces cerevisiae[J]. Genetics, 2012, 190(4): 1 157-1 195.
[21] 王子朝. 以甘油为底物发酵生产黄原胶及其特性和应用研究[D]. 无锡:江南大学, 2017.
WANG Z C. Production of xanthan gum with glycerol and the study of its properties and application[D]. Wuxi: Jiangnan University, 2017.
[22] DHAR R, SÄGESSER R, WEIKERT C, et al. Yeast adapts to a changing stressful environment by evolving cross-protection and anticipatory gene regulation[J]. Molecular Biology and Evolution, 2012, 30(3):573-588.
[23] DAKAL T C, SOLIERI L, GIUDICI P. Adaptive response and tolerance to sugar and salt stress in the food yeast Zygosaccharomyces rouxii[J]. International Journal of Food Microbiology, 2014, 185: 140-157.
[24] RUFFING A M, CASTRO-MELCHOR M, HU W S, et al. Genome sequence of the curdlan-producing Agrobacterium sp. strain ATCC 31749[J]. Journal of Bacteriology, 2011, 193(16): 4 294-4 295.
[25] 李晶. 热凝胶制备β-1, 3-葡聚寡糖及其诱导抗真菌功能的研究[D]. 无锡: 江南大学, 2014.
LI J. Preparation of β-1, 3-glucooligosaccharides from curdlan and the elicitation effect against fungi[D]. Wuxi: Jiangnan University, 2014.
[26] 王磊, 詹晓北, 朱一晖, 等. pH控制对热凝胶发酵的影响[J].生物工程学报, 2002,18(5): 634-637.
WANG L, ZHAN X B, ZHU Y H, et al. Influence of pH control on the production of curdlan by Alcaligenes faecalis strain[J]. Chinese Journal of Biotechnology, 2002, 18(5): 634-637.
[27] COLUSSI F, GARCIA W, ROSSETO F R, et al. Effect of pH and temperature on the global compactness, structure, and activity of cellobiohydrolase Cel7A from Trichoderma harzianum[J]. European Biophysics Journal, 2012, 41(1): 89-98.
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