KR20100117189A - Method for producing biofuel using sea algae - Google Patents

Abstract

PURPOSE: A method for producing bio fuel using sea algae is provided to improve ingredient supply and to economically produce bio fuel. CONSTITUTION: A method for producing bio fuel using sea algae comprises: a step of pulverizing sea algae; a step of extracting polysaccharide from the sea algae; and a step of fermenting pulverized material. The bio fuel is alcohol of C1-C4 or ketone of C2-C4. The polysaccharide is gelidium jelly, starch, carrageenan, alginic acid or cellulose.

Description

Method for Producing Biofuel Using Seaweed {Method for Producing Biofuel Using Sea Algae}

The present invention relates to a method for producing biofuel, and more particularly, to a method for producing biofuel using seaweed.

Biofuel is a generic term for energy obtained by using biomass as a raw material, and is obtained through direct combustion, alcohol fermentation, methane fermentation, and the like.

Biomass, which is a raw material of biofuel, is divided into sugar-based (sugar cane, sugar beet, etc.), starch-based (corn, potato, sweet potato, etc.), and wood-based (wood, rice straw, waste paper, etc.).

In the case of carbohydrates, raw materials can be converted into biofuels immediately through a relatively simple pretreatment followed by a fermentation process.In the case of starch and woods, bioconcentration is achieved through fermentation using an appropriate pretreatment and saccharification process. Fuel can be produced.

Wood-based materials can use waste wood in the form of urban waste or forest by-products scattered throughout the forest as raw materials, and there is no useful value as food, so the stability of supply and demand of raw materials can be secured, but the lignin removal pretreatment process must be accompanied in the process. Due to the increase in the process cost, due to the crystalline structure consisting of hydrogen bonds, which is a characteristic of the wood-based cellulose substrate, there is a disadvantage that the economic efficiency is low due to low saccharification yield.

The successful commercialization of biofuels as an alternative fuel for transportation lies in securing the price competitiveness of biofuels such as bioethanol over gasoline. In general, the ratio of raw material costs and process costs among biofuel manufacturing costs varies greatly depending on the type and process of raw materials. For example, in case of sugar system using sugar cane or sugar beet, raw material cost: process cost is about 75: 25, while starch system of corn, potato, cassava, etc. is about 50: 50, and in case of wood system, about 25: 75 to be.

However, except for the wood-based biofuel production technology that is currently commercialized, it uses the sugar- or starch-based raw materials that humans can use as food. Supply-demand problems can occur, and economically, using grains is a problem in terms of raw material costs. Corn cultivation also requires significant amounts of pesticides and nitrogen fertilizers, as well as environmental disadvantages that severely corrode the soil compared to other crops.

Algae are largely divided into macroalgae and microalgae, and large algae include red algae, brown algae, green algae, and microalgae, chlorella and spirulina. Seaweed production is estimated at around 14 million tonnes per year worldwide and is expected to increase to more than 22 million tonnes by 2020. This production amount corresponds to about 23% of the total aquaculture production, and more than 90% is composed of brown algae such as seaweed and kelp, and red algae such as laver, seaweed, and stalks. The production of algae farming in Korea is now about 500,000 tons, which is somewhat lower than about 700,000 tons in the mid-90s, but the total area of the farms is about 70,000 ha, which is higher than about 60,000 ha in the mid-90s.

Algae have much better growth than other biomasses (4 to 6 times a year in subtropical regions), and because of the large seas available, the cultivation area is large, and the cost of fresh water, land, and fertilizer is high. It has the advantage of using less resources. In addition, in the case of seaweed, since there is no lignin component that must be removed from the woody system, the biofuel manufacturing process is simple and the total energy conversion yield is high. In addition, algae have an annual uptake of carbon dioxide at 36.7 tons per ha, which is 5 to 7 times higher than that of wood. Assuming that E20 (20% ethanol-added gasoline) is used, the annual greenhouse gas reduction rate is about 27%. In turn, this translates into a carbon tax reduction of around W300bn.

