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.

白腐菌Pleurotus eryngii-Co007产木质素降解酶条件的优化

木质素降解酶是参与降解木质素的酶类，广泛存在于白腐真菌中。为提高白腐菌Pleurotus eryngii—Co007的产木质素降解酶的活力，研究了该菌株的产酶曲线、发酵参数及培养基组分对其产木质素降解酶的影响。单因素试验结果表明该菌株发酵第8天时发酵液中木质素降解酶酶活最高；较优的发酵参数是pH5．0—6．0、发酵温度25—27℃，接种量10％～15％，其中pH影响最显著；麦芽糖和有机氮源有利于木质素降解酶的表达。但考虑到酶表达的增加量和生产成本的经济性选择3％的葡萄糖、0．2％酵母粉和0．2％牛肉膏组合氮源；诱导剂中秸杆、Cu^2＋和吐温80诱导产酶效果明显。

响应面法优化白腐菌Pleurotus eryngii_(-Co007)产木质素降解酶条件

为提高Pleurotus eryngii-Co007产木质素降解酶能力,考察了初始pH、秸秆浓度、Cu2+浓度、吐温-80含量对其产木质素降解酶的影响。采用单因素试验和响应面分析法的Central Composite进行试验设计,得到Pleurotus eryngii-Co007产木质素降解酶的最佳条件:培养基初始pH 5.63、秸杆加量1.12%、Cu2+浓度4.00mmol/L、吐温-80含量0.83g/L。在此条件下,第8天摇瓶发酵液中木质素降解酶活性达到241.40U/ml,与回归模型的预测值相对误差仅2.18%。

Quality Evaluation and Standards Development of Liquid Spawn of Pleurotus eryngii

With four Pleurotus eryng# varieties as test materials, the mycelium pelletdensity, dry weight and diameter, fermentation broth pH value, and deoxidized sugar content were determined during their liquid fermentation to investigate the growth of mycelia and changes in fermentation broth. In addition, the sensory, biochemical and microbiological indicators of excellent P. eryngii mycelia were determined by sensory tests, microscope morphology observation and mycelium pellet back-inoculation tests. The results showed that the standards for high-quality liquid spawn of P. eryngii were as follows： bacterial liquid was light yellow and clarifiedwith rich bitter almond flavor but no bacterial contamination; mycelium pellets were visible, white and uniform with burrs on the surface, diameter 〈2.5 mm and concentration of 80%-100%; mycelium pellets were stout, and there were 3--4 clamp connectionsper visual field under the microscope; fermentation time was generally 120-144 h.

Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

A versatile peroxidase (VP-Peco60-7 ) was generated and purified from the liquid culture of Pleurotus eryngii. The purification procedure included ammonium sulfate precipitation, ion exchange chromatography, and gel chromatography. The molecular weight and isoelectric point (pI) of VP-Peco60-7 were determined to be approxi-mately 40 kDa and 4.1, respectively. By N-terminal sequence determination and peptide mapping analysis, VP-Peco60-7 was found to be similar to the versatile peroxidase isoenzyme VPL1, which was previously isolated from liquid cultures of the same species. However, the molecular weight and pI of VP-Peco60-7 were different from those of versatile peroxidases of liquid cultures, implying that the VP-Peco60-7 in this study is of a novel type. With 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as a substrate, the maximal enzyme activity was obtained at 50 °C and pH 3.0. The catalysis of ABTS by VP-Peco60-7 was expressed by the Michaelis-Menten equa-tion. At 50 °C and pH 3.0, the maximum velocity (V max ) was 188.68 U·mg-1 and the michaelis constant (K m ) was 203.09 μmol·L-1 .

Effects of Pleurotus eryngii Residue on Weight Gain of Chick and Duck

In the present study, Pleurotus eryngii residue was added into conventional feed of Chai chicks and meat-type ducks at different ratios to measure its effects on the production performance of chicks and ducks. The results revealed that the growth performance of chicks and ducks was the best when 20% Pleurotus eryngii residue was added into the feed. Under such condition, the daily weight gain of 8 - 11 and 12 - 15-week-old Chai chicks was increased by 6.8% and 13.5%, and that of 8 - 11 and 12 - 15-week-old ducks was increased by 11.5% and 19.3%, respectively.

