KR101438549B1 - Method of producing hydrolysate from sea algae using acidic ionic liquid catalysts - Google Patents

Abstract

The present invention relates to a method for producing a saccharified liquid derived from algae using an ionic liquid as a saccharification catalyst and a method for producing biofuel. More particularly, the present invention relates to a method for preparing a polysaccharide material extracted from seaweed basidiomycetes or seaweeds by treating an ionic liquid with a saccharification catalyst A method for producing a saccharified liquid comprising the step of producing a monosaccharide; And a method of fermenting the monosaccharide with a microorganism. Since the method of producing biofuel according to the present invention uses marine algae as a raw material for biomass, it is possible to drastically improve the problem of raw material supply and demand, and to suppress or minimize 5-hydroxymethylfurfural (5-HMF) . In addition, since the lignin removing step, which is essential for the use of conventional woody raw materials, is not required, the process cost can be lowered, which is economical and environmentally advantageous.

Seaweeds, ionic liquids, biofuels, 5-HMF, fermentation inhibitors

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing a saccharide-derived saccharide liquid from a seaweed using an acidic ionic liquid as a catalyst,

More particularly, the present invention relates to a method for producing a 5-hydroxymethylfurfural (5-HMF) inhibitory substance when fermenting marine algae as a raw material, The present invention relates to a method for producing a saccharified liquid using an ionic liquid as a catalyst.

Biofuels are energy obtained by using biomass as a raw material and obtained by direct combustion, alcohol fermentation, methane fermentation, and the like. Biomass, which is a raw material for biofuels, is mainly divided into carbohydrates (sugar cane, beet), starchy maize (corn, potatoes, sweet potatoes) and woody plants (wood, rice straw, However, in the case of starch and woody materials, biofuels are produced through a fermentation process using an appropriate pretreatment process and a glycosylation process after saccharification process. can do. The woody system can be used as a raw material for waste materials in the form of municipal waste or forestous by-products scattered throughout the forests. However, since it is not worth using as food, the stability of raw material supply can be secured. However, lignin removal pretreatment And a crystalline structure composed of hydrogen bonds, which is characteristic of a woody cellulose substrate, has a disadvantage in that the saccharification yield is low and the economical efficiency is low.

Successful commercialization of biofuels as alternative fuels for transportation is to secure price competitiveness of biofuels such as bioethanol compared to gasoline. Generally, the proportion of raw material costs and process costs in biofuel production costs varies greatly depending on the type and process of biomass. For example, in case of saccharides using sugar cane or sugar beet, the ratio of raw material: process is about 75:25, while that of starch such as corn, potato, and cassava is about 50:50 and that of wood is about 25:75 to be.

However, with the exception of lignocellulosic, biofuels production technology that is currently commercialized is not only a matter of using food as an energy source because humans use carbohydrates or starchy materials that can be used as food, Supply and demand problems can arise. In terms of economy, the use of grain is a problem in terms of raw material costs. In addition, corn cultivation requires not only a considerable amount of pesticide and nitrogen fertilizer, but also environmental disadvantages such as severe erosion of soil or emission of carbon dioxide compared with other crops.

As of 2006, bioethanol has been produced in about 51.3 billion liters worldwide. The global production of biofuels, specifically bioethanol, using saccharides is about 18.7 billion liters (2006), with Brazil, India, and Taiwan as major producers, followed by Brazil with 17.8 billion liters (Global Bioenergy Partnership (GBEP), 2006). Brazil is actively producing bioethanol for transporting sugarcane, which is an abundant resource, as raw materials. Actually, various types of ethanol-mixed gasohol are spreading. In 2003, FFV (Flexible Fuel Vehicle), which can be operated even when the ethanol and gasoline contents have changed, has begun to be marketed. As of May 2005, it occupied about 50% of total passenger car sales.

Worldwide production of corn-based bioethanol is about 19.8 billion liters (2006), with the major producers being the US, Europe and China, with the United States leading the world to produce 18.5 billion liters (see Table 1). The US has enacted the Energy Tax Act in 1978, shortly after the oil crisis, and is expanding its supply by giving $ 4 per gallon of federal tax breaks for gasoline containing less than 10% ethanol. As such, the United States is actively producing biofuels for transporting large crops and corn as a source of abundant resources, and is developing advanced energy technologies that will escape from the oil-dependent economy through the development of new and renewable energy technologies. As part of its efforts to develop ethanol production technology, the use of ethanol for corn-based ethanol production is on the rise.

There is no observed industry trend because biofuel production technology using woody system has not reached commercialization stage yet. However, in Canada, Iogen is actively developing wood biomass manufacturing technology. In order to commercialize technology for extracting ethanol from agricultural wastes and plant raw materials as next generation biomass by 2012, the US budget is set at $ 150 million , Which is in the process of replacing 30% of the total transport fuel with ethanol.

Seaweeds are divided into macroalgae and microalgae. Major algae include red algae, brown algae, green algae, and microalgae such as chlorella and spirulina. Seaweeds are much better than other conventional biomass (4 to 6 times a year in the case of subtropical regions) and can be exploited in the wider seas, so the available cultivation area is wide and the cost of freshwater, land, fertilizer, etc. The use of high resources is advantageous. In addition, since the lignin component, which must be removed in the case of wood, is not present, the production process of the biofuel is simple and the total energy conversion yield is high. In addition, seaweed has an annual carbon dioxide absorption of 36.7 tonnes per hectare, which is 5 to 7 times higher than woody, and assuming that E20 (20% ethanol-added gasoline) is used, annual greenhouse gas reduction rate is about 27% , Which translates into a carbon tax savings of about KRW 300 billion (Table 1).

[Table 1] Characteristics of land plants and marine plants

division Athletic plants Marine plant Sugar / starch family Woody Seaweed Raw material Sugarcane, corn Timber Cockatoo, Cocktail, Kotoni Raw material production cycle 1-2 times a year At least 8 years 4 to 6 times a year Raw material production per unit area (tonne / ha) 180 9 565 CO ₂ absorption per unit area (ton / ha) 5-10 4.6 36.7 Manufacture process simple Complexity (lignin removal) Simple (lignin-free) Cultivation environment Sunlight, CO ₂ , fresh water, land, fertilizer Sunlight, CO ₂ , fresh water, land, fertilizer Sunlight, CO ₂ , seawater

However, seaweeds have so far been mainly used in fine chemical materials such as electrophoresis reagents, fertilizers, emulsifiers, anticancer agents, and medical materials, or as health foods such as edible and medicinal drugs, and there is no research on biofuel development using them to be.

