TWI537387B - Napier grass and uses thereof - Google Patents

Description

Pennisetum and its use

The present application relates to culture media, compositions and methods for culturing cellulolytic microorganisms for the production of cellulases.

In the past few decades, much of the research on cellulase systems has focused primarily on cellulolytic biomass conversion for the production of energy-rich materials such as ethanol, hydrogen and methane. Therefore, many related technologies have been developed (Sheehan and Himmel, 1999). The key to such biotechnology is to achieve the practical goal of microbial and enzymatic cellulose hydrolysis (Lynd et al., 2002). However, biotechnological methods currently available for the conversion of cellulose to biofuels are relatively promising but relatively lacking in-depth research. In addition, the use of naturally occurring lignocellulose (from wood, grass, forestry waste, waste paper, municipal waste and agricultural residues such as corn stalks and straws) to produce cellulase trends has become significant worldwide. increase. Among all the raw materials, agricultural biomass (consisting of more than 50% cellulose and hemicellulose) is an excellent carbon source for the production of microbial enzymes due to extremely low operating costs. Reducing the cost of microbial enzyme production has become an important research target for the development and production of biofuels.

Due to the wide application potential of cellulase in industrial processes such as animal feed, starch processing, koji and wine making, grain alcohol fermentation, fruit and vegetable processing, pulp and paper, and textiles (Bhat, 2000), extensive research Focus on cellulases. Many microorganisms that produce various cellulolytic enzymes have been studied for decades (Lynd et al., 2002). Culture methods for increasing cellulase production have been widely reported, including immersion and solid fermentation (Grajek, 1987), different types and concentrations of substrates (Pavarina and Durrant, 2002), using carbon and nitrogen sources. Modulation (Lockington et al., 2002), co-cultivation to produce cellulase (Wang et al., 2006) and strain improvement (Adsul et al., 2007). Based on the consideration of the need to use cellulase for saccharification of cellulose-based materials, screening for high cellulase activity, leaving no residue in the hydrolysate (eg cellobiose) and having a thermo-pH stability enzyme Microorganisms will be beneficial.

The cost issue is critical in the development and production of cellulases with applicability, high activity and high productivity. The chemical substance in which the medium is prepared is a major expense for growing cellulolytic microorganisms for producing cellulase. There is a need for a cost effective medium.

The present invention relates to a cost effective medium and a method of growing a cellulose decomposing microorganism for producing a cellulase.

Thus, one aspect of the invention features a medium containing a wolftail (Napier; NP) grass ( Pennisetum purpureum ). In one example, the medium comprises from about 0.1% to 5% (w/v) NP grass debris. The medium may contain an N source. In one example, the medium is sterilized. In another example, the medium further contains an antibiotic.

In a second aspect, the invention features a method of culturing a cellulolytic microorganism. The method comprises the steps of: obtaining the above medium; and culturing the cells in the medium under conditions that allow growth of cells of the cellulolytic microorganism. Examples of cellulolytic microorganisms include, but are not limited to, Fusarium spp., Aspergillus spp., and Neurospora spp. Preferred cellulolytic microorganisms to be used may be fungi, such as Aspergillus niger , Candida pelliculosa , Kluyveromyces fragilis , Phanerochaete. Chrysosporium), Schizophyllum (Schizopphyllum commune) and Trichoderma reesei (Trichoderma reesei); bacteria, such as Agrobacterium (Agrobacterium) ATCC 21400, Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), cyclic Bacillus (Bacillus circulans), Bacillus subtilis ( Bacillus subtilis ), Bacteroides succinogenes , Cellumonas fimi , Cellulomonas uda , Clostridium thermocellum , non-decarboxyl (Escherichia coli) Escherichia adecarboxylata), Erwinia chrysanthemi (Erwinia chrysantemi), Agaricus small Agaricus fungus (Microbispora bispora) and high temperature sp YX (Thermonospora YX); or obtained from sources such as Reticulitermes (Reticulitermes flavipes), food wooden maggots ( Xylophaga ), Australian ridged termites ( Coptotermes lacteus ), black breasts Termites ( Nasutitermes walkeri ), termites ( Leucotermes speratus ) ( Reticulitermes ), snails ( Helix pomatia ), Cryptocercus punctulates , Panesthia cribrata , mulberry Other microbes of animals of Calolampra elegans and Geoscapheus dilatatus .

