WO2009005390A1 - Procédé de production de bioéthanol à partir de lignocellulose - Google Patents

Procédé de production de bioéthanol à partir de lignocellulose Download PDF

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WO2009005390A1
WO2009005390A1 PCT/RU2007/000365 RU2007000365W WO2009005390A1 WO 2009005390 A1 WO2009005390 A1 WO 2009005390A1 RU 2007000365 W RU2007000365 W RU 2007000365W WO 2009005390 A1 WO2009005390 A1 WO 2009005390A1
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treatment
hydrolysis
stock
lignocellulose
cellulose
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PCT/RU2007/000365
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English (en)
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Oleg Ivanovich Lomovsky
Kirill Georgievich Korolev
Anatoly Aleksandrovich Politov
Olga Viktorovna Bershak
Tatyana Fedorovna Lomovskaya
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'arter Technology Limited'
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Priority to PCT/RU2007/000365 priority Critical patent/WO2009005390A1/fr
Publication of WO2009005390A1 publication Critical patent/WO2009005390A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/06—Ethanol, i.e. non-beverage
    • C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
    • C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

  • the present invention relates to ethanol from Iignocellulose materials.
  • Production of ethanol from cellulose enjoys immense popularity due to a large available quantity of cellulose-containing waste because it is inadvisable to incinerate or burry it, besides ethanol-based fuel is environment friendly.
  • the process of production of carbohydrates from cellulose materials is employed already to output bioethanol by sugar fermentation. The majority of proto- types of this process were tried during WW2 in Germany, Japan, and Russia after fuel prices leapt. Initially these processes were linked to acid hydrolysis, but their technology and equipment design were rather intricate they were vulnerable to slightest variations of parameters, such as temperature, pressure and acid concentration. Comprehensively these early processes and some contemporary methods are discussed in "Production of Sugars from Wood Using High ⁇ pressure Hydrogen Chloride", Biotechnology and Bioengineering, 1983, vol. XXV, pp. 2757-2773.
  • biomass such as wood, agricultural waste, grassy crops and solid rural waste are considered as a stock suitable to produce ethanol.
  • These materials consist basically of cellulose, hemicellulose, and lignin.
  • the present invention relates to conversion of polysaccharides contained in the lignocellulose stock into ethanol.
  • the present invention does not cover the well-known process of producing ethanol from the starch containing stock when the starch is converted into glucose by acid and/or fermentation hydrolysis and then fermented into ethanol.
  • This method permits to process many types of the lignocellulose stock into the liquid fuel.
  • the main types of this stock are grain crops, quick-growing trees, agricultural waste, wooden waste, and cellulose fibers from solid rural waste and paper waste. It is preferable that these vegetable materials be in the form of small particles, like sawdust, chips, or pulverized biomass.
  • the lignocellulose stock suitable for this method to produce bioethanol includes, without limitations, the following types: agricultural plants, corn stocks, corn ears, wheat, oat straw, rice straw, sugar cane stocks (bogassa), flax straw (boon), soya been stems, groundnut stems, pea stems, sugar beat stems, sorghum stems, tobacco stems, maize, barley straw, buckwheat straw, cassava stems, potato stems, bean stems, cotton and its stems, inedible parts of plants, grain shells (husk); wood of fir, pine, silver fir, cider, larch, oak, ash, birch, aspen, poplar, beech, maple, nut-tree, cypress, elm, chestnut, alder, hickory, acacia, platan, pep- peridge, butternut, apple-tree, pear-tree, plum-tree, cherry-tree, cornel, catal
  • the waste of agricultural plants containing cellulose can be crushed into fine particles and used in the present invention.
  • the commercial waste containing cellulose such as paper, cotton fabric, timber can also be used in the present invention.
  • Partially decomposed vegetable materials such as mowed grass, humus, peat, can be used in the present invention.
  • the biomass of vegetable materials consists of five major components: cellulose, hemi- cellulose, lignin, protein, and inorganic matter.
  • the cellulose, hemicellulose, and lignin are the most essential for ethanol production.
  • Cellulose is a linear polysaccharide consisting of elementary links of anhydro-D-glucose and represents a poly- ⁇ -l,4-D-glucopyranosyl-D-glucopyranose.
  • the cellulose macromole- cule can in addition to the anhydroglucose contain remnants of other monosacharrides (pen- tose and hexose) and uronic acids. The nature and the concentration of these remnants are determined by the conditions of biochemical synthesis.
  • the degree of polymerization of the native cellulose can amount to over ten thousand monomeric units; the degree of polymerization of majority of grassy plants does not exceed one and a half thousand units.
  • Cellulose is the main component of the cellular walls of higher plants. It plays together with the accompanying substances the role of the skeleton bearing the main mechanical loading.
  • Cellulose has a complex super molecular structure resulting from the ordering of its molecules.
  • the smallest cellulose super molecular link is the primary fibril in which groups of arranged in parallel macromolecules are linked together by numerous hydrogen bonds.
  • the cellulose macromolecules in the primary fibrils form highly ordered crystalline zones that al- ternate with inhomogeneous, less ordered amorphous zones.
  • the crystalline zones in the primary fibrils stretch for 15 nm; their cross section is 3-7 nm.
  • the primary fibrils in the cellulose are linked together with hydrogen bonds into microfibrils that are the main links of the fibrous cellulose structure.
  • microfibrils that are the main links of the fibrous cellulose structure.
  • Freigh-Wissling model a significant role in formation of the microfibrils is played by the occluded water and lignin and hemicellulose found in between the primary fibrils.
  • Such specific cellulose morphological structure makes its stable when exposed to significant mechanical loads.
  • the cellulose is also quite stable to enzymes and microorganisms.
  • the structural strength is because natural cellulose is a composite material with the crystalline matrix and amorphous fillers, hemicellulose and lignin acting as adhesives.
  • the intricacy of the process of conversion of the lignocellulose stock into the bioethanol is that it transforms stable cellulose into glucose. The latter is known to ferment easily by yeast into ethanol.
  • Hemicellulose are polysaccharides in the composition of the plan tissue cellular walls that together with the cellulose and lignin are branched polymers of different structures, the main monomeric units of the hemicellulose being galactose, glucose, mannose, xylose, ara- binose, uronic acids.
  • Hemicellulose differs from cellulose by better solubility in alkaline solutions and the capability to be hydrolyzed quickly by the solutions of cellulosolytic enzymes and weak solu- tions of acids.
  • the degree of polymerization of the hemicellulose is, as a rule, inferior to that of the cellulose.
  • the monosaccharide units are usually combined by ⁇ -l,4-links, the latter having frequently lateral links of another type.
  • the main component of the hemicellulose is xylose (50-70 % monomeric links); the main class of the hemicellulose is xylane.
