Structural characterization and anti-inflammatory activities of novel polysaccharides obtained from Pleurotus eryngii

Natural polysaccharides named PEP-0.1-1,PEP-0-1 and PEP-0-2 from edible mushroom species Pleurotus eryngii were obtained in the present study.Results showed that molecular weights of these polysaccharides were 3235,2041 and 23933 Da,respectively.Further,structural characterization revealed that PEP-0.1-1 had a→4-α-D-Glcp-1→backbone and contained→4)-α-D-Glcp and→4)-β-D-Glcp reducing end groups.PEP-0-1 backbone contained→4-α-D-Glcp-1→and→6-α-3-O-Me-D-Galp-1→,and the side chains containedα-D-Glcp,β-D-Manp-1→andα-D-Glcp-3→.However,PEP-0-2 backbone consisted of→4-α-DGlcp-1→and→6-α-3-O-Me-D-Galp-(1→6)-α-D-Galp-1→while the side chains containedα-D-Glcp andβ-D-Manp-1→.Biological activity analysis was then carried out and found that all these polysaccharides could significantly suppress the relative mRNA expression of toll-like receptor 4,nitric oxide(NO),tumor necrosis factor-α,interleukin(IL)-1βand IL-6 in lipopolysaccharide(LPS)-induced inflammation of RAW264.7 cells,as well as the over secretion of the above cell cytokines.Moreover,Western blotting analysis revealed that all these purified fractions displayed significant inhibition effects on the expression of c-Jun N-terminal kinases protein induced by LPS in mitogen activated protein kinase pathway,along with the relieving on the inhibition effect of LPS on IκB-αprotein expression.In summary,the information generated by the present study could provide a theoretical basis for the exploration of novel healthy food materials from edible mushroom with antiinflammation activities.

虎奶菇Pleurotus tuber-regium深层发酵的研究

报道了虎奶菇Pleurotus tuber-regium深层发酵的最适培养基和摇瓶发酵条件。试验结果表明,Pleurotus tuber-regium对单糖、双糖、多糖和糖醇都有一定的利用,但葡萄糖、麦芽糖等对菌丝生长效果较好;酵母粉、黄豆饼粉等蛋白质含量高的氮源对菌丝的生长较为有利;通过L9(34)正交试验确定了Pleurotus tuber-regium发酵的最适培养基为:5%葡萄糖,0.4%酵母粉,0.1%KH2PO4,0.3%MgSO4.7H2O,0.001%VB1及微量NaCl;一定浓度内Na+对菌丝体生长有促进效果。最适的摇瓶发酵条件为:培养基起始pH7.0,培养温度32℃,接种量15%,摇床转速150r/min,250mL三角瓶最适装液量为70mL,发酵周期为6d。

Pleurotus nebrodensis子实体多糖提取工艺的研究

对Pleurotus nebrodensis子实体多糖酶法提取工艺条件中酶用量、酶解温度、酶解时间、pH值4因子的最优化组合问题进行了研究,建立了具有良好预测性能的Pleurotus nebrodensis子实体多糖提取条件模型,并对工艺条件的最优组合及双因素效应进行分析。结果表明,当酶用量为3.06%、酶解温度44.9℃、酶解时间3.8 h、pH值6.7时,多糖得率最高可达12.37%。

黄白侧耳(Pleurotus cornucopiae(paul.ex. pers)Roll)子实体多糖的研究

黄白侧耳(Pleurotus cornucopiae(paul.ex.pers)Roll)子实体经热水提取,乙醇沉淀得到多糖粗制品P_Ⅲ。再经去蛋白,透析,DEAE—SephadexA—25纯化得P_(FⅢ—1)、P_(FⅢ—2)、P_(FⅢ—3)三个多糖组分。其含量比约为146:2:1。P_(FⅢ—1)用SephadexG—200凝胶层析及醋酸纤维素薄膜电泳证明为均一体。平均分子量W=2.32×10~5,比旋度[α]_D^20=+168°(C=0.01,H_2O)。经P. L. C和G. L. C分析表明,P_(FⅢ—1)是一种含果糖的杂多糖。由葡萄糖、果糖、甘露糖组成,其摩尔比为1:0.76:0.286。元素分析表明P_(FⅢ—1)不含N,C:39.91％,H:6.13％,在3300—3490cm^(-1),1645cm^(-1),1195cm^(-1)处有红外吸收。动物药理实验表明:P_Ⅲ、P_(FⅢ—1)具有增强机体免疫功能,对Lewis肺癌,C_(27)结肠癌,Hep肝癌等多种肿瘤均有抑制作用。

