FR2963009A1 - PROCESS FOR PRODUCING SUGARS FROM LIGNOCELLULOSIC BIOMASS PRETREATED WITH HYDRATES INORGANIC SALTS - Google Patents

Abstract

The present invention relates to a process for converting lignocellulosic biomass to sugars comprising at least three steps. The first step is a step of cooking the lignocellulosic biomass in the presence of at least one hydrated inorganic salt. The second step is a step of separating at least one solid fraction having undergone the firing step and the third step is a step of enzymatic hydrolysis of said solid fraction to convert the polysaccharides to monosaccharides. The sugars thus obtained are then fermentable in alcohols.

Description

Field of the Invention The present invention is part of a process for producing so-called "second generation" alcohol from lignocellulosic biomass. Background Art In the face of increasing pollution and global warming, numerous studies are currently being conducted to utilize and optimize renewable bio-resources, such as lignocellulosic biomass. Lignocellulosic biomass is composed of three main polymers: cellulose (35 to 50%), hemicellulose (23 to 32%) which is a polysaccharide essentially consisting of pentoses and hexoses and lignin (15 to 25%) which is a polymer of complex structure and high molecular weight, from the copolymerization of phenylpropenoic alcohols. These different molecules are responsible for the intrinsic properties of the plant wall and are organized in a complex entanglement. Cellulose, the majority of this biomass, is thus the most abundant polymer on Earth and the one with the greatest potential for forming materials and biofuels. However, the potential of cellulose and its derivatives has not, for the moment, been fully exploited, mainly because of the difficulty of extracting cellulose. Indeed, this step is made difficult by the very structure of the plants. The identified technological barriers related to the transformation of cellulose include its accessibility, its crystallinity, its degree of polymerization, the presence of hemicellulose and lignin. It is therefore essential to develop new methods of pretreatment of lignocellulosic biomass for easier access to cellulose and allow its transformation.

Biofuel production is an application requiring pretreatment of biomass. Indeed, the second generation of biofuel uses as load vegetable or agricultural waste, such as wood, wheat straw, or plantations with high growth potential such as miscanthus. This raw material is perceived as an alternative, sustainable solution with little or no impact on the environment and its low cost and high availability make it a solid candidate for biofuel production. 1 The principle of the process of converting lignocellulosic biomass into biofuel uses a step of enzymatic hydrolysis of the cellulose contained in plant material to produce glucose. This glucose is then fermented into ethanol, the biofuel.

However, the cellulose contained in the lignocellulosic biomass is particularly refractory to enzymatic hydrolysis, especially since the cellulose is not directly accessible to the enzymes. To overcome this refractory nature, a pretreatment step upstream of the enzymatic hydrolysis is necessary. There are many methods of treating materials rich in cellulose, chemical, enzymatic, microbiological to improve the subsequent step of enzymatic hydrolysis.

These methods are for example: steam explosion, organosolv process, dilute or concentrated acid hydrolysis or AFEX ("Ammonia Fiber Explosion") process. These techniques can still be improved and suffer in particular from costs that are still too high, problems of corrosion, low yields and difficulties of extrapolation at the industrial level (F. Talebnia, D. Karakashev, I. Angelidaki Biores, Technol 2010, 101 , 4744-4753).

In recent years, a new type of pretreatment consisting of using ionic liquids has been studied. Ionic liquids are salts consisting solely of liquid ions at temperatures of less than or equal to 100 ° C and provide highly polar media. They are thus used as solvents or as reaction media for treating cellulose or lignocellulosic materials (WO 05/17252; WO 05/23873). But like other pretreatments, ionic liquids have significant cost issues related to the price of ionic liquids, their often difficult recyclability, and their limited availability. It appears necessary to limit the pretreatment costs, in particular by using readily available and inexpensive reagents. It is in this context that the present invention falls.