  • Lignin Lignin is an amorphous cross-linked phenol polymer that only vascular plants have and it account for up to 30 % of their mass. Microorganisms capable to produce ethanol do not digest lignin and so it is useless for ethanol production. Lignin remnant can be used as fuel for production facilities in order, for instance, steam and power generation. Lignin can be oxidized into a number of useful chemical substances, but so far there are no well-developed processes and they have not yet gained broad application.
  • the plant biomass consists of cellulose macrofibers coated with a hemicellulose layer. These layers are embedded into the lignin matrix. The diameter of the cellulose macrofibers is about 1-4 ⁇ m. Thus, the mechanical separation of cellulose from lignin can be achieved by crushing the material to particles 1-4 ⁇ m. Pulverization of the vegetable materials into a pow- der of the same size with the efficiency acceptable from the industrial viewpoint is linked with large difficulties.
  • amorphous cellulose and hemicellulose parts of lignocellulose materials are easily hydrolyzed yielding water-soluble carbohydrates in the process called saccharification leaving lignin and unhydrolyzed crystalline cellulose.
  • the process of saccharification implies hy- drolytic decomposition of the cellulose in the presence of a catalyst.
  • the sulfuric acid is a common chemical catalyst.
  • the residue of saccharification by sulfuric acid contains lignin and unhydrolyzed cellulose.
  • the common biochemical catalysts are cellulose enzymes that are obtained, as a rule, as a complex preparation by ultrafiltration of cultural. fluids of definite microorganisms.
  • the en- zymes in the composition of these cellulosolytic complexes have inherent specialization: some of them hydrolyze effectively internal glycoside links between monosaccharide units (endopolymerases, endoglucanases, endoenzymes); others split preferably the external glycoside links at the ends of the polysaccharide chain (exodepolymerase, exogluconases, exoen- zymes); still another glucosidases perform hydrolysis of glycoside links of di- and oligosac- charides.
  • Acid hydrolysis is attractive because it occurs quite quickly. However, it needs special acid resistant equipment; moreover, the acids during hydrolysis of carbohydrates and lignin produce by-products that are toxic for majority of the microorganisms generating ethanol. Therefore application of the acid hydrolysis to produce bioethanol demands special techniques of cleaning hydrolates leading to a considerably higher cost of the end product. Utilization of the acidic waste and regeneration of acids also complicate the process. There is a risk to personnel health and environment contamination; • The enzyme hydrolysis of polysaccharides in the lignocellulose stock evolves with larger selectivity and larger yields characterize it. Until recently, the application of enzymes was limited by the duration of the hydrolysis processes and their costliness.
  • Another unresolved problem of the acid process is to obtain the lignin from the lignocellulose stock free of sulfuric acid impurities. Only this lignin can serve as an environmentally friendly fuel and as a component in the formulas to feed animals.
  • Enzyme hydrolysis Usually treatment with enzymes is performed during mixing of the substrate (the lignocellulose material) with water to obtain 5-12 % suspensions of the cellulose mass, afterwards the enzymes are added. Hydrolysis is conducted during 24-150 hours at 37-50 0 C, pH 4.5-5. Once the hydrolysis is over the soluble monosacharrides are in the liquid, unhydrolyzed portion of cellulose, lignin and other insoluble components of the substrate remain in the solid portion of glucose molasses. They are extracted by filtering the suspensions, the solid residue is washed through to increase the glucose yield.
  • the glucose molasses are fermented into ethanol by yeast; ethanol is purified by distillation or other method. The ethanol fermentation and purification are a well-known process applied in alcohol production.
  • the effectiveness of enzyme hydrolysis depends on the specific features and the mechanism of action of the enzymes. For instance, the cellulase T. longibrachiatum bonds strongly to the cellulose resulting in a reversible inactivation of the enzyme (Brooks, T. A., and In- gram, L. O., Conversion of Mixed Office Paper to Ethanol by Genetically Engineered Klebsiella oxytoca Strain P2, 1995, Biotechnol. Prog., vol. 11, no. 6, pp. 619-625). The degree of bonding is governed by the stirring intensiveness (Kaya, F., J. A. Heitmann, Jr., and T. W.
  • An effective pre-treatment method should combine the advantages of the known methods, including a high degree of cellulose processing, low yield of side-products and frugal consumption of cellu- losolytic enzymes.
  • the effect of the pre-treatment method is characterized by the degree of transformation of cellulose components into soluble sugars and the amount of the enzyme consumed to convert a definite amount of cellulose into glucose.
  • Pre-treatment in the present invention combines the known approaches to acceleration of enzymatic hydrolysis:
  • the pre-treatment by all the methods employs steam energy, mechanical energy, and energy of radiaton.
  • One or several types of pre-treatment are used to increase the rate and degree of hydrolysis.
  • the effect of pre-treatment is commonly explained by the fact that it in- creases the availability and the surface area of hydrolyzed polysaccharides, destroys the physical and molecular structure of the original material and splits up (reduces sharply the intermolecular interactions between the macrostructural components) lignocellulose materials into lignin , hemicellulose and cellulose components.
  • steam treatment steam explosive treatment (steam explosion, powerful steam extrusion, i.e. feeding steam under pressure into the stock and its destruction due to a sharp pres- sure drop when steam passes through the outlet hole);
  • Acids catalyze hydrolysis of polysaccharides into soluble carbohydrates, monosacharrides primarily.
  • the hydroxides of alkaline metals serve to delignify the polysaccharides, then the polysaccharides undergo the acid hydrolysis into soluble carbohydrates, the sulfuric acid is used, as a rule.
  • biomass is first wetted by the solution of the alkaline metal hydroxide, and then it is stirred in order to distribute the catalyst over the substrate and destroy interactions between lignin and polysaccharides.
  • the hydroxides of alkaline metals are introduced in the amount sufficient to initiate thermal reactions. The latter release carbon dioxide from the cellulose carbohydrates and modify the lignin.
  • the carbohydrates formed by the present method can serve to produce bioethanol as animal feed or in the synthesis, for instance, to synthesize high-molecular alcohols.
  • Steam treatment is one of the main methods of pre-treatment of the lignocellulose stock (U.S. Patent # 4,461,648).
  • the biomass is charged into a vessel, the so-called steam gun.
  • a solution of acids (up to 1 %) is added into the vessel with the biomass.
  • the vessel is filled up rapidly with steam and kept under high pressure during the assigned time.
  • the pressure in the vessel is rapidly released, the treated biomass is thrown out, hence the method is called «steam explosion».
  • the pre-treatment effect depends on the time of exposure under treatment, temperature, the concentration of acids and particle sizes in the stock.
  • the steam pressure ranges between 17 and 72 atm, the temperature between 208 and 285 0 C.
