复合酶解法提取香菇多糖蛋白的研究

存在于香菇中的多糖物质,具有增强人体非特异性免疫功能的作用。本文报道采用复合酶解和热水浸提法分离纯化香菇中多糖蛋白的综合程序。与单纯热水浸提法等其它方法相比,本法能显著提高香菇有效成分的浸提效果,总氨基酸与必需氨基酸百分比含量均提高２倍以上,香菇多糖含量提高了４倍,一些特殊成分及较高分子量的葡聚糖物质含量也明显提高。     

香菇多糖体内抗氧化活性研究

目的：探究香菇多糖的体内抗氧化活性。方法：以灌胃生理盐水的小鼠为正常对照组,将香菇多糖药物组分为低、中、高3种剂量组。小鼠在灌胃3w后,由眼眶取血,按照试剂盒要求测定小鼠血清中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）活力以及丙二醛（MDA）含量。结果：香菇多糖可显著提高小鼠血清中SOD和GSH-Px的活力,同时减少血清中MDA含量。结论：香菇多糖对小鼠体内抗氧化活性的提高具有显著作用。     

香菇不同发育阶段子实体多糖的理化性质及体外免疫活性研究

香菇多糖是从香菇菌丝体或子实体中提取纯化得到的高分子葡聚糖,作为香菇主要的生物活性物质,具有提高免疫力、抗病毒、抗氧化和抗真菌等生物活性。本研究对幼菇期、菌褶期、采收期、成熟期和开伞期5个不同发育阶段香菇粗多糖含量、理化性质和体外免疫活性进行比较分析。结果表明,其粗多糖含量、理化性质和体外免疫活性具有明显差异。粗多糖得率呈现先增加后降低的趋势,菌褶期得率最高;粗多糖含量也呈现先增加后降低的趋势,成熟期含量最高;多糖的分子量因发育期不同而有较大差异,发育后期所得粗多糖分子量千万级以上组分的比例增加,分子量百万级和十万级的多糖组分比例降低;不同发育阶段香菇子实体多糖的单糖组成均包含果糖、半乳糖、葡萄糖和甘露糖4种单糖,且都以葡萄糖为主;5个阶段的多糖组分均有较好的体外免疫活性,其中幼菇期粗多糖在浓度500μg/mL时表现出最高的体外免疫活性。本研究探讨了香菇不同发育阶段子实体多糖的理化性质及体外免疫活性,为香菇采收期的确定及香菇多糖的制备与利用提供理论依据。     

微波辅助提取灰树花多糖工艺研究

采用提取时间、微波功率、液料比的单因素试验和正交试验法优化微波辅助提取灰树花多糖条件.结果表明,以净多糖得率为指标,影响微波辅助提取灰树花多糖的主次因素为:提取时间＞微波功率＞液料比,并且提取时间和微波功率的影响达到了极显著水平.灰树花多糖最佳提取工艺条件为:提取时间为10 min,微波功率为80%(全功率为800 W),液料比为25∶1.创立了一种用苯酚-硫酸法测定多糖时排除蛋白质干扰的方法.     

酶法提取香菇柄多糖

比较了菇柄多糖的多种提取方法。结果表明,利用复合酶解法结合热水浸提法提取菇柄多糖,提取率达７．１３％（克多糖／１００克菇柄干品）,是热水浸提法的３．４３倍,酶解的最适作用条件为ｐＨ４．５,温度４６℃,时间８０ｍｉｎ。     

两种方法提取香菇多糖的比较研究

采用正交试验比较研究热水浸提法和微波提取法提取香菇多糖。热水浸提法的最佳工艺为:提取温度70℃、提取时间4h、料液比1∶15,提取率3.243%;微波提取法的最佳工艺为:微波功率560W、微波处理时间60s、料液比1∶10,提取率4.771%。微波提取法效率高、时间短,是理想的香菇多糖提取方法。     

香菇多糖提取分离的研究

采用DEAE—cellulose柱层析和Sephadex G-200柱层析从香菇子实体热水浸提的粗多糖中分离出两个多糖组分Lenl—A、Len2-A,经旋光度法、Sepharose 4B柱层析、红外光谱等方法检测确定为均一多糖组分,且均为α-构型的吡喃糖,苯酚硫酸法测得其糖质量分数分别为91．5％和92．3％,旋光度为＋5．6°、＋11．2°,相对分子质量为375×10^3、718×10^3。     

