Structure and immunomodulatory activity of Lentinus edodes polysaccharides modified by probiotic fermentation

Plant-based fermentations provide an untapped source for novel biotechnological applications.In this study,a probiotic named Lactobacillus fermentum 21828 was introduced to ferment Lentinus edodes.Polysaccharides were extracted from fermented and non-fermented L.edodes and purified via DEAE-52 and Sephadex G-100.The components designated F-LEP-2a and NF-LEP-2a were analyzed by FT-IR,HPGPC,HPAEC,SEM,GC-MS and NMR.The results revealed that probiotic fermentation increased the molecular weight from 1.16×10^(4) Da to 1.87×10^(4) Da and altered the proportions of glucose,galactose and mannose,in which glucose increased from 45.94%to 48.16%.Methylation analysis and NMR spectra indicated that F-LEP-2a and NF-LEP-2a had similar linkage patterns.Furthermore,their immunomodulatory activities were evaluated with immunosuppressive mice.NF-LEP and F-LEP improved immune organ indices,immunoglobulin(Ig G and Ig M)and cytokines concentrations;restored the antioxidation capacity of liver;and maintained the balance of gut microbiota.F-LEP displayed better moderating effects on the spleen index,immunoglobulin,cytokines and the diversity of gut microbiota than NF-LEP(200,400 mg/kg).Our study provides an efficient and environment-friendly way for the structural modification of polysaccharides,which helps to enhance their biological activity and promote their wide application in food,medicine and other fields.

Fermented Lentinus edodes extract containingα-glucan ameliorates concanavalin A-induced autoimmune hepatitis in mice

Autoimmune hepatitis(AIH)is a chronic inflammatory liver disease that threatens human health worldwide.The aim of this study was to detect the protective effect of a fermented Lentinus edodes extract containingα-glucan(FLA),in a concanavalin A(Con A)-induced AIH mouse model and to determine the underlying liver-protective mechanism.The results showed that compared with the model group,the level of proinflammatory cytokines in serum of FLA pretreated mice was significantly decreased,and the degree of inflammatory cell infiltration in liver,thymus and spleen was significantly reduced.Quantitative polymerase chain reaction,immunohistochemistry,and Western blotting showed that FLA pre-treatment inhibited the Con A-induced apoptosis of hepatocytes by down-regulating the expression of BAX and up-regulating the expression of BCL-2.Further research found that FLA may improve liver injury in mice by activating NRF2 signaling pathway and inhibiting TRAF6/NF-κB signaling pathway.Thus,FLA may improve liver injury in mice by shifting gut microbial composition to reduce the release of inflammatory cytokines in the serum and prevent the necrosis of hepatocytes.Up-regulation of NRF2 signaling pathway,down-regulation of TRAF6/NF-κB signaling pathway,and an increase in the relative abundance of Lactobacillus_johnsonii and Ligilactobacillus_murinus play a protective role in liver.

Bioremediation of Textile Azo Dyes Amido Black 10B, Reactive Black 5, Reactive Blue 160 by Lentinus squarrosulus AF5 and Assessment of Toxicity of the Degraded Metabolites

Bioremediation is an eco-compatible and economical approach to counter textile dye menace. The isolated Lentinus squarrosulus AF5 was assessed for decolourization of textile azo dyes, and had shown ~93%, 88% and 70% decolorization of Reactive blue 160 (RB160), Reactive black 5 (RB5) and Amido black 10B (AB10B) respectively. Further analysis using UV-vis, HPLC, and FTIR, 1H NMR had shown the degradation of the dyes. Toxicity analysis of the metabolites was performed using seed germination and plant growth on two agriculturally important plants Guar (Cyamopsis tetragonoloba) and wheat (Triticum aestivum) as well as cytotoxicity analysis using the human keratinocyte cell line (HaCaT). The dye mix appeared inhibitory for seed germination (20% - 40%), whereas metabolites were non-inhibitory for germination. Treatment of HaCaT cells with of dye mix and metabolites led into 45% and ~100% of cell viability of HaCaT cells respectively. Therefore, metabolites following degradation of the dye mix were observed to be non-toxic.

PEG-mediated Transformation of Lentinus edodes

Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, PEG-mediated transformation of protoplast of L. edodes was studied. Mixed with PEG-purified plasmid DNA, the protoplasts of L. edodes were treated with PEG solution and cultured on CYM regeneration plate containing 40 mug/mL bialaphos. Bialaphos-resistant and GUS-positive transformants were obtained using this transformation system. Although the transformation efficiency was relatively low, the protocols release large expenses on expensive instrument and restriction enzymes, providing a simple and economical method for mushroom breeding at the molecular level.