However, seaweeds have been used mainly for fine chemicals and medical materials such as electrophoretic reagents, fertilizers, emulsifiers, and anticancer agents, or have been used only as health foods such as edible and medicinal products. In Korea, a technology for producing bioethanol by separating monosaccharides from seaweed and fermenting them has been developed (public patent 10-2009-0025221).

Anaerobic cellulolytic bacteria have been isolated from various habitats (eg, soils, sediments, wetlands, mammalian gut, etc.) (Madden, et al ., (1982) Int J Syst Bacteriol 32, 87-91; Madden; Murray et al . , (1986) Syst Appl Microbiol 8, 181-184; He et al ., (1991) Int J Syst Bacteriol 41, 306-309; Monserrate et al ., (2001) Int J Syst Evol Microbiol 51, 123-132. ).

A representative anaerobic cellulose-degrading bacterium, Clostridium phytofermentans cells (American type culture collection 700394 ^T ), is isolated from the wet slits at the bottom of the intermittent streams of the tree district near the Qubin reservoir in Massachusetts, USA. It became.

In general, Clostridium phytofermentans cells are long, thin, straight locomotor rods (3.0 to 15.0 μm, 0.5 to 0.8) that form round terminal spores (0.9 to 1.5 μm in diameter). Additional features of Clostridium phytofermentans cells are described in Warnick et al., Int. J. Systematic and Evol. Microbiology, 52, 1155-1160 (2002).

Clostridium phytofermentans can ferment a broad spectrum of materials with high efficiency. Usefully, waste may be produced using waste, for example, lactose, waste paper, leaves, grass cuts, and / or sawdust (published patent 10-2008-0091257).

An object of the present invention is to improve the problems of the conventional biofuel manufacturing method as described above, by using seaweed instead of sugar, starch or wood based raw materials used as raw materials of conventional biofuel, instability of raw material supply, low An object of the present invention is to provide a method for producing biofuel using raw algae or algae as a raw material which has improved problems such as saccharification efficiency.

According to the present invention, there is provided a method for producing a biofuel using seaweed including the step of grinding the algae raw or algae, and the step of fermenting the ground by the microorganism.

Preferably, further comprising the step of extracting a polysaccharide material from the vinegar or seaweed between the grinding step and the microbial fermentation step.

Preferably, the biofuel is selected from the group consisting of alcohols of C1 to C4 and ketones of C2 to C4.

Preferably, the polysaccharide material is selected from the group consisting of radish, starch, carrageenan, alginic acid and fibrin.

Preferably, the seaweed is a large algae selected from red algae, brown algae and green algae, or a microalgae selected from chlorella and spirulina.

Preferably, the fermentation step, by fermenting the pulverized product containing Clostridium phytofermentans cells, the second microorganism and carbohydrate in the medium.

The biofuel is preferably methanol, ethanol, propanol, butanol or acetone, but is not limited thereto.

As the red algae, wood starfish, laver, kotoni, dog gambling, round stone laver, ox radish, buckwheat, green grass, walnut, jindubal, sesame gourd, spiny radish, silk grass, sweet leaf, rock star, stone tree, jinari But it is not limited thereto, and among them, it is preferable to use a stump.

The most popular species of red algae are the most diverse species, and the growth is excellent. About 15-25% of cellulose, which is a cellulose component, and about 50-70% of radishes, which are the main component of galactan, are dry weight. It consists of less than 15% protein and less than 7% lipid.

As the brown algae, seaweed, kelp, barn horse, folk eggplant, shellfish, hooked seaweed, seaweed, Ecklonia cava, gompi, rhubarb, iron seaweed cousin, mabanban, hoesan mabanban, jichungyi, 톳 and the like may be used. It doesn't happen. Brown algae are multicellular bodies and are best differentiated among algae.

The green algae may be used, but are not limited to Cheongtae, Hakkham, blue, auditory, bead hearing, jade, salt-jumping, and the like. Green algae have chlorophyll and make starch by photosynthesis.

Looking at the components of brown algae and green algae, brown algae contain about 30-40% of alginic acid and about 5-6% of fibrin, and green algae contain about 40-50% of starch, the main component of carbohydrate, and 5% of fiber. It contains less than.