SUMMARY OF THE INVENTION The present invention relates to a process for converting lignocellulosic biomass to sugars comprising at least three steps. The first step is a step of cooking the lignocellulosic biomass in a medium comprising at least one hydrated inorganic salt. The second step is a step of separating at least one solid fraction having undergone the firing step and the third step is a step of enzymatic hydrolysis of said solid fraction to convert the polysaccharides to monosaccharides. The sugars thus obtained are then fermentable in alcohols. Description of the Figures

FIG. 1 represents the kinetics of the enzymatic hydrolysis of wheat straw according to the process of the present invention, using a step of cooking the biomass in the presence of LiCl.sub.2H.sub.2 at 140.degree. FIG. 2 represents the kinetics of enzymatic hydrolysis of wheat straw according to the method of the present invention, implementing a step of cooking the biomass in the presence of LiCl.sub.2.2H.sub.2O at 140.degree. FIG. 3 represents the kinetics of the enzymatic hydrolysis of the wheat straw according to the process of the present invention, implementing a step of cooking the biomass in the presence of LiCl.sub.2H.sub.2O containing 5% by weight of LiCH.sub.3000 at 140.degree. . FIG. 4 represents the kinetics of the enzymatic hydrolysis of wheat straw according to the process of the present invention, using a step of cooking the biomass in the presence of LiBr 2H 2 O at 120 ° C. Figure 5 shows the kinetics of enzymatic hydrolysis of wheat straw on a native wheat straw, having not undergone any pretreatment. Figure 6 shows the kinetics of enzymatic hydrolysis of wheat straw when pretreated by steam explosion prior to enzymatic hydrolysis.

DETAILED DESCRIPTION OF THE INVENTION The process for converting lignocellulosic biomass into monosaccharides according to the present invention comprises at least: a) a step of cooking the biomass, at a temperature of between 120 and 250 ° C., in the presence or in the presence of absence of an organic solvent, in a medium comprising one or more hydrated inorganic salts of formula (1): MXn.n'H2O wherein X is an anion and M a metal selected from groups 1 and 2 of the periodic table, n is an integer of 1 or 2 and n is between 0.5 and 6; B) a step of separating a solid fraction having undergone the firing step; c) a step of enzymatic hydrolysis of said solid fraction. This process transforms lignocellulosic biomass into fermentable sugars with excellent yields. It also has the advantage of using inexpensive reagents, widely available and recyclable, thus obtaining a low pretreatment cost, particularly compared to a process using ionic liquids. This technology is also simple to implement and makes it easy to envisage an extrapolation at the industrial level.

Thanks to the process according to the present invention, the transformation of the lignocellulosic biomass into fermentable sugars is carried out with an excellent yield. The firing step carried out according to the process of the present invention makes it possible to reduce the duration of enzymatic hydrolysis with respect to the processes described in the prior art. The method according to the present invention makes it possible to achieve high glucose yields with short enzymatic hydrolysis times, allowing a significant gain in terms of equipment productivity since it becomes possible to use smaller amounts of enzymes and / or to reduce the size of the enzymatic hydrolysis tanks. Preferably, the medium in which the firing step is carried out consists of one or more hydrated inorganic salts of formula (1).

This firing step is carried out in the presence or absence of an organic solvent. The lignocellulosic biomass, or lignocellulosic materials used in the process according to the invention is obtained from wood (hardwood and softwood), raw or treated, by agricultural products such as straw, plant fibers, cultures. forestry, residues of alcoholic, sugar and cereal plants, residues of the paper industry, or products of transformation of cellulosic or lignocellulosic materials. The lignocellulosic materials can also be biopolymers and are preferably rich in cellulose. Preferentially the lignocellulosic biomass used is wood, wheat straw, wood pulp, rice straw or corn stalks.

According to the process of the present invention, the different types of lignocellulosic biomass can be used alone or in mixture.