SUMMARY OF THE INVENTION The present invention has been made in order to solve the problems of the conventional biofuel production method. By using seaweed instead of saccharide, starch or lignocellulosic material used as a raw material of conventional biofuel, it is possible to overcome the low saccharification efficiency in the case of using raw materials and to use the ionic liquid having acidic group as a saccharification catalyst, It is an object of the present invention to provide a method for producing a glycosylated liquid capable of inhibiting or minimizing the formation of 5-HMF (5-hydroxymethylfurfural) as an inhibitory substance, and a method for producing biofuel using the glycosylated liquid.

The present invention provides a method for producing a saccharified liquid, which produces a monosaccharide by treating an acidic ionic liquid catalyst with a polysaccharide material extracted from algae seeds or seaweeds.

The present invention also provides a process for producing a saccharified liquid characterized in that the acidic ionic liquid catalyst is selected from the group consisting of imidazolium, phosphonium, ammonium, morpholinium and choline represented by the following Chemical Formulas 1 to 5 to provide:

Wherein R ¹ , R ² , R ³ and R ⁴ are each independently C ₁ -C ₈ alkyl, phenyl, benzyl or fluorinated alkyl, A is SO ₄ , SO ₃ , BH ₃ , n ₃ ) (n = 1 or 2), CF ₃ COO, CH ₃ COO, HPO ₄ , p-CH ₃ (C ₆ H ₄ ) SO ₃ .

The present invention also relates to a process for the preparation of the acidic ionic liquid wherein the acidic ionic liquid catalyst is selected from the group consisting of [Rmim] [HSO ₄ ] (1-alkyl-3-methylimidazolium hydrogenesulfate), [Hmin] [HSO ₄ ] (1-methylimidazolium hydrogensulfate) (1-alkylimidazolium triflouroacetate), [Rmim] [H ₂ PO ₄ ] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al ₂ Cl ₆ ] ₁ to C ₈ ). The present invention also provides a method for producing a saccharified liquid.

The present invention also relates to a process for the preparation of the compound of formula I wherein the extraction solvent is selected from the group consisting of H ₂ SO ₄ , HCl, HBr, HNO ₃ , CH ₃ COOH, HCOOH, HClO ₄ , H ₃ PO ₄ , PTSA and conventional solid acids The present invention also provides a method for producing a saccharified liquid.

Also, the present invention provides a method for producing a saccharified liquid, wherein the polysaccharide material is selected from the group consisting of horseradish, starch, carrageenan, alginic acid and fibrin.

The present invention also provides a method for producing a saccharified liquid, wherein the algae are large algae or microalgae.

The present invention also provides a method for producing a saccharified liquid, wherein the large algae is selected from the group consisting of red algae, brown algae and green algae.

The present invention also relates to a method for producing the red algae according to the present invention, wherein the red algae are selected from the group consisting of Cryptomeria japonica, Kottoni, dog gambling, rosemary, round starch, Wherein the saccharide is selected from the group consisting of saccharides.

In addition, the present invention relates to the above-mentioned composition, wherein the brown algae is selected from the group consisting of seaweed, kelp, horses, horsetail, lobster, horsetail, vinegar, horse mackerel, Wherein the saccharification solution is selected from the following.

In addition, the present invention provides a method for producing a saccharified liquid, wherein the green algae are selected from the group consisting of chrysanthemum, sunflower, parasitic, audible, beaded, octagonal and beaded.

The present invention also provides a method for producing a saccharified liquid characterized in that the monosaccharide is selected from the group consisting of galactose, a galactose derivative, 3,6-anhydrogalactose, glucose, fucose, rhamnose, xylose and mannose to provide.

The present invention also relates to a method for producing a saccharified liquid, which comprises reacting the monosaccharide with an acidic ionic liquid catalyst at a concentration of 0.05 to 30% to provide.

The present invention also relates to a process for producing a saccharified liquid characterized by that the monosaccharide is produced by reacting cellulose with an acidic ionic liquid catalyst at a concentration of 0.05 to 50% at a temperature of 80 to 300 ° C. for more than 0 to 6 hours .

The present invention also relates to a method for producing a saccharified liquid, which comprises reacting the monosaccharide with an acidic ionic liquid catalyst at a concentration of 0.05 to 50% based on the seaweed base at a temperature of 60 to 300 ° C. for more than 0 to 6 hours .

In addition, the present invention relates to a method for producing a starch-containing starch, which comprises reacting the monosaccharide with an acidic ionic liquid catalyst at a concentration of 0.05 to 50% based on the seaweed starch at a temperature of 60 to 300 ° C. for more than 0 to 6 hours, Wherein the saccharide is produced by a secondary or tertiary saccharification reaction under the reaction conditions.

The present invention also relates to a method for treating a polysaccharide material extracted from a seaweed or seaweed to produce a monosaccharide by treating an acidic ionic liquid catalyst; And

The present invention also provides a method for producing a biofuel comprising the step of fermenting the monosaccharide with a microorganism.

The present invention also provides a method for producing a biofuel, wherein the biofuel is selected from the group consisting of C ₁ to C ₄ alcohols and C ₂ to C ₄ ketones.

The present invention also relates to a method for producing a fermented microorganism, wherein the fermenting microorganism is selected from the group consisting of Saccharomyces cerevisiae, Sarsinia ventriculi, Kluyveromyces flajilis, Zygomomonas mobilis, Kluyveromyces maxinus IMB3, , Clostridium acetyobutylicum, Clostridium bi-lingki, Clostridium aurantibutylicum and Clostridium tetanomorphi. The present invention also provides a method for producing a biofuel comprising the steps of: .

Hereinafter, the present invention will be described in detail.

The method for producing a saccharified liquid of the present invention or the method for producing a biofuel,

Treating an acidic ionic liquid catalyst to a polysaccharide material extracted from a seaweed source or seaweed to produce a monosaccharide; And

And fermenting the monosaccharide with a microorganism.

In the present invention, the biofuel may include C ₁ to C ₄ alcohols, C ₂ to C ₄ ketones, and the like, preferably methanol, ethanol, propanol, butanol or acetone. It is not. Examples of the polysaccharide material include horseradish, starch, fibrin, carrageenan, and alginic acid, but are not limited thereto.