Preparation of Rice Seedling-raising Substrate with Spent Mushroom Substrate of Pleurotus eryngii

This study aimed to explore the effect of fermented spent mushroom substrate(SMS) of Pleurotus eryngii as a basic material on rice seedling-raising substrate. The physical and chemical indices of the SMS indicated that the fermented SMS was fully composted and was very suitable for preparing rice seedling-raising substrate. The fermented SMS effectively regulated the bulk density, total porosity, aeration porosity and water-holding porosity of rice seedling-raising substrate. With the increased addition amount of fermented SMS, the bulk density of rice seedling-raising substrate was decreased, but the total porosity, aeration porosity and water-holding porosity were increased. Compared with those in the substrates of 100% soil and 100% SMS,the height, chlorophyll content, 100-shoot dry weight, 100-root dry weight, root activity, nitrogen content, phosphorus content and potassium content of rice seedlings in the substrate composed of spent mushroom substrate of P. eryngii and soil were higher.The quality of rice seedlings in the substrate composed of 20% soil and 80% SMS was best, followed by that in the substrate composed of 30% soil and 70% SMS.

Key Technology for Liquid Spawn Production and Green and High-yield Cultivation of Pleurotus eryngii

[Objectives]Pleurotus eryngii is currently the second largest edible fungus variety cultivated in China,and it mainly adopts the bag cultivation mode.This study aims to effectively solve the problems of high cost of solid spawn,long spawn production cycle,low application level of green and high-yield cultivation technology and equipment,and unstable output and quality in the factory production process and to further promote industrial upgrading.[Methods]Since June,2018,a series of research and technical optimizations had been carried out by a technical research team consisting of personnels from Vegetable Office of Guannan County,Institute of Edible Fungi of Shanghai Academy of Agricultural Sciences and Jiangsu Xiangru Biotechnology Co.,Ltd.,around the production of liquid spawn,introduction of excellent varieties of P.eryngii,medium pretreatment,autoclaving,clean room inoculation,digital cultivation,harvesting,packaging,etc.[Results]The key technology of industrial production,liquid spawn production and green and high-yield cultivation of P.eryngii are summarized.[Conclusions]The economic benefit of liquid spawn of P.eryngii is extremely considerable compared with liquid spawns such as branch spawn.

Effects of Spent Mushroom Substrate of Pleurotus eryngii as Fermentation Bed Padding on Growth Performance, Intestinal Microflora and Immunity of Weaning Piglets

[Objective] The paper was to examine the effects of spent mushroom substrate of Pleurotus eryngii as fermentation bed padding on growth performance, intestinal microflora and immunity of weaning pigs. [Method] A total of 120 weaning piglets （DurocxLandracexYorkshire） with the average initial body weight of （8.0±0.5） kg were allocated to five dietary treatments in a randomized complete block design for 42 d, each of which was replicated three times with eight piglets per replicate （ half male and half female）. The padding for control group was 50% sawdust ＋50% rice husk; experimental group I, 100% spent mushroom substrate; experimental group II, 15% sawdust ＋15% rice husk ＋70% spent mushroom sub- strate; experimental group III, 25% sawdust ＋25% rice husk ＋50% spent mushroom substrate; and experimental group IV, 35% sawdust ＋35% rice husk ＋30% spent substrate. [Result] Except for experimental group IV, the other three experimental groups had higher average daily gain compared to the control group （P〈0.05）. The average daily feed intake in experimental group I increased obviously compared to the control group（P〈0.05）. Except for experimental group I, the diarrhea rate of weaning piglets in experimental groups II, III and IV significantly decreased compared to the control group（P〈0.05）. The number of lactobacilli and bifidobacteria in colon and cecum in experimental groups I, II and III increased distinctly （P〈 0.05）, while the number of Escherichia coli and Salmonella decreased remarkably compared to the control group （P〈0.05）. The positive rates of T and B lymphocytes in peripheral blood of weaning piglets in four experimental groups were significantly higher than that in control group at 21 and 42 d post weaning （P〈0.05）. The IgA content of intestinal mucous in piglets was significantly improved in experimental groups II and III （P〈0.05）. [Conclusion] It enhances the production performance when improving immunity and reducing diarrhea rate of piglets by using spent mushroom substrate of P. eryngii as the fermentation bed padding. Experimental group III （25% sawdust ＋25% rice husk ＋50% spent mushroom substrate） is the optimal proportion of spent mushroom substrate.

