Monitoring the polysaccharide quality of Agaricus blazei in submerged culture by examining molecular weight distribution and TNF-α release capability of macrophage cell line RAW 264.7

A method for monitoring the biological activity of broth polysaccharides of Agaricus blazei (AB-BP) in a submerged culture is described. The TNF-alpha releasing capability of AB-BP on the macrophage cell line, RAW 264.7, correlated with the molecular weight of AB-BP. The quality of polysaccharide in a submerged culture of Agaricus blazei was indirectly monitored by analyzing the distribution of its molecular weight within 1 h. The harvest time of the maximum polysaccharide production did not coincide with that of the maximum biological characteristics in the batch culture.     

姬松茸的研究与进展

概述了大型真菌姬松茸的生物学特性、保健功能以及近年来的研究进展,并对其主要功效物质如多糖等成分、姬松茸深层发酵技术的现状和菌丝体应用情况作了介绍。     

Anti-tumor polysaccharide from the mycelium of liquid-cultured Agaricus blazei mill.

Anti-tumor active polysaccharide against Sarcoma 180 was isolated by DEAE-Sepharose CL-6B and Sepharose 4B column chromatography from the hot-water soluble fraction of the mycelium of liquid-cultured Agaricus blazei mill. This polysaccharide did not react with antibodies of anti-tumor polysaccharides such as lentinan, gliforan, and FIII-2-b which is one of anti-tumor polysaccharides from Agaricus blazei. Moreover, the analyses of 13C-NMR and GC-MS suggested that this polysaccharide was preliminarily glucomannan with a main chain of beta-1,2-linked D-mannopyranosyl residues and beta-D-glucopyranosyl-3-O-beta-D-glucopyranosyl residues as a side chain. This polysaccharide was completely different from the anti-tumor polysaccharide from fruiting body of Agaricus blazei, beta-1,6-glucan.     

基于姬松茸深层发酵的Plackett-Burman筛选

对姬松茸液体深层发酵培养工艺条件进行单因素实验：应用Minitab15软件设计Plackett—Burman筛选实验,筛选出蔗糖及酵母膏质量浓度、培养温度、微量元素配比4个显著性影响因素,通过正交试验对其进行优化。确定最佳培养工艺条件：蔗糖质量浓度45g/L,酵母膏质量浓度3g／L,KH2PO4质量浓度2．5g／L,MgSO4质量浓度1．25g／L,培养温度27℃,培养时间7d,多糖产量可达4．64g／L。     

巴西蘑菇菌体深层发酵培养条件的优化及成分分析*

对巴西蘑菇深层发酵条件进行了详细的研究,确定了巴西蘑菇菌体发酵的最优方案是:7%葡萄糖,1.5%酵母膏,0.3%磷酸氢二钾,0.1%硫酸镁,VB1 10mg/100ml,自然pH,接种量为15%,装液量为100ml/250ml三角瓶,150r/min,25℃恒温培养9天,菌丝干重达到1.8g每100ml发酵液。通过对巴西蘑菇子实体与深层培养菌丝体中蛋白质营养成分及多糖含量进行了分析比较,发现它们均是极好的营养食品之源。     

巴西蘑菇多糖研究进展

巴西蘑菇是一种味道鲜美、富含生物活性物质的珍稀食 (药 )用菌。巴西蘑菇多糖作为一种极有开发前景的生物活性物质已得到国内外的普遍重视。综述了国内外巴西蘑菇多糖化学分析和药理研究方面的进展情况。     

原生质体紫外诱变选育姬松茸新菌株

对姬松茸原生质体进行紫外诱变处理,选育出在液体培养条件下生物量明显高于原始菌株的诱变株。经10代传代培养及摇瓶培养,表明该菌株稳定性良好,与出发菌株相比生物量提高5.5%,胞外多糖产量提高6.1%。     

姬松茸菌丝体中各组分多糖的抗肿瘤免疫活性

对深层发酵培养的姬松茸菌丝体依次采用热水、冷碱、热碱提取,得到了四种主要的多糖组分。其中冷碱提取的水不溶性多糖(CAASP)得率最高,达8.5 8g/ 10 0g干菌,且不同的多糖组分具有不同的单糖组成。其中热水提取的水溶性多糖(HWSP)及冷碱提取的水溶性多糖(CAWSP)在2 0mg/kg·d的剂量下对小鼠S180 移植瘤显示出较高的抗肿瘤活性,抑瘤率分别高达5 9.84 %和6 4 .6 6 %。同时发现在四种多糖在发挥抗肿瘤作用过程中,小鼠的脾脏指数和胸腺指数有不同程度的提高。并且各多糖活性组分在体外均具有显著的刺激脾淋巴细胞增殖的作用     