响应面优化超声辅助提取香菇多糖的工艺研究(英文)

为确定香菇多糖的最佳提取工艺,利用响应面分析法对香菇多糖的提取工艺进行优化。在单因素实验的基础上,以超声时间、超声功率、浸提温度和浸提时间为响应因素,多糖提取得率为响应值,根据正交旋转组合试验设计原理进行四因素三水平的响应面分析。实验结果表明,采用超声功率174.94 W,超声时间为18.94min,在80.71℃下提取3.01 h,得到的香菇多糖提取率最高,为9.61%。当香菇多糖的浓度为3 mg/mL,其·OH清除率为52.1%。     

香菇活性成分提取工艺研究及药理学研究进展CSCD

香菇是一种具有丰富的营养价值同时兼有良好的药用价值的真菌,可食药两用。追溯到我国古代,就已有相关记载提示其有增强脑细胞功能的作用:服食饵菌类可以"增智慧""益智开心"(《神农本草》)。香菇作为亚洲传统医药,含有多种活性成分,如香菇多糖、香菇嘌呤、香菇蛋白等。这使得其在医学领域得到越来越多的应用,尤其在抗肿瘤、调节免疫力、抗氧化、抗病毒及降血脂等方面都具有较好的治疗效果。本文就香菇主要活性成分的提取方法、化学结构及药理作用作一综述。     

龙眼壳黄酮的微波提取及体外抗氧化活性研究

研究了微波辅助提取龙眼壳黄酮的工艺条件,并对其进行了抗氧化性能的研究。正交试验结果表明较佳提取工艺为:V(70%乙醇)∶m(龙眼壳粉)=30 m L∶1 g,微波功率480 W处理20 s后,于水浴30℃浸提30min,龙眼壳黄酮得率为3.465%。抗氧化试验表明龙眼壳黄酮对亚硝酸盐具有良好的清除能力,其效果优于Vc,对卵黄脂蛋白脂质过氧化及羟基自由基具有明显的抑制作用,其效果分别弱于对照物2,6-二叔丁基-4-甲基苯酚(BHT)和Vc。     

氯化两面针碱的提取分离、全合成及药理活性的研究进展

两面针是我国民间常用的传统中药之一,从根部提取分离到的氯化两面针碱是两面针植物多种生物活性的主要有效成分。当前,直接从植物中提取分离有效成分的方法不仅回收率低、成本高,而且氯化两面针碱存在水溶性差和生物利用度低的问题,因此大多学者致力于改造其结构以提高活性和利用度,同时探索高效经济的全合成方法以满足研究和生产的需求。该文以国内外相关文献为基础,对氯化两面针碱的提取、分离纯化、结构化合物的全合成路线以及药理活性机制的研究进展做一综述,总结目前研究中存在的不足,并对今后的研究重点做出展望,旨在为今后深入研究两面针碱及衍生物活性改造、解析作用机制、开发基于此结构的创新药物分子等提供理论依据。     

银杏叶多酚的机械力辅助提取及其体外抗氧化活性研究

为优化银杏叶多酚提取工艺,通过单因素试验考察填充率、球磨转速、球磨时间、乙醇浓度、料液比、提取温度、提取时间七个因素对机械力辅助提取银杏叶多酚得率的影响,以银杏叶多酚得率为响应值,采用Box-Benhnken三因素三水平响应面设计优化工艺,同时比较了4种提取方法对银杏叶多酚提取得率和抗氧化活性的差异。结果表明,机械力辅助提取银杏叶多酚的最佳工艺条件为:填充率26%、球磨转速为400rpm、球磨时间为15min。在此条件下,银杏叶多酚的得率为7.33%。机械力辅助乙醇提取银杏叶多酚得率低于碱水提取法,但是抗氧化活性高于碱水法提取的银杏叶多酚;抗氧化活性与乙醇回流法提取的银杏叶多酚相当,但是提取得率高于乙醇回流法。此提取工艺高效可行,具有一定的参考价值。     