巴西革耳Lentinus striguellus深层发酵的研究

报道了巴西革耳Lentinusstriguellus深层培养的适宜发酵培养基、摇瓶发酵条件。试验结果表明 ,Lentinusstriguellus对单糖、双糖、多糖和糖醇都有一定的利用 ,但葡萄糖等单糖对菌丝生长效果较好 ;酵母粉等蛋白质含量高的氮源对菌丝生长较为有利 ;通过L9(3 4 )正交试验确定了Lentinusstrigullus发酵的最适培养基为 :5 %葡萄糖 ,0 .7%酵母粉 ,0 .2 %KH2 PO4,0 .3 %MgSO4·7H2 O ,0 .0 0 1%VB1 ;一定浓度内 ,Fe2 + 、Ca2 + 对菌丝体生长有促进效果。适宜的摇瓶发酵条件为 :培养基起始 pH5 .0～ 6.0、培养温度 2 6℃、接种量 10 % ,摇床转速为 2 0 0r/min ,2 5 0mL三角瓶最适装液量为 80mL ,发酵周期为 6d。

香菇(Lentinus edodes)对重金属镉(Cd)的吸收规律及控制技术研究

通过在香菇培养料中投放一定浓度的重金属镉(Cd),研究香菇子实体对Cd的吸收富集规律。结果表明,香菇子实体对培养料中Cd的吸收富集规律为y=541.77x2.938(2R2=0.9135)(x、y分别为培养料、香菇中Cd的含量),富集系数达到10.39～18.00,当香菇子实体中Cd的含量为y=1.5mg·kg-1时,培养料中Cd的临界含量值x=0.1347mg·kg-1。分别选择3个香菇主栽品种、两种不同栽培方式进行比较试验,研究香菇对Cd的积累特性。结果表明,Cd低富集的品种为"庆元9015",比对镉积累最多的"庆科20"Cd含量降低33.19%,Cd低富集的栽培方式为"离地层架栽培方式"。据此形成了香菇中重金属Cd的综合控防技术,为香菇质量安全控制提供依据。

香菇属(Lentinus)真菌的研究进展兼论中国香菇属的种类资源

对香菇属 (LentinusFr .)的分类地位、分类学进展及中国香菇属的种类、生态地理分布进行了综述。我国香菇属真菌共有 31种 (包括变种 ) ,全国各地均有分布。

Mathematical Modeling and Effect of Various Hot-Air Drying on Mushroom(Lentinus edodes)

An experimental study was performed to determine the characteristics and drying process of mushroom （Lentinus edodes） by 6 different hot-air drying methods namely isothermal drying, uniform raise drying, non-uniform raise drying, uniform intermittent drying, non-uniform intermittent drying and combined drying. The chemical composition （dry matter, ash, crude protein, crude fat, total sugars, dietary fiber, and energy）, color parameters （L, a＊, b＊, c＊, and h~） and rehydration capacities were determined. Among all the experiments, non-uniform intermittent drying reached a better comprehensive results due to the higher chemical composition, better color quality associated with high bright （26.381＋5.842）, high color tone （73.670＋2.975）, low chroma （13.349a：3.456） as well as the highest rehydration （453.76% weigh of dried body）. Nine kinds of classical mathematical model were used to obtained moisture data and the Midili-kucuk model can be described by the drying process with the coefficient （R2 ranged from 0.99790 to 0.99967）, chi-square （X2 ranged from 0.00003 to 0.00019） and root mean square error （RMSE ranged from 0.000486 to 0.0012367）.

STRUCTURE,MOLECULAR WEIGHT AND BIOACTIVITIES OF(1→3)-β-D-GLUCANS AND ITS SULFATED DERIVATIVES FROM FOUR KINDS OF LENTINUS EDODES

