首页分享羊肚菌胞外多糖液态发酵培养基配方优化及其体外降血糖活性

羊肚菌胞外多糖液态发酵培养基配方优化及其体外降血糖活性

来源：花匠小妙招时间：2025-06-16 13:12

摘要: 本研究以甘肃省野生三地羊肚菌为研究对象，探究羊肚菌胞外多糖液态发酵培养基最佳方案与体外降血糖活性。采用单因素实验比较红糖、玉米粉等7种添加物对羊肚菌液态发酵过程中胞外多糖含量的影响，在此基础上采用响应面分析法优化羊肚菌胞外多糖液态发酵培养基配方。通过测定羊肚菌胞外多糖对α-淀粉酶和α-葡萄糖苷酶抑制率检测其体外降血糖活性。结果表明：当红糖添加量2.20 g/L、尿素添加量3.18 g/L、玉米粉添加量2.40 g/L时，实际得到羊肚菌胞外多糖的含量可达到1.20 g/L。羊肚菌胞外多糖在浓度为1.0 mg/mL时对α-葡萄糖苷酶和α-淀粉酶的抑制率则分别达到73.46%和36.37%，羊肚菌胞外多糖具有较好的降血糖活性。

Abstract: In this study, strains of wild Morchella eohespera in Gansu province were selected as the research object to explore the optimal solution of liquid fermentation medium of M. eohespera extracellular polysaccharide and its hypoglycemic activity in vitro. A single factor experiments were conducted to compare the effects of seven additives, including brown sugar and corn meal, on the content of M. eohespera extracellular polysaccharide during liquid fermentation. On this basis, response surface methodology was used to optimize the liquid fermentation medium formula of M. eohespera exopolysaccharides. The hypoglycemic activities of extracellular polysaccharide from M. eohespera on α-amylase and α-glucosidase were determined in vitro. The results showed that when the amounts of addition of brown sugar was 2.20 g/L, urea was 3.18 g/L and corn flour was 2.40 g/L, the actual content of extracellular polysaccharide of M. eohespera could reach 1.20 g/L. At the concentration of 1.0 mg/mL, effects of extracellular polysaccharide of M. eohespera on α-glucosidase and α-amylase showed that the inhibition rates for these enzymes were 73.46% and 36.37%, respectively. The results showed that extracellular polysaccharide of M. eohespera was with hypoglycemic activity.

图 1 不同添加物浓度对羊肚菌胞外多糖的影响

Figure 1. Effects of different contents of additives on exopolysaccharides from Morchella

图 2 不同因素间交互作用对羊肚菌液态发酵产胞外多糖的影响

Figure 2. Effect of interaction between different factors of exopolysaccharides from Morchella

图 3 羊肚菌胞外多糖的α-葡萄糖苷酶抑制活性

注：不同小写字母表示同一指标差异显著，P<0.05；图4同。

Figure 3. Inhibitory activity of α-glucosidase of MEP

图 4 羊肚菌胞外多糖的α-淀粉酶抑制活性

Figure 4. Inhibitory activity of α-amylase of MEP

表 1 羊肚菌胞外多糖Plackett-Burman试验设计与水平

Table 1 Plackett-Burman experiment design and level of exopolysaccharide in Morchella

因素水平 −1 +1 A红糖添加量（g/L）1.82.2B玉米粉添加量（g/L）2.02.8C黄豆粉添加量（g/L）1.11.3D葡萄糖添加量（g/L）1.62.4E硫酸铵添加量（g/L）3.03.4F蛋白胨添加量（g/L）2.02.8G尿素添加量（g/L）3.03.4

表 2 羊肚菌胞外多糖Box-Behnken试验的因素和水平

Table 2 Factors and levels of Box-Behnken experiment of exopolysaccharides from Morchella

因素水平−101 A红糖添加量（g/L）1.82.02.2B尿素添加量（g/L）3.03.23.4C玉米粉添加量（g/L）2.02.42.8

表 3 Plackett-Burman试验设计与结果

Table 3 Design and results of Plackett-Burman experiment

编号因素低水平（−1）高水平（+1）Prob>F显著性排序 A红糖−110.00221B玉米粉−110.00412C黄豆粉−110.22416D葡萄糖−110.49027E硫酸铵−110.14715F蛋白胨−110.05874G尿素−110.00743R2=0.9682，R2Adj=0.9126