  • U.S. Patent # 4,461,648 Other researchers who tested different substrates and equipment later confirmed the opti- mum pre-treatment conditions disclosed in U.S. Patent # 4,461,648.
  • U.S. Pat # 4,237,226 describes pre-treatment of oak, poplar wood, newspaper and corn straw in an impact flow-through continuous type reactor resembling an extruder. Rotating screws force the suspended stock through a small hole and the stock structure is destroyed mechanically at the outlet.
  • Modern publications study the pre-treatment mechanisms that improve the enzyme hydrolysis of the lignocellulose substrate.
  • U.S. Patent # 5,628,830 describes pre-treatment of the lignocellulose material by steam explosion in order to destroy the hemicellulose followed by the cellulose hydrolysis. Knappert et al.
  • Mechanical grinding and amorphization usually implies application of impact, shear, pressure, grinding, mixing, compression/expansion or other types of mechanical effects.
  • U.S. Patent # 5,366,558 describes an improved method of producing glucose when the stock is subjected to mild hydrolysis during which the hemicellulose splits without any substantial cellulose hydrolysis.
  • the solid residue containing cellulose and lignin is subjected to fine grinding, for instance, by the method disclosed in U.S. Patent # 4,706,903.
  • the ground substrate is subjected to acid hydrolysis under tougher conditions until the glucose solution is obtained.
  • U.S. Patent # 5,268,830 the fine ground solid residue resulting from the biomass after hydrolysis of the lignocellulose stock, as disclosed in U.S.
  • Patent # 5,366,558 is subjected to enzymatic hydrolysis by cellulosolytic enzymes producing an aqueous glucose solution that is later fermented into ethanol. Hydrolysis of polysaccharides into monosachar- rides and their fermentation into ethanol can be conducted simultaneously in the presence of cellulosolytic enzymes and special microorganisms, yeast or bacterial fermenting monosa- charrides into ethanol .
  • the ethanol yield can be increased compared with the method by which the residue after removal of easily hydrolyzed polysaccharides (the hemicellulose) serves as the substrate without any further fine grinding.
  • Azuma J. et al. (Journal of Fermentation Technology, 1984, vol. 62, no. 4, pp. 377-384, and U.S. Patent No 5,196,069) proposed a method of microwave pre-treatment for enzymatic hydrolysis of polysaccharides in the lignocellulose stock.
  • the enzymatic hydrolysis rate of polysaccharides accelerates in case of microwave pre-treatment of the stock at 160 0 C; the maximum effect is reached at 223-228 0 C.
  • Such treatment enables to obtain 77-84 % of the reducing carbohydrates from the total concentration of polysaccharides in the lignocellulose stock.
  • U.S. Patent # 6,333,181 considers the improved method of enzymatic hydrolysis of polysaccharides from the lignocellulose stock.
  • the method is based on the ultrasound treatment of the lignocellulose stock in the presence of water and enzymes ensuring further hydrolysis of polysaccharides.
  • the duration and conditions of the ultrasound treatment are selected such as to prevent heating of the mixture to the temperature at which a considerable portion of enzymes denaturizes. It is taken into account that ultrasound treatment leads to a considerable destruction of the cellulose crystalline structure. This method saves consumption of enzymes two-three times versus the common methods.
  • Blotkamp P. J. et al. (American Institute of Chemical Engineering (AIChE) Symposium 1981, Series # 181, vol. 74) described the process of combined saccharification of cellulose and fermentation of sugars into ethanol (CAF) using the enzymes of the fungi Trichoderma reesea and the yeast Saccharomyces cerev.
  • CAF ethanol
  • the rate of hydrolysis of the cellulose stock increases in comparison to the process of consecutive stages of saccharification and fermentation due to reduction of the rivaling inhibition of enzymes by glucose and other soluble carbohydrates. 7. Fermentation
  • Any suitable method is applicable to fermentation of carbohydrates to produce ethanol according to the present method.
  • Any yeast capable to induce conversion of carbohydrates into ethanol can be added to the aqueous solution of carbohydrates obtained under the present method. The mixture is fermented until the carbohydrates are fully consumed. Ethanol is pu- rified by distillation.
  • Another method can be used, like microbic conversion, or combined saccharification and fermentation.
  • yeast used to produce ethanol on industrial scale, like Montrachet, Pasteur Chalmmpagne, Cote des Blancs, Pasteur Red, Lalvin Kl-V-1 116 and Lalvin 71 B-1 122. 8. Mechanical activation and mechanochemical treatment
  • the mechanical activation is rated an effective method of modification of physicochemical properties of solid phases.
  • the mechanical activation implies enhance of the reactivity due to stable changes in the structure of a substance under the effect of mechanical loading.
  • the mechanical and activated solid differs by the fact that its deformation process and physicochemical consequences of deformation are divided by the time insufficient for the relaxation processes to complete (Avakumov, E.G., Mechanical Methods of Activation of Chemical Processes, Novosibirsk: Nauka, Siberian Branch, 1986, 303 pp.).
  • the mechanical strain applied to the solid can relax through several ways.
  • the mechanical energy is expended primarily for formation of new surface and defects in the crystalline structure. These processes increase the free energy in the solid resulting in its enhanced reac- tivity. The latter circumstance has general nature. So, the mechanical and activated solid phases are characterized by higher dissolution rates and easier react chemically with gases and liquids compared with the non-activated phases.
  • a more general term of mechanochemical treatment is applied to heterogeneous systems that have a complex phase composition and consist of numerous components.
  • the mechanochemical treatment like mechanical activation, induces stable changes in the system's phys- icochemical properties. Stronger reactivity affects, as a rule, most of the phases and components of the system.
  • the chemical reactions can evolve resulting from stronger mobility of the components and their larger free energy under effect on mechanical loading (Boldyrev, V.V., Mechanochemistry and Mechanical Activa- tion, Materials Sci. Forum, 1996, vol.225-227, pp. 51 1-520).
  • the mechanoenzymatic treatment is the mechanochemical treatment in which enzymes participate. This type of effect is applicable to plant stock, natural polymers and organic materials.
  • the mechanoenzymatic treatment is conducted in order to increase the substrate reactivity; in number of cases, chemical reactions can evolve catalyzed by enzymes directly at the time of treatment.
  • the reactivity of solid phases is restricted by low mobility of the components making up these phases. Under the effect of intensive mechanical loading the components mix up, arrange directly close one to another so that the paths of diffusion are reduced sharply.
  • the mechanical treatment of the mixture of solid phases intensifies the mobility of the components in the time of treatment and increases mobility due to disordering of the crystalline structure of solid phases.
  • the solid components can accumulate defects and amorphize resulting in stronger reactivity of both the components and the system in general.
  • the mechanical energy from the viewpoint of economics is an «expensive» type of energy. It should be consumed effectively.
