Isolation, purification and structure characterization of the polysaccharide from the fermentation broth of Phlebopus portentosus

Authors

PAN Zhangchao, Key Laboratory of Tropical Translational Medicine of Ministry of Education, Haikou, Hainan 571199, China;Hainan Provincial Key Laboratory for Research and Development on Tropical Herbs, Haikou, Hainan 571199, China;School of Pharmacy, Hainan Medical University, Haikou, Hainan 571199, China
WANG Yong, Key Laboratory of Tropical Translational Medicine of Ministry of Education, Haikou, Hainan 571199, China;Hainan Provincial Key Laboratory for Research and Development on Tropical Herbs, Haikou, Hainan 571199, China;School of Pharmacy, Hainan Medical University, Haikou, Hainan 571199, ChinaFollow
WU Li, Key Laboratory of Tropical Translational Medicine of Ministry of Education, Haikou, Hainan 571199, China;Hainan Provincial Key Laboratory for Research and Development on Tropical Herbs, Haikou, Hainan 571199, China;School of Pharmacy, Hainan Medical University, Haikou, Hainan 571199, China
ZENG Niankai, Key Laboratory of Tropical Translational Medicine of Ministry of Education, Haikou, Hainan 571199, China;Hainan Provincial Key Laboratory for Research and Development on Tropical Herbs, Haikou, Hainan 571199, China;School of Pharmacy, Hainan Medical University, Haikou, Hainan 571199, China

Corresponding Author(s)

王勇（1982—），男，海南医学院研究员，硕士。E-mail: wangyong1982_2004@163.com

Abstract

[Objective] This study aimed to efficiently utilize the resources of Phlebopus portentosus and develop its polysaccharide functional food. [Methods] The mycelium of P. portentosus was cultured and fermented, and the combined fermentation broth was concentrated at low a temperature and reduced pressure. Then, precipitated by ethanol, separated, purified by DEAE-52 cellulose column chromatography, and the polysaccharide was finally refined by dialysis to remove the small molecular impurities. The total sugar content was determined by phenol-sulfuric acid colorimetry, the monosaccharide composition was determined by liquid chromatography-mass spectrometry (LC-MS), and the impurities such as nucleic acid and protein were detected by ultraviolet spectrophotometer. The structure of polysaccharides was analyzed preliminarily by infrared spectroscopy (IR) and nuclear magnetic resonance spectroscopy (NMR). [Results] The content of total sugar was 84%. The main monosaccharides of the polysaccharide were glucose, mannose, galactose, and arabinose, the molar ratio of which was 2.07∶1.95∶1.00∶1.58. Obvious polysaccharide absorption peaks were shown in the infrared spectrum and nuclear magnetic resonance spectra, through which an α-type pyranose can be inferred in the polysaccharide. [Conclusion] The polysaccharides prepared from the fermentation broth of P. portentosus were mainly composed of four monosaccharides, the structure of which contained α-pyranose.

Publication Date

9-11-2024

First Page

12

Last Page

16,62

DOI

10.13652/j.spjx.1003.5788.2023.81084

Recommended Citation

Zhangchao, PAN; Yong, WANG; Li, WU; and Niankai, ZENG (2024) "Isolation, purification and structure characterization of the polysaccharide from the fermentation broth of Phlebopus portentosus," Food and Machinery: Vol. 40: Iss. 7, Article 3.
DOI: 10.13652/j.spjx.1003.5788.2023.81084
Available at: https://www.ifoodmm.cn/journal/vol40/iss7/3