In a third aspect, the invention features a method of producing a cellulase. The method comprises the steps of: obtaining the above medium and cultivating a cellulolytic microorganism containing a nucleic acid encoding a cellulase (such as selected from the group consisting of Fusarium, Aspergillus, and Neurospora) in the medium under conditions permitting expression of cellulase. a cell of a fungus of the genus; and a cellulase purified from the cultured cells or culture medium.

In a fourth aspect, the invention features a composition having the above-described medium and cells of a cellulolytic microorganism such as a fungus selected from the group consisting of Fusarium, Aspergillus, and Neurospora. The above methods and compositions are useful for producing saccharable sugars and energy from lignocellulosic matter.

Thus, in a fifth aspect, the invention features a method of producing a succulent sugar from a lignocellulosic material. The method comprises providing a composition having cells of the above medium and cellulolytic microorganisms; and contacting the composition with a lignocellulosic material to produce a sucrose. In one example, the medium contains from about 0.1% to 5%, such as 1.0% (w/v) NP grass debris. The soluble sugar can be glucose, xylose, arabinose, galactose, mannose, rhamnose, sucrose, fructose, lactose, maltose, trehalose, cellobiose, oligosaccharide (such as fiber oligomer or xylose). Oligomers) or any combination thereof. Examples of lignocellulosic materials include cellulosic animal waste, municipal solid waste, waste paper, yard waste, agricultural residues, forestry residues, and any combination thereof. The method can further comprise the step of converting the fermentable sugar to a fermentation product. This transformation step can be carried out by microbial fermentation or enzymatic treatment.

In a sixth aspect, the invention features a method of generating energy from a lignocellulosic material. The method comprises: providing the above composition; contacting the composition with a lignocellulosic material to produce a sucrose; converting the sucrose to produce a flammable fermentation product; and burning the flammable fermentation product or hydrolyzing solid waste Object or residue to produce energy.

In a seventh aspect, the invention features a method of processing feed, biopulping or composite processing of feed or food additives. The method comprises providing the above composition comprising a medium and a cell of a cellulolytic microorganism; and contacting the composition with a substance to be processed or treated.

In an eighth aspect, the invention features a bioreactor comprising a lignocellulosic material and the above composition.

The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and appended claims.

The present invention is based at least in part on the surprising discovery that Pennisetum can be used as an effective and inexpensive primary carbon source for growing cellulolytic enzymes that produce cellulase.

Wolftail (NP) grass (grass), which is commonly found in tropical and subtropical regions, has been widely used as a feed for ruminants and its composition, nature and genetic information have been well studied; however, no reports have been reported on NP grass. The use of microbial cellulase preparation. The present invention provides a new application of NP grass as an effective and inexpensive C source for growing cellulolytic fungi (i.e., Fusarium, Aspergillus, and Neurospora) isolated from straw or bagasse compost. The results showed that when 1% (w/v) NP grass was used as the main C source of the three fungi, the total cellulase activity (FPase) was significantly increased (for Fusarium, Aspergillus and Neurospora, respectively). It increased from 0.008 to 0.085, increased from 0 to 0.085, and increased from 0.002 to 0.124 U/ml (Fig. 1). After optimizing all individual culture conditions, the optimal cellulase yields of Fusarium and Neurospora were 0.18 to 0.22 U/ml and 0.14 to 0.16 U/ml, respectively. Among the C sources studied (i.e., glucose, CMC, and NP grass), the medium containing 1% NP grass not only achieved optimal cellulolytic activity, but was the cheapest combination among all tested C sources. This makes NP grass a major C source for the improvement of biofuels that must be produced by cellulolytic microorganisms to produce cellulase, the textile industry and other industries.