The large algae include red algae, brown algae, and green algae, and microalgae include chlorella and spirulina. The algae used in the biofuel production method of the present invention may be any of large algae or microalgae. As the red algae, there may be used red ginseng, red ginseng, red ginseng, round ginseng, red ginseng, green ginseng, green ginseng, green ginseng, true gambling, However, the present invention is not limited to this, and it is preferable to use the mugwort. On the dry weight basis, cellulose is about 15 ~ 25%, and galactan is about 50 ~ 70% of the main component. In addition to less than 15% protein and less than 7% consists of lipids. The brown algae may be selected from the group consisting of seaweed, kelp, horsetail, marine horses, lobster, horsetail, seagrass, mackerel, rhubarb, rhubarb, cumin, mosquito, It is not. Brown algae are multi-cellular bodies and are best differentiated among algae. Examples of the green algae include, but are not limited to, chrysanthemum, chestnut, chestnut, hearth, beard, jade, salt, and the like. The green algae have chlorophyll and make the whole classification by photosynthesis. The brown algae contain about 30 to 40% of alginic acid and about 5 to 6% of fibrin. The green algae contain about 40 to 50% of carbohydrates, about 5 to 10% of fibrin, .

Galactan is mainly composed of galactanose polymer composed of galactose polymer, and galactan can be converted into a monosaccharide such as galactose and 3,6-anhydrogalactose through an appropriate low molecular weight process. Fibrin is a substance made of cellulose. In the case of Fagus crenata, it accounts for about 15 to 25% of the total components. The cellulose may be converted to glucose, a monosaccharide, through a saccharification process using an appropriate enzyme or an acid catalyst. The above-described galactose and glucose are used as precursors that can be converted into biofuels through a fermentation process.

Starch, also called starch, is a carbohydrate that is made by photosynthesis in chloroplasts of green plants and is a polysaccharide composed of glucose as a constituent unit. The starch may be converted to glucose, a monosaccharide, through a glycosylation process using an appropriate enzyme or acid catalyst.

The method for extracting a polysaccharide substance such as hyaline, fibrin, starch, carrageenan, alginic acid and the like from the seaweed is not particularly limited, and any method known in the art can be used. According to a preferred embodiment, seaweeds are immersed in an aqueous alkali solution for a certain period of time, washed with water, and the washed seaweeds are immersed in an extraction solvent consisting of acidic chemicals for a certain period of time to extract components such as hyaluronidase, carrageenan and alginic acid, Through the step of collecting the whole classification, it is possible to extract horseradish, carrageenan, alginic acid component and starch or fibrin. At this time, the extraction temperature is not particularly limited, but is preferably in the range of 80 to 150 ° C. Examples of the acidic chemicals include H ₂ SO ₄ , HCl, HBr, HNO ₃ , CH ₃ COOH, HCOOH, HClO ₄ (perchloric acid), H ₃ PO ₄ (phosphoric acid), PTSA (para-toluene sulfonic acid) And the like. However, the aqueous alkali solution is not limited to potassium hydroxide, sodium hydroxide, calcium hydroxide, aqueous ammonia, and the like, but is not limited thereto.

A monosaccharide can be obtained by treating a polysaccharide substance extract such as starch, starch, cellulose, carrageenan, alginic acid and the like with a hydrolysis catalyst containing an appropriate acidic ionic liquid and saccharifying it. Examples of the monosaccharide include, but are not limited to, galactose, 3,6-anhydrogalactose, glucose, fucose, rhamnose, xylose, mannose and the like.

The saccharification process of the present invention can be roughly classified into a direct saccharification method and an indirect saccharification method, and the following two saccharification processes and a saccharified liquid obtained by the method will be described.

First, the indirect saccharification method is a method of extracting a polysaccharide substance from a seaweed source and then converting the polysaccharide substance into a monosaccharide. As an example of a saccharification method using an indirect saccharification method, a method of saccharification using a succinate as a starting material will be described. Galactan, which is a galactose polymer, accounts for most of the components, and the galactan can be converted to galactose or 3,6-anhydrogalactose, which is a monosaccharide capable of fermentation through a suitable saccharification process (low molecular weight process). In this case, the hydrolysis method using an acidic ionic liquid catalyst is used as a method used in the saccharification process. Examples of usable catalysts include [Rmim] [HSO ₄ ] (1-alkyl-3-methylimidazolium hydrogenesulfate) _{HSO 4] (1-methylimidazolium hydrogensulfate} ), [Rmin] [TFA] (1-alkylimidazolium triflouroacetate), [Rmim] [H 2 PO 4] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al 2 Cl ₆ ] (1-alkyl-3-methylimidazolium chloroaluminate) (R = C ₁ -C ₈ ). At this time, it is preferable that the saccharification yield of galactose is maximized by setting the concentration of the acid used, the reaction temperature and the reaction time, and at the same time, the produced galactose is over-cleaved so that 5-HMF is not produced. According to a preferred embodiment of the present invention, the monosaccharide is selected from the group consisting of [Rmim] [HSO ₄ ] (1-alkyl-3-methylimidazolium hydrogenesulfate), [Hmin] [HSO ₄ ] (1-methylimidazolium alkylimidazolium triflouroacetate, [Rmim] [H ₂ PO ₄ ] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al ₂ Cl ₆ ] (1-alkyl- 3-methylimidazolium chloroaluminate (R = C ₁ to C ₈ ) at 60 to 200 ° C for 0 to 6 hours.

As a saccharification method using an indirect saccharification method, a method of saccharification using cellulose as a starting material will be described. Fibrin is a substance made of cellulose and can be hydrolyzed and converted to glucose using an acidic ionic liquid catalyst. As a catalyst for hydrolyzing cellulose, [Rmim] [HSO ₄ ] (1-alkyl-3-methylimidazolium hydrogenesulfate), [Hmin] [HSO ₄ ] (1-methylimidazolium hydrogensulfate), [Rmin] [TFA] etc. triflouroacetate), [Rmim] [H2PO4 ] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al 2 Cl 6] (1-alkyl-3-methylimidazolium chloroaluminate) (R = C 1 ~ C 8) In this case, it is preferable to maximize the saccharification yield of glucose by controlling the concentration of the used acid, the reaction temperature and the reaction time, and at the same time, to prevent the produced glucose from being overdispersed to produce 5-HMF. According to one preferred embodiment of the present invention, the monosaccharide comprises [Rmim] [HSO ₄ ] (1-alkyl-3-methylimidazolium hydrogenesulfate), [Hmin] [HSO ₄ ] alkylimidazolium triflouroacetate, [Rmim] [H ₂ PO ₄ ] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al ₂ Cl ₆ ] (1- alkyl-3-methylimidazolium chloroaluminate (R = C ₁ -C ₈ )) at a temperature of 80 to 300 ° C. for more than 0 to 6 hours. According to another preferred embodiment of the present invention,