  • the mechanochemical treatment of the solid mixture can be suspended at an early stage of transformation of agents and full chemical transformation is achieved with other, energy-saving processes involving, as a rule, liquid phases. In case of this approach, the mechanochemical treatment is achieved:
  • the mechanocomposites are products of the mech- anochemical treatment of solid heterogeneous mixtures and they represent a system, having the physicochemical properties significantly different from the original mixture and they are determined by substantial changes in the morphology of the components, the developed interface phases with pronounced interphase surface interaction.
  • the interphase material possesses the physicochemical characteristics that are different for any of the original components or individual phases.
  • the mechanochemical treatment of the systems containing enzymes requires considering the fact that their complex structural set-up governs the catalytic activity of the enzymes. Secondary and tertiary structures can change in the enzymes under the mechanical effect affecting their activity. For instance, the mechanochemical synthesis of the immobilized enzymatic catalyst turned out a failure (Trevan, M.D., Immobilized Enzymes, Chichester - New York: John Wiley, 1982, 213 pp.). The mechanical treatment of the substrate inactivated the enzyme irreversibly.
  • the conditions are proposed in the present invention under which the lignocellulose stock can undergo mechanoenzymatic treatment turning it into heterogeneous systems consisting of just solid phases and systems containing water. These conditions enable to achieve the tech- nological effect that comprises:
  • the main criterion of effectiveness of the mechanoenzymatic treatment in the present process is enhancing of hydrolysis of polysaccharides from the lignocellulose stock as promotion of yield of water-soluble carbohydrates.
  • the technical task of the invention is to develop a method of production of bioethanol that would enable to introduce idle biorenewable sources of polysaccharides, predominantly the lignocellulose stock fit to be processed in other spheres of chemical and biochemical technologies;
  • the present invention is based on application of preliminary and/or intermediate mechanoenzymatic treatment that enhances noticeably the hydrolysis of polysaccharides, promotes the yield of fermentable carbohydrates, and reduces material and energy cost of the process of production of bioethanol from the lignocellulose stock.
  • the preliminary (intermediate) treatment implies that the mechanochemical effect of certain intensity and duration acts on the mixture of the lignocellulose stock and enzymes (the solution of the en- zymes ) in the mechanochemical reactor (the caviation or ultrasound device).
  • the inventors have discovered that implementation of definite conditions, such as utilization of the soft lignocellulose materials as the raw stock, the optimum intensity and duration of the mechanical effect that ensure formation of mechanocomposites and preservation of the activity of the enzymes, is sufficient and necessary for effective conversion of the polysac- charides from the lignocellulose stock into bioethanol .
  • the preliminary enzyme hydrolysis is performed instead of the above stage to achieve the degree of conversion of polysaccharides 20-40 %
  • the hydrolysisate - solid residue system is treated in the caviation devices in the presence of solid residue, ethanologenic microorganisms are introduced to perform the process of saccharification and combined fermentation (SSCF);
  • the preliminary and/or intermediate mechanoenzymatic treatment increase the degree of conversion of the cellulose raw stock to 90%, saves considerably the consumption of the cel- lulosolytic enzymes needed for hydrolysis of the polysaccharides in comparison with the known methods.
  • Application of the claimed method makes production of ethanol from ligno- cellulose materials much cheaper.
  • the mechanochemical treatment during conversion of the polysaccharides from the Hg- nocellulose raw stock into ethanol is a significant improvement of the known processes.
  • An additional advantage of the method is that the unconverted residue of the lignocellulose stock contains no unhydrolyzed polysaccharides, etc., or traditional impurities, sulfur in the first place, that inhibit use of this residue for production process needs, for instance, for combustion in order to generate heat, steam and power.
  • Microorganisms that the biomass contains are usually used to obtain additional products, such as feed protein or feed additives with biologically active properties for agricultural animals. This method can be optimized further by changing the types of treatment, the intensity, and duration of the effects.
  • the present invention embodies the method of producing ethanol from the ligno- cellulose raw stock, namely the method that is applicable in production of bioethanol from lignocellulose plant materials that comprises the mechanoenzymatic treatment of the material in the mechanochemical reactor in the presence of cellulosolytic enzymes with or without of additional water that follows hydrolysis of the polysaccharides or combined performance of fermentation of resulting carbohydrates into ethanol with the help of suitable etha- nologenic microorganisms .
  • the ethanologenic microorganisms can be special strains of bacteria or yeast, including recombinant strains that are capable to ferment the main monomeric units of the polysaccharides from the lignocellulose raw stock, namely xylose and glucose that hydrolysates contain.
  • the preferable ethanol-producing microorganisms include Saccharomyces, Zymomonas, Er- winia, Klebsiella, Xanthomonas, Escherichia, etc.
  • Fig. 1 Mechanochemical introduction of the enzyme into the lignocellulose stock mass - into the reaction zone (right), for comparison, left - addition of the substrate into the aqueous solution of the enzyme.
  • Fig. 2. The chromatographic splitting of carbohydrates. The hydrolate of the wheat straw.
  • Fig. 3 The chromatographic splitting of carbohydrates. The artificial mixture of carbohydrates as a reference.
  • Fig. 4 Dependence of the degree of transformation of microcrystalline cellulose into soluble sugars on the duration of enzymatic hydrolysis (the lower curve - without mech- anoenzymatic treatment, the upper curve - with mechanoenzymatic treatment).
  • Fig. 5 Diagram of hydrolysis processes in the counterflow mode using several reactors.
  • Fig. 1 illustrates its technological sense.
  • the mechanical treatment of the solid mixture of the substrate and the enzyme does not affect the structure and the activity of the introduced enzyme; it permits to distribute the enzyme molecules in the substrate volume.
  • aqueous solution into the substrate shown in Fig. 1,2 for comparison, left
  • a major portion of the enzyme molecules appears outside the substrate and cannot be used effectively.
  • the lignocellulose stock implies any raw stock that can be used in the processes of conversion of cellulose and attendant polysaccharides into ethanol.
  • the raw stock contains at least 20-35 % cellulose; most of it is hydro lysable into glucose.
  • the concentration of water in the so-called “air-dry" stock dehydrated without vacuum at the temperature below 50-100 0 C is 8-15 % of the stock mass.
  • concentrations of lignin, starch, protein, or inorganic compounds in the raw stock can be wood, grass, straw, waste of agricultural crops.
  • Conversion into ethanol means conversion of at least 90 % of cellulose and hemicellulose into glucose and other soluble carbohydrates intended for further fermentation into ethanol.
  • Hemicellulose contains different monosacharrides. Different publications from different sources
  • Osources indicate the composition of the hemicellulose obtained by different methods.
  • the composition of the stock can be determined approximately.