表 4 Box-Behnken响应面试验设计与结果

Table 4 Design and results of Box-Behnken response surface methodology

实验号ABC多糖含量（g/L） 10001.29201−11.0730001.284−1010.465−10−10.7960−110.68710−10.738−1−100.3690110.64100001.27111−100.92121100.90131010.89140001.05150−1−10.7916−1100.83170001.26

表 5 Box-Behnken响应面试验结果方差分析

Table 5 Analysis of variance of Box-Behnken response surface methodology

方差来源平方和自由度均方F值Prob>F显著性模型1.290.1313.130.0013**A0.1210.1212.090.0103*B0.05710.0575.60.0498*C0.06210.0626.070.0432*AB0.0610.065.890.0456*AC0.05910.0595.770.0473*BC0.02710.0272.690.145A20.3310.3332.20.0008**B20.1710.1716.660.0047**C20.2310.2322.950.002**残差0.07170.01失拟项0.03130.011.030.4703纯误差0.0440.01总和1.2716R2=0.9441，R2Adj = 0.8722 注：**表示极显著（P < 0.01）；*表示显著（P <0.05）。 [1]

ZHANG T, ZHAO W, XIE B, et al. Effects of Auricularia auricula and its polysaccharide on diet-induced hyperlipidemia rats by modulating gut microbiota[J]. Journal of Functional Foods,2020,72:104038. doi: 10.1016/j.jff.2020.104038

[2]

XIONG C, LI Q, CHEN C, et al. Neuroprotective effect of crude polysaccharide isolated from the fruiting bodies of Morchella importuna against H2O2-induced PC12 cell cytotoxicity by reducing oxidative stress[J]. Biomedicine & Pharmacotherapy,2016,83:569−576.

[3] 杨芳, 王新风, 翁良, 等. 两种羊肚菌胞内多糖体外抗氧化性[J]. 食品科学,2010,31(23):76−78. [YANG F, WANG X F, FENG L, et al. Antioxidant activity of intracellular polysaccharides (IPS) from Morchella esculenta L

J]. Food Science,2010,31(23):76−78.

[4] 张强, 王松华, 孙玉军, 等. 羊肚菌蛋白的硒化修饰及其体外抗氧化活性[J]. 精细化工,2017,34(11):1252−1259. [ZHANG Q, WANG S H, SUN Y J, et al. Selenylation modification of Morchella esculenta proteins and their antioxidant activity in vitro[J]. Fine Chemicals,2017,34(11):1252−1259. doi: 10.13550/j.jxhg.2017.11.009 [5] 罗果, 保玉心, 李三华, 等. 羊肚菌发酵粗提产物的抗氧化活性鉴定[J]. 生物化工,2018,4(2):42−45. [LUO G, BAO Y X, LI S H, et al. The antioxidant activity of crude extraction from fermented Morchella esculenta[J]. Biochem Technol,2018,4(2):42−45. doi: 10.3969/j.issn.2096-0387.2018.02.011 [6] 王峰, 陶明煊, 程光宇, 等. 4种食用菌提取物自由基清除作用及降血糖作用的研究[J]. 食品科学,2009,30(21):343−347. [WANG F, TAO M X, CHENG G Y, et al. Free radical scavenging activities and hypoglycemic effects of aqueous extracts of four species of edible fungi[J]. Food Science,2009,30(21):343−347. doi: 10.3321/j.issn:1002-6630.2009.21.080 [7] 明建, 曾凯芳, 赵国华, 等. 羊肚菌水溶性多糖PMEP-1降血脂作用研究[J]. 食品科学,2009,30(17):285−288. [MING J, TAO M X, ZHAO G H, et al. Hypolipidemic activity of water soluble polysaccharide pmep-1 from Morchella esculenta (L.) pers[J]. Food Science,2009,30(17):285−288. doi: 10.3321/j.issn:1002-6630.2009.17.066 [8] 刘华晶, 赵妍. 羊肚菌液体培养基最适碳源氮源优化研究[J]. 南方农业,2019,13(26):136−137,145. [LIU H J, ZHAO Y. Optimization of carbon and nitrogen sources for Morchella liquid culture medium[J]. Southern Agriculture,2019,13(26):136−137,145. [9] 刘梅森, 陈海晏, 孙红斌. 液态发酵技术培养食用菌的研究概况[J]. 江西科学,1997(3):193−198. [LIU M S, CHEN H Y, SUN H B. A study of studies on edible fungi by submerged culture technology[J]. Jiangxi Science,1997(3):193−198. [10] 吴宽, 米伟丽, 吴云锋. 云芝糖肽发酵条件的响应面法优化试验[J]. 陕西农业科学,2020,66(1):55−58. [WU K, MI W L, WU Y F. Optimization of saccharopeptide fermentation conditions of Ganoderma formosa by response surface methodology[J]. Shanxi Agricultural Sciences,2020,66(1):55−58. doi: 10.3969/j.issn.0488-5368.2020.01.014 [11] 冮洁, 盖萌. 羊肚菌菌丝体液体培养产胞外多糖条件的研究[J]. 食用菌,2010,32(3):11−13. [GANG J, GAI M. Study on extracellular polysaccharide production conditions of Morchella mycelium by liquid culture[J]. Edible Fungi,2010,32(3):11−13. doi: 10.3969/j.issn.1000-8357.2010.03.007 [12] 杨恒, 赵萍, 刘裕慧, 等. 响应面优化发酵磨盘柿褐变条件[J]. 食品工业科技,2019,40(22):187−191. [YANG H, ZHAO P, LIU Y H, et al. Optimization of browning condition of fermented mopan persimmon by response surface methodology[J]. Science and Technology of Food Industry,2019,40(22):187−191. [13]