In the firing step, the lignocellulosic biomass is present in an amount of between 0.5% and 40% by weight of the total mass of the lignocellulosic biomass / hydrated inorganic salt mixture, preferably in a quantity of between 3% and 25% by weight. . Preferably, the anion X is a halide anion chosen from Cl, F, Br and I, a perchlorate anion (ClO4), a thiocyanate anion (SCN), a nitrate anion (NO3) or an acetate anion (CH3000). The metal M is preferably selected from lithium, magnesium, calcium, potassium or sodium. Preferably, in the formula MX 3 H 2 O (1) of the hydrated inorganic salt, is between 0.5 and 3, preferably between 0.5 and 2. According to a preferred embodiment, the Inorganic hydrated salt is a salt of the formula LiX, nH2O, in which the anion X is chlorine or bromine. Even more preferably, the hydrated inorganic salt is a salt of the formula LiX.n'H 2 O, wherein the anion X is chlorine or bromine and is between 0.5 and 2. Even more preferably the anion X is chlorine and, by way of example, the inorganic hydrated salts used are LiCl.sub.2H.sub.2 or LiCl.sub.2.2H.sub.2O. The medium in which the biomass cooking step is carried out may consist of a mixture of different hydrated inorganic salts having the formula (1). According to the present invention, the inorganic salt corresponding to the formula (1) can be prepared in situ by the combination of a salt composed of a cation of groups 1 and 2 of the periodic table and a carbonate anion, hydrogencarbonate or hydroxide with an acid. The acid can be used pure or in aqueous solution. As examples of hydrated inorganic salts that can be used for the firing step according to the present invention, mention may be made of CaCl 2 · 6H 2 O, LiBr 2 H 2 O, CaBr 2 · 6H 2 O, KF 2H 2 O, NaCH 3 000 · 3H 2 O, LiCH 3 000 · 3H 2 O, LiCl.2H2O, LiCl.H2O. According to the process of the present invention, several successive firing steps can be carried out in a medium consisting of one or more hydrated inorganic salts corresponding to formula (1). Preferably, for the firing step a), the firing temperature is preferably between 120 and 160 ° C.

The duration of the cooking is between 0.5 minutes and 168 hours, preferably between 5 minutes and 4 hours and even more preferably between 20 minutes and 2 hours.

The step of cooking the lignocellulosic biomass can be carried out in the presence of an organic solvent, chosen from alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol and isobutanol. or tert-butanol, diols and polyols such as ethanediol, propanediol or glycerol, amino alcohols such as ethanolamine, diethanolamine or triethanolamine, ketones such as acetone or methyl ethyl ketone, dimethylformamide , dimethylacetamide, dimethylsulfoxide, acetonitrile, aromatic solvents such as benzene, toluene, xylenes, alkanes.

According to another embodiment, the step of cooking the lignocellulosic biomass can be carried out in the absence of an organic solvent.

The second step b) of the process according to the invention consists in separating a solid fraction which has undergone the firing step a) described above. This separation is generally carried out by adding at least one anti-solvent which causes the precipitation of the solid fraction. The separation of this precipitated solid fraction and a liquid fraction containing the hydrated inorganic salt and the anti-solvent can be carried out by the usual solid-liquid separation techniques. For example, the separation of the solid fraction having undergone the firing step a) described above can be carried out by filtration or by centrifugation. The solid fraction having undergone the firing step a) described above may optionally be subjected to additional treatments before the enzymatic hydrolysis step. These additional treatments may in particular be intended to eliminate the traces of hydrated inorganic salts in this solid fraction. These additional treatments may be washings with the anti-solvent. The solid fraction which has undergone the baking step a) described above may optionally be dried or pressed to increase the percentage of dry matter contained in the solid.

The anti-solvent used is a solvent or a mixture of solvents chosen from water, alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol or tert-butanol, diols and polyols such as ethanediol, propanediol or glycerol, amino alcohols such as ethanolamine, diethanolamine or triethanolamine, ketones such as acetone or methyl ethyl ketone, dimethylformamide, dimethylacetamide, dimethylsulfoxide, acetonitrile. Preferably, the anti-solvent is selected from water, methanol or ethanol.