  • Mechanoenzymatic treatment is the mechanochemical treatment of the lignocellulose stock in the presence of enzymes or a solution of the enzymes. This type of effect is applicable both to the lignocellulose raw stock and to individual polysaccharides.
  • the mechanoenzymatic treatment is conducted in order to increase the reactivity of the substrate, namely, to accelerate the hydrolysis and to hydrolyze more polysaccharides in the stock. Without mechanoenzymatic treatment consumption of the cellulosolytic enzymes grows considerably when it is necessary to achieve 90 % conversion of polysaccharides by the reaction of enzymatic hydrolysis.
  • the mechanoenzymatic treatment of the mixture (90-98 % - the lignocellulose stock with the concentration of natural moisture 0.5-15 % of the stock mass; 0.2-2.0 % - the cellulosolytic enzymatic preparations containing the optimum ratio of endo-l,4- ⁇ -glucanase, exo- 1 ,4- ⁇ -glucanase, exo-l,4- ⁇ -glycosidase and ⁇ -glycosidase; 00-8 % - inorganic salt; 0.0-1.0 - surfactant) is performed during 0.5-10 min in the ball mill with the acceleration of balls 60- 400 m/s 2 or in the rotary mill with the speed of rotors 10-120 m/s or in the pneumatic vortex mill with the rate of the gas flow 10-120 m/s.
  • the product of the mechanoenzymatic treatment of the solid mixture of the lignocellulose stock and enzymes is a mechanocomposite containing the polysaccharides more reactive in respect to the enzymatic hydrolysis.
  • the preliminary mechanoenzymatic treatment forms a qualitatively new product, or mechanocomposite, the polysaccharides in which are hydrolyzed at a faster rate than by the known methods; it is characterized by a high degree of transformation into monosacharrides and smaller consumption of the enzymes.
  • the pre-treatment of the lignocellulose stock proposed in the present invention is preferably a part of a more complex process of conversion of the lignocellulose stock into the etha- nol.
  • the general process comprises pre-treatment of the substrate, enzyme hydrolysis of polysaccharides into monosacharrides, fermentation of the latter into ethanol and ethanol purifica- tion.
  • Complex preparations of cellulosolytic enzymes are preferable for mechanoenzymatic treatment of the lignocellulose stock and subsequent hydrolysis.
  • a smaller portion of the cellulose is hydrolyzed during pre-treatment, a larger portion is hydrolyzed in the process of saccharification.
  • the method of implementation of the fer- mentative hydrolysis is not limited by the invention, but the following conditions are prefer- able.
  • the hydrolysis of the product of the mechanoenzymatic treatment is conducted in the water suspension with the hydromodulus 5-10, pH 4.5-5 at a temperature 50 0 C.
  • compositions of enzymes separated by ultrafiltration from the cultural fluid of Trichoderma viride (reesei) and/or Aspergillus awamori and/or Bacillus subtilis can serve as cellulosolytic preparations directly obtained during production of bioethanol
  • ⁇ -glycosidase can be added into the enzymatic complexes to en- sure fuller conversion of cellobiose into glucose.
  • the following mass-produced preparations of enzymes with the ⁇ -glycosidase activity were used: Novozym 188 produced by Novo Nordisk and/or Glucolux produced by Sibbiopharm.
  • the quantity of the enzymes in the hydrolytic process determines the time of hydrolysis, the yield of fermentable carbohydrates and their concentration..All these values influence the profitability of the processes and can vary in response to the technology.
  • the usual dosage of the enzymes is 1-50 U/g of the substrate for 12-128 hours.
  • the preferable dosage of the enzymes was 1 - 10 U/g of the cellulose. Examples 2 and 3 describe the cellulose hydrolysis in more detail.
  • the invention is illustrated with detailed examples showing its preferable embodiments, but they do not limit the method that can be used to produce carbohydrates and ethanol.
  • the preferable embodiment is to mix up the enzyme with the lignocellulose material followed by hydrolysis to produce fermentable sugars.
  • the invention ensures effective enzyme hydrolysis of polysaccharides in the hemicellulose stock to produce fermentable carbohydrates and the water insoluble solid residue.
  • the invention relates to the method accelerating the fermentative hydrolysis of polysaccharides in the lignocellulose stock; it comprises the mechanoenzymatic treatment of the Hg- nocellulose stock, mixing of the product with water under the conditions sufficient for the hydrolysis of polysaccharides.
  • the aqueous suspension of the lignocellulose stock can be subjected to the mechanoenzymatic treatment too.
  • the mechanoenzymatic treatment is performed with the help of the known equipment.
  • the mechanochemical reactors applicable for the purposes of the invention should possess definite working parameter, such as the effect intensity and duration of the operation.
  • the examples of the relevant mechanochemical reactors include: • mechanochemical reactors, such as a planetary ball mill or a vibration ball mill in which the intensity of the mechanical effect is characterized by the acceleration of the balls.
  • the optimum range of the acceleration of the balls is 60-400 i ⁇ /s 2 .
  • the coefficient of the acceleration of the balls in the usual gravitation mill is about 10 m/s 2 .
  • the standard vibration mills of the series VCM and CEM produced by Novic, Russia, or Tribochem, Ger- many, are applicable for these processes ;
  • Vortex or jet mills in which particles of the original material are accelerated by the flow of air or gas up to 10-120 m/s. The material is crushed by collision of the particles with deflecting obstacles.
  • the following mills can be used: Vortex Mills of Hydan Technologies, USA, or Jet" Micronizers of Sturtevent, Inc., USA, vortex mills VIT, of VIT Ltd., Novosibirsk, Russia.
  • the mechanochemical treatment can be conducted within a broad range of intensities that yield close results.
  • the duration and intensity of the mechanochemical treatment can be se- lected such as to avoid conditions when a considerable quantity of enzymes is denaturized.
  • Usually the mechanochemical treatment lasts 1-10 minutes.
  • Continuous and discrete modes of treatment The continuous mode is characterized by the fact that the material can be delivered into the working chamber of the activator during indefinite time (tens and hundreds of seconds).
  • the mass of the treated material is determined by the speed of passage through the working chamber.
  • the discrete mode is characterized by the fact that the material is charged into the working chambers in a quantity the working chamber can accommodate while the activator is off.
  • the solid mixture after the mechanoenzymatic treatment and after adding of water or the solution of enzymes can be further subjected to the enzymatic hydrolysis.
  • the cellulases can be used unpurified or as suspensions produced by filtrating the cultural fluid of the relevant producers.
  • the suitable sources of the cellulases comprise standard cellulase preparations like SpezymeTM CP, CytolaseTM M 104, and MultifectTM CL (Genencor International), Glucolux (Sibbiopharm, Russia).
