玉米秸秆Lentinula edodes发酵液清除·OH主效物质分析

对Lentinula edodes在玉米秸秆木质纤维素基质中液体培养的胞外液清除羟自由基(.OH)主效物质及其清除能力进行了研究。结果表明:L.edodes在木质纤维素基质上的培养胞外液具有较高的.OH清除作用,其清除率最高达69.32%;粗多糖为最主要的胞外.OH清除物质,其清除率最高可达54.19%,占整个胞外液.OH清除效率的作用比例为78.17%;粗蛋白为另一重要的胞外.OH清除物质,其清除率最高可达27.17%,占整个胞外液.OH清除效率的作用比例为39.38%。粗多糖及粗蛋白成分共同构成L.edodes胞外清除.OH的主效物质。

Comparison of hydroxyl radical-scavenging abilities between white rot fungus Lentinula edodes and five mildews on lignocellulose substance

The hydroxyl radical（-OH）-scavenging ability of culture filtrates from submerged culture of Lentinula edodes AX3 and five mildews on lignocellulose substance was analysed. Only L. edodes AX3 showed significant -OH-scavenging ability which reached 52.2% at about the 48th hour. All mildews could hardly scavenge -OH under the experimental conditions. -OH-scavenging ability is considered related to the mode and ability of lignocellulose degradation of a strain. The degradation or bioconversion products might be the substance base for.this effect.

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.

Intergenic spacer 1(IGS1) polymorphism map: A marker for the initial classification of cultivated Lentinula edodes strains in China

China is currently the world＇s leading producer of Lentinula edodes and owns many cultivated strains of this species. This study was performed in order to investigate intergenic spacer I （IGS1） polymorphism and classification among 49 popular cultivated strains. The great majority of the 49 strains possessed two different IGS1 sequences, with distinct lengths and homologies. Based on the length and homology of the IGS1 sequences of the 49 strains, the strains were classified into two groups： A and B. Group A was subdivided into six subgroups. Forty-seven strains were homozygous or heterozygous among these six subgroups in group A, Cr01 was heterozygous between A and B, and Guangxiang 9 was homozygous in group B. An IGS1 polymorphism map of each cultivated L. edodes strain is reported for the first time and could be used as a marker for the initial classification and management of cultivated L. edodes strains in China.

Genetic Analysis of Distorted Segregation Ratio of Mating Types AmongBasidiospores in Lentinula edodes

This study prepared 17 strains of Lentinula edodes, including wild and cultivated strains as materials, and statistically analyzed the ratios of spores from different aspects via mating types＇ analysis and the OWE-SOJ technique. The results from this study first systematically identified skewed expected distribution of mating-type factors segregation in Lentinula edodes spores has commonly statistical meanings in wild and cultivated strains. Genetic analysis of positive and negative parental-recombined fruiting showed that the nuclear type of F1 progeny spores among those strains segregated through theoretical distribution mainly depended on the combined state of parental dikaryons, and the predominant spores were those with the mating type identical to the dikaryotic parent, indicating that the genetic basis of segregation distortion of spores is different from that of protoplast monokaryons in which the B factor takes predominant responsibility for that phenomenon, and it cooperates A factor with B factor to influence the ratio of spores.

Lentinula edodes extract inhibits matrix metalloproteinase expression and increases typeⅠprocollagen expression via the p38 MAPK/c-Fos signaling pathway in ultraviolet A and B-irradiated HaCaT keratinocytes

Objective:To determine the effect of Lentinula edodes extract on ultraviolet(UV)A and UVB-induced changes in matrix metalloproteinase(MMP)and type I procollagen expression using human immortalized HaCaT keratinocytes.Methods:Lentinula edodes ethanol extract(LEE)was obtained by extraction with 80%ethanol for 4 h at 80℃.Effect of LEE on UVinduced alteration on the expression and production of MMPs and type I procollagen in keratinocytes was investigated using ELISA,RT-PCR,and Western blotting assay.To determine the underlying mechanism of LEE-mediated effects,mitogen-activated protein kinase(MAPK)and activator protein 1 signaling pathways were analysed by Western blotting assay.Results:LEE significantly inhibited the expression of MMP-1 and MMP-9 and increased the expression of type I procollagen in UVA and UVB-irradiated HaCaT keratinocytes.The phosphorylation levels of p38 were significantly inhibited by LEE whereas it did not affect c-Jun N-terminal kinase and extracellular signal-regulated kinase phosphorylation.Suppression of p38 phosphorylation was also accompanied by downregulation of UVA and UVB-induced increase in c-Fos.Conclusions:LEE effectively inhibits the expression of MMP-1 and MMP-9 and increases type I procollagen production through the p38 MAPK/c-Fos signaling pathway in UVA and UVB-irradiated HaCaT keratinocytes.This findings suggest that Lentinula edodes may be developed as a cosmetic material to suppress UV exposuremediated skin aging.