热风干燥对刺芹侧耳多糖-蛋白质复合物加工特性及抗氧化性的影响

对刺芹侧耳(Pleurotus eryngii)子实体进行热风干燥处理,检测其色差值、褐变指数、中间指数、紫外可见吸收光谱,分析其美拉德反应程度;提取刺芹侧耳多糖-蛋白质复合物,分析其基本成分、加工特性(溶解性、相对黏度、起泡性、乳化性、硬度、内聚性、弹性、咀嚼性和凝胶强度)和抗氧化性。结果表明:新鲜刺芹侧耳的色差值最高,干制24 h的最低;新鲜刺芹侧耳的褐变指数与中间指数最低,干燥24 h的褐变指数与中间指数最高。新鲜刺芹侧耳多糖-蛋白质复合物溶解所需时间最长,干燥18、24 h的溶解时间较短;干燥24h的相对黏度最高,新鲜刺芹侧耳的最低;干燥24h的乳化性、起泡性均最好,干燥12、18、24h的乳化稳定性较好,干燥18 h的起泡稳定性较好;干燥24 h的硬度、内聚性、弹性、咀嚼性和凝胶强度均最高;干燥18、24h的对DPPH自由基清除率较高,干燥24h的对羟基自由基清除率最高,干燥24h的对超氧阴离子清除率较高,干燥18、24 h的对多酚氧化酶抑制率较高。研究结果可为刺芹侧耳的进一步开发利用提供参考。

杏鲍菇多糖水提物新型护肤乳配方工艺及抗氧化功效

探究了以杏鲍菇多糖水提物为主要成分的护肤乳配比及功效。通过提取物的不同比例来设计配方,以质量分数0、25%、50%和75%的杏鲍菇多糖水提物添加量制备出4种具有抗氧化效果的护肤乳,并通过感官评定、耐热耐寒测试等试验,测定其理化性质及稳定性,同时结合DPPH自由基与超氧阴离子清除试验考察其氧化活性。结果表明4种产品均符合行业标准;由体外抗氧化试验可知,随着多糖水提物添加量的增加,护肤乳对DPPH和超氧阴离子的清除能力也增强。其中,含质量分数75%提取物的护肤乳效果最佳,皮肤水分含量增加8%,DPPH自由基和超氧阴离子能力的最大清除率分别为30.52%和51.86%。杏鲍菇多糖水提物具有良好的抗氧化效果和保湿性,可作为新型化妆品中的天然抗氧化活性物质。

杏鲍菇多糖对小鼠急性化学性肝损伤的抗炎及抗氧化作用

为探究杏鲍菇多糖(Pleurotus eryngii polysaccharides,PEP)对CCl_(4)诱导小鼠急性肝损伤的干预作用,首先分析PEP体外清除DPPH自由基及ABTS^(+)自由基的能力,之后将小鼠随机分为对照组、模型组、PEP低、中、高剂量组(100、200、400 mg/kg),连续灌胃干预21 d后,腹腔注射CCl_(4)以诱导小鼠急性肝损伤,最后测定小鼠血清谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST)活性、总胆红素(total bilirubin,TBIL)、总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)含量,分析肝组织丙二醛(malondialdehyde,MDA)含量、过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性并检测肝脏组织形态学变化。结果表明,PEP对ABTS^(+)自由基及DPPH自由基清除率分别可达(70.50±0.63)%和(63.25±0.56)%。与模型组比较,PEP各剂量组肝脏、肾脏及脾脏指数呈不同程度降低。血清ALT、AST活性、TBIL、TC、TG、TNF-α、IL-6含量显著下降,肝组织SOD、CAT、GSH-Px活性显著增高,MDA含量显著下降(P<0.05),且呈剂量依赖性。PEP各剂量组肝组织病理损伤不同程度减轻。因此PEP能够通过抗氧化及抗炎对CCl_(4)所致的小鼠肝损伤发挥保护作用。