Effects of Metal Ions on the Activity of Cellulase Produced by Pleurotus ostreatus and Pleurotus eryngii

[ Objectivel The study aims to discuss the effects of metal ions on the activity of Pleurotus ostreatus and Pleurotus eryngii cellulose. [ Method] 0.5 mmol/L FeSO4, CuSO4, MnSO4 and ZnSO4 solutions were added to the basal culture media containing silkworm excrement and rice husk, and the effects of metal ions on activity of FPA and Cx enzyme produced by P. ostreatus and P. eryngiiwere analyzed. [ Resultl The addi- tion of Zn2. and Mn~＊ to the basal culture media containing silkworm excrement and rice husk was beneficial to the improvement of the activity of FPA and Cx enzyme produced by P. ostreatus and P. eryngii, while the addition of Fe2＋ and Cu2. inhibited the activity of the cellulase. EConclu- sion I The research could provide theoretical references for improvement of utilization rate of cellulase in silkworm excrement and rice husk.

Study on the Variation of Three Parameters during Industrial Cultivation of Liquid Spawn to Pleurotus eryngii

In order to effectively guide industrial cultivation of liquid spawn to Pleurotus eryngii, the changes of the mycelial amount, CO2 concentration and pH value during the industrial cultivation of liquid spawn to Pleurotus eryngii were studied. The results showed that during the culture of mycelium, the changes of my- celium amount and CO2 concentration accorded with the S type growth curve, and the 4^th -8^th d was the logarithmic phase with the growth amount of mycelium 0.05 g/100 ml per day. After 9 d, the mycelium growth tended to slow down, and by tile end of the mycelium cultivation, mycelium began to slightly decrease. The pH value of medium changed slightly in 0.5 during the mycelium cultivation. The three groups of parameters showed that the liquid strain fermentation time should be 8 to9d.

Effect of Fermentation Bed with Spent Mushroom Substrate of Pleurotus eryngii on Weaning Piglets

[Objective] The paper was to investigate effects on fermentation bed temperature,growth performance,diarrhea rate and digestive en-zyme activity of weaning piglets by using spent mushroom substrate of Pleurotus eryngii as padding.[Method] A total of 120 weaning piglets（Duroc × Landrace ×Yorkshire） with average initial body weight of（8.0 ±0.5）kg were allocated to five dietary treatments in a randomized complete block design for 42 d,each of which was replicated three times with eight piglets per replicate（half male,half female）.The padding for control group was（50% sawdust ＋50% rice husk）;experimental group Ⅰ 100% spent mushroom substrate;experimental group Ⅱ（15% sawdust ＋15% rice husk ＋70% spent mushroom substrate）;experimental group Ⅲ（25% sawdust ＋25% rice husk ＋50% spent mushroom substrate）;experimental group Ⅳ（35% sawdust ＋35% rice husk ＋30% spent substrate）.[Result] There was no significant difference in surface temperature of fermentation bed between experimental groups and control group（P〉0.05）.Compared with the control group,the temperature under 20 cm of fermentation bed in ex-perimental groups I,Ⅱ and Ⅲ increased significantly（P〈0.05）.Except for experimental group Ⅳ,other three experimental groups had higher aver-age daily gain（P〈0.05） and experimental group Ⅰ had higher average daily feed intake（P〈0.05） compared to the control group.The diarrhea rate and mortality of weaning piglets in experimental groups Ⅱ,Ⅲ and Ⅳ were significantly decreased compared with the control group（P〈0.05）.Compared with the control group,other three experimental groups had higher digestive enzyme activity in duodenal contents except for experimental group Ⅳ（P〈0.05）.[Conclusion] Spent mushroom substrate of P.eryngii can be used as fermentation bed padding,and the optimal proportion was experimental group Ⅲ.