Lentinan samples,(1→3)-β-D-glucans containing 4.6-15.2 wt% proteins,coded as L-I_1 L-I_2 L-I_3 and L-I_4(L-I)were isolated from four kinds of Lentinus edodes.These glucans were treated with acetone to remove the protein in orderto obtain free protein glucans coded as LNP-I_1,LNP-I_2.LNP-I_3 and LNP-I_4(LNP-I).The free-protein polysaccharideswere sulfated to give derivatives(S-LNP-I)with degree of substitution(DS)from 0.4-0.8.The structural features andweight-average molecular weight(M_w)of the samples were investigated by using infrared spectroscopy,elemental analysis,^(13)C-NMR,size exclusion chromatography combined with laser light scattering(SEC-LLS)and viscometry.The effects ofstructure and conformation of the polysaccharides on antitumor activities were assayed in vivo(Sarcoma 180 solid tumors)and in vitro(Sarcoma 180,HL-60,MCF-7 and Vero tumors).The results indicated that the predominant species of thesamples L-I and LNP-I in 0.2 mol/L NaCl aqueous solution existed as triple-helical chains with high rigidity and in dimethylsulfoxide(DMSO)as single-flexible chains.Interestingly,the antitumor activities of LNP-I are lower than those of the nativeglucans(L-I),whereas their sulfated derivatives have higher inhibition ratio against Sarcoma 180 than LNP-I.The resultsreveal that the binding of protein,sulfated modification and the triple helix conformation are important factors in theenhancement of the antitumor activities of polysaccharides on the whole.

Optimization of medium components using response surface methodology (RSM) for mycelium biomass and exopolysaccharide production by <i>Lentinus squarrosulus</i>

The interaction between sucrose, yeast extract and initial pH was investigated to optimize critical medium components for mycelium biomass and production of exopolysaccharide (EPS) of Lentinus squarrosulus using Response Surface Methodology (RSM). A central composite design (CCD) was applied and a polynomial regression model with quadratic term was used to analyse the experimental data using analysis of variance (ANOVA). ANOVA analysis showed that the model was very significant (p Lentinus squarrosulus are as follows: sucrose concentration 114.61 g/L, yeast extract 1.62 g/L and initial pH of 5.81;sucrose concentration 115.8 g/L, yeast extract of 3.39 g/L and initial pH of 6.44 respectively.

虎皮得菇（Lentinus tigrinus）菌丝原生质体的分离、培养与再生

以虎皮香菇（Ｌｅｎｔｉｎｕｓｔｉｇｒｉｎｕｓ）为材料，以酶解法分离原生质体，进行培养，获得再生菌丝。本研究主要结论是：（１）适合于分离原生质体的菌丝菌龄以２４～４８ｈ为佳，以１００目筛过滤得到的菌丝片段进行继代培养较不处理的菌丝体为好；（２）酶解温度确定在３３℃左右情况下，采用１％Ｌｙｗａｌｌｚｙｍｅ和１％Ｃｅｌｌｕｌａｓｅ，以０．６５ｍｏｌ／ＬＭｇＳＯ４为稳渗剂，酶解４ｈ，采用４００目不锈钢网除去未被酶解的菌丝片段；每克湿菌丝的原生质体产量可达４．５×１０７个／ｍＬ；（３）原生质体再生是一个间歇的过程。

吉林省香菇属(Lentinus Fr)的订正

整理出吉林省香菇属真菌１３种，对每一种的名称做文献考证，并按ＰｅｇｌｅｒＤ．Ｎ．（１９８３）的分类系统编排名录。同时，对Ｌｅｎｔｉｎｕｓ、Ｌｅｎｔｉｎｕｌａ和Ｌｅｎｔｉｎｅｌｕｓ三属的中文译名问题进行了讨论。

Nutrient Requirements and Fermentation Conditions for Mycelia and Crude Exo-Polysaccharides Production by <i>Lentinus squarrosulus</i>

Lentinus squarrosulus Mont. is an emerging tropical white rot basidiomycete, with nutritional and medicinal benefits. Low levels of commercial cultivation of the mushrooms limit their availability for use as food and medicine. Mycelia from submerged fermentation are a suitable alternative to the mushroom from L. squarrosulus. Three strains, 340, 339 and 218, were studied to determine optimum growth conditions for mycelia mass and crude exo-polysaccharides (CEPS) production. The experiments were conducted in a completely randomized design (CRD) with a factorial structure. Nutrients involving 8 carbon and 8 nitrogen sources were screened, and concentrations of the best sources were optimized. Optimized nutrients, interaction between strains and other parameters such as agitation and medium volume were investigated to obtain optimum fermentation conditions for biomass and CEPS production. Biomass yield varied among strains depending on carbon or nitrogen nutrient sources. Starch and yeast extract at 30 and 25 g/L were identified as the most important nutrients in mycelia and CEPS production. Nutrient optimization resulted in a 3-fold increase in mycelia mass: 12.8, 10.0 and 15.3 g/L in strains 340, 339 and 218 respectively. There was a significant interaction between strain, agitation, and volume (p p = 0.02). Static conditions favored more polysaccharide production. Optimized fermentation conditions resulted in very high increase in biomass: 238.1, 266.9 and 185.0 g/L in strains 340, 339 and 218 respectively. Results obtained could be useful in modeling fermentation systems for large-scale production of mycelia mass, CEPS and other bio-products from L. squarrosulus.