As a saccharification method using an indirect saccharification method, a method of saccharification using starch as a starting material will be described. Starch is an acidic ionic liquid catalyst which is made of glucose and hydrolyzes starch [Rmim] [HSO ₄ ] (1-alkyl-3-methylimidazolium hydrogenesulfate), [Hmin] [HSO ₄ ] (1-methylimidazolium hydrogensulfate) , [Rmin] [TFA] ( 1-alkylimidazolium triflouroacetate), [Rmim] [H 2 PO 4] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al 2 Cl 6] (1-alkyl-3- methylimidazolium chloroaluminate (R = C ₁ ~C ₈ ), etc., and by controlling the concentration of acid used, the reaction temperature and the reaction time, the yield of glucose is maximized and glucose produced is overdispersed So that 5-HMF is not produced.

The saccharification method using the direct saccharification method can be carried out by using a seaweed starch containing both fibrin and / or hyaluronic acid, carrageenan or starch and / or alginic acid and fibrin as starting materials and separately extracting the above substances without extracting the polysaccharide material And then directly saccharified with a monosaccharide. At this time, a hydrolysis method using an acidic ionic liquid as a catalyst can be used. An acidic ionic liquid catalyst system used in the indirect saccharification step as described above can be used. At this time, the conditions under which the saccharification yield of glucose and galactose are maximized by appropriately adjusting the concentration of the acid catalyst, the reaction temperature, and the reaction time, and the reaction conditions that the generated monosaccharide is excessively decomposed to prevent 5-HMF from being generated It is important. In case of saccharification using seaweed as a starting material, the collected seaweeds are washed with impurities and washed through a washing process, then completely dried using a hot-air drier or a natural drying method, and the dried seaweeds are finely pulverized It is preferable to use after the particles are crushed and converted into a fine powder form. According to a preferred embodiment of the present invention, the monosaccharide is selected from the group consisting of [Rmim] [HSO ₄ ] (1-alkyl-3-methylimidazolium hydrogenesulfate), [Hmin] [HSO ₄ ] alkylimidazolium triflouroacetate, [Rmim] [H ₂ PO ₄ ] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al ₂ Cl ₆ ] (1- alkyl-3-methylimidazolium chloroaluminate (R = C ₁ -C ₈ ) at a temperature of 60 to 300 ° C. for more than 0 to 6 hours. According to another preferred embodiment of the present invention, the monosaccharide is selected from the group consisting of [Rmim] [HSO ₄ ] (1-alkyl-3-methylimidazolium hydrogenesulfate), [Hmin] _{[HSO 4] (1-methylimidazolium} hydrogensulfate), [Rmin] [TFA] (1-alkylimidazolium triflouroacetate), [Rmim] [H 2 PO 4] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al 2 Cl ₆ ] (1-alkyl-3-methylimidazolium chloroaluminate) (R = C ₁ -C ₈ ) acidic ionic liquid at a temperature of 60 to 300 ° C for more than 0 to 6 hours And then subjecting the residual fibrin or starch to a secondary or tertiary saccharification reaction under the above reaction conditions.

The saccharified liquid containing galactose, 3,6-anhydrogalactose, glucose, or a mixture thereof produced as described above can be converted into a bioalcohol using a strain for biofuel fermentation, such as yeast. A yeast for fermentation can be used in the present invention is Clostridium acetonitrile unit Tilikum (Clostridium acetobutylicum ), C lostridium beijerinckii), Clostridium brother is unsurpassed Tilikum (Clostriduim aurantibutylicum) or Clostridium Te Gaetano know fume (Clostridium tetanomorphum ), but are not limited thereto, and they are more preferable for fermentation of butanol and acetone. In addition, Saccharomyces < RTI ID = 0.0 > cerevisiae , Sarcina < RTI ID = 0.0 > ventriculi), Cluj Vero My process infrastructure jilriseu (Kluyveromyces fragilis , Zygomomonas mobilis ) or Kluyveromyces marxianus ) IMB3, Brettanomyces ( Brettanomyces custersii ) and the like, which are more preferable for ethanol fermentation.

In particular, bio-butanol having similar components to gasoline among biofuels is a bio-butanol which satisfies the criteria of good fuel, including characteristics such as energy density, volatility control, sufficient octane number, low impurity, etc., Is very similar to gasoline fuel, and the energy density of biobutanol is close to that of unleaded gasoline. Unlike bioethanol, biobutanol does not cause phase separation even in the presence of water, and has a low oxygen content, which makes it possible to mix high-concentration biobutanol with gasoline.

5-hydroxymethylfurfural (5-HMF), which is known as an inhibitor of subsequent microbial fermentation, is used as a saccharification solution by using an ionic liquid according to the present invention as a saccharification catalyst, as compared with a saccharification method using acidic hydrolysis of algae using conventional sulfuric acid. It is possible to maximize the production yield of the biofuel using the existing saccharified liquid. In addition, since the lignin-removing step, which is essential for the use of conventional wood-based raw materials, is not required, the process cost can be lowered. In addition to glucose, most of sugar components contained in seaweed such as galactose, 3,6-anhydrogalactose, It can not only solve the energy resource problem but also can absorb CO _{2 in} seaweeds, contributing to the reduction of GHGs in the whole country and actively coping with international environmental regulations. Economical and environmentally advantageous.

Hereinafter, the present invention will be described in detail with reference to examples.

However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

In this embodiment, experiments were carried out using the following saccharification apparatuses, experimental materials and analytical methods.

One. Glycation  Device

A glycosylation system composed of a reactor and a control box for performing glycation experiments is shown in Fig. The reactor was cylindrical with an internal volume of 12.5 cm and an internal diameter of 7 cm with a capacity of 500 ml (effective volume: 400 ml), and a temperature jacket was installed to reach the set reaction temperature. A thermocouple was installed to measure the internal temperature of the reactor, and cooling water was circulated outside the reactor to prevent overheating. To facilitate sampling during the reaction, high-pressure N ₂ gas can be introduced from the outside of the reactor, an N ₂ gas reservoir and a sample port are mounted, a control box is equipped with an RPM meter, a digital temperature controller and a pressure gauge .