Very preferably, the anti-solvent is water.

The hydrated inorganic salt contained in the liquid fraction can be separated from the anti-solvent and recycled. This separation can be carried out by any method known to those skilled in the art, such as, for example, evaporation, precipitation, extraction, passage over ion exchange resin, electrodialysis, chromatographic methods, solidification. hydrated inorganic salt by lowering the temperature or adding a third body, reverse osmosis.

The liquid fraction containing the hydrated inorganic salt and the antisolvent may also contain products derived from biomass. For example, the liquid fraction may contain hemicellulose (or products derived from hemicellulose) and lignin. The biomass-derived products contained in the liquid fraction can be separated before or after the hydrated inorganic salt-anti-solvent separation. The products derived from biomass may for example be extracted by addition of a solvent immiscible with the hydrated inorganic salt or with the mixture inorganic salt hydrated - anti-solvent. Products derived from biomass can also be precipitated by modifying conditions (temperature, pH, etc.) or by adding a third body.

The inorganic salt or salts thus recovered can be purified by heating at high temperatures, higher than 400 ° C., to eliminate by combustion the organic products derived from biomass potentially still present.

The third step c) of the process according to the invention is the enzymatic hydrolysis step of the cellulose-enriched solid fraction. The solid fraction which has undergone the firing step, also referred to below as pre-treated substrate is subjected to enzymatic hydrolysis to convert the polysaccharides to monosaccharides under the usual conditions normally applied in these conversion processes.

Typically, the pretreated substrate is placed in an aqueous medium to reach dry matter concentrations of between 0.5% and 40%, preferably between 1% and 20% by weight. The enzymatic hydrolysis is carried out under mild conditions, at a temperature of the order of 40 to 60 ° C, at a pH of between 4.5 and 5.5. Very preferably, the pH is between 4.8 and 5.2. If a pH adjustment is necessary, it is carried out prior to the enzymatic hydrolysis step, when the pretreated substrate is placed in an aqueous medium, in particular by adding a buffer solution.

Enzymatic hydrolysis is carried out by means of enzymes produced by a microorganism. Microorganisms, such as fungi belonging to the genera Trichoderma, Aspergillus, Penicillum or Schizophyllum, or anaerobic bacteria belonging for example to the genus Clostridium, produce these enzymes, notably containing cellulases and hemicellulases, suitable for the extensive hydrolysis of cellulose and hemicelluloses.

The duration of step c) hydrolysis varies between 1h and 150h, preferably between 2h and 72h, preferably between 4h and 24h. At the end of the enzymatic hydrolysis step, the glucose formed is soluble in water while the optionally unconverted cellulose, lignin or other products remain insoluble. The aqueous glucose solution is recovered by filtration. The monosaccharide thus obtained is easily converted into an alcohol by fermentation with yeasts such as, for example, Saccharomyces cerevisiae. The fermentation must obtained is then distilled to separate the vinasses and the alcohol produced. EXAMPLES

The substrate used in these examples is a wheat straw. The compositional analysis carried out according to the NREL protocol TP-510-42618 indicates that the composition of the raw substrate is the following: 37% cellulose, 28% hemicellulose and 20% lignin.

Examples 1 to 4 are in accordance with the invention. Example 5 is given for comparison as enzymatic hydrolysis is performed on a native wheat straw. Example 6 is a comparative example, the pretreatment carried out consisting of a steam explosion, currently known as one of the best technologies for obtaining fermentable sugars. EXAMPLE 1 Pretreatment of wheat straw by LiCl.sub.2H.sub.2 O at 140.degree. C. and enzymatic hydrolysis 38 g of LiCl.sub.2H.sub.2 O and 2 g of wheat straw (500-1000 .mu.m) are placed in a 170 ml flask and shaken mechanically by pale at 140 ° C for 1 hour in a Tornado shaker equipped with a carousel 6 positions (Radleys). After this cooking (step a) of the process according to the invention), the heating is stopped and 80 ml of distilled water are rapidly added to the mixture: the pretreated straw precipitates. The suspension containing the inorganic salt, water and biomass is placed in a centrifuge tube and stirred at 9500 rpm for 10 minutes. The supernatant containing the inorganic salt is then separated from the solid. The operation is repeated three times by adding 80 ml of distilled water to the solid part still present in the centrifugation tube. 3.89 g of solid, with a solids content of 35%, are recovered.