For example, cellulases produce saccharable sugars and energy from lignocellulosic matter. The term "lignocellulosic material" as used herein refers to a substance containing cellulose and/or hemicellulose. In general, such materials also contain xylan, lignin, proteins and carbohydrates such as starch and sugar. For example, in the stems, leaves, shells, skins and cobs of plants or the leaves, branches and xylem of trees. The process of converting complex carbohydrates, such as starch, cellulose or hemicellulose, into saccharable sugars is also referred to herein as "saccharification." A saccharable sugar as used herein refers to a monosaccharide such as glucose, xylose, arabinose, galactose, mannose, rhamnose, sucrose, fructose, lactose, maltose, trehalose or cellobiose. Lignocellulosic materials can include virgin plant biomass and/or non-primitive plant biomass, such as agricultural biomass, commercial organic matter, construction and destructive debris, municipal solid waste, waste paper, and yard waste. Common forms of lignocellulosic material include trees, shrubs and grasses, wheat, wheat straw, bagasse, corn, corn husks, corn kernels, including grains from grains, ground such as corn, rice, wheat, and barley (including wet Grinding and dry grinding of the products and by-product fibers, as well as municipal solid waste, waste paper and yard waste. Lignocellulosic materials can also be, but are not limited to, herbal materials, agricultural residues, forestry residues, and paper mill residues. "Agricultural biomass" includes branches, shrubs, stalks, corn and corn husks, energy crops, forests, fruits, flowers, grains, grasses, herbaceous crops, leaves, bark, needles, logs, roots, Seedlings, short-rotation woody crops, shrubs, switch grass, trees, vegetables, peels, vines, beetroots, wheat bran, oat hulls, hardwoods and cork (excluding wood containing harmful substances), agriculture Organic waste generated by the process (including cultivation and forestry activities, including forestry wood waste, in particular) or mixtures thereof.

Examples of lignocellulosic materials used in this method include, but are not limited to, orchard pruning, shrubs, factory waste, municipal wood waste, municipal waste, felling waste, forest tending thinning, short rotation wood Crops, industrial waste, wheat, wheat straw, oat straw, straw, barley straw, rye straw, flax straw, soybean hulls, rice husks, oat hulls, sugar cane, corn, corn stalks, corn stalks, corn gluten feed, Ears of corn, corn husks, corn kernels, grain fibers, stalks, gamagrass, foxtail, beet mites, citrus fruit pulp, seed shells, cellulosic animal waste, lawn pruning Products, cotton, seaweed, trees, shrubs, grasses, bagasse, wet and dry grinding products and by-products, municipal solid waste, waste paper, yard waste, herbal materials, agricultural residues, forestry residues , municipal solid waste, waste paper, pulp, paper mill residues, branches, bushes, stalks, corn, corn husks, energy crops, forests, fruits, flowers, Cereals, grasses, herbaceous crops, leaves, bark, needles, logs, roots, seedlings, shrubs, switchgrass, trees, vegetables, peels, vines, beetroots, wheat bran, oat hulls, hardwoods and cork, agricultural processes Organic waste produced, forestry wood waste or a combination thereof.

In this study, fungal strains of Fusarium, Aspergillus and Neurospora were cultured in a medium containing Wolftail (NP) grass (Ceramic), CMC or glucose as the main carbon source to produce extracellular cellulose. Enzyme. The results showed that when three isolated fungi were cultured using 1% (w/v) NP grass, total cellulase activity (based on FPase measurements) increased significantly for Aspergillus, Fusarium, and Neurospora Increased from 0 (no NP) to 0.085 U/ml (1% NP), increased from 0.008 to 0.085 U/ml, and increased from 0.002 to 0.124 U/ml; that is, cultured with 1% (w/v) CMC. The total cellulase activity is about 1 to 23 times. Optimal cellulase activity was achieved under optimized culture conditions (0.18 to 0.22 U/ml and 0.14 to 0.16 U/ml for Aspergillus and Neurospora, respectively). In addition, cellulases collected from 1% NP batches have good pH and temperature operating ranges and are stable over a wide range of pH values. Among all the C sources tested, the medium containing NP grass not only achieved the best cellulolytic activity, but also the cheapest combination. This finding makes NP grass a major carbon source for the improvement of biofuels that must be produced by cellulolytic fungi to produce cellulase, the textile industry and other industries.

Thus, the culture media, compositions, and methods disclosed herein can be used in feed or food additive processing, biopulping, or composites in the manner described in U.S. Patent Application Nos. 20,100,047, 869 and 2008, 013, 958, and U.S. Patent Nos. 5,232,851, 5,671,151, 6, 061, 063, 6, 602, 00 In the processing, the contents of each case are incorporated herein by reference.

The following specific examples are to be construed as illustrative only and not limiting in any way. Without further elaboration, the skilled artisan can make the most of the present invention based on the description herein. All publications cited herein are hereby incorporated by reference in their entirety. In addition, any mechanism proposed below does not limit the scope of the claimed invention in any way.

1 Introduction

In this study, three cellulase producing strains, Fusarium, Aspergillus, and Neurospora were used. The culture conditions of the three fungal strains were optimized to achieve optimal cellulase activity. Three different natural or artificial carbon sources (i.e., Pennisetum, CMC or glucose) were supplemented to the medium to assess their potential to increase cellulase production at lower cost.