Effects of Penicillium spp. and Trichoderma spp.on Pleurotus ostreatus Growth and Screening of Effective Disinfectants

[Objective] This study was conducted to investigate the effects of Penicillium spp. and Trichoderma spp. on the growth of Pleurotus ostreatus （oyster mushroom） and to screen effective disinfectants to control them and other pathogenic microbes. [Method] Six strains or species of Penicilliurn and Trichoderma were inocu- lated into PDA plates growing with P. ostreatus to observe the growth of their mycelia. And the inhibitory effects of hydrogen peroxide, bromogeramine bromide, 84 disinfectant, lysol, potassium permanganate, and 75% medical alcohol on Trichoderma pleuroticola, Trichoderma viride, Trichoderma harzianum, Penicillium citrinum, Penicillium thomii and Penicillium brevicompactum were detected. [Result] Confrontation test revealed that Penicillium spp. and Trichoderma spp. competed for nutrients with P. ostreatus mycelia, antagonized against the growth of P. ostreatus mycelia, invaded and twined around P. ostreatus mycelia, and also produced toxins poisoning P. ostreatus mycelia. The six disinfectants exhibited different inhibitory effects against different species of Penicillium and Trichoderma. Among them the in- hibitory effects of bromogeramine bromide against the six species of Penicillium and Trichoderma were significantly different. However, the inhibitory effects of 3% hydrogen peroxide, 84 disinfectant, 5% lysol, potassium permanganate and 75% medical alcohol were significantly different among Penicillium spp., but did not among Tri- choderma spp. [Conclusion] Bromogeramine bromide can be used to control the bacteria and fungi in mushroom production for it has significant inhibitory effects on the six species of both Penicillium and Trichoderma.

榆黄蘑(Pleurotus citrinopileutus)原生质体分离和再生的研究

试验从影响原生质体制备的诸多方面进行了研究，探索榆黄蘑原生质体制备的优化系统。试验时取鲜嫩的菌丝，用ｐＨ值为６０的ＫＣｌ、蔗糖、甘露醇做酶解稳定液，３０℃下水浴酶解，酶解时间为４～５ｈ，此时榆黄蘑原生质体释放量最大，达到１０７个／ｍＬ。

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。

红平菇(Pleurotus djamor)的镉耐性及其对镉去除能力的研究

食用菌富集重金属是生物修复的一个重要研究方向。在培养料中投放不同浓度的重金属镉(0、1.0、2.0、10.0、20.0、100.0mg.kg-1Cd)并做栽培试验,研究了Cd对红平菇菌丝体生长、子实体产量的影响及红平菇对培养料中Cd的去除能力。结果显示,培养料中初始Cd浓度越大,菌丝体的生长速度越慢,尽管两潮菇子实体平均产量有显著下降,但仍具有较好的Cd耐性。红平菇具有很强的Cd富集能力,并且Cd浓度越高,富集量越高,当Cd浓度从1 mg.kg-1增加到100 mg.kg-1时,一、二潮菇的子实体中Cd富集量分别从3.58 mg.kg-1和9.51 mg.kg-1增加到了18 mg.kg-1和85.61 mg.kg-1。红平菇对培养料中Cd的去除率随Cd浓度的增加而降低,当Cd浓度为1 mg.kg-1和2 mg.kg-1时总去除率高达69.78%和36.61%,100 mg.kg-1时去除率为6.04%。红平菇的Cd耐性较好,并对培养料中低浓度的Cd有较好的去除能力。