  • the conditions of the enzymatic hydrolysis are usually selected taking into account the source of the cellulases, i.e. bacteria or fungi.
  • the cellulases of the fungi are usually more effective at temperatures 30-48 0 C and pH 4.0-6.0 within the action range 30-60 0 C and pH 4.0-8.0.
  • the microorganisms capable to ferment sugars or oligosaccharides into ethanol comprise yeast and bacteria.
  • the microorganisms are capable to secrete one or more that individually or together convert sugars into ethanol.
  • Saccharomyces such as S. cere- visiae
  • Other similar microorganisms comprise the following types: Schizosaccharomyces (such as S. pombe), Zymomonas (including Z. mobilis), Pichia (P. stipitis), Candida (C. shehatae) and Pachysolen (P. tannophilus.
  • the genetically modified strains of E. coli can also serve to convert carbohydrates into ethanol.
  • the preferable example of the microorganisms capable to produce ethanol comprises the microorganisms secreting alcohol dehydrogenase and decarboxylase pyruvate; for instance, Zymomonas mobilis (see U.S. Patents Nos. 5,000,000; 5,028,539; 5,424,202; and 5,482,846)
  • microorganisms in the processes fermentation capable to ferment into ethanol, pentoses, and hexoses that produce one main enzyme and an additional complex of enzymes.
  • the examples of such microorganisms include those disclosed in U.S. Patents Nos. 5,000,000; 5,028,539; 5,424,202; 5,482,846; 5,514,583; and Ho et al., WO 95/13362.
  • the microorganisms including Klebsiella oxytqca P2 and Escherichia coli KOl 1 are specifically preferable.
  • the conditions of conversion of sugars into ethanol are usual conditions disclosed in the quoted patents; mainly the temperature is 30-40 0 C and pH 5.0-7.0.
  • Nutritive substances and/or cofactors for microorganisms and/or enzymes are added to optimize the conversion. It is also desirable to add digestible carbon, nitrogen, and sulfur to accelerate proliferation of the microorganisms.
  • Numerous nutritive media for growth of microorganisms are known, in particular, Luria broth (LB) (Luria and Delbruk, 1943).
  • Membrane filtration can be applied at any stages of the claimed process.
  • the systems of membrane filters are selective to the molecular weight or size of molecules.
  • the membrane filter is used at the stage of saccharification, at the stage of reversion of side products and at the stage of fermentation trapping enzymes, carbohydrates, salt, yeast and allowing to water and ethanol molecules to penetrate through the membrane.
  • Application of the membrane fil- tration enables to use side products, such as glycerol, lactic acid and others and to reduce the quantity of solid substances reaching the evaporator.
  • the process enables to save the cost and increase profitability of ethanol production.
  • the waste heat boiler serves to evaporate the remaining liquid from the lignin, and then the organic substances are incinerated to generate heat and steam, the combustion products are reduced into the environmentally tolerable condition.
  • the wheat straw, surface portion of corn without ears and microcrystalline cellulose were used as the raw stock. All vegetable raw stock was harvested in the Novosibirsk region, Russia.
  • the microcrystalline cellulose complied with TU 6-09-10-1818-87, had the index of crystallinity equal to 86 % (Segal, L., Tripp,V.U., Determination of Cellulose Crystallinity. In: Cellulose and its Derivatives. Ed. by N. Bicles, L. Seagull, Moscow: 1974, vol. 1, pp. 214-235.).
  • the concentrations of moisture and volatile components, lipids, water soluble substances, water soluble carbohydrates, easily hydrolysable and hardly hydrolysable polysac- charides, lignin and ash were determined in the original stock .
  • the stock was roughly crushed in the disintegrator to the size of particles 500 ⁇ m. Then the crushed stock was kept at a temperature 15-25 0 C in sealed packs. The humidity was checked by drying to a constant weight at the temperature 100 0 C. The moisture content in the specimens was 5-10 %. The ash content was determined by the residue after the specimens were baked in porcelain crucibles at the temperature 560 0 C during 3-4 hours.
  • the lipids were separated by exhaustive extraction of the dry stock with hexane in the Sockslet apparatus.
  • the solvent was removed from the extract in a rotary evaporator with the vacuum in the water-jet pump at the temperature 50 0 C.
  • the extracted substance and the stock extracted with hexane were dehydrated in the vacuum dessicator to the constant weight.
  • the water-soluble substances were determined by triple aqueous extraction of the crushed, degreased, and dehydrated stock. The extraction was conducted in the ultrasound bath at the room temperature and the hydromodulus equal to 20, during 20 minutes.
  • the solid residue was rinsed, filtered through a fine-pore glass filter, the aqueous extracts and rinsed water were combined; water was eliminated in the rotary evaporator in the vacuum in the water-jet pump at the temperature 50 0 C.
  • the obtained residue was dehydrated in the vacuum dessicator to the constant weight.
  • the solid residue of the plant stock was also dehydrated in the vacuum dessicator and served to determine further the easily hydrolysable polysaccharides. Free disaccharides, hexoses, pentoses, and oligosaccharides were determined in the water-soluble substance. The disaccharides, hexoses, and pentoses were determined- with the method of HPLC, as described below.
  • the concentration of oligosaccharides was determined from the difference between the carbohydrates in hydrolysates and the sum of free di- and monosacharrides.
  • the method of acid hydrolysis is described below in the section relating to determination of easily hydrolysable polysaccharides.
  • the analysis was performed in the isocratic mode (25 % methanol in the aqueous 0.001 M solution of the chloral acid containing 2 % lithium perchlorate with the analytic chromatographer Milichrom A-02 equipped with a microcolumn with the inverted phase (ProntoSil C-18. 5 ⁇ m, 2x70 mm) and a spectrophotometric detector.
  • the instrument was calibrated with the solution containing known quantity of carbohydrates (lactose, cellobiose, glucose, mannose, and xylose).
  • Fig. 2.3 exemplifies the calibration chromatogram and chromatogram recorded with the wheat straw hydrolysate.
  • the activity of the enzymatic complex Cellolux is described as an example.
  • the activity was determined by hydrolysis of the filtering paper Whatman # 1 with a partially modified the method (Ghose, T.K., Measurement of Cellulase Activity, Pure Appl. Chem., 1987, vol. 59, pp. 257-268).
  • the shredded filtering paper was placed into a plastic reactor (5.0 ml), the acetate buffer (2:0 ml) and thermostatted at 50 0 C periodically until a suspension.
  • the activity of several specimens was measured with solutions of the complex with different concentrations within the range 0.6-5 mg/ml.
  • the substrate was hydrolyzed during 60 minutes lightly shaking the reactors meanwhile, and then the reactors were heated in the water bath to 70 0 C to inactivate the enzymatic complex.