The polysaccharide isolated from Pleurotus nebrodensis(PN-S) shows immune-stimulating activity in RAW264.7 macrophages

A novel Pleurotus nebrodensis polysaccharide(PN-S) was purified andcharacterized,and its immune-stimulating activity was evaluated in RAW264.7 macrophages.PN-S induced the proliferation of RAW264.7 cells in a dose-dependent manner,as determined by the MTT assay.After exposure to PN-S,the phagocytosis of the macrophages was significantly improved,with remarkable changes in morphology being observed.Flow cytometric analysis demonstrated that PN-S promoted RAW264.7 cells to progress through S and G2/M phases.PN-S treatment enhanced the productions of interleukin-6(IL-6),nitric oxide(NO),interferon gamma(INF-γ),and tumor necrosis factor-α(TNF-α) in the macrophages,with up-regulation of m RNA expressions of interleukin-6(IL-6),inducible nitric oxide synthase(i NOS),interferon gamma(INF-γ) and tumor necrosis factor-α(TNF-α) being observed in a dose-dependent manner,as measured by q RT-PCR.In conclusion,these results suggest that the purified PN-S can improve immunity by activating macrophages.

Pleurotus nebrodensis polysaccharide(PN-S) enhances the immunity of immunosuppressed mice

In the present study, the effects of Pleurotus nebrodensis polysaccharide(PN-S) on the immune functions of immunosuppressed mice were determined. The immunosuppressed mouse model was established by treating the mice with cyclophosphamide(40 mg/kg/2d, CY) through intraperitoneal injection. The results showed that PN-S administration significantly reversed the CY-induced weight loss, increased the thymic and splenic indices, and promoted proliferation of T lymphocyte, B lymphocyte, and macrophages. PN-S also enhanced the activity of natural killer cells and increased the immunoglobulin M(Ig M) and immunoglobulin G(Ig G) levels in the serum. In addition, PN-S treatment significantly increased the phagocytic activity of mouse peritoneal macrophages. PN-S also increased the levels of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), interferon-γ(INF-γ), and nitric oxide(NOS) in splenocytes. q RT-PCR results also indicated that PN-S increased the m RNA expression of IL-6, TNF-α, INF-γ, and nitric oxide synthase(i NOS) in the splenocytes. These results suggest that PN-S treatment enhances the immune function of immunosuppressed mice. This study may provide a basis for the application of this fungus in adjacent immunopotentiating therapy against cancer and in the treatment of chemotherapy-induced immunosuppression.

Polysaccharides from edible fungi Pleurotus spp.:advances and perspectives

Pleurotus spp.was one of the most precious and common edible fungi,polysaccharides were the main active component.The structural,biological activities and structure-activity relationship(SAR)of polysaccharides from Pleurotus spp.were systematically reviewed.On the basis of structure and biological activity,structure-activity relationships were also analyzed and discussed to look forward to its future research direction and application prospect.In the past 5 years,about 30 kinds of polysaccharides were isolated from Pleurotus spp.,and their preliminary structures were studied,but the fine structures seldomly reported.The polysaccharides showed the activities of immunomodulatory,hypoglycemic,anti-inflammatory,antioxidant,anti-ageing,hepatoprotective,anti-tumor,hypolipidemic and regulating intestinal flora,but the mechanism was needed further study.There were few studies on the SAR of polysaccharides from Pleurotus,and the polysaccharides of P.eryngii were studied.It was found that the biological activities of polysaccharides were affected by molecular weight,monosaccharide composition,sugar chain structure and configuration.In addition,sulfonation and selenization could significantly increase the bioactivities of polysaccharides.These findings might help to better understand the research status of polysaccharides from Pleurotus spp.and provided a scientific basis for their application as functional foods.