Daikon is the main component of galactan composed of galactose polymer, and galactan can be converted into monosaccharides such as galactose and 3,6-anhydrogalactose through proper low molecular weighting process.

Fibrin is a substance composed of cellulose and occupies about 15 to 25% of the total components of the mantle.

The cellulose may be converted into 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 the fermentation process.

Starch, also called starch, is a carbohydrate that is made and stored photosynthesically in the chloroplasts of green plants. It is a polysaccharide that contains glucose as a structural unit. The starch may be converted into glucose, a monosaccharide, through a saccharification process using an appropriate enzyme or an acid catalyst.

The method for extracting polysaccharide material such as radish, cellulose, starch, carrageenan, alginic acid and the like from the seaweed is not particularly limited, and any method known in the art may be used.

As a preferred example, the seaweeds are immersed in an aqueous alkali solution for a certain time and washed with water, and the washed seaweeds are immersed in an extraction solvent consisting of an acidic chemical for a predetermined time to extract the radish, carrageenan, and alginic acid components, and then the remaining cellulose and starch are removed. The collection step may extract the radish, carrageenan, alginic acid and starch or fiber.

At this time, the extraction temperature is not particularly limited, but is preferably in the range of 80 to 150 ° C. The acidic agent may be 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) or a commercial solid. Acids and the like, but is not necessarily limited thereto, and the alkali aqueous solution may include potassium hydroxide, sodium hydroxide, calcium hydroxide, aqueous ammonia solution, but is not necessarily limited thereto.

Clostridium phytofermentans cells ferment the fermentable material to produce flammable fuels such as ethanol and / or hydrogen. Other useful products and by-products can also be produced. Other products include organic acids, or their bases.

Clostridium phytofermentans cells are cultured in an anaerobic environment, which by bubbling a substantially oxygen-free gas through a bubbler containing a gas outlet submerged below the surface of the medium. Acquired and / or maintained. Excess effluent and gas from the reaction in the medium fills the headspace and eventually exits through a gas outlet hole formed in the vessel wall. Gases that can be used to maintain anaerobic conditions include N ₂ , N ₂ / CO ₂ (80:20), N ₂ / CO ₂ / H ₂ (83: 10: 7), and Nobel gases (eg , Helium and argon). To obtain and / or maintain homogeneity, the medium may be stirred. Homogeneity can also be maintained by skating or vibrating vessels.

The algae raw or seaweed fermentable material may be or include one or more low molecular weight carbohydrates. The low molecular weight carbohydrate may be, for example, monosaccharides, disaccharides, oligosaccharides, or mixtures thereof. The monosaccharides can be, for example, galactose, galactose derivatives, 3,6-anhydrogalactose, glucose, fucose, rhamnose, xylose and mannose or mixtures thereof.

In one embodiment, the low molecular weight carbohydrate is obtained by breaking down high molecular weight polysaccharides (eg, radish, starch, carrageenan, alginic acid and fibrin). As another example, the cleavage is done as a separate process and then low molecular weight carbohydrates are used.

As another example, the high molecular weight carbohydrate can be added directly to the medium, cleaved in situ into low molecular weight carbohydrates, and also chemically cleaved by oxidation, base hydrolysis, and / or acid hydrolysis. It may be. Chemical hydrolysis is described in Bjerre, Biotechnol. Bioeng., 49: 568, 1996, and Kim et al., Biotechnol. Prog., 18: 489, 2002.

The low molecular weight carbohydrate may be an enzyme or enzymes (e.g., β-agarase, β-galactosidase, β-glucosidase, endo-1,4-β-glucanase, α-amylase, β-amylase). , Glucoamylase and cellulase) can be cleaved into polysaccharides. These enzymes can be added to the polysaccharide source as an enzyme preparation, or can be produced in situ by the organism. Enzymatic cleavage is described by T. Juhasz, Food Tech. Biotechnol. (2003), 41, 49.

