The white-rot Agaricomycetes species Phlebia radiata is an efficient wood-decaying fungus degrading all wood components, including cellulose, hemicellulose, and lignin. We cultivated P. radiata in solid state cultures on spruce wood, and extended the experiment to 6 weeks to gain more knowledge on the time-scale dynamics of protein expression upon growth and wood decay. Total proteome and transcriptome of P. radiata were analyzed by peptide LC–MS/MS and RNA sequencing at specific time points to study the enzymatic machinery on the fungus’ natural growth substrate. According to proteomics analyses, several CAZy oxidoreductase class-II peroxidases with glyoxal and alcohol oxidases were the most abundant proteins produced on wood together with enzymes important for cellulose utilization, such as GH7 and GH6 cellobiohydrolases. Transcriptome additionally displayed expression of multiple AA9 lytic polysaccharide monooxygenases indicative of oxidative cleavage of wood carbohydrate polymers. Large differences were observed for individual protein quantities at specific time points, with a tendency of enhanced production of specific peroxidases on the first 2 weeks of growth on wood. Among the 10 class-II peroxidases, new MnP1-long, characterized MnP2-long and LiP3 were produced in high protein abundances, while LiP2 and LiP1 were upregulated at highest level as transcripts on wood together with the oxidases and one acetyl xylan esterase, implying their necessity as primary enzymes to function against coniferous wood lignin to gain carbohydrate accessibility and fungal growth. Majority of the CAZy encoding transcripts upregulated on spruce wood represented activities against plant cell wall and were identified in the proteome, comprising main activities of white-rot decay. Our data indicate significant changes in carbohydrate-active enzyme expression during the six-week surveillance of P. radiata growing on wood. Response to wood substrate is seen already during the first weeks. The immediate oxidative enzyme action on lignin and wood cell walls is supported by detected lignin substructure sidechain cleavages, release of phenolic units, and visual changes in xylem cell wall ultrastructure. This study contributes to increasing knowledge on fungal genetics and lignocellulose bioconversion pathways, allowing us to head for systems biology, development of biofuel production, and industrial applications on plant biomass utilizing wood-decay fungi.

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Time-scale dynamics of proteome and transcriptome of the white-rot fungus Phlebia radiata: growth on spruce wood and decay effect on lignocellulose

From plant phenols to novel bio-based polymers

Comparative transcriptomics of Pleurotus eryngii reveals blue-light regulation of carbohydrate-active enzymes (CAZymes) expression at primordium differentiated into fruiting body stage

Comparative secretomic analysis of lignocellulose degradation by Lentinula edodes grown on microcrystalline cellulose, lignosulfonate and glucose.

Pleurotus eryngii (P. eryngii) can secrete large amount of hydrolytic and oxidative enzymes to degrade lignocellulosic biomass. In spite of several researches on the individual lignolytic enzymes, a direct deconstruction of lignocellulose by enzyme mixture is not yet possible. Identifying more high-performance enzymes or enzyme complexes will lead to efficient in vitro lignocelluloses degradation. In this report, secretomic analysis was used to search for the new or interesting enzymes for lignocellulose degradation. Besides, the utilization ability of P. eryngii to ramie stalk substrate was evaluated from the degradation of cellulose, hemicellulose, and lignin in medium and six extracellular enzymes activities during different growth stages were discussed. The results showed that a high biological efficiency of 71% was obtained; cellulose, hemicelluloses, and lignin decomposition rates of P. eryngii were 29.2, 26.0, and 51.2%, respectively. Enzyme activity showed that carboxymethyl cellulase, xylanase, laccase, and peroxidase activity peaks appeared at the primordial initiation stage. In addition, we profiled a global view of the secretome of P. eryngii cultivated in ramie stalk media to understand the mechanism behind lignocellulosic biomass hydrolysis. Eighty-seven nonredundant proteins were identified and a diverse group of enzymes, including cellulases, hemicellulases, pectinase, ligninase, protease, peptidases, and phosphatase implicated in lignocellulose degradation were found. In conclusion, the information in this report will be helpful to better understand the lignocelluloses degradation mechanisms of P. eryngii.