Contamination Identification of Lentinula Edodes Logs Based on Improved YOLOv5s

In order to improve the accuracy and efficiency of Lentinula edodes logs contamination identification,an improved YOLOv5s contamination identification model for Lentinula edodes logs(YOLOv5s-CGGS)is proposed in this paper.Firstly,a CA(coordinate attention)mechanism is introduced in the feature extraction network of YOLOv5s to improve the identifiability of Lentinula edodes logs contamination and the accuracy of target localiza-tion.Then,the CIoU(Complete-IOU)loss function is replaced by an SIoU(SCYLLA-IoU)loss function to improve the model’s convergence speed and inference accuracy.Finally,the GSConv and GhostConv modules are used to improve and optimize the feature fusion network to improve identification efficiency.The method in this paper achieves values of 97.83%,97.20%,and 98.20%in precision,recall,and mAP@0.5,which are 2.33%,3.0%,and 1.5%better than YOLOv5s,respectively.mAP@0.5 is better than YOLOv4,Ghost-YOLOv4,and Mobilenetv3-YOLOv4(improved by 4.61%,5.16%,and 6.04%,respectively),and the FPS increased by two to three times.

Cloning of a Putative Pheromone Receptor Gene Fragment from Lentinula edodes

A 508 bp fragment of a putative pheromone receptor gene from Lentinula edodes was amplified by PCR using a pair of degenerate primers designed according to conserved sequences of pheromone receptor genes reported in other Basidiomycetes.The fragment encodes 117 amino acids and contains three introns,56 bp,51 bp and 50 bp in size.Alignment analysis revealed that the deduced amino acid sequence of the L.edodes fragment exhibited 65% identity and 82% similarity with the corresponding segments of pheromone receptors reported in Coprinus cinereus and Schizophyllum commune.

A Review on General Nutritional Compounds and Pharmacological Properties of the Lentinula edodes Mushroom

Lentinula edodes?is a macrofungus with great potential for therapeutic applications and serves as a model for investigating functional fungi properties and isolating pure compounds for pharmaceutical use. Mushrooms have a great nutritional value and present medicinal molecules including polysaccharides, terpenoids, sterols and lipids, that participate actively in several human disorders and modulate mechanisms involved in the immune system regulation. This review will focus on general nutritional compounds and pharmacological properties of L. edodes.

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.

In vitro bone inducing effects of Lentinula edodes (shiitake) water extract on human osteoblastic cell cultures

The effect of Lentinula edodes water extract(LE)on two osteoblastic cell cultures(HOS 58 and Saos-2)was investigated to determine if this edible medicinal mushroom has osteoinductive properties.Activity of alkaline phosphatase and mineralization were used as indicators for the vitality and maturation of the bone cells.Cultivation of human osteosarcoma cells HOS 58 for five days in presence of a serial dilution of the aqueous extract of L.edodes(0.8μg/mL-125μg/mL)resulted in a significant elevation of alkaline phosphatase activity(ALP)of the cells in comparison to untreated cells.Saos-2 cells,incubated with LE(20μg/mL)andb-glycerol phosphate(2 mM)for 21 days,displayed a 2 fold level of mineralization than cells cultured soley with the positive control,b-glycerophosphate.The obtained results clearly indicate the activity of LE as a bone inducing agent in vitro.Therefore,the shiitake mushroom(L.edodes)deserves attention as a supportive dietary treatment or nutraceutical in the case of diseases accompanied with bone disorder,such as osteoporosis,osteopenia,and late complication of diabetes.

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.

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。

固定香菇(Lentinus edodes)废弃菌柄对溶液中Cd^(2+)的吸附

为开发廉价实用的吸附材料,利用聚乙烯醇-海藻酸钠(PVASA)固定香菇废弃菌柄作为生物吸附剂吸附溶液中的Cd^(2+),吸附过滤后的溶液用原子吸收光谱仪(AAS)测定Cd^(2+)的浓度。结果表明:对溶液中Cd^(2+)的吸附7 h达到平衡,平衡吸附量为0.200 8 mg/g;模拟二级方程能很好地拟合吸附过程,决定系数R^2为0.994 6;Cd^(2+)适宜的吸附pH值范围为4～7,比自由态香菇粉广;随着Cd^(2+)初始质量浓度的增加,溶液中Cd^(2+)的吸附量逐渐增加;与Cd^(2+)共存于同一溶液中的Cu^(2+)或Pb^(2+)明显降低了Cd^(2+)的吸附率;分别用Langmuir、Freundlich、Dubinin-Radushkevich(D-R)和Langmuir-Freundlich等温吸附模型对Cd^(2+)的吸附过程进行拟合,结果表明Langmuir吸附模型能很好地拟合溶液中Cd^(2+)的等温吸附过程,R^2达0.998 1。微观形态研究表明,PVA-SA香菇小球吸附Cd^(2+)后粗糙多孔的表面变得密实,表面有Cd沉积和Cd晶体存在。红外光谱(FTIR)研究发现,在PVA-SA香菇小球吸附Cd^(2+)的过程中香菇细胞壁上的—OH、—CO、—CO—NH及PVA发挥了作用。用10 mmol/L的HCl作为解吸剂,3次解吸后PVASA小球对Cd^(2+)的吸附率仍可以达到64.17%,表明香菇废弃菌柄的PVA-SA固定技术在重金属生物吸附的实际应用中是可行的。

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%。

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.

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℃.