2. Method

2.1 quality

NP grass was obtained from the Taiwan Livestock Research Institute (Hsin-hua, Tainan, Taiwan). Carboxymethylcellulose (CMC), glucose, β-p-nitro-phenyl-glycoside (β-pNPG), Whatman No. 1 paper, and all chemicals used in this study were purchased from Sigma. . CMC, Waterman No. 1 paper and β-pNPG were the substrates for CMCase, β-glucosidase and FPase measurements, respectively. Among the three fungi, CMC, glucose, and NP grass were used as a carbon source at a concentration of 1% (w/v) to study the production of cellulase. The NP grass was air dried, cut into small pieces using a chopper, ground into smaller particles, and finally separated by passing through a 0.45 mm screen. The medium is prepared using a portion that passes through the screen and a commercially available chemical.

2.2 inoculation and culture conditions

The Fp and RS-N strains were maintained on PDA agar for 5 to 7 days at 28 ° C, whereas the RS-A line was maintained at 35 ° C, at which time sporulation was in good condition. Fresh hyphae from this culture were used as inoculum. With the sterilized with 5 ml H ₂ O and washed slant culture prepared conidium suspension. The spore suspension was counted to 10 ⁸ spores/ml using a hemocytometer. A 2 ml spore suspension of Fusarium, Aspergillus and Neurospora was grown in a 250 ml flask containing 100 ml of MR medium (without 0.1% soy peptone) using 1% NP grass, 1% CMC, respectively. Or 1% glucose as a carbon source to study cellulase production. Cellulase activity of three fungi grown in different carbon sources was recorded during 5 days of culture. Regarding the optimal concentration of NP grass, different percentages of NP were used and cultured under MR medium (excluding 0.1% soy peptone) under the optimal growth conditions.

2. 3 crude fiber enzyme preparations of fungi

The culture solution of Fusarium, Aspergillus, and Neurospora cultured in 1% NP grass was centrifuged at 8,000 × g for 20 minutes to remove hyphae and unused coat. The supernatant was filtered through a Waterman No. 1 paper and a 0.45 μm membrane, and then concentrated by filtration through an Amicon concentrator equipped with a 3 kDa membrane (Vivaspin 20), and the FPase activity loss of the filtration process was less than 3%. 0.01% (v/w) sodium azide (NaN ₃ ) was added to the supernatant to prevent microbial growth. The concentrated crude enzyme preparation containing 0.01% NaN ₃ and 0.01% phenylmethanesulfonate (PMSF, a serine protease inhibitor) can be stored at 4 ° C and retains 80% activity after 1 week. Enzyme characteristics were studied using crude enzyme preparations, including optimal pH, pH stability, optimal temperature and thermal stability.

2.4 Cellulase (CMCase , FPase and β-glucosidase) activity assay

For the carbon source activity test, the corresponding 1% substrate described in the method was used to measure the activity of CMCase, β-glucosidase and FPase of the crude enzyme extract of Fusarium at 40 ° C, but for Aspergillus spp. The genus Neurospora was measured at 50 °C. The reducing sugar released from 1% CMC was measured in 10 minutes in 200 μl of the reaction mixture (100 μl of crude enzyme and 100 μl of 2% substrate in 100 mM NaOAC, 100 mM NaCl (pH 5.0)). CMCase. FPase activity was determined using 3 mg No. 1 (Watman) filter paper as the substrate in a 200 μl reaction mixture (as above) for 1 hour. After the addition of 200 μl of dinitrosalicylic acid (DNS) and heating at 100 ° C for 10 minutes, the total release reducing sugar was determined using the modified DNS method (Wood et al., 1988). One unit (IU) of CMCase and FPase activity is defined as the amount of enzyme that releases 1 μmol of glucose per minute (as reducing sugar equivalent) under assay conditions. Regarding β-glucosidase activity, a β-pNPG (final concentration: 1 mM) was mixed with a 10 μl enzyme solution prepared in a sodium acetate buffer (100 mM NaOAC, 100 mM NaCl, pH 5.0). A 150 μl working volume was used to perform three fungal assays. After 10 minutes of incubation, the reaction was stopped by the addition of 50 μl of 2 M sodium carbonate. The absorbance of the reaction mixtures of the three fungi was measured at 400 nm using a spectrophotometer. The β-glucosidase activity was calculated based on the 4-nitrophenol calibration curve. One unit of enzyme activity is defined as the release of 1 μmol of 4-nitrophenol equivalent per minute.