羊肚菌多糖提取及其抗氧化活性研究

为了探讨碱法提取羊肚菌多糖的工艺条件并测定其抗氧化活性,该研究以四川北川羊肚菌为原料,采用碱法提取羊肚菌多糖,利用苯酚-硫酸法对羊肚菌多糖得率进行测定,并通过单因素探讨提取温度(70、80、90、100℃)、提取时间(2、4、6、8 h)、碱液浓度(0.4、0.6、0.8、1.0 mol·L~(-1))、料液比(1∶15、1∶20、1∶25、1∶30 g·mL~(-1))对羊肚菌多糖得率的影响,同时采用正交试验优化提取工艺,对其抗氧化活性进行测定。结果表明:在提取温度90℃、提取时间5 h、碱液浓度0.7 mol·L~(-1)、料液比1∶20(g·mL~(-1))条件下得到的羊肚菌多糖得率为5.39%。羊肚菌多糖具有较强的清除DPPH自由基、羟自由基、超氧阴离子的能力和较好的还原能力,其IC50分别为0.468、0.208、0.022、0.014 mg·mL~(-1),抗氧化能力依次为还原能力超氧阴离子清除能力羟自由基清除能力DPPH自由基清除能力。优化后的羊肚菌多糖提取工艺合理、可行,且羊肚菌多糖具有较强的抗氧化活性。     

Effects of excluded volume and polydispersity on solution properties of lentinan in 0.1 M NaOH solution

Seven lentinan fractions of various weight-average molecular weights (M(w)), ranging from 1.45 x 10(5) to 1.13 x 10(6) g mol(-1) were investigated by static light scattering and viscometry in 0.1M NaOH solution at 25 degrees C. The intrinsic viscosity [eta] - M(w) and radius of gyration s(2)(z) (1/2) - M(w) relationships for lentinan in 0.1M NaOH solution were found to be represented by [eta] = 5.1 x 10(-3)M(w) (0.81) cm(3) g(-1) and s(2)(z) (1/2) = 2.3 x 10(-1)M(w) (0.58) nm, respectively. Focusing on the effects of the M(w) polydispersity with the Schulz-Zimm distribution function, the data of M(w), s(2)(z) (1/2), and [eta] was analyzed on the basis of the Yoshizaki-Nitta-Yamakawa theory for the unperturbed helical wormlike chain combined with the quasi-two-parameter (QTP) theory for excluded-volume effects. The persistence length, molecular weight per unit contour length, and the excluded-volume strength were determined roughly to be 6.2 nm, 980 nm(-1), and 0.1, respectively. Compared with the theoretical value calculated by the Monte Carlo model, the persistence length is longer than that of the single (1 --> 3)-beta-(D)-glucan chain. The results revealed that lentinan exists as single-stranded flexible chains in 0.1M NaOH solution with a certain degree of expansion due to the electrostatic repulsion from the interaction between the OH(-) anions and lentinan molecules.     

A primer on cancer immunology and immunotherapy

The role of immunity in cancer has been abundantly demonstrated in murine tumor models as well as in man. Induction of clinically effective antitumor immune responses, based on this information, in patients with cancer however, remains elusive. This is not because tumors lack recognizable antigens [in fact there is evidence that there are thousands of potential novel targets in each tumor cell] but rather due to the fact that the induction of responses is not adequate nor particularly well understood. Tumors seem to be rather effective at limiting immune responses. Many of the molecularly defined antigens that have been detected on tumor cells are derived from self-proteins and as such are subject to tolerizing mechanisms. Such tumors have also developed escape mechanisms capable of evading or suppressing immune responses. Understanding the role of dendritic cells during the effector phase of the immune response and the complex interactions of stromal, immune, and tumor cells in the tumor microenvironment represent the next challenges to be understood for tumor immunology.This is a summary of the work presented at the First Cancer Immunology and Immunotherapy Summer School, 8–13 September 2003, Ionian Village, Peloponnese, Greece     

Therapeutic effects of lentinan on inflammatory bowel disease and colitis‐associated cancer

In this study, we investigated the therapeutic potential of lentinan in mouse models of inflammatory bowel disease (IBD) and colitis‐associated cancer (CAC). Lentinan decreased the disease activity index and macroscopic and microscopic colon tissue damage in dextran sulphate sodium (DSS)‐induced or TNBS‐induced models of colitis. High‐dose lentinan was more effective than salicylazosulfapyridine in the mouse models of colitis. Lentinan decreased the number of tumours, inflammatory cell infiltration, atypical hyperplasia and nuclear atypia in azoxymethane/DSS‐induced CAC model. It also decreased the expression of pro‐inflammatory cytokines, such as IL‐13 and CD30L, in IBD and CAC model mice possibly by inhibiting Toll‐like receptor 4 (TLR4)/NF‐κB signalling and the expression of colon cancer markers, such as carcinoembryonic antigen, cytokeratin 8, CK18 and p53, in CAC model mice. In addition, lentinan restored the intestinal bacterial microbiotal community structure in IBD model mice. Thus, it shows therapeutic potential in IBD and CAC model mice possibly by inhibiting TLR4/NF‐κB signalling‐mediated inflammatory responses and disruption of the intestinal microbiotal structure.     