Physicochemical Characterization of Two Species of Wild Edible Mushrooms: <i>Lentinus brunneofloccosus pegler</i>and <i>Auricularia auricularia judae</i>

In tropical Africa, mushrooms play an important role for people in rural areas as a source of food, medicine and income. In order to promote the mycology sector in Côte d’Ivoire, a study was carried out on the determination of the biochemical parameters of the mushroom Lentinus brunneofloccosus pegler and Auricularia auricularia judae found in the localities of Daloa and Yamoussoukro. The samples were purchased in the markets of Daloa and Yamoussoukro. The dry matter, ash and lipid contents were determined by the AOAC method. Those of proteins, total sugars, reducing sugars were carried out respectively by the methods of Kjeldahl, Bernfeld and Dubois, and six minerals (potassium, phosphorus, calcium, magnesium, iron and sodium) were quantified by atomic absorption spectrophotometry (AAS). The results showed that the ash (3.46 - 7.74%) and protein (6.10 - 12.40%) levels were high, but the lipid contents (4% to 6.60%), total sugars (1.34 - 5.16%) and reducing sugars (0.01 to 0.27%) are low. The major minerals are calcium (898 to 1006 mg/100g), potassium (1182 to 2433 mg/100g) and phosphorus (2091 - 4375 mg/100g). Magnesium (270 - 433 mg/100g) is represented in average proportion. Iron (2.83 to 46.17 mg/100g) and sodium (4.87 to 5.99 mg/100g) are in low amounts. The mushrooms Lentinus brunneofloccosus pegler and Auricularia auricularia judae are low in fat, high in ash, and are good sources of protein and minerals.

Antioxidant Properties and Glucan Compositions of Various Crude Extract from <i>Lentinus squarrosulus</i>Mycelial Culture

Various crude extracts of Lentinus squarrosulus mushroom mycelium and culture broth were prepared for antioxidant studies, glucan and carbohydrate compound analysis. Crude extract from ethanol, cold and hot water extraction was tested on several complementary test systems namely DPPH free radical scavenging, β-carotene linoleic acid oxidation, total phenolic compound, total flavonoid compound, total carbohydrate content and total glucan compositions. The antioxidant activity of the crude extracts of Lentinus squarrosulus mushroom mycelium and culture broth was determined by 2,2’ diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method and β-carotene linoleic acid assay. Crude extracted from culture broth EPS EE recorded the highest radical scavenging effects at 20 mg/mL (85%) which is closer to the control antioxidants (ascorbic acid 97%;BHT 92.4%;BHA 93.6%). Both the ethanol extract (IPS EE and EPS EE) exhibited the highest β-carotene linoleic acid oxidation by recording IC50 values of 0.6 and 0.65 mg/mL respectively. IPS EE (0.59 mg/g) and EPS WE (0.58 mg/g) showed high flavonoid content whereas total phenol content was high in EPS WE (0.85 mg/g) and IPS EE (0.84 mg/g). Total carbohydrate concentrations were high in IPS HWE and IPS CWE with 19.1 and 16.3 mg/mL respectively and these data correlate with the β-glucan content in both extracts. The highest β-glucan content found in IPS CWE and IPS HWE were 12.2 and 11.4% w/w respectively.

固定香菇(Lentinus edodes)菌柄废弃物吸附Cd和Pb后的解吸试验研究

利用6种不同的解吸剂对吸附了Pb^(2+)和Cd^(2+)的固定香菇小球进行解吸。结果表明,HCl溶液做解吸剂最佳,可以分别在90 min和60 min内将大量的Pb^(2+)和Cd^(2+)从聚乙烯醇-海藻酸钠(PVA-SA)固定香菇小球上解吸出来。利用模拟二级动力学方程描述HCl溶液对PVA-SA固定香菇小球上Pb^(2+)和Cd^(2+)的解吸过程,决定系数R^2分别为0.996 9和0.998 9。Cd^(2+)的解吸率在HCl浓度为0.1 mol·L^(-1)时达到平衡,解吸率为80%;Pb^(2+)的解吸率在HCl浓度为1.0 mol·L^(-1)时达到最大,为99.01%。微观形态研究表明,未吸附重金属的PVA-SA固定香菇小球表面粗糙多孔,小球通过物理性吸附或形成无机沉淀将重金属沉积在细胞壁上后,表面变得密实。经HCl解吸后的PVA-SA固定香菇小球表面粗糙度和孔隙度都恢复到吸附前的状态。PVA-SA固定香菇小球对Pb^(2+)和Cd^(2+)进行吸附后,利用HCl进行解吸,再吸附,如此3次循环使用后,其吸附率略有降低,但仍可达到85.24%和69.07%。