3. Results and discussion Carbon (C) source effect

Culture factors such as media complexity and complexity of the structure of the substrate may affect the secretion of hydrolase by filamentous fungi (Kurchenko et al., 2001). In this study, it was estimated that different C sources (ie, NP grass and CMC) improved the cellulase secretion potential of Fusarium, Aspergillus, and Neurospora. The results showed that the total cellulase activity (FPase) of the three fungi grown in the medium without the C source supplement was extremely low. Once in the presence of 1% (w/v) NP grass (but cultured in the absence of organic N) (Table 1 and Figure 1), FPase activity is significantly increased by about 60 to 105 times (ie for Fusarium, Aspergillus) For the genus and Neurospora, they increased from 0 to 0.062 U/ml, from 0.008 to 0.085 U/ml and from 0.002 to 0.124 U/ml, respectively, indicating that NP grass can induce cellulase production in large quantities. . CMCase and β-glucosidase activities also showed similar increases (Table 1), and the activity was increased to a greater extent than FPase. In addition, the FPase activity of the batch cultured with 1% NP grass was about 1 to 23 times that of the batch cultured with 1% CMC. These results indicate that NP grass can be used as an ideal C source to improve the production of cellulase enzymes of the cellulolytic fungi inoculated in this study.

In addition, the optimal NP grass concentration produced by cellulase was further evaluated. In general, for Fusarium and Neurospora, FPase activity is enhanced with increasing amounts of NP grass, except for 5% NP (Figure 2b and Figure 2c). Fusarium, Aspergillus, and Neurospora showed FPase maximal after 4 to 5 days of incubation under 2%, 5%, and 3% NP grasses; this value was even better than the 1% NP value. The NP grass-based medium developed in this study is expected to save 50% of the culture cost, while the C source cost is usually more than one-half of the total medium cost (Table 2). It is concluded that NP grass can be a better source of quality for economic cellulase production.

Knot s

When NP grass was used as the main carbon source, the three fungi achieved FPase activity of 1.1 to 23 times the FPase activity when using 1% CMC as the main carbon source; and 60 to 105 times the FPase activity in the absence of NP grass. Among the C sources studied (i.e., glucose, CMC, and NP grass), the medium using the optimal NP grass concentration not only achieved optimal cellulolytic activity, but was the cheapest combination among all C sources tested. The above properties indicate that cellulases derived from RS-A, Fp and RS-N can be used as key enzymes for the preparation of bioethanol from cellulose.

Other embodiments

All of the features disclosed in this specification can be combined in any combination. Each feature disclosed in this specification can be replaced by alternative features for the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, the disclosed features are only one example of a series of generally equivalent or similar features.

A number of embodiments of the invention have been described. It will be understood, however, that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims.

references:

Bhat, MK, 2000. Cellulases and related enzymes in biotechnology. Biotechnol Adv , 18, 355-83.

Grajek, W., 1987. Comparative studies on the production of cellulases by thermophilic fungi in submerged and solid-state fermentation. Appl Microbiol and Biotechnol , 26, 126-129.

Kurchenko, IM, Zhdanova, NM, Sokolova, OV, 2001. Study of availability of some hydrolytic and redox enzymes in strains of Fusarium oxysporum (Schlecht.) Snyd. and Hans. isolated from different habitats. Mikrobiol Z , 63, 34-43 .

Lee, M., Hwang, S., Chiou, PW 2000. Metabolizable energy of roughage in Taiwan. Small Rumin Res , 36(3), 251-259.

Lockington, RA, Rodbouurn, L., Barnett, S., Carter, CJ, Kelly, JM, 2002. Regulation by carbon and nitrogen sources of a family of cellulases in Aspergillus nidulans. Fungal Genet Biol , 37, 190-6.

Lynd, LR, Weimer, PJ, van Zyl, WH, Pretorius, IS, 2002. Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev , 66, 506-77, catalog.

Pavarina, EC, Durrant, LR, 2002. Growth of lignocellulosic-fermenting fungi on different substrates under low oxygenation conditions. Appl Biochem Biotechnol , 98-100, 663-77.

Prakash, B., Dhali, A., Mondal, M., Sangtam, M., Khate, K., Rathore, SS, Rajkhowa, C. 2008. Effect of feeding Lagerstroemia speciosa and conventional fodder based rations on nutrient utilization, ruminal Metabolites and body weight gain in mithun (Bos frontalis). J Anim Physiol Anim Nutr (Berl), 92(5), 591-6.