References

[1] 曹旸, 纪光燕, 罗顺珍, 等. 暗褐网柄牛肝菌人工驯化研究的回顾与前瞻[J]. 菌物学报, 2021, 40（12）: 3 064-3 080. CAO Y, JI G Y, LUO S Z, et al. Domestication and artificially cultivation of Phlebopus portentosus: retrospect and prospect[J]. Mycosystema, 2021, 40（12）: 3 064-3 080.
[2] KUMLA J, DANELL E, BUSSABAN B, et al. Suitable growth conditions and nutrition factors on in vitro culture of Phlebopus portentosus （Boletales） [J]. Chiang Mai Journal of Science, 2011, 38（1）: 156-159.
[3] 秦如冰, 叶丽, 尹嘉梁, 等. 响应面法优化暗褐脉柄牛肝菌粗多糖提取工艺[J]. 食品工业, 2022, 43（6）: 199-203. QIN R B, YE L, YIN J L, et al. Optimization of extraction process for crude polysaccharide from the fruiting bodies of Phlebopus portentosus based on response surface methodology[J]. The Food Industry, 2022, 43（6）: 199-203.
[4] CHUANKID B, SCHREY H, THONGBAI B, et al. Secondary metabolites of Phlebopus species from northern Thailand[J]. Mycological Progress, 2020, 19: 1 525-1 536.
[5] KUMLA J, HOBBIE ERIK A, SUWANNARACH N, et al. Theectomycorrhizal status of a tropical black bolete, Phlebopus portentosus, assessed using mycorrhizal synthesis and isotopic analysis[J]. Mycorrhiza, 2016, 26: 333-343.
[6] 胡生华, 朱志钢, 李文佳, 等. 暗褐网柄牛肝菌研究进展[J]. 食用菌, 2018, 40（1）: 6-8. HU S H, ZHU Z G, LI W J, et al. Research progress of Phlebopus portentosus[J]. Edible Fungi, 2018, 40（1）: 6-8.
[7] ZHANG C X, HE M X, LIU J, et al. Brief introduction to a unique edible bolete: Phlebopus portentosus in southern China[J]. Journal of Agricultural Science and Technology B, 2017, 7（6）: 386-394.
[8] SUN Z C, HU M G, SUN Z H, et al. Pyrrole alkaloids from the edible mushroom Phlebopus portentosus with their bioactive activities[J]. Molecules, 2018, 23（5）: 1 198.
[9] KAEWNARIN K, SUWANNARACH N, KUMLA J, et al. Phenolic profile of various wild edible mushroom extracts from Thailand and their antioxidant properties, anti-tyrosinase and hyperglycaemic inhibitory activities[J]. Journal of Functional Foods, 2016, 27: 352-364.
[10] 王振娟, 刘思婕, 王杰. 黑牛肝菌黑色素抗氧化活性和稳定性及染发效果[J]. 食用菌学报, 2023, 30（4）: 67-77. WANG Z J, LIU S J, WANG J. Antioxidant activity, stability and application of melanin obtained from Phlebopus portentosus[J]. Acta Edulis Fungi, 2023, 30（4）: 67-77.
[11] TIAN R, LIANG Z Q, WANG Y, et al. Analysis of aromatic components of two edible mushrooms, Phlebopus portentosus and Cantharellus yunnanensis using HS-SPME/GC-MS[J]. Results in Chemistry, 2022, 4: 100282.
[12] 钱正明, 黄琦, 沈千汇, 等. 暗褐网柄牛肝菌尿苷和麦角甾醇含量测定及指纹图谱分析[J]. 菌物研究, 2019, 17（2）: 103-109. QIAN Z M, HUANG Q, SHEN Q H, et al. Content determination and HPLC fingerprint analysis of uridine and ergosterol in Phlebopus portentosus[J]. Journal of Fungal Research, 2019, 17（2）: 103-109.
[13] KAEWNARIN K, SUWANNARACH N, KUMLA J, et al. Characterization of polysaccharides from wild edible mushrooms from Thailand and their antioxidant, antidiabetic, and antihypertensive activities[J]. International Journal of Medicinal Mushrooms, 2020, 22（3）: 221-233.
[14] KARNCHANATAT A, APHICHART S, SONGCHAN P, et al. Antioxidation and antiproliferation properties of polysaccharide-protein complex extracted from Phaeogyroporus portentosus （Berk. & Broome） McNabb[J]. African Journal of Microbiology Research, 2013, 7（17）: 1 668-1 680.
[15] 李燕平, 袁永俊, 张黎明, 等. 苯酚—硫酸法测定牛肝菌多糖含量[J]. 西华大学学报（自然科学版）, 2009, 28（3）: 76-78, 84. LI Y P, YUAN Y J, ZHANG L M, et al. Determination of the contents of polysaccharides in Boletus by phenol-sulfuric acid method[J]. Journal of Xihua University （Natural Science）, 2009, 28（3）: 76-78, 84.
[16] 江飞凤, 谭晓辉, 胡鹏刚, 等. 超声—微波协同提取柚子皮多糖工艺优化及单糖组成、结构和抗氧化活性分析[J]. 食品与发酵工业, 2021, 47（2）: 196-204. JIANG F F, TAN X H, HU P G, et al. Optimization of ultrasonic-microwave extraction process of pomelo peel polysaccharide and analysis of monosaccharide composition, structure and antioxidant activity[J]. Food and Fermentation Industries, 2021, 47（2）: 196-204.
[17] 周蓉, 尹军华, 翁榕安, 等. 黑柄炭角菌水溶性多糖XNW-1的分离纯化与结构分析[J]. 湖南师范大学自然科学学报, 2011, 34（5）: 75-79. ZHOU R, YIN J H, WENG R A, et al. Isolation, purification and structure analysis of water-soluble polysaccharides of Xylaria nigripes[J]. Journal of Natural Science of Hunan Normal University, 2011, 34（5）: 75-79.
[18] 彭慧群, 徐小刚, 陈昭月, 等. 红缘拟层孔菌多糖分离纯化、结构表征及其免疫活性分析[J]. 食品与生物技术学报, 2019, 38（3）: 152-159. PENG H Q, XU X G, CHEN Z Y, et al. Isolation, purification, structural characterization and immunological activity of Fomitopsis pinicola polysaccharides[J]. Journal of Food Science and Biotechnology, 2019, 38（3）: 152-159.
[19] 粟思源. 美网柄牛肝菌的鉴定及其多糖（BRS-X）的结构鉴定、免疫活性及抗肿瘤活性机制的研究[D]. 南充: 西华师范大学, 2019: 14-15. SU S Y. Identification of Boletus reticulatus Schaeff and studies on structure identification, immunological activity and the mechanism of anti-tumor activity of the polysaccharide （BRS-X） [D]. Nanchong: China West Normal University, 2019: 14-15.
[20] 伍燕, 朱家豪, 汪伟, 等. 暗褐网柄牛肝菌多糖AHP分离纯化和结构研究[J]. 食品与发酵工业, 2020, 46（8）: 92-96. WU Y, ZHU J H, WANG W, et al. Isolation, purification and structural elucidation of AHP polysaccharide from Phlebopus portentosus[J]. Food and Fermentation Industries, 2020, 46（8）: 92-96.