DUBOIS M, GILLES K, HAMILTON J K, et al. A colorimetric method for the determination of sugars[J]. Nature,1951,168(4265):167.

[14] 赵萍, 曹心张, 张新国, 等. 冬虫夏草菌丝体发酵液中多糖的抗氧化活性研究[J]. 中医药学报,2014,42(5):27−29. [ZHAO P, CAO X Z, ZHANG X G, et al. Antioxidant activity of polysaccharide in fermentation broth of Cordyceps sinensis[J]. Acta Chinese Medicine and Pharmacology,2014,42(5):27−29. doi: 10.3969/j.issn.1002-2392.2014.05.010 [15] 王珍珍, 官月, 刘洋, 等. 六妹羊肚菌多糖的提取工艺优化及结构表征和抗氧化活性[J]. 菌物学报,2019,38(9):1548−1558. [WANG Z Z, GUAN Y, LIU Y, et al. Extraction process optimization, structural characterization and antioxidant activities of polysaccharide from Morchella sextelata[J]. Mycosystema,2019,38(9):1548−1558. doi: 10.13346/j.mycosystema.190067 [16] 刘璐, 兰阿峰, 郭素芬, 等. 羊肚菌胞外多糖提取工艺研究[J]. 生物技术,2019,29(4):382−386. [LIU L, LAN A F, GUO S F, et al. Study on the extraction technology of extracellular polysaccharide from morel[J]. Biotechnology,2019,29(4):382−386. [17] 罗倩, 邹荣灿, 刘明月, 等. 羊肚菌多糖的提取分离纯化及保健功效研究进展[J]. 食品研究与开发,2019,40(15):211−216. [LUO Q, ZOU C R, LIU M Y, et al. Research progress of extraction, isolation, purification and health benefits of polysaccharides in Morchella[J]. Food Research and Development,2019,40(15):211−216. doi: 10.12161/j.issn.1005-6521.2019.15.035 [18]

LU X, BRENNAN M A, NARCISO J, et al. Correlations between the phenolic and fibre composition of mushrooms and the glycaemic and textural characteristics of mushroom enriched extruded products[J]. LWT,2020,118:108730. doi: 10.1016/j.lwt.2019.108730

[19] 陈树俊, 崔云. 甘薯渣多糖提取, 结构鉴定及体外功能研究[J]. 中国粮油学报,2021(9):67−73. [CHEN S J, CUI Y. Extraction, structure identification and in vitro function study of polysaccharides from sweet potato residue[J]. Journal of the Chinese Cereals and Oils Society,2021(9):67−73. doi: 10.3969/j.issn.1003-0174.2021.09.012 [20] 殷伟伟, 张松, 吴金凤. 尖顶羊肚菌活性提取物降血脂作用的研究[J]. 菌物学报,2009,28(6):873−877. [YIN W W, ZHANG S, WU J F. Hypolipidemic effect of the bioactive extract from Morchella conica[J]. Mycosystema,2009,28(6):873−877. [21] 李井雷, 刘玉婷, 宗帅, 等. 羊肚菌胞外多糖体外降血糖降血脂活性研究[J]. 食品研究与开发,2020,41(16):39−45. [LI J L, LIU Y T, ZONG S, et al. In vitro hyperglycemic and hypolipidemic activity of Morchella esculenta extracellular polysaccharides[J]. Food Research and Development,2020,41(16):39−45. doi: 10.12161/j.issn.1005-6521.2020.16.006 [22]