  • the obtained hydrolysates were centrifuged, the solid residue in the hydrolysate was discarded, the concentration of carbohydrates (converted into glucose ) with the phenol-sulfur oxide method (Ghose, T.K., Measurement of Cellulase Activity, Pure Appl.
  • the enzymatic preparations were diluted with the 50 mM citrate buffer to the concentra- tions equivalent to those used in the processes of separation of sugars from paper 250 FPU SpezymeTM, CP/L and 50 unit/1 Novozyme 188.
  • the solutions contained 0.5 g/1 thymol, 40 Mg/1 chloramphenicol to prevent proliferation of bacteria.
  • the enzymatic mixture was stirred during 15 minutes with the speed 120 r.p.m. until full distribution of the enzyme. Stirring continued for during 48 hours. Samples were taken every 0, 12, 24, 36, 48 h.
  • the enzymatic activity was deter- mined with the above described the method. The effect on the structure
  • the RFA was performed with a diffractometer DRON-5 ( Russia) in the CuK-alpha emission.
  • Example 1 Acceleration of microcrystalline cellulose hydrolysis.
  • the enzyme hydrolysis of microcrystalline cellulose samples was conducted and the initial hydrolysis rate was measured as a function of cellulose pre-treatment conditions.
  • the obtained mixture was hydro- lyzed while stirring in a magnetic mixer in a glass reactor at a temperature 51 ⁇ 1 0 C. The hydrolysis lasted for 8 hours. Then the reactors were rapidly heated to 70 0 C in order to inactivate the enzymatic complex.
  • the mechanical treatment of the cellulose jointly with the enzyme accelerates substantially the hydrolysis rate.
  • the effect is achieved by producing the mechanocomposite consisting of amorphized cellulose particles with the enzyme distributed over its surface and in its body.
  • the enzyme turns out introduced directly in the zone in which it should function rather than being distributed in the entire solution volume. This condi- tion accelerates the reaction rate.
  • the hydrolysis rate is accelerated additionally by increasing the share of the amorphous cellulose in the substrate.
  • Water is known to stimulate cellulose re- crystallization processes that evolve both during preservation of the mechanically activated cellulose and in the course of mechanical activation.
  • the mechanical treatment is conducted in the presence of dry inorganic salts capable to absorb water and produces complexes with carbohydrates, thus inhibiting the process of re-crystallization. Dry carbonate or calcium chlo- ride served as these agents. The latter is because it is practically harmless for the pH in subsequent hydrolysis.
  • the obtained mixture was hydrolyzed while stirring in a magnetic mixer in a glass reactor at a temperature 51 ⁇ 1 0 C. The hydrolysis lasted for 6 days. After the first, second and third days of hydrolysis, fresh enzyme doses were added into the reaction mixture in the amounts 30, 20 and 10 unit per gram of the substrate, respectively.
  • Fig. 3 shows the results of the enzymatic hydrolysis of the microcrystalline cellulose after mechanochemical treatment. According to the obtained data, the conversion is 87 percent. The results of the comparative experiments indicate that the effect is achieved due to the combination of the intensive mechanical treatment and subsequent separate addition of the enzymatic complex.
  • Fig. 4 shows how the degree of transformation of microcrystalline cellulose into soluble sugars depends on the duration of enzymatic hydrolysis (the lower curve - without mechanoenzymatic treatment, the upper curve - with mechanoenzymatic treatment).
  • the stirring of the reactive mass drives the exogluconases into the solution so that their concentration under fhe surface drops.
  • Addition doses of the enzyme are introduced into the reactor in order to compensate this lower concentration of exogluconases and the total reduction of the concentration of the enzymes due to their natural inactivation during the first three days.
  • Example 3 Enzyme hydrolysis of the lignocellulose stock exposed to mech- anoenzymatic treatment.
  • the dried and pre-crushed to particle size under 0.5 mm plant stock was mixed with calcium chloride (97:3 by mass); the cellulosolytic complex was added for 15 mg
  • the hydrolysis was performed in a series of consecutive reactors.
  • the hydrolysis scheme envisaged that the substrate remains in each reactor for 12 hours and then the substrate would transferred for hydrolysis into the neighboring reactors in the flow through mode, as Fig. 5 shows it.
  • the fresh substrate produced by mechanoen- zymatic treatment of the lignocellulose stock comes into the first reactor.
  • the substrate contacts for 12 hours the hydrolysate coming from reactor 2.
  • This reactor receives the substrate that is free already of the amorphous cellulose and other polysaccharides eas- ily hydrolysable by the enzyme.
  • This substrate is subjected to treatment with the fresh solution of the enzymatic complex.
  • the lignocellulose stock is hydrolyzed in each reactor at a temperature 51 ⁇ 1 0 C and the hydromodulus 7-10.
  • the pH of the reaction mixture is maintained 4.5.
  • the solution of the enzymatic complex delivered into reactor 10 contains 15 units of the com- plex per gram of carbohydrates.
  • Reactors 3 and 6 receive fresh doses of the enzymatic complex in the amount 15 units per gram of carbohydrates.
  • the solutions of the enzymes delivered into reactors 3, 6, and 10 were based on the cultural fluid of Tricho- derma viride (reesei) as a cellulosolytic complex producer.
  • the yield of monosacharrides under the conditions in Example 2 was determined from the obtained data conversion of polysaccharides amounting to 90 %.
  • Example 4 The effect of the surfactant additive during mechanoenzymatic treatment on the subsequent hydrolysis rate.
  • the plant stock was hydrolyzed in the reactors under periodic action during 8 hours while stirring in a magnetic mixer 600 1/min.
  • the hydrolysis conditions were the same in all the alternatives: the temperature
  • Example 5 The stepwise enzyme hydrolysis and intermediate ultrasound treatment
  • the mechanoenzymatic pre-treatment was conducted under the conditions of Ex- ample 3.
  • the enzyme hydrolysis of polysaccharides was performed in steps.
  • the cultural fluid ⁇ Aspergillus awamori and/or Bacillus subtilis) enriched with exogluconases was added into the reactor. Addi- tion was made from the calculation of 10 units per gram of the substrate.
  • the suspension was exposed to ultrasound with the frequency 22 kHz for 5-15 minutes at a temperature 50-90 0 C, then the solid phase was separated from the hydrolysate.
  • the bi-step hydrolysis with intermediate ultrasound treatment yields 90-92 % conversion of the polysaccharides into water-soluble monosa- charrides.
  • Example 6 Enzymee hydrolysis and combined fermentation of the lignocellu- lose stock after its mechanoenzymatic treatment.
  • the required quantity of the preparation TWEEN-20 was introduced directly into the zone of contact between the crushing bodies and the vegetable stock.
  • the preparation TWEEN-20 would increase the effectiveness of mechanoenzymatic treatment due to the adsorption of the surfactant on the lignin thus preventing partial inactivation of enzymes.