In another embodiment, lactose may be used as the carbohydrate. Lactose is produced in large quantities by the cheese industry. For example, Elliott, Proceedings of the 38th Annual Marschall Cheese Seminar (2001) estimates that about 470 million pounds are produced annually by the US cheese industry and 742 million pounds are produced in Europe. do. Lactose is used, for example, as a growth substrate for Clostridium phytofermentans cells alone, or along with other growth substrates, in a seed fermenter fed to a fermenter, for example a main fermenter. Lactose may be added to the fermentation vessels to enhance fermentation of low molecular weight carbohydrates and / or to promote degradation and fermentation of cellulose or other high molecular weight carbohydrates.

In addition, the fermentable material may be or include one or more high molecular weight carbohydrates. Examples of the high molecular weight carbohydrates include radish, starch, carrageenan, alginic acid and fibrin or mixtures thereof.

Raw materials including seaweeds can be reduced in size, for example, by shearing with a rotary knife cutter or by grinding with a grinder. When reduced in size using a rotary knife cutter, the resultant material is typically fibrous in nature and has a ratio of length to substantial diameter (e.g., greater than 5/1, greater than 10/1, or 15 /). Greater than 1, greater than 20/1, or greater than 25/1). If a grinder is used, typically, the resulting material is in powder form, and typically, for example, having a diameter of less than 5 microns (e.g., less than 4 microns, less than 2.5 microns, less than 1 micron). Have spherical particles. Alternatively, it may be desirable to use a fibrous material having a relatively high surface area and / or relatively high porosity. High surface area and / or high porosity can increase hydrolysis rate and / or efficiency.

As another example, a fermentor including a medium in which Clostridium phytofermentans is dispersed may be configured to continuously remove fermentation products such as ethanol.

The concentration of the desired product in the fermentation is kept substantially constant or, for example, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, or 10 hours of fermentation with an initial concentration of about 10 mM to 25 mM. When measured later, it can be maintained within about 25 percent of the average concentration.

In the present invention, the raw algae raw material or the algae pulverized product or its polysaccharide substance material or mixture can be continuously supplied to the fermentor.

Clostridium pie stopper menthane seuyong medium buffer (for _{example, NaHCO 3, NH 4 Cl,} NaH 2 PO 4 · H 2 O, K 2 HPO 4 and KH ₂ PO _4), electrolytes (such as, KCl and NaCl), growth Additional components such as factors, surfactants, and chelating agents. As growth factors, for example, biotin, folic acid, pyridoxine-HCl, riboflavin, urea, yeast extract, thymine, tryptone, adenine, cytosine, guanosine, uracil, nicotinic acid, pantothenic acid, B12 (cyanocobalamin), p -Aminobenzoic acid, and thioctic acid. Examples of the minerals include MgSO ₄ , MnSO ₄ · H ₂ O, FeSO ₄ · 7H ₂ O, CaCl ₂ · 2H ₂ O, CoCl ₂ · 6H ₂ O, ZnCl ₂ , CuSO ₄ · 5H ₂ O, AlK (SO ₄ ) _2. 12H ₂ O, H ₃ BO ₃ , Na ₂ MoO ₄ , NiCl ₂ · 6H ₂ O, and NaWO ₄ · 2H ₂ O. As a chelating agent, nitrilo triacetic acid is mentioned, for example. As surfactant, polyethyleneglycol (PEG), polypropylene glycol (PPG), copolymer of PEG and PPG, and polyvinyl alcohol are mentioned, for example.

As fermentation conditions, the temperature of the medium may be maintained below about 45 ° C., for example below 42 ° C. (eg, about 34 ° C. to 38 ° C., or about 37 ° C.). In some cases, the medium is maintained at a temperature higher than about 5 ° C., for example higher than 15 ° C.

As fermentation conditions, the pH of the medium can be maintained at less than about 9.5, for example, about 6.0 to 9.0, or about 8 to 8.5. In general, during fermentation, the pH of the medium typically does not change much by more than 1.5 pH units. For example, if the fermentation starts at about pH 7.5, typically, at the end of the fermentation, it will not be lower than pH 6.0, which is within the growth range of the cells.