Effects on microbial activity by extraction of indigenous cells from soil slurries

Abstract: Possible effects on the physiological activity and culturability of soil microorganisms by different soil dispersion procedures, and effects on activity caused by extracting bacteria from soil, were investigated. There was no apparent difference in cfu's with dispersion of a silty loam soil and a loamy sand soil with pyrophosphate as compared to dispersion in NaCl. Substrate-induced respiration was reduced in the silty loam soil, and methanol oxidation was reduced in the loamy sand soil with dispersion in pyrophosphate, and the soil pH was irreversibly increased by the treatment. Extracted bacterial fractions had lower numbers of culturable cells as percentage of the total number of bacteria in each fraction, lower respiration rates and no methanol oxidation activity as compared to the soil slurry both before and after extraction. The physiological activity was apparently not affected by the number of cells extracted. This indicates that the increased extraction rate of indigenous soil bacteria obtained by effective disruption of aggregates and detachment of cells from surfaces, only results in increased extraction of cells that have been physiologically changed as a result of the extraction process.     

Immunological analysis and clinical effects of intraabdominal and intrapleural injection of lentinan for malignant ascites and pleural effusion

Twenty effusions in sixteen patients with malignant peritoneal and/or pleural effusions were treated with intracavitary injection of lentinan. Lentinan was injected at a dosage of 4 mg/week for 4 weeks. In total, sixteen (80%) of twenty lesions demonstrated clinical responses. Performance status was improved in seven patients. The average survival time in responders was 129 days, while, in non-responders, it was 49 days. Serious toxicities were not observed.NK activity of PBMC significantly decreased after lentinan injection. NK activity of PEC in responders was augmented significantly. Anti-Daudi and lymphokine activated killer activity were also augmented or maintained after lentinan injection.     

辣椒素的提取及抑菌活性研究

以干红辣椒为原料,选取95%的乙醇和正己烷作为提取溶剂,利用索氏提取法提取其中的辣椒素。结果表明,以95%的乙醇作为提取溶剂的效果好于正己烷,辣椒素的提取率最高可达1.77%,高于正己烷的1.12%的提取率。用滤纸片法研究辣椒素对大肠杆菌等8种常见食品腐败菌的抑菌活性。结果表明:辣椒素对金黄色葡萄球菌、枯草芽孢杆菌有明显的抑菌效果,对大肠杆菌抑菌作用较弱,对黑曲霉、青霉、保加利亚乳酸杆菌和噬热链球菌没有作用。辣椒素经过高温灭菌后,抑菌作减弱,培养时间对抑菌圈的影响不大。以正己烷为助溶剂,确定辣椒素对枯草芽孢杆菌的最低抑菌浓度(MIC)为50mg/mL。     

菊芋粕菊粉的酶法提取、组成及抗氧化活性研究

本研究拟应用酶法提取技术解决菊芋菊粉工业化生产中的废弃物——菊芋粕再利用程度低的问题,并评价菊芋粕菊粉的抗氧化活性功效。对菊芋粕菊粉的果胶酶酶法提取的最佳条件通过响应面法进行了优化,并对菊芋初次水提菊粉(primary water-extracted inulin,PWI)和二次酶提菊粉(secondary enzymatic-extracted inulin,SEI)的组成成分和抗氧化活性进行了比较分析。响应面法优化确定的菊芋粕菊粉最佳酶法提取条件为:pH4.5、提取温度50 ℃、酶底比7.5 U/g、提取时间2 h,该提取方法所得菊芋粕菊粉的得率为35.30%±0.85%,与传统热水浸提法相比菊粉得率提高38.16%。组成分析结果显示,SEI的总糖和菊粉含量均显著高于PWI( P <0.05)。在菊粉聚合度方面,PWI中蔗果三糖和蔗果四糖含量较高,而SEI中蔗果五糖、蔗果六糖及其以上聚合度菊粉的含量较高。此外,SEI的抗氧化活性优于PWI。因此,果胶酶辅助提取方法有望为菊芋粕菊粉的再利用难题提供新的解决思路。