YUHAN D, YANRAN Q, MIN L, et al. Antioxidant, anti-hyperlipidemia and hepatic protection of enzyme-assisted Morehella esculenta polysaccharide[J]. International Journal of Biological Macromolecules,2018:120.

[23]

ZHONG Q, ZHOU T, QIU W, et al. Characterization and hypoglycemic effects of sulfated polysaccharides derived from brown seaweed Undaria pinnatifida[J]. Food Chemistry,2021,341:128148. doi: 10.1016/j.foodchem.2020.128148

[24]

LIU Y, SUN J, LUO Z, et al. Chemical composition of five wild edible mushrooms collected from Southwest China and their antihyperglycemic and antioxidant activity[J]. Food and Chemical Toxicology,2012,50(5):1238−1244. doi: 10.1016/j.fct.2012.01.023

[25] 金朝霞, 王云龙, 王培忠, 等. 羊肚菌液体发酵培养条件优化[J]. 大连工业大学学报,2014(6):416−419. [JING Z X, WANG Y L, WANG P Z, et al. Optimization of liquid fermentation conditions of Morchella[J]. Journal of Dalian Polytechnic University,2014(6):416−419. doi: 10.19670/j.cnki.dlgydxxb.2014.06.007 [26]

LI Y, YUAN Y, LEI L, et al. Carboxymethylation of polysaccharide from Morchella angusticepes peck enhances its cholesterol-lowering activity in rats[J]. Carbohydrate Polymers,2017,172:85−92. doi: 10.1016/j.carbpol.2017.05.033

[27]

CHEN J, LI L, ZHANG X, et al. Structural characterization of polysaccharide from Centipeda minima and its hypoglycemic activity through alleviating insulin resistance of hepatic HepG2 cells[J]. Journal of Functional Foods,2021,82:104478. doi: 10.1016/j.jff.2021.104478

[28]

WU M, LI W, ZHANG Y, et al. Structure characteristics, hypoglycemic and immunomodulatory activities of pectic polysaccharides from Rosa setate×Rosa rugosa waste[J]. Carbohydrate Polymers,2021,253:117190. doi: 10.1016/j.carbpol.2020.117190

[29]

TAKEWAKI F, NAKAJIMA H, TAKEWAKI D, et al. Habitual dietary intake affects the altered pattern of cut microbiome by acarbose in patients with type 2 diabetes[J]. Nutrients,2021,13(6):2107. doi: 10.3390/nu13062107

[30]

NSARI P, FLATT P R, HARRIOTT P, et al. Insulinotropic and antidiabetic properties of eucalyptus citriodora leaves and isolation of bioactive phytomolecules[J]. Journal of Pharmacy and Pharmacology,2021,73(8):1−13.

[31]

YE L H, SUN H, ZHANG X, et al. A novel polysaccharide from Pleurotus citrinopileatus mycelia: Structural characterization, hypoglycemic activity and mechanism[J]. Food Bioscience,2020,37:100735. doi: 10.1016/j.fbio.2020.100735

[32] 王梦雅, 赵喆禛, 薛娇, 等. 桦褐孔菌纯化多糖体外降血糖活性研究[J]. 食品工业科技,2020,41(10):6. [WANG M Y, ZHAO Z Z, XUE J, et al. Hypoglycemic activity of purified polysaccharides from Inonotus obliquus in vitro[J]. Science and Technology of Food Industry,2020,41(10):6. [33] 柏秋月, 邓百万, 杨学英, 等. 4株羊肚菌胞外多糖含量及其生物活性的研究[J]. 食品研究与开发,2020,41(8):25−31. [BAI Q Y, DENG B W, YANG X Y, et al. Study on extracellular polysaccharide content and biological activity of four Morchella strains[J]. Food Research and Development,2020,41(8):25−31. doi: 10.12161/j.issn.1005-6521.2020.08.004