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.

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 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.

Characterization of insoluble dietary fiber from Pleurotus eryngii and evaluation of its effects on obesity-preventing or relieving effects via modulation of gut microbiota

The purpose of this study was to evaluate the obesity preventing or relieving effect of insoluble dietary fiber(IDF)from Pleurotus eryngii through its modulations on intestinal microbiota.The IDF was extracted and structurally characterized through FTIR,X-Ray and thermogravimetric(TG)analysis.The mice for in vivo experiments were divided into obesity preventing group(feeding high-fat diet while gavage IDF simultaneously)and obesity relieving group(IDF intervention after the establishment of the obese model)to evaluate its hypolipidemic effect.Results:The yield of P.eryngii IDF extracts was 56.49%(on P.eryngii dry weight base)with IDF content of 85.15%in the extracts.The in vivo results showed that the body weight of the obesity-preventing groups was still increasing after intervention with IDF,but the growth rate was lowered,and the abnormal lipid metabolism and inflammation syndromes were alleviated;the acetic acid content of the medium and low dose dietary fiber intervention groups was significantly higher than that of the normal group,and the structure of the intestinal flora was improved.The mice weight from the obesity-relieving group decreased significantly.The abnormal lipid metabolism and inflammation were alleviated.In conclusion,The IDF intervention decreased the abnormal lipid metabolism and inflammation in both obesity-preventing and relieving experiments.The intestinal flora structure was obviously improved.P.eryngii IDF is a good candidate as hypolipidemic agent.

Polysaccharide, Adenosine and Crude Triterpenoid Contents of Pleurotus Eryngfi and Its Immunostimulant Capacity in vitro

Purpose of the study was to demonstrate the ability of Pleurotus eryngii （PE） extracts to stimulate the production of nitric oxide （NO） and TNF-α as well as to evaluate the cell viability by the use of chicken peripheral blood mononuclear cells （PBMCs） and RAW 264.7 macrophages in vitro. The PE contained 341.0 ± 14.5 mg/g DW, 78,2 ± 4.1 Bg/g DW and 38.4 ＋ 3.3 mg/g DW of crude polysaccharide, adenosine and crude triterpenoid contents, respectively. The results of cell viability assay showed that 95,0% of chicken PBMCs and RAW 264.7 macrophages were survived under the treatment of 9.6 mg/mL of PE extracts by microculture tetrazolium assays （MTAs）. Moreover, the cell inhibitory activity did not show the 50% cyto-toxicity effect （ICs0）. In addition, no production by stimulated chicken PBMCs and RAW 264.7 macrophages exhibited individually the linear relations with the concentration of PE extracts, and they were approximately 65 μM and 45 p.M in 1600 μg/mL, respectively. The tendency with the release of TNF-α by RAW 264.7 maerophages also corresponded with concentration of PE extracts. It is concluded, PE extracts act as an immunostimulant for both tested cells but have no serious effect on inhibiting chicken PBMCs viability, and therefore it may be suggested as immune supplements for broilers.

玉米芯作为杏鲍菇栽培原料的可行性研究

以甘肃省丰富的玉米芯和玉米芯发酵料为栽培主料,以玉米面、麦麸、高粱壳等为辅料,探索不同主料以及不同基质配比对杏鲍菇(Pleurotus eryngii Quel.)产量及生物学转化率的影响,结合经济效益分析,筛选出了适合杏鲍菇生产的最佳培养料配方。结果表明,利用玉米芯为主料或以玉米芯添加高粱壳的培养基均可以获得杏鲍菇较好的出菇特性,而其中以配方Ⅴ处理(玉米芯40%、高粱壳48%、玉米面10%、石膏粉2%、基质含水量65%)为最佳栽培基质配方,菌丝生长速度最快,可达到0.48 cm/d,子实体产量高,生物转化率可达93.82%。

杏鲍菇菌糠发酵生物肥成分分析及其效果评价

测定了杏鲍菇(Pleurotus eryngii Pholiotanameko)菌糠的重金属含量及发酵处理前后的营养成分,并分析和评价了其在盆栽试验中对作物生长的影响,为利用菌糠制作农用肥从而实现废物资源再利用提供相关理论依据。结果表明,杏鲍菇菌糠的重金属含量符合行业标准NY 525—2012的相关要求,菌糠发酵后粗蛋白质和粗纤维含量下降,氨基酸和有机质含量则大幅增加。与单施复合肥相比,杏鲍菇菌糠发酵后的有机肥和复合肥施用使得香姜(Alpinia coriandriodora D.Fang)、玉米(Zea mays L.)和小白菜(Brassica chinensis L.)的叶长、株高、茎粗、叶绿素含量、过氧化物酶活性和可溶性蛋白质含量均明显增加。