Secretome analysis of Pleurotus eryngii reveals enzymatic composition for ramie stalk degradation.

The invention relates to a sisal hemp-plant leaf cream production method. According to the production method, sisal hemp-plant leaves are produced into fibers to obtain sisal hemp-plant leaf juice, the juice is fermented, sediments are collected, leuconostoc mesenteroides, acclimated ethyl alcohol candida and lactobacillus delbrueckii are mixed to prepare fermentation inoculants for fermenting, acclimation is performed by the aid of a gradual acclimation method, the proportion of sisal hemp-plant leaf juice in acclimation culture solution is gradually increased, and the ethyl alcohol candida and the lactobacillus delbrueckii are respectively and gradually inoculated into high-proportion culture solution from low-proportion culture solution to obtain the acclimated ethyl alcohol candida and the acclimated lactobacillus delbrueckii. According to the method, a fermentation period is shortened to 4-5 days, yield of the sisal hemp-plant leaf cream is greatly improved, diosgenin content is improved by 20-30%, and difficulties further extracting diosgenin are reduced. The method changes a traditional production mode and solves the problem that a traditional method is large in occupied area, long in settlement period, low in diosgenin content and the like.

Sisal hemp-plant leaf cream production method

The invention relates to a pleurotus geesteranus culture medium and a preparation method and application thereof. The pleurotus geesteranus culture medium is prepared from, by weight, 32-45% of hibiscus cannabinus side products, 32-48% of mixed saw dust, 5-15% of vegetable seed peel, 5-15% of bran and 1-3% of gypsum and lime, wherein the sum of the vegetable seed peel and the bran is 19-21%. The hibiscus cannabinus side products and the vegetable seed peel are matched for culture of pleurotus geesteranus, the problem of shortages of culture raw materials at present can be relieved, the edible fungus culture raw material source can be enriched, raw material localization is achieved, an industry chain is prolonged, the production cost is reduced, the economic benefits are improved, the economic income is increased for farmers, the problem of environment pollution caused by random abandonment or incineration of agricultural wastes can be avoided, and food safety can be ensured.

Pleurotus geesteranus culture medium and preparation method and application thereof

The invention provides a ramie grass block and a method for breeding beef cattle by the ramie grass block, and relates to a processing and application method of a high-quality feed raw material. The method is characterized in that the following raw materials are uniformly mixed and pressed into a grass block with the density of 500kg/m to 600kg/m in percentage by weight: 90 percent to 95 percent of a ramie raw material with the moisture content of 25 percent to 30 percent, 2 percent to 3 percent of starch, 1 percent to 3 percent of air-dried vinasse, 0.25 percent to 0.5 percent of propionic acid, 0.1 percent to 0.2 percent of table salt and 1 percent to 5 percent of alfalfa meal. The ramie grass block which accounts for 1.5 percent to 2 percent of the weight of the beef cattle, concentrated feed supplementary feed which accounts for 0.5 percent to 0.75 percent of the weight of the beef cattle and green and coarse forage which accounts for 1.5 percent to 2 percent of the weight of the beef cattle are fed to the beef cattle every day, and the ramie raw material with relatively high moisture content can be sufficiently utilized to be prepared into the ramie grass block with a good nutrient structure, stable grass block and good palatability; 50 percent of the concentrated feed supplementary feed or more and 20 percent of the forage or more can be reduced; feed grain is remarkably saved, and the breeding benefits are increased.

Li Zhimin

Papers

Secretome analysis of Pleurotus eryngii reveals enzymatic composition for ramie stalk degradation.

Sisal hemp-plant leaf cream production method

Pleurotus geesteranus culture medium and preparation method and application thereof

Ramie grass block and method for breeding beef cattle by ramie grass block