Sheehan, J., Himmel, M., 1999. Enzymes, Energy, and the Environment: A Strategic Perspective on the US Department of Energy's Research and Development Activities for Bioethanol. Biotechnol Prog , 15, 817-827.

Wood, TM, Willis, AWaSTK, 1988. Preparation of crystalline, amorphous, and dyed cellulase substrates. Method Enzymol . Academic Press, pp. 19-25.

Figures 1a to 1c show the use of 1% (w/v) for 3 fungi (a) Fusarium, (b) Aspergillus and (c) Neurospora compared to 1% CMC. A map of NP grass enhancing total cellulase production (FPase activity). All strains were cultured in MR medium in the absence of soy peptone and with 1% NP or 1% CMC as the main carbon source. The strain cultured in the same medium without the above C source was designated as a control experiment (i.e., no NP). The general conditions were pH 7.0, 25 ° C to 35 ° C, and 150 rpm with 1% NP grass as a control experiment.

Figures 2a to 2c show that NP grass significantly enhances total cellulase production (FPase) for the three fungi (a) Fusarium, (b) Aspergillus, and (c) Neurospora. The map of the concentration. All strains were cultured in MR culture medium in the absence of soy peptone with different percentages of NP as the main carbon source under the respective optimal growth conditions. The general conditions were pH 7.0, 25 ° C to 35 ° C, and 150 rpm with 1% NP grass as a control experiment.

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Claims (17)

A method for cultivating a cellulolytic microorganism, the method comprising: obtaining a medium comprising a wolftail (Napier; NP) grass ( Pennisetum purpureum ); and a condition for allowing cell growth of the cellulolytic microorganism in the medium The cells are cultured under the action wherein the cellulolytic microorganism is one or more fungi selected from the group consisting of Fusarium spp., Aspergillus spp., and Neurospora spp. The method of claim 1, wherein the medium comprises from about 0.1% to 5% (w/v) of wolftail (NP) grass (grass) fragments. The method of claim 2, wherein the medium comprises from about 1% to 3% (w/v) of wolftail (NP) grass (grass) fragments. The method of any one of items 1 to 3, wherein the medium is sterilized. The method of any one of claims 1 to 3, wherein the medium further comprises an antibiotic. A method for producing a cellulase, the method comprising: obtaining a medium as defined in the method of any one of claims 1 to 5; and cultivating the nucleic acid encoding the cellulase under conditions permitting expression of the cellulase Cellulolytic cell of a microorganism, wherein the cellulolytic microorganism is one or more fungi selected from the group consisting of Fusarium, Aspergillus, and Neurospora; and the cellulase is purified from the cultured cell or the culture medium . A composition comprising the method of any one of claims 1 to 5 A defined medium and a cell of a cellulolytic microorganism, wherein the cellulolytic microorganism is one or more fungi selected from the group consisting of Fusarium, Aspergillus, and Neurospora. A method of producing a succulent sugar from a lignocellulosic material, the method comprising: providing a composition of claim 7; contacting the composition with a lignocellulosic material to produce a sucrose. The method of claim 8, wherein the saccharable sugar is selected from the group consisting of glucose, xylose, arabinose, galactose, mannose, rhamnose, sucrose, fructose, lactose, maltose, trehalose, cellobiose, and oligosaccharide. a group of sugars and any combination thereof. The method of claim 9, wherein the lignocellulosic material is selected from the group consisting of cellulosic animal waste, municipal solid waste, waste paper, garden waste, agricultural residues, forestry residues, and any combination thereof. The method of any one of claims 8 to 10, further comprising converting the saccharable sugar into a fermentation product. The method of claim 11, wherein the converting step is carried out by microbial fermentation or enzymatic treatment. A method of producing energy from a lignocellulosic material, the method comprising: providing a composition according to claim 7; contacting the composition with the lignocellulosic material to produce a sucrose; converting the sucrose to produce a combustible fermentation product; and burning the combustible fermentation product or hydrolyzing solid waste or residue for production Raw energy. A bioreactor comprising a lignocellulosic material and a composition of claim 7. A method for processing a feed or food additive, providing a composition as claimed in claim 7; and contacting the composition with a substance to be processed or treated. A method for biopulping, comprising the composition of claim 7; and contacting the composition with a substance to be processed or treated. A method for complex treatment, comprising the composition of claim 7; and contacting the composition with a substance to be processed or treated.