CRITICAL CONCENTRATIONS OF α(1→3)-D-GLUCAN FROM LENTINUS EDODES IN NaOH AQUEOUS SOLUTION

Critical concentrations of α-(1→3)-D-glucan L-FV-Ⅱ from Lentinus edodes were studied by viscometry andfluorescence probe techniques. The dependence of the reduced viscosity on concentration of the glucan in 0.5 mol/L NaOHaqueous solutions with or without urea showed two turning points corresponding to the dynamic contact concentration c_s andthe overlap concentration c~* of the polymer. The values of c_s and c~* were found to be 1×10^(-3) g cm^(-3) and 1.1×10^(-2) g cm^(-3),respectively, for L-FV-Ⅱ in 0.5 mol/L NaOH aqueous solutions. The two critical concentrations of L-FV-Ⅱ in 0.5 mol/LNaOH aqueous solutions were also found to be 1.2×10^(-3) g cm^(-3) fbr c_s and 9.2×10^(-3) g cm^(-3) for c~* from the concentrationdependence of phenanthrene fluorescence intensities. The overlap concentration c~* of L-FV-Ⅱ in 0.5 mol/L NaOH aqueoussolutions was lower than that of polystyrene with same molecular weight in benzene, owing to the fact that polysaccharidetends to undergo aggregation caused by intermolecular hydrogen bonding. A normal viscosity behavior of L-FV-Ⅱ in 0.5 mol/L urea/0.5 mol/L NaOH aqueous solutions can still be observed in an extremely low concentration range at 25℃.

Lentinus squarrosulus an edible macro-fungus reported from Pakistan

Lentinus squarrosulus(Mont.)Singer:an edible macro-fungus is reported from the moist temperate Himalayan forests of Pakistan.The sample was studied based on morphological characteristics and nucleotide sequence analyses of the ITS region.Studies based on morpho-anatomical and molecular data are consistent with formerly reported specimens.Lentinus squarrosulus represents the first report for Pakistan.

Study of the Production of Lentinus crinitus(L.)Fr.Lignolytic Enzymes Grown on Agro-Industrial Waste

Exclusively Brazilian, the Caatinga biome has been thus far a place of very few studies on the Basidiomycetes fungi. Due to its semiarid climate, fungi found in the region are likely to carry lignolytic enzymes which hold biotechnological potential to be used in industrial processes of agro-industrial residue bioconversion. This study performed a response surface statistical planning to optimize the secretion of enzymes such as laccase (Lac), lignin peroxidase (LiP) and manganese peroxidase (MnP) by Lentinus crinitus. Three variables were under analysis: different concentrations of barley and cassava residue, pH and temperature. MnP enzyme showed the highest enzymatic activity rate (23.5 IU/L). Additionally, MnP had the best results of enzyme secretion for substrate composition of 50% barley and 50% cassava, at pH 7 and temperature at 28°C for a 28-day incubation period. However, further studies are pivotal to test the efficiency in lignin bioconversion by the enzymes synthesized in this work and also to establish their usage pattern on a large scale.

Flavor changes in Lentinus Edodes enzymatic hydrolysate maillard reaction products with different sugars by gas chromatography‐ion mobility spectrometry

The characteristic flavor of the enzymatic hydrolysate of Lentinus edodes,which was involved in the Maillard reaction by xylose,fructose,glucose,sucrose,mannose,maltose,vitamin C(VC)and L-arabinose,was determined and analyzed by gas chromatography-ion mobility spectroscopy(GC-IMS).The characteristic flavor fingerprints of Maillard reaction products(MRPs)of L.edodes enzymatic hydrolysate were constructed.The differences in the characteristic flavors of Maillard reaction with different reducing sugars and VC were compared.The main characteristic flavor,characteristic peak,characteristic marker substance and content were determined,a principal component analysis(PCA)of volatile organic compounds(VOCs)was carried out.The results showed that there were 42 kinds of monomers and some dimers of volatile compounds in the samples including 17 aldehydes,5 alcohols,7 ketones,2 esters,2 acids,7 other compounds and 2 ethers.The volatile VOCs of the sample with added sucrose,VC and L-arabinose were significantly higher than those of the enzymatic hydrolysate of L.edodes.After the Maillard reaction,the flavor of the hydrolysate was significantly improved,and the main components changed significantly.