2. Experimental material

2.1. temperament

In this example, Moroccan mugwort, Jeju mugweed, Cocktail, and Kotoni were used as the red algae, and audible as green algae and kelp as brown algae.

In this example, the direct saccharification method using direct sugarcane as a raw material and the indirect sugarcake method using raw materials of fibrin and hyaluronic acid separated / extracted from an eelgrass were conducted. For the direct glycation method, the mugwort was washed with distilled water, dried at 40 ° C and pulverized, and then pulverized into 106 and 300 mesh. For the indirect saccharification method, Uumu was dipped in KOH aqueous solution for a certain period of time, washed with distilled water, semi-dried, extracted with distilled water or ethyl alcohol or methyl alcohol, dried and pulverized at 40 ° C., After the bamboo fiber was extracted, the remaining fiber was bleached twice with O ₃ (1 h / 1 time), bleached twice with CIO ₂ (1.5 h / 1 time) at 60 ° C., and then washed with H ₂ O ₂ 2 times (1 h / once) bleached.

2.2. Strain and medium

In this example, Saccharomyces cerevisiae DKIC413 and Brettanomyces custersii kccm 11490 were used. YPD medium (yeast extract 10 g / l, peptone 20 g / l, dextrose 20 g / l) was used. The medium was used in a high pressure reactor (Woosung Scientific co., Korea) after sterilization at 121 ° C for 15 minutes.

3. Analysis method

3.1. Party analysis

Carbopac PA 1 (4250 ㎜, Dionex Co., USA) and Carbopac PA 1 (450 ㎜, Dionex Co., USA) were used for analysis of the saccharified solution using HPLC equipped with current detector (ICS-3000, Dionex Co., USA) , USA) was used as a column. The mobile phase was a 16 mM NaOH solution, the flow rate was 1 ml / min, and the column temperature was 30 ° C. Concentrations of glucose and galactose were quantitatively analyzed using calibration curves of standard materials, and the yields of glucose and galactose were calculated as the yields of glucose and galactose produced relative to the total carbohydrates contained in the dry ingredients according to Equation 1. [

C = concentration of glucose or galactose (g / l)

V = Total amount of solvent used in saccharification (l)

S = Amount of total carbohydrate contained in the dry ingredients (g)

3.2. Protein analysis ( Semi - micro Kjeldahl method)

In order to analyze the protein, 0.5 g of the sample was taken into a proteolytic tube, and 20 ml of sulfuric acid and a decomposition accelerator (K ₂ SO ₄ : CuSO ₄ .5H ₂ O = 9: 1) was added to the reaction solution to decompose the protein. After the decomposition, 70 ml of distilled water was added, and 75 ml of 32% NaOH was added to the distiller, followed by distillation using a protein distillation apparatus. Ammonia generated by distillation was collected with 100 ml of 3% boric acid, titrated with 0.1 N HCl, and the total nitrogen content was calculated according to Equation 2.

V ₀ = 0.1 N HCl consumption of the blank (ml)

V ₁ = 0.1 N HCl consumption (ml) of this sample,

f = Factor of 0.1 N HCl

N = Nitrogen Coefficient

s = amount of sample (mg)

0.0014: Amount of nitrogen (g) equivalent to 1 ml of 0.1 N HCl

3.3. Batch analysis (dry painting method)

The crucible was heated in a painting furnace at 550 DEG C until the weight became constant, and then cooled in a desiccator and weighed. 2 g of the sample was placed in a weighed crucible, and the mixture was stirred at 550 ° C until the white or off-white material remained. The mixture was allowed to cool to 200 ° C in a painting furnace and cooled to room temperature. The ash content (%) was calculated according to Equation 3.

W ₁ = Constant weight of container (g)

W ₀ = Container after painting + amount (g)

S = sample weight (g)

3.4. Cell density measurement

The cell concentration was measured at 600 nm using a spectrophotometer (Genesys 10-S, Thermo electron corp., USA). The cell culture was centrifuged at 3,500 rpm for 10 minutes using a centrifuge (VS-150FN, Vision Science Co., LTD., Korea), washed with distilled water and centrifuged. And dried at 50 DEG C for 24 hours. The dry cell weight in the case of Saccharomyces cerevisiae was 0.3135 absorbance + 0.1811 (correlation coefficient = 0.994) in the case of Saccharomyces cerevisiae, the cell dry weight in the case of Bretania maizeuscusterius = 0.1292 absorbance + 0.8554 = 0.999).

3.5. Ethanol analysis

The concentration of ethanol in the culture was analyzed using HPLC (Breeze HPLC system, Waters Co., USA) equipped with RI detector. The column was Aminex HPX-87H (3007.8 mm, Bio-rad). The mobile phase used was a 5 mM aqueous solution of sulfuric acid, the flow rate was 0.6 ml / min, the temperature of the column and RI detector was set to 50 ° C, and the ethanol concentration was quantitatively analyzed using a calibration curve of the standard material.

Example  1. Fibers and seeds according to seaweed type Galactan  Component analysis

0.3 g of seaweed (Mugwort mugwort, Jeju mugwort, Cocktail, Kotoni, hearing, seaweed, sea tangle) and 3 ml of aqueous sulfuric acid were placed in a glass tube and reacted at 30 ° C for 2 hours (primary hydrolysis). At this time, 72% sulfuric acid concentration was used for quantitative determination of glucose and D-galactose, and 1% sulfuric acid concentration was used for quantitative determination of 3,6-anhydrogalactose to prevent overdosing. After the reaction was completed, the reaction mixture was placed in a 250-ml bottle, and 84 ml of distilled water was added thereto. The mixture was hydrolyzed at 121 ° C for 1 hour using a high-pressure reactor (VS-150FN, Vision Science Co., Car hydrolysis). After the hydrolysis was completed, the bottle was taken out at a temperature of 50 ° C in a high-pressure reactor. The bottle was left at room temperature, cooled, and 1 ml of the solution was neutralized with CaCO ₃ and centrifuged (VS-150FN, Vision Science Co., LTD. Korea) and centrifuged at 8,000 rpm for 10 minutes. Cellulose and galactan components were analyzed.

As a result, as shown in Table 2, depending on the kind of seaweeds, the contents of the components were different depending on the sampling sites, and the carbohydrate content was the highest (70-80%) in the mugwort (Morocco and Jeju) , And seaweed (41%). In addition, the content of non-carbohydrate (protein, lipid and other) was highest in seaweed (59%) and the lowest (20 ~ 28%) in mugwort (Morocco and Jeju) Respectively. Therefore, in the subsequent experiments, the carbohydrate / fermentation experiment was carried out by selecting Moroccan bean sprout which has a relatively high carbohydrate content.