The Interplanting Technology of Pleurotus ostreatus and Dendrobium officinale

In summer, Cha No. 39 of 292 bags and 205 of 244 bags were inter- planted with Dendrobium officinale on cultivation bed, respectively, whose yields and biological efficiencies reached 102.46 kg and 103.98%, and 96.77 kg and 113.32%, accordingly. The seedlings of Dendrobium officinale interplanted with Pleurotus os- treatus grew by 2.78 cm during 10 weeks on cultivation beds, and the seedlings grew by 2.68 cm in the cultivation beds without interplanting arrangement. These demonstrated that the interplanting of Dendrobium officinale and Pleurotus ostreatus has none adverse effects on growth of Dendrobium officinale.

Pleurotus ostreatus对线虫作用机理的研究

本文详细地研究了粗皮侧耳 ( Pleurotus ostreatus)对线虫 ( Panagrellus redivivus)的致病过程、线虫感病症状及菌丝与线虫的相互作用。探明了粗皮侧耳对线虫致病的实质是杀主寄生 ,菌丝分泌的毒素首先将线虫击倒并杀死 ,然后菌丝从线虫体孔侵入 ,或靠酶和机械力的作用从表皮侵入 ,最后消解、吸收和利用线虫。一般菌丝与线虫接触 1～ 5min线虫被击倒 ,2 0～ 30 min卵和幼虫解体 ,4～6h成虫死亡 ,12 h后菌丝侵入虫体营腐生生活。菌丝也可不侵染线虫 ,仅靠毒素将线虫杀死 ,在酶的协同作用下将线虫分解 ,直接吸收利用线虫。若菌丝分泌的毒素不足以杀死线虫 ,纵然菌丝与线虫充分接触也不能侵染活线虫。试验还表明 ,该菌平板培养产生的毒素 ,除去菌丝也能杀死线虫。

平菇(Pleurotus ostreatus)对稻草中木质素的生物降解及降解产物分析

变色圈试验证明平菇可以选择性优先降解稻草中的木质素,培养15d后,平菇对稻草中的木质素降解率为17.86%,对综纤维素降解率为2.44%,选择性指数为9.79。生料栽培平菇后,稻草中的木质素被降解50.24%。用气—质色谱(GC/MS)和红外光谱(IR)对木质素降解产物分析结果表明,平菇对稻草中木质素的降解效果十分明显,降解产物中检测出了大量含有苯环的小分子,证明木质素聚合体的降解首先发生在单体的侧链及单体间的连键上,发生Cα-Cβ、β-O-4等断裂,形成了单体。在进一步的降解过程中,平菇表现了其自身特有的降解机制,取代苯环单体上的甲氧基为甲基,而后发生苯环的开裂,这与报道的白腐菌降解过程有所不同。红外光谱分析中,平菇对木质素的降解明显,降解产物中含有很多木质素单体所特有的基团,如紫丁香基、愈创木基等,说明木质素的降解首先发生的是侧链的氧化反应。

糙皮侧耳(Pleurotus ostreatus)的AFLP指纹图谱分析

在对糙皮侧耳 (Pleurotusostreatus)的AFLP分析条件进行优化的基础上 ,利用该技术建立了 14株产自不同地区的糙皮侧耳DNA指纹图谱 ,并进行了数据分析。结果表明 ,在合成的 14条引物的不同组合中 ,引物对E 3 M 3可以产生较多的DNA多态片段 ,E AGC M CAT引物对的扩增效果最好 ,共获得 184条DNA扩增带 ,其中多态性条带 10 1条 ,占 5 4 89%。利用UPGMA法对所获数据进行聚类分析 ,计算得到糙皮侧耳菌株之间的遗传距离 ,发现不同品种间遗传距离差异较大 ,从 0 192到 0 75 4 ,说明糙皮侧耳的遗传多样性比较丰富。绘制的指纹分析树状图表明 ,14个糙皮侧耳菌株被分为 6个组群 ,其中P17和杂 3的相似性系数最高 ,达到了 81 2 % ,而侧