不同干燥方法对杏鲍菇结构及品质特性的影响

为探讨不同干燥方法对杏鲍菇干制品组织结构和品质的影响,以干制品微观形貌、收缩率、复水比、色泽、氨基酸含量、多糖含量、感官评价为评价指标,选取热风干燥、微波真空干燥、间歇微波真空干燥、热风-微波真空联合干燥、真空冷冻干燥5种方法对杏鲍菇进行干燥。结果表明,组织结构完整性保持最好的是真空冷冻干燥,其收缩率最小(30.2%),复水比最大(3.03),复原效果最好,多糖含量最高(13%)。热风-微波真空联合干燥氨基酸含量最高(560.94 mg/100 g),感官评价最高(9.4分),收缩率(55.43%)、复水比(1.83)、色差(76.29)、多糖(11.8%)仅次于真空冷冻干燥。热风干燥多糖含量最低(5.4%),感官评价最低(4.4分)。综合外观、营养成分及感官评价,热风-微波真空联合干燥是最适合于杏鲍菇等一类菌丝体构成组织、含水量大的食用菌的一种干燥方法。

一株野生盖囊侧耳的鉴定与驯化栽培

以一株野生食用菌菌株为研究对象,探究其分类地位及驯化栽培特性,为我国野生资源的开发利用提供参考。通过形态学特征和ITS序列分析鉴定,将采自广州市天河区的一株野生食用菌鉴定为盖囊侧耳(又称鲍鱼菇),并通过测定菌丝生长速度、子实体商品性等对其开展生物学特性和驯化栽培研究。结果表明,该菌株最适碳源是麦芽糖,最适氮源是牛肉膏,适宜生长的pH范围是7.0~9.0,适宜生长温度是25~30℃。在栽培配方为棉籽壳50%、玉米芯34%、麸皮15%、石灰1%、含水量62%的人工培养基中,在25℃条件下,液体菌种接种30 d左右满包,开袋7 d左右出原基,15 d左右可采收,子实体单丛鲜重95~195 g,一潮菇生物转化率约为27%,子实体多糖含量为5.97%,三萜含量为1.12%。本研究可为盖囊侧耳的人工栽培及推广应用提供依据,也为后续选育广温、耐高温型食用菌品种提供优良种质资源。

杏鲍菇液体发酵沛县黄皮牛蒡对多糖含量影响的研究

沛县黄皮牛蒡营养物质丰富,可有效地利用杏鲍菇中的糖类、氨基酸等营养成分,促进多糖合成。利用液体发酵技术,以沛县黄皮牛蒡作为培养基,液体培养杏鲍菇,通过响应面试验优化单因素,采用水提醇沉法提取发酵液多糖,经Sevag法纯化后,用苯酚-硫酸法检测多糖含量,利用高效液相色谱法检测多糖中单糖组成。结果表明,当培养条件为沛县黄皮牛蒡含量占总装液量的3%,温度28℃,装液量100 mL时,得到多糖提取量为2.41 mg/mL。发酵液多糖由甘露糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、木糖和阿拉伯糖组成,其中葡萄糖占比最高。

一种榆黄蘑菌丝体多糖的结构表征及其降血糖作用

为探究榆黄蘑(Pleurotus citrinopileatus)菌丝体多糖的结构及降血糖作用,首先从榆黄蘑菌丝体中分离纯化得到水溶性多糖(water-soluble polysaccharide from P.citrinopileatus,PCP-W),采用采用高效分子排阻色谱、红外光谱等方法表征其结构;然后建立2型糖尿病小鼠模型,灌胃PCP-W 8周后,检测小鼠空腹血糖水平、糖耐量和空腹胰岛素水平,计算定量胰岛素敏感性检查指数(quantitative insulin sensitivity check index,QUICKI);检测小鼠血脂、氧化应激水平和肝脏己糖激酶(hexokinase,HK)、丙酮酸激酶(pyruvate kinase,PK)活力。结果表明,PCP-W分子质量为5238 Da,含D-吡喃葡萄糖,单糖组成为甘露糖、核糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖和岩藻糖,其物质的量比为0.003∶0.010∶0.011∶0.015∶0.001∶0.944∶0.013∶0.001。治疗8周后,PCP-W能显著降低糖尿病小鼠空腹血糖水平,改善葡萄糖耐受,升高空腹胰岛素水平,改善QUICKI(P<0.01);PCP-W能显著改善糖尿病小鼠的血脂代谢和氧化应激水平,提高PK和HK的活力(P<0.01)。因此,PCP-W具备开发成降糖药物的潜力。