[Table 2] Chemical composition of seaweeds

Seaweed cellulose
(%) Galactan
(%) (carbohydrate)
(%) protein
(%) Etc
(Lipid, ash) (%) Red algae Mugwort
(Morocco) 16.6 58.6
(Gal: 25.6%, < RTI ID =
AHG: 33.0%) 75.2 17.9 6.9 Woo 0 88.5
(Gal: 37.5%, < RTI ID =
AHG: 51.0%) 88.5 7.1 4.4 fibrin 99.7 0 99.7 0.2 0.1 Mugwort
(Jeju) 23.0 56.4 79.4 11.8 8.8 Croaker 19.7 54.4 74.1 11.0 14.9 Kotoni 7.1 43.4 50.5 4.9 44.6 Green algae ear 10.9 47.8 58.7 34.7 6.6 Brown algae Seaweed 2.4 38.7 41.1 24.2 34.7 Kelp 6.7 40.0 46.7 12.2 38.1

Example  2. Sulfuric acid catalysts and ionic liquids Glycation  Experimental fertilizer

2.1. Indirect glycosylation

2.1.1. Effect of type of catalyst

The yield of glycation was determined by using the separated dry wool as a substrate. Since the substrate is bound, the monosaccharides that can be formed are galactose and 3,6-anhydrogalactose (3,6-AHG). [Bmim] [HSO4] (Abdol R. Hajipour et al., Catalyst communication 9, pp. 921-9) with a substrate 5 g (S / L = 5%) or 10 g (S / L = 10%) and 10.7 and 13.8 wt% 89-96, 2008) or sulfuric acid and water (100 ml) were placed in a 250 ml Erlenmeyer flask and reacted at 121 ° C for 15 minutes. After completion of the reaction, the saccharified solution lowered to room temperature was neutralized and analyzed by HLPC (ICS-3000, Dionex Co.). , USA) where S / L is the solid / liquid ratio. Table 3 compares the yields of galactose, 3,6-anhydrogalactose and 5-HMF produced at 121 ° C for 15 minutes depending on the type of catalyst. The yields were calculated as the maximum convertible concentration calculated on the basis of the compositional analysis results of 37.5% of D-galactose and 51% of 3,6-anhydrogalactose (Table 2), and when the S / L ratio was 5% Galactose 36.41 g / L, 3,6-anhydrogalactose 49.52 g / L in the case of S / L 10%, and 37.1 g / L of 3,3-anhydrogalactose As a yield of 100%.

When the sulfuric acid was used as a catalyst at S / L = 5%, the amount of galactose and 3,6-anhydrogalactose was increased from 10.7 wt% to 24.3 g / L (16.6 g / L + 7.70 g / L The amount of [bmim] [HSO ₄ ] (1-butyl-3-methylimidazolium hydrogenesulfate) was 13.8 wt% at 20.04 g / L (17.81 g / L + 2.23 g / (13.8 g / L + 22.65 g / L) from 13.8 wt% of the catalyst, as well as [bmim] The amount of 5-HMF, which is a fermentation inhibitor, was significantly lower than that of sulfuric acid (4.29 ~ 5.10 g / L → 0.05 ~ 0.08 g / L) when HSO ₄ was used. This tendency was the same when S / L = 10% (Tables 3 and 4).

[Table 3] Comparison of the amounts of monosaccharides and 5-HMF produced using a sulfuric acid catalyst and an ionic liquid catalyst as a substrate (S / L = 5%)

catalyst Yield,% (g / L) Kinds wt% D-galactose 3,6-L-AHG 5-HMF H ₂ SO ₄ 10.7 91.4 (16.60) 31.1 (7.70) 10.0 (4.29) 13.8 97.9 (17.81) 9.0 (2.23) 11.9 (5.10) [bmim] [HSO ₄ ] 10.7 58.8 (10.70) 83.6 (20.70) 0.1 (0.05) 13.8 73.5 (13.38) 91.5 (22.65) 0.2 (0.08)

[Table 4] Comparison of the amounts of monosaccharides and 5-HMF produced using sulfuric acid catalyst and ionic liquid catalyst as substrate (S / L = 10%)

catalyst Yield,% (g / L) Kinds wt% D-galactose 3,6-L-AHG 5-HMF H ₂ SO ₄ 10.7 88.2 (32.11) 30.8 (11.44) 11.5 (9.88) 13.8 89.6 (32.62) 8.8 (3.27) 13.8 (11.86) [bmim] [HSO ₄ ] 10.7 61.0 (22.21) 75.6 (28.08) 0.3 (0.23) 13.8 71.8 (26.14) 77.6 (28.82) 0.4 (0.37)

2.1.2. Influence of acidic ionic liquid type

[Bmim] [HSO ₄ ] (1-butyl-3-methylimidazolium hydrogenesulfate), [Hmin] [HSO ₄ ] (1-methylimidazolium hydrogensulfate) as a saccharification catalyst, [bmim] [HSO ₄ ] (1-butyl-3-methylimidazolium hydrogenesulfate) was performed in a high pressure reactor at S / L = 10% and 121 ° C for 15 minutes (Table 5).

[Table 5] Comparison of the yields of monosaccharides and 5-HMF using sulfuric acid catalyst and ionic liquid catalyst as substrate (S / L = 10%)

catalyst Yield,% (g / L) Kinds wt% D-galactose 3,6-L-AHG 5-HMF [bmim] [HSO ₄ ] 10.7 61.0 (22.21) 75.6 (37.44) 0.3 (0.26) [Ammonium] [HSO ₄ ] 62.3 (22.68) 76.6 (37.93) 0.2 (0.17) [Phosphonium] [HSO ₄ ] 63.0 (22.94) 77.6 (38.43) 0.3 (0.26) [choline] [HSO ₄ ] 60.8 (22.14) 74.8 (37.04) 0.2 (0.17)

2.2. Fibrin Indirect glycosylation

The saccharification yields of different types of catalysts were compared using the separated dry fiber as a substrate. Since the substrate is osmotic, the monosaccharide that can be produced is glucose. [Bmim] [HSO ₄ ], or 400 ml of sulfuric acid and water, of 20 g of substrate (S / L = 5%) and 10 wt% of the raw material was placed in a 500 ml high-pressure reactor and reacted at 210 ° C for 15 minutes. The saccharified solution lowered to room temperature after neutralization was neutralized and analyzed by HLPC (ICS-3000, Dionex Co., USA). Table 6 compares the yield of produced 5-HMF with glucose according to the type of catalyst. The yield of D-glucose was calculated as 100%, which is the maximum convertible concentration of 48 g / L calculated on the basis of 99.7% of D-glucose, which is the result of analysis of constituents in the cellulose (Table 2). The yield of D-glucose was 22.79 g / L, which was 47.5% when saccharified using 50% sulfuric acid as a catalyst. The yield of D-glucose when using [bmim] [HSO ₄ ] was 28.96 g / L, the yield of 5-HMF was 6.8 g / L when sulfuric acid was used as a catalyst and 0.41 g / L when [bmim] [HSO ₄ ] was used (Table 6).