不同培养料和发酵次数栽培巴氏菇比较

分别以玉米秸秆和稻草为培养料栽培巴氏菇,对菌丝生长性状进行对比;同时,在我国传统发酵栽培方法的基础上,将三次发酵法与二次发酵法进行了对比。试验结果表明:利用玉米秸秆栽培的巴氏菇在发菌速度及子实体质量和产量上均优于稻草;培养料经过3次发酵更适宜巴氏菇生长。经过3次发酵的玉米秸秆培养料栽培的巴氏菇产量和生物学效率分别为7.7 kg/m2和27.5%,子实体粗蛋白含量44.89%,可溶性糖含量44.01%,18种氨基酸总量31.70%,明显高于2次发酵的玉米秸秆培养料及2次、3次发酵的稻草培养料。     

Agaricus blazei Murill Polysaccharides Protect Against Cadmium-Induced Oxidative Stress and Inflammatory Damage in Chicken Spleens

Biological Trace Element Research - Agaricus blazei Murill polysaccharide (ABP) has exhibited antioxidant and immunoregulatory activity. The aim of this study was to investigate the effect of ABP...     

Influence of drying methods over in vitro antitumoral effects of exopolysaccharides produced by Agaricus blazei LPB 03 on submerged fermentation

Agaricus blazei is a mushroom that belongs to the Brazilian biodiversity and is considered as an important producer of bioactive compounds beneficial to human health. Studies have demonstrated that these compounds present immuno-modulatory, antioxidant and antitumor properties. In order to compare the most used method for fungal polysaccharide drying, lyophilization with other industrial-scale methods, the aim of this work was to submit A. blazei LPB 03 polysaccharide extracts to vaucum, spray and freeze drying, and evaluate the maintenance of its antitumoral effects in vitro. Exopolysaccharides produced by A. blazei LPB 03 on submerged fermentation were extracted with ethanol and submitted to drying processes. The efficiency represents the water content that was removed during the drying process. The resultant dried products showed water content around 3% and water activity less than 0.380, preventing therefore the growth of microorganisms and reactions of chemical degradation. Exopolysaccharide extracts dried by vacuum and spray dryer did not showed any significant cytotoxic effect on cell viability of Wistar mice macrophages. Content of total sugars and protein decrease after drying, nevertheless, 20 mg/ml of exopolysaccharides dried by spray dryer reached 33% of inhibition rate over Ehrlich tumor cells in vitro.     

姬松茸原生质体诱变育种研究

以姬松茸原生质体为诱变材料,用不同诱变剂紫外线（UV）,钴60（Co^60）,亚硝基胍（NTG）进行多次反复诱变处理,获得2株活性多糖产量较高,遗传稳定的变异株C811/N516。与出发菌株各项发酵指标比较,突变株其他各发酵特性变化不明显,而单位发酵液活性多糖含量分别提高260%和300%以上。     

巴氏蘑菇多糖对铅中毒大鼠脾脏细胞因子mRNA表达的影响

为了研究巴氏蘑菇Agaricus blazei多糖对铅中毒大鼠脾脏细胞因子mRNA表达的影响,进而探索巴氏蘑菇多糖对铅中毒大鼠的免疫调节作用。选用45日龄SD大鼠,随机分为6组,每组8只,雌雄各半。分别为正常对照组、多糖组、铅中毒模型组、铅+50mg/kg·d多糖试验组、铅+100mg/kg·d多糖试验组、铅+200mg/kg·d多糖试验组。铅中毒模型组、铅+多糖试验组分别给予含0.2%醋酸铅饮水,自由饮用。饲养60d后,采集脾脏,荧光定量RT-PCR对其细胞因子IL-1β、IL-2、IL-6、TNF-α     

巴氏蘑菇微卫星序列的分离及其对Co60辐射诱变菌株的鉴别

根据链霉素磁珠和生物素特异结合的特性,用生物素标记的二聚核苷酸重复序列探针从巴氏蘑菇的基因组中分离微卫星序列。将结合于链霉素磁珠上的标记探针同两端连接已知序列人工接头的巴氏蘑菇DNA酶切片段杂交。洗脱未杂交DNA片段后,用磁珠富集的片段建立微卫星文库。挑取522个菌落用对应重复序列为引物进行PCR筛选,得到48个阳性克隆,经测序有32个菌落含微卫星序列。微卫星富集效率为阳性克隆数的67%,总克隆数的6%。除去重复或无效的微卫星序列,在设计出的12对用于鉴别85个巴氏蘑菇的Co60辐射变异株微卫星引物中,有4对引物总共扩增出明显的变异菌株17个。证明有些微卫星位点可用于巴氏蘑菇辐射变异品种的指纹筛选与鉴别。     

Activation of the alternative complement pathway by Agaricus blazei Murill.

Components of Agaricus blazei Murill have been demonstrated to have a wide range of immunopotentiating activities. The present study was designed to evaluate the effect of A. blazei Murill upon activation of the complement system in human serum in vitro. Additional studies were performed to determine the cytotoxic effect of complement-opsonized particles of A. blazei Murill against human tumor cells in culture. A fine particle of A. blazei Murill (ABP), prepared by mechanical disruption, was used throughout the experiments. ABP activated the human complement system via the alternative pathway in human serum. Activation of the alternative pathway was both time- and dose-dependent. When the particles from fruiting bodies of A. blazei Murill (ABP-F) were reacted with human serum, the formation of complement-opsonized ABP, iC3b-ABP-F complexes, and binding of the complexes to human peripheral blood monocytes, were demonstrated in vitro by immunofluorescence. Further, the resident human peripheral nucleated cells incubated in the presence of iC3b-ABP-F complexes inhibited the proliferation of human tumor cell line TPC-1 in vitro.     