  • the cultural fluid ⁇ Aspergillus awamori and/or Bacillus subtilis) enriched with exogluconases was added into the reactor. Its introduction was made 10 units per gram of the substrate.
  • the suspension of the plant stock and hydrolysate was pumped by the cavitators into the reactor for the following saccharification and com- bined fermentation (SSCF).
  • SSCF saccharification and com- bined fermentation
  • Utilization of the cavitators as a pumping device altered the rheological characteristics of the pulp positively. This operation would reduce the viscosity of the solution 2-3 times, the optical microscopy in Fig. 6 shows the disintegration of the conglomerates of particles in the substrate, where: the sequence of structures a-c - formation of conglomerates of particles in the course of preliminary hydrolysis , the structure d - the result of treatment in the cavitations device.
  • the SSCF process is conducted for 7 days at the hydromodulus 8 (dilution by introduction of the components of the nutritive medium).
  • the enzymes are added separately calculated in units per gram of the substrate: 10 units of the cellu- losolytic complex Trichoderma viride (reesei) after two and four days, 5 units of the en- zymatic complex Aspergillus awamori and/or Bacillus subtilis after three and six days.
  • the mechanoenzymatic treatment, preliminary enzymatic hydrolysis, and SSCF- process push to 90 % the conversion of polysaccharides from the stock into water-soluble carbohydrates.
  • the obtained hydrolysate contained 2.8-3.2 % ethanol that was separated by distil- lation.
  • 1000 kg corn straw containing 66 % carbohydrates can yield 307 liters of ethanol; the wheat straw can yield 330 liters.
  • the invention provided a method of producing bioethanol enabling to utilize unused bio-renewable sources of polysaccharides, predominantly the lignocellulose inapplicable in'other spheres of chemical and biochemical technologies.
  • the present invention is embodied with multipurpose equipment extensively employed by the industry.

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Abstract

L'invention concerne un procédé destiné à produire du bioéthanol à partir de matériaux de lignocellulose d'origine végétale ; il est fondé sur le traitement mécanochimique (mécanoenzymatique) d'un mélange solide du substrat de lignocellulose et de complexes enzymatiques cellulosolytiques, et la fermentation des glucides obtenus par des microorganismes éthanologènes. Le traitement mécanoenzymatique est un traitement mécanique du mélange solide du substrat de lignocellulose et de complexes enzymatiques qui assure une capacité de réaction élevée de l'hydrolyse enzymatique qui permet de préserver l'activité des enzymes et également d'économiser leur consommation.
PCT/RU2007/000365 2007-07-04 2007-07-04 Procédé de production de bioéthanol à partir de lignocellulose WO2009005390A1 (fr)

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102146211A (zh) * 2011-01-04 2011-08-10 聊城大学 提高糖化效率的纤维素组合物及制糖方法
US8890395B2 (en) 2010-05-28 2014-11-18 Koninklijke Philips N.V. Beamshaping optical stack, a light source and a luminaire
KR101554874B1 (ko) 2013-10-28 2015-10-22 한국세라믹기술원 리그노셀룰로스계 바이오매스를 이용한 당화합물의 제조방법
WO2015126463A3 (fr) * 2014-02-19 2015-10-29 Xyleco, Inc. Traitement de la biomasse
JP2016013550A (ja) * 2009-02-11 2016-01-28 キシレコ インコーポレイテッド バイオマスの加工方法
WO2018157248A1 (fr) * 2017-03-01 2018-09-07 The Royal Institution For The Advancement Of Learning/Mcgill University Procédé de saccharification enzymatique d'un polysaccharide
EP4051804A4 (fr) * 2019-10-28 2023-12-06 The Royal Institution for the Advancement of Learning / McGill University Dégradation mécano-enzymatique de polymères
WO2024057333A1 (fr) 2022-09-16 2024-03-21 Rohit Khaitan Procédé de préparation de bioproduits à partir de la bomasse dans le cadre d'une économie à faible émissions de carbone

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US4321328A (en) * 1980-12-05 1982-03-23 Hoge William H Process for making ethanol and fuel product
RU2162103C1 (ru) * 2000-02-08 2001-01-20 Московский Государственный Университет пищевых производств Способ производства этилового спирта из зернового сырья
US20060084156A1 (en) * 2002-02-08 2006-04-20 Oreste Lantero Methods for producing ethanol from carbon substrates
US20060177917A1 (en) * 2005-02-09 2006-08-10 Michel Warzywoda Process for the production of cellulolytic and hemicellulolytic enzymes using distillation residues from the ethanolic fermentation of enzymatic hydrolyzates of (ligno)cellulosic materials

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4321328A (en) * 1980-12-05 1982-03-23 Hoge William H Process for making ethanol and fuel product
RU2162103C1 (ru) * 2000-02-08 2001-01-20 Московский Государственный Университет пищевых производств Способ производства этилового спирта из зернового сырья
US20060084156A1 (en) * 2002-02-08 2006-04-20 Oreste Lantero Methods for producing ethanol from carbon substrates
US20060177917A1 (en) * 2005-02-09 2006-08-10 Michel Warzywoda Process for the production of cellulolytic and hemicellulolytic enzymes using distillation residues from the ethanolic fermentation of enzymatic hydrolyzates of (ligno)cellulosic materials

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016013550A (ja) * 2009-02-11 2016-01-28 キシレコ インコーポレイテッド バイオマスの加工方法
US8890395B2 (en) 2010-05-28 2014-11-18 Koninklijke Philips N.V. Beamshaping optical stack, a light source and a luminaire
CN102146211A (zh) * 2011-01-04 2011-08-10 聊城大学 提高糖化效率的纤维素组合物及制糖方法
CN102146211B (zh) * 2011-01-04 2012-11-07 聊城大学 提高糖化效率的纤维素组合物及制糖方法
KR101554874B1 (ko) 2013-10-28 2015-10-22 한국세라믹기술원 리그노셀룰로스계 바이오매스를 이용한 당화합물의 제조방법
WO2015126463A3 (fr) * 2014-02-19 2015-10-29 Xyleco, Inc. Traitement de la biomasse
WO2018157248A1 (fr) * 2017-03-01 2018-09-07 The Royal Institution For The Advancement Of Learning/Mcgill University Procédé de saccharification enzymatique d'un polysaccharide
EP4051804A4 (fr) * 2019-10-28 2023-12-06 The Royal Institution for the Advancement of Learning / McGill University Dégradation mécano-enzymatique de polymères
WO2024057333A1 (fr) 2022-09-16 2024-03-21 Rohit Khaitan Procédé de préparation de bioproduits à partir de la bomasse dans le cadre d'une économie à faible émissions de carbone

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