[Table 6] Comparison of the amounts of monosaccharides and 5-HMF produced by using sulfurous acid catalyst and ionic liquid catalyst as a substrate (S / L = 5%)

catalyst Yield,% (g / L) Kinds wt% D-glucose 5-HMF H ₂ SO ₄ 10 47.5 (22.79) 14.1 (6.8) [bmim] [HSO ₄ ] 60.3 (28.96) 0.9 (0.41)

2.3. Multi-stage using primordial Glycation

The saccharification yields were compared according to the type of catalyst using Morocco mugwort as a substrate. Since the substrate is the primrose starch, the monosaccharides that can be produced are D-galactose and 3,6-anhydrogalactose (3,6-AHG), D-gluconose. [Bmim] [HSO ₄ ] or 100 ml of sulfuric acid and water in an amount of 10 g (S / L = 10%) of the substrate and 10 wt% of the raw material was placed in a 250 ml Erlenmeyer flask and reacted at 121 ° C for 60 minutes. The saccharified solution lowered to room temperature after neutralization was neutralized and analyzed by HLPC (ICS-3000, Dionex Co., USA). Table 7 shows the yields of galactose, glucose, 3,6-anhydrogalactose and 5-HMF produced at 121 ° C for 60 minutes according to the type of catalyst. The yield of the monosaccharide was determined to be 16.6% for D-glucose, 25.6% for D-galactose and 33.0% for 3,6-anhydrogalactose, galactose, 23.8 g / L of D-galactose, and 30.7 g / L of 3,6-anhydrogalactose in a yield of 100%. The yields of D-glucose and D-galactose and 3,6-anhydrogalactose were 0.19 g / L, 19.1 g / L and 6.23 g / L, respectively, when saccharified using sulfuric acid as a catalyst, while [bmim] [HSO ₄ ], the yields were about twice as high as 0.17 g / L, 20.1 g / L and 27.8 g / L, respectively, and the [bmim] [HSO ₄ ] The amount of HMF produced is also reduced by 97% (7.35 g / L → 0.21 g / L) compared with the use of sulfuric acid (Table 7). However, in the case of D-gluconose, the yield was as low as about 1% because the saccharification temperature did not satisfy the fibrin glycosylation condition (about 200-220 ° C.). Therefore, the unreacted residual fibrin was subjected to secondary saccharification The results are shown in Table 8. As a result, as shown in Table 8, the yield of [bmim] [HSO ₄ ] was 27.91 g / L, which was 57.9%, and the yield of 5-HMF was 7.85 g / L, and when [bmim] [HSO ₄ ] was used, it was reduced to 0.34 g / L by 96%.

[Table 7] Comparison of the amounts of monosaccharides and 5-HMF produced using a sulfuric acid catalyst and an ionic liquid catalyst as a substrate (S / L = 5%)

catalyst Yield,% (g / L) Kinds wt% D-glucose D-galactose 3,6-L-AHG 5-HMF H ₂ SO ₄ 4.8 1.2 (0.19) 80.3 (19.1) 20.3 (6.23) 10.5 (7.35) [bmim] [HSO ₄ ] 4.8 1.1 (0.17) 84.6 (20.1) 90.6 (27.8) 0.3 (0.21)

[Table 8] Comparison of the yields of monosaccharides and 5-HMF using residual fibrin as a substrate with sulfuric acid catalyst and ionic liquid catalyst (S / L = 5%)

catalyst Yield,% (g / L) Kinds wt% D-glucose 5-HMF H ₂ SO ₄ 10 40.6 (19.60) 16.3 (7.85) [bmim] [HSO ₄ ] 57.9 (27.91) 0.7 (0.34)

Example  3. Ethanol production culture

3.1. In the saccharification method  Identification of growth characteristics of fermentation strains

Experiments were conducted to compare the growth pattern of the saccharified solution prepared from the sulfuric acid prepared in Example 2 and the ethanol production strain Bretanomyces custerius against the saccharified solution using the ionic liquid of the present invention. 3 is a graph showing the growth curves of Bretanomia suscusterii against a saccharified solution using sulfuric acid and a saccharified solution using [bmim] [HSO ₄ ]. Hydrolyzate with sulfuric acid (5-HMF concentration: 4.29 g / L) In the case of there and the concentration of cells showed a lower growth rate of 1.4 (OD600 nm), [bmim ] [HSO 4] for using Hydrolyzate (5-HMF Concentration of 0.05 g / L) showed a high growth rate of 4 (OD600 nm) at the concentration of the cells in the experiment. Therefore, the fermentation inhibition effect of 5-HMF- It was confirmed that the growth of the strain was inhibited by the presence of 5-HMF, which is a substance, whereas the glycation solution using an ionic liquid showed that the strain grows well without inhibiting the fermentation.

3.2. The saccharified solution  Fermentation using

Figs. 4 and 5 show the results of fermentation using a saccharification solution using sulfuric acid and a saccharification solution using [bmim] [HSO ₄ ], which is a catalyst of the present invention, as a strain of Bretania maiasiscus tercus. In the case of the saccharified solution (5-HMF concentration: 4.29 g / L) using sulfuric acid, the sum of the concentrations of D-glucose and galactose (D-galactose + 3,6-AHG) was 50 g / (5-HMF concentration: 0.05 g / L) using [bmim] [HSO ₄ ] was used as a test solution in the experiment using sulfuric acid. The concentration of monosaccharide was increased from 50 g / L to 3.2 g / L after 48 hours, and the amount of ethanol was twice as high as 12.3 g / L. In the case of the saccharified solution using 5-HMF, the inhibition of the growth of the strain due to the presence of 5-HMF, which is an inhibitory substance of fermentation, was confirmed to reduce the production of ethanol, In the case of the saccharified liquid using the sex liquid, it was confirmed that ethanol was produced without such inhibitory action.