The solid recovered after precipitation and washing is subjected to enzymatic hydrolysis. Half of the recovered solid is placed in a 100 ml Schott flask. 5 ml of acetate buffer, 10 ml of a 1 wt% solution of NaN 3 in water are added and the mixture is then made up to 100 g with distilled water. This solution is then left overnight at 50 ° C for "activation" with stirring of 550 rpm in a STEM dry water bath. Then added to the solution are known amounts of enzyme: -Cellulases XL508, 10FPU per gram of dry matter - R-glucosidases NOVOZYM 188, 25 CBU per gram of dry matter The solution is then stirred at 400 rpm at 50 ° C. ° C, always in dry water baths, and samples are taken after 1 hour, 4 hours and 7 hours. These samples are placed in centrifuge tubes and rapidly placed for 10 minutes in an oil at a temperature of 103 ° C. to neutralize the enzymatic activity. Centrifuge tubes are stored in a refrigerator at 4 ° C pending measurement of glucose. They are then diluted by 5 with distilled water before being assayed using the Analox GL6 analyzer apparatus, called glucostat, which measures by enzymatic assay the glucose concentration in aqueous solutions. The result of the enzymatic hydrolysis is shown in FIG. 1. It is expressed in glucose yield, defined as the ratio of the glucose concentration in the solution to the theoretical maximum concentration according to the cellulose content of the native wheat straw. . This glucose yield therefore represents the percentage of the cellulose actually converted into glucose.

EXAMPLE 2 Pretreatment of Wheat Straw by LiCl 2 H 2 O at 140 ° C. and Enzymatic Hydrolysis The protocol is identical to that of Example 1 except that 38 g of LiCl 2 H 2 O are used in place of 38 g. LiCI.H2O. The result of the enzymatic hydrolysis is shown in FIG.

EXAMPLE 3 Pretreatment of Wheat Straw at 140 ° C. by LiCl.sub.2H.sub.2 Containing 5% by Weight of LiCH3OOO.3H2O and Enzymatic Hydrolysis

The protocol is identical to that of Example 1 except that 38 g of LiCl.sub.2H.sub.2 containing 5% by weight of LiCH.sub.3000.sub.3H.sub.2O.sub.2 are used in place of 38 g of LiCl.sub.2H.sub.2O. The result of the enzymatic hydrolysis is shown in FIG.

EXAMPLE 4 Pretreatment of Wheat Straw at 120 ° C. by LiBr.2H2O and Enzymatic Hydrolysis The protocol is identical to that of Example 1 except that 38 g of LiBr.2H2O are used in place of 38 g of LiCl.H2O and that the firing step is carried out at 120 ° C. The result of the enzymatic hydrolysis is shown in FIG.

Example 5 (not in accordance with the invention): Enzymatic hydrolysis of native wheat straw (without pretreatment)

The protocol used for the enzymatic hydrolysis is identical to that described in Example 1. The wheat straw is used native, without any pretreatment. The result of the enzymatic hydrolysis is shown in FIG. 5. Example 6 (not in accordance with the invention): Enzymatic hydrolysis of wheat straw resulting from a pretreatment by steam explosion.

The protocol used for enzymatic hydrolysis was identical to that described in Example 1. The wheat straw used was pretreated by steam explosion, pretreatment whose three steps are described below. 1. Impregnation of wheat straw with 0.1 N sulfuric acid for a minimum of 8 hours followed by draining and pressing (100 bar) of the straw to obtain a solid at about 30% dry matter . 2. Pretreatment at 210 ° C (18 bar) of the straw for 2.5 minutes then decompression. 3. Final treatment by pressing at 100 bar. The result of the enzymatic hydrolysis is shown in FIG.