榆黄菇多糖提取工艺优化及其免疫调节活性评价

以榆黄菇为研究对象,对榆黄菇多糖(PCP)提取工艺进行优化、纯化、测定单糖组成和分子量,再对其体外免疫活性进行评价。在单因素基础上通过响应面法确定榆黄菇多糖最优提取工艺为:提取温度59.81℃,提取时间2.40 h,液料比29.91 mL/g,在此条件下,预测得率为18.82%,实际得率为18.60%。经DEAE-Sepharose Fast Flow柱层析从榆黄菇多糖中分离纯化出3个多糖组分(PCP-1、PCP-2、PCP-3),采用离子色谱法和高效凝胶渗透色谱法分析多糖成分和分子量,得出PCP-1由半乳糖组成,分子量为1.90×10^(4)u;PCP-2由半乳糖和葡萄糖组成,摩尔比为4.36:5.64,分子量2.76×10^(4)u;PCP-3由岩藻糖、盐酸氨基葡萄糖、半乳糖、葡萄糖和葡萄糖醛酸组成,其摩尔比为0.07:0.11:0.51:7.46:1.85,分子量为4.81×10^(4)u。通过体外试验评估PCP对巨噬细胞RAW264.7免疫调节活性。结果表明,PCP-1、PCP-2和PCP-3多糖质量浓度在25~200μg/mL范围内对巨噬细胞RAW264.7无毒性并具有一定增殖作用、显著提高了NO释放量并增强了巨噬细胞RAW264.7的吞噬能力。当PCP-1、PCP-2和PCP-3多糖质量浓度为200μg/mL时,巨噬细胞RAW264.7 NO释放量达到最大值,分别为11.40、11.56、11.76μmoL/L。该研究结果可为榆黄菇的精深加工综合利用提供理论依据。

响应面优化超声波辅助提取杏鲍菇多糖的工艺及其抗疲劳活性测定研究

本试验以杏鲍菇为原料,采用超声波辅助提取杏鲍菇多糖,并对其抗疲劳作用进行研究。以杏鲍菇多糖提取率为指标,在单因素基础上进行响应面优化提取工艺,得到最佳工艺条件为:超声功率420 W、超声温度50℃、超声时间50min,在此条件下,杏鲍菇多糖提取率为14.71%。观察小鼠的爬杆和负重游泳时间、血清生化指标、肝糖原、肌糖原等指标考察杏鲍菇多糖的抗疲劳作用。结果表明,杏鲍菇多糖能明显延长爬杆和游泳时间,提高SOD活性、降低LA含量,提高肝糖原和肌糖原含量,表明杏鲍菇多糖具有抗疲劳作用。

超声辅助酶解法提取杏鲍菇多糖工艺研究

以杏鲍菇为原料,利用超声波及纤维素酶进行处理,以蒸馏水为提取剂,采用苯酚-硫酸法测定杏鲍菇多糖。以多糖得率为优化参数,通过单因素试验及正交试验得到酶解法对杏鲍菇多糖的提取率最适宜的工艺条件。结果表明,超声提取的效果最好为料液比1∶30,pH值6.0,超声时间20 min;酶解法提取的最佳条件为酶添加量0.8%,酶解时间90 min,酶解温度60℃。在此条件下杏鲍菇中多糖的提取率可达28.06%,高于传统水提法。