(B) a sampling port, (c) a pressure gauge, (d) an N ₂ gas regulator, (e) an N ₂ gas reservoir, (f) a control box , (g) N ₂ gas storage tank.

2 is a chemical structural formula showing the binding structure of the agarose.

Fig. 3 shows the growth curves of Bretania mieasis catheters against a saccharified solution using sulfuric acid and a saccharified solution using [bmim] [HSO ₄ ] as a catalyst of the present invention.

Figs. 4 and 5 show the results of ethanol fermentation when the saccharification solution using sulfuric acid and the saccharification solution using [bmim] [HSO ₄ ] were used as the strain Bretania maiasiscusterii. In the figure, glucose represents D-glucose and galactose represents the sum of D-galactose and 3,6-AHG.

Claims (18)

A method for producing a saccharified liquid which produces a monosaccharide by treating an acidic ionic liquid catalyst with a polysaccharide material extracted from a seaweed starch or seaweed, Wherein the acidic ionic liquid catalyst is selected from the group consisting of imidazolium, phosphonium, ammonium and choline represented by the following Chemical Formulas 1 to 3 and Chemical Formula 5: [Chemical Formula 1]

(2)

(3)

[Chemical Formula 5]

Wherein R ¹ , R ² , R ³ and R ⁴ are each independently C ₁ -C ₈ alkyl, phenyl, benzyl or fluorinated alkyl, A is SO ₄ , SO ₃ , BH ₃ , n ₃ ) (n = 1 or 2), CF ₃ COO, CH ₃ COO, HPO ₄ and p-CH ₃ (C ₆ H ₄ ) SO ₃ . . delete The method according to claim 1, The acidic ionic liquid catalyst _{[Rmim] [HSO 4] (} 1-alkyl-3-methylimidazolium hydrogenesulfate), [Hmin] [HSO 4] (1-methylimidazolium hydrogensulfate), [Rmin] [TFA] (1-alkylimidazolium triflouroacetate _{), [Rmim] [H 2} PO 4] (1-alkyl-3-methylimidazolium dihydrogenphosphate), [Rmim] [Al 2 Cl 6] (1-alkyl-3-methylimidazolium chloroaluminate) (R = C 1 ~ C 8) ≪ RTI ID = 0.0 > 1, < / RTI > The method according to claim 1, Wherein the extraction solvent at the time of extraction is selected from the group consisting of H ₂ SO ₄ , HCl, HBr, HNO ₃ , CH ₃ COOH, HCOOH, HClO ₄ , H ₃ PO ₄ , PTSA, ≪ / RTI > The method according to claim 1, Wherein the polysaccharide material is selected from the group consisting of wool, starch, carrageenan, alginic acid and fibrin. The method according to claim 1, Wherein the algae are large algae or microalgae. The method of claim 6, Wherein the large algae is selected from the group consisting of red algae, brown algae and green algae. The method of claim 7, The red algae are selected from the group consisting of Cryptomeria japonica, Kotoni, Dog Gambling, Kim, Round Bud, Doguwu, Sambaru, Sambuca, Wherein the saccharified solution is selected from the group consisting of alumina and silica. The method of claim 7, The brown algae are characterized in that the brown algae are selected from the group consisting of seaweed, kelp, horseshoe, leek, lobster, horsetail, seagrass, kelp, myphpathia, rhubarb, By weight. The method of claim 7, Wherein the green algae are selected from the group consisting of chrysanthemum, chestnut, chestnut, hearing, bearded helix, jade, and rosewood. The method according to claim 1, Wherein the monosaccharide is selected from the group consisting of galactose, galactose derivatives, 3,6-anhydrogalactose, glucose, fucose, rhamnose, xylose and mannose. The method according to claim 1, Wherein the monosaccharide is produced by reacting the monosaccharide at an acidic ionic liquid catalyst at a concentration of 0.05 to 30% relative to the maleic acid at a temperature of 60 to 200 DEG C for more than 0 to 6 hours. The method according to claim 1, Wherein the monosaccharide is produced by reacting cellulose with an acidic ionic liquid catalyst at a concentration of 0.05 to 50% at a temperature of 80 to 300 DEG C for more than 0 to 6 hours. The method according to claim 1, Wherein the monosaccharide is produced by reacting the seaweed starch with an acidic ionic liquid catalyst at a concentration of 0.05 to 50% at a temperature of 60 to 300 DEG C for more than 0 to 6 hours. The method according to claim 1, The monosaccharide is reacted with the acidic ionic liquid catalyst at a concentration of 0.05 to 50% based on the seaweed starch at a temperature of 60 to 300 ° C. for more than 0 to 6 hours and then the residual fibrin or starch is subjected to the second Or a tertiary saccharification reaction. ≪ RTI ID = 0.0 > 21. < / RTI > Treating an acidic ionic liquid catalyst to a polysaccharide material extracted from a seaweed source or seaweed to produce a monosaccharide; And A step of fermenting the monosaccharide with a microorganism, the method comprising the steps of: Wherein the acidic ionic liquid catalyst is selected from the group consisting of imidazolium, phosphonium, ammonium and choline represented by the following Chemical Formulas 1 to 3 and Chemical Formula 5: [Chemical Formula 1]

(2)

(3)

[Chemical Formula 5]

Wherein R ¹ , R ² , R ³ and R ⁴ are each independently C ₁ -C ₈ alkyl, phenyl, benzyl or fluorinated alkyl, A is SO ₄ , SO ₃ , BH ₃ , n ₃ ) (n = 1 or 2), CF ₃ COO, CH ₃ COO, HPO ₄ and p-CH ₃ (C ₆ H ₄ ) SO ₃ . . 18. The method of claim 16, Wherein the biofuel is selected from the group consisting of C ₁ to C ₄ alcohols and C ₂ to C ₄ ketones. 18. The method of claim 16, The fermenting microorganism may be selected from the group consisting of Saccharomyces cerevisiae, Sarsinia ventricculia, Kluyveromyces flajilis, Zygomomonas mobilis, Kluyveromyces maximans IMB3, Bretanomyces custerius, Clostridium aceto Wherein the biofuel is selected from the group consisting of Butylicum, Clostridium Bayerinki, Clostridium aurantibutylicum and Clostridium Tetanomorphi.

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