From the various representative figures of each of the examples, it appears that the process according to the present invention makes it possible to obtain rapidly (after 1 hour of enzymatic hydrolysis) glucose yields higher than those obtained without pretreatment, or even after pretreatment type steam explosion, since in this case, the glucose yield at one hour is only about 20%. In the same manner, the glucose yields obtained after 7 hours of enzymatic hydrolysis are improved compared to those of the reference technology (steam explosion), when the firing step is carried out according to the process described in present invention.

Claims (13)

REVENDICATIONS1. A process for converting lignocellulosic biomass into monosaccharides comprising at least: a) a step of cooking the biomass, at a temperature between 120 and 250 ° C, in the presence or absence of an organic solvent, in a medium comprising one or several hydrated inorganic salts of formula (1) MXn.n'H2O wherein X is an anion and M a metal selected from groups 1 and 2 of the Periodic Table, n is an integer of 1 and 2 and n 'is between 0.5 and 6; b) a step of separating a solid fraction having undergone the firing step a); c) a step of enzymatic hydrolysis of said solid fraction. 2. Conversion process according to the preceding claim wherein the medium in which the firing step is carried out consists of one or more hydrated inorganic salts of formula (1). 3. Method according to one of the preceding claims wherein the anion X is a halide anion selected from Cl, F, Br, I, a perchlorate anion, a thiocyanate anion, a nitrate anion or an acetate anion. 25 4. Method according to one of claims 1 to 3 wherein the metal M is selected from lithium, magnesium, calcium, potassium or sodium. 5. Method according to one of the preceding claims wherein the hydrated inorganic salt has the formula (1) wherein n 'is between 0.5 and 3, preferably between 0.5 and 2. 6. Method according to one of claims 1 to 5 wherein the hydrated salt is a salt of formula LiX.n'H2O, wherein the anion X is chlorine or bromine. 35 7. Method according to one of claims 1 to 6 wherein the lignocellulosic biomass is present in an amount between 0.5% and 40% by weight of the totalmass of the lignocellulosic biomass / hydrated inorganic salt mixture, preferably in an amount between 3% and 25% weight. 8. Method according to one of claims 1 to 7 wherein the firing step a) is carried out in a medium consisting of a mixture of different hydrated inorganic salts of formula (1). 9. Method according to one of claims 1 to 8 wherein the firing step is carried out at a temperature between 120 and 160 ° C. 10. Process according to claims 1 to 9 wherein the cooking time is between 0.5 min and 168 h, preferably between 5 minutes and 2 hours. 11. Method according to one of claims 1 to 10 wherein the step of firing the lignocellulosic biomass is carried out in the presence of an organic solvent selected from alcohols such as methanol, ethanol, n- propanol, isopropanol, n-butanol, isobutanol or tert-butanol, diols and polyols such as ethanediol, propanediol or glycerol, amino alcohols such as ethanolamine, diethanolamine or triethanolamine ketones such as acetone or methyl ethyl ketone, dimethylformamide, dimethylacetamide, dimethylsulfoxide, acetonitrile, aromatic solvents such as benzene, toluene, xylenes, alkanes. 12. Method according to one of claims 1 to 11 wherein the step of separating the cellulose enriched solid fraction is carried out by precipitation by addition of at least one anti-solvent selected from water, alcohols such as methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol or tert-butanol, diols and polyols such as ethanediol, propanediol or glycerol, amino alcohols such as ethanolamine, diethanolamine or triethanolamine, ketones such as acetone or methyl ethyl ketone, dimethylformamide, dimethylacetamide, dimethylsulfoxide, acetonitrile. 13. Method according to one of claims 1 to 12 wherein the enzymatic hydrolysis is carried out at a temperature of the order of 40 to 60 ° C, at a pH between 4.5 and 5.5 for a period of time. varying from 1 to 150h. 35