杏鲍菇多糖及其消化产物对淀粉消化酶的抑制及相互作用

【目的】以杏鲍菇多糖(PEP)为原料,探究其基本理化性质和对葡萄糖扩散和吸附能力的影响。利用体外模拟消化模型制备PEP模拟消化产物(D-PEP),探究PEP和D-PEP对与糖代谢相关的消化酶活性的影响,并进一步评价其与α-葡萄糖苷酶的相互作用。【方法】首先基于前人的研究方法对PEP的基本理化性质进行评价;然后采用DNS法测定PEP和D-PEP对α-淀粉酶活力的抑制效果,采用对硝基苯-α-D-吡喃葡萄糖苷(PNPG)法测定对α-葡萄糖苷酶抑制率;最后采用荧光光谱法研究PEP和D-PEP与α-葡萄糖苷酶的相互作用。【结果】PEP具备良好的溶解/溶胀性、持水/持油性,且其对葡萄糖扩散和吸附能力具有一定的抑制作用。PEP对麦芽糖酶和α-葡萄糖苷酶具有显著的抑制作用,对α-淀粉酶没有抑制作用,当PEP浓度为4 mg·mL^(-1)时,其对麦芽糖酶和α-葡萄糖苷酶的抑制率分别为(77.20±2.71)%和(78.91±0.51)%。然而消化产物D-PEP对3种酶都有显著的抑制作用,4 mg·m L^(-1)的D-PEP对α-淀粉酶、麦芽糖酶和α-葡萄糖苷酶的抑制率分别为(84.08±1.79)%、(20.58±1.20)%和(95.58±0.12)%。荧光光谱法结果表明,随着PEP和D-PEP浓度增大,α-葡萄糖苷酶的内源荧光逐渐减弱,且其对α-葡萄糖苷酶的内源荧光淬灭以静态淬灭为主,其结合位点大于等于1。【结论】与PEP相比,D-PEP不仅能抑制麦芽糖酶和α-葡萄糖苷酶活性,对α-淀粉酶活力也有显著的抑制效果,D-PEP对淀粉消化酶表现出更好的抑制效果,可在一定程度上影响机体糖代谢。

HepG2细胞胰岛素抵抗模型建立及在筛选天然有效多糖中的应用

探讨高糖、地塞米松、胰岛素诱导肝细胞HepG2胰岛素抵抗模型(IR-HepG2)的建立方法,检测杏鲍菇多糖、红枣多糖和枸杞多糖体外改善肝细胞胰岛素抵抗的作用,评估3种多糖的降糖效果.选择不同浓度的地塞米松、胰岛素及两者联合作用,在高糖培养条件下诱导HepG2胰岛素抵抗,MTT法检测不同诱导剂对HepG2细胞活性的影响,葡萄糖氧化酶法测定细胞葡萄糖消耗量,同时检测去除诱导剂后,IR-HepG2细胞模型的稳定性.以3种天然多糖处理IR-HepG2细胞,检测其对细胞活力及胰岛素抵抗的影响.结果表明:在高糖培养条件下,诱导剂为地塞米松(3.75μmol/L)联合胰岛素(0.001μmol/L)可成功诱导IR-HepG2细胞模型,且对细胞活性无显著影响,模型能够在72 h内保持稳定.枸杞多糖具有更好的降糖效果.

响应面法优化白灵菇菌丝体多糖运动饮料配方及其抗疲劳研究

本研究探讨了白灵菇菌丝体多糖运动饮料的最佳工艺配方。研究在单因素实验的基础上,以白灵菇菌丝体多糖使用量、木糖醇使用量、柠檬酸使用量作为试验因素,以饮料的感官评分作为响应面值,应用响应面设计方法建立数学模型并进行分析。结果表明,白灵菇菌丝体多糖运动饮料的最佳工艺配方为:白灵菇菌丝体多糖使用量40%、木糖醇使用量8%、柠檬酸使用量0.2%。按照上述工艺配方,实际感官评分为87.1。通过抗疲劳试验证明,白灵菇菌丝体多糖运动饮料能够延长小鼠负重竭力游泳时间,控制小鼠血清尿素氮含量增长,促进小鼠机体抗疲劳能力的提升。