Secretion of TNF-alpha, IL-8 and nitric oxide by macrophages activated with Agaricus blazei Murill fractions in vitro

Water extracts of the mycelial culture and fruiting bodies of Agaricus blazei Murill were fractionated by ethanol precipitation using various ethanol concentrations. Original water extracts from mycelia (Fraction A-0) and fruiting bodies (Fraction B-0) induced TNF-alpha secretion by macrophages derived from rat bone marrow. Fractions B-4 and B-5 obtained from ethanol precipitation of fruiting bodies using 44% and 50% ethanol, respectively, and Fraction B-6 obtained from the supernatant at 50% ethanol markedly induced TNF-alpha secretion. Similar effects were observed in IL-8 secretion by macrophages. Regarding nitric oxide (NO), Fraction B-5 induced a significant increase in NO secretion and Fractions B-4 and B-6 induced slightly NO secretion. Northern blot analysis showed that the increases in cytokine- and NO secretion were due to an increase in cytokine mRNAs or NO synthase mRNA. Therefore, it is concluded that Agaricus blazei Murill components which activate macrophages result in the induction of cytokine- and NO secretion in vitro.     

A polysaccharide from Agaricus blazei inhibits proliferation and promotes apoptosis of osteosarcoma cells

Many reports have proved that traditional Chinese herbal medicines (TCM) have become popular used in disease prevention and as alternatives to cancer chemotherapy. In this study, we purified a polysaccharide (ABP-Ia) from the fruiting bodies of Agaricus blazei and identified its molecular weight to be 4.2×10(5)Da. ABP-Ia was a heteropolysaccharide fraction consisting of glucose, mannose, and galactose in a molar ratio of 1:1:1, along with trace of rhamnose. The effect of ABP-Ia at three concentrations of 100, 200 and 400 μg/mL on the cell growth and apoptosis was evaluated in osteosarcoma cell lines HOS and a normal human osteoblast cell line NHOst. ABP-Ia had a significant inhibitory effect against the growth of HOS cells, whereas a mild cytotoxicity to the HOS cells mediated by ABP-Ia was observed, which was in accordance with the results that ABP-Ia substantially induced apoptosis in a dose-dependent fashion in the HOS cells. However ABP-Ia had no or minor inhibitory and cytotoxic effects on the viability of NHOst cells even at the high concentration of 400 μg/mL. Base on all the observations, we could conclude that ABP-Ia had an evident inhibitory effect on the growth of HOS cells mainly through induction of apoptosis, with a minor toxicity to normal human osteoblast cell.     

采用SCGE和SCE分析法研究姬松茸菌体多糖(Ab-Mp)对DNA损伤的抑制作用

热水浸提法提取姬松茸菌丝体多糖,每天以10mg/(kg bw)和20mg/(kg bw)两种剂量对小鼠进行灌胃,连续15d.腹腔注射环磷酰胺(CP),运用单细胞凝胶电泳(SCGE)法和姐妹染色体交换(SCE)分析法,研究Ab-Mp对染色体损伤的保护作用.结果表明,Ab-Mp降低了CP损伤后彗星细胞的比例,缩短了彗星尾长;降低了CP损伤的姐妹染色单体交换率.Ab-Mp对环磷酰胺诱发的DNA损伤具有拮抗作用.     

Purification and characterization of a novel polysaccharide involved in the pellicle produced by a thermotolerant Acetobacter strain

Acetobacter strains able to produce a thick pellicle at 37 degrees C were screened among many thermotolerant strains isolated from fruits in Thailand. As a result, Acetobacter sp. SKU 1100 was selected as the producer of a relatively thick pellicle even when cultured at higher temperatures such as 37 degrees C or 40 degrees C. This strain could produce a pellicle polysaccharide in a shaking submerged culture as well as under static culture conditions. The polysaccharide was found to be attached to the bacterial cells. Although the polysaccharide production was higher at 30 degrees C than at 37 degrees C in shaking submerged culture, the productivity in static culture was not decreased even at higher temperatures. The membrane-attached polysaccharide was purified from the SKU 1100 strain by cell disruptions using either ultrasonic treatment or lysozyme treatment, followed by ultracentrifugation, enzyme treatments, dialysis against SDS, DEAE-cellulose column chromatography, alcohol precipitation, and gel filtration chromatography. The polysaccharide purified by the sonic treatment and also by the mild conditions using lysozyme treatment had the same average molecular mass of 120 kDa. The purified polysaccharide was composed of three different monosaccharides; glucose, galactose, and rhamnose, in an approximately equimolar ratio of 1:1:1.