Clostridium phytofermentans cells adapt to relatively high concentrations of ethanol, such as at least 7% by weight (eg 12.5% by weight). Clostridium phytofermentans cells can adapt to higher concentrations of ethanol (eg, 20% ethanol). By growing in an ethanol-rich environment (eg 7% ethanol) before fermentation, Clostridium phytofermentans are continuously started in high concentrations of ethanol (eg 2% ethanol, then 5% ethanol, then Can be adapted to 10% ethanol).

According to the invention, products other than ethanol can be produced. More generally, fermentation products include alcohols such as ethanol, n-propanol, isopropanol, n-butanol, or mixtures thereof, and fuels such as hydrogen. Other products include organic acids (eg, formic acid, lactic acid, acetic acid, or mixtures thereof) or their bases (eg, formate, lactate or acetate ions) or salts thereof.

Clostridium phytofermentans alone or in yeast or fungi (e.g., Saccharomyces cerevisiae, Pichia stipitis, Trichoderma species, Aspergillus species or other bacteria (e.g., Zymommonas moblis, Klebsiella oxytoca, Escherichia coli, Clostridium acetobu) Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium papyrosolvens, Clostridium cellulolyticum, Clostridium josui Clostridium termitidis, Clostridium cellulosi, Clostridium celerecrescens, Clostridium celerecrescens Host lithium popul retina (Clostridium populeti), Clostridium cellulose Robo lance (Clostridium cellulovorans)) may be used in combination with one or more other micro-organisms and the like.

For example, cellulolytic Clostridium (strain C7) is 2.5 times higher than Clostridium alone in ethanol yield when grown by coculture with Zymonas mobilis in a medium containing cellulose as a growth substrate. Known (Leschine and Canale-Parola, Current Microbiology, 11: 129-136, 1984).

The microbial mixture may be provided as a solid mixture (eg, lyophilized mixture) or as a liquid dispersion of microorganisms and may be grown in coculture with Clostridium phytofermentans, or before addition of Clostridium phytofermentans Alternatively, the microorganisms may be added sequentially to the culture medium by adding another microorganism later.

In addition, any of the algae raw or algae pulverized products in the present invention may be treated with microorganisms by one or more of the above-mentioned in a sequential or co-treatment manner.

For example, the algae vinegar or algae crushed may be treated (co-cultured) simultaneously with the microbial mixture, or the algae vinegar or the algae crushed may be initially treated with a first microorganism or a first mixture of microorganisms (eg, one or more). Yeast, fungi or other bacteria), and the resulting algae vinegar or algae pulverization can be treated with one or more strains of Clostridium phytofermentans. Alternatively, the algae primitive or algae pulverization is initially treated with one or more Clostridium phytofermentans strains, and the resulting algae primitive or algae pulverization is treated with one or more other microorganisms (any one or a mixture thereof). Can be handled with

Hereinafter, the method applied to the Example of this invention mentioned later is demonstrated briefly.

1.Large-scale production of ground algae or ground algae

For the production of ground products from algae raw or algae, Moroccan loot, Jeju loot, stalk, kotoni were used as green algae, auditory was used as green algae, and kelp was used as brown algae. Very simple method was washed with distilled water, dried at 40 ℃ and pulverized (Universal Cutting Mill), using a pulverized powder of less than 1mm with a Vibratory Sieve Shaker.

2. Production of polysaccharides from seaweeds or seaweeds

In the production of polysaccharides from seaweeds or seaweeds, woody larvae were used as raw materials, or polysaccharides and vinegar were used as raw materials. After washing with distilled water, it was dried and pulverized at 40 ℃, and used by powdering.

The radish was immersed in KOH solution for a certain time, washed with distilled water and semi-dried Moroccan loot was extracted with distilled water or ethyl alcohol or methyl alcohol, dried and pulverized at 40 ° C. Bleached with ₃ and bleached with CIO ₂ at 60 ° C., and then bleached with H ₂ O ₂ at 80 ° C. was used.

At this time, the carbohydrate content was different depending on the type of seaweed, and even the same species, depending on the place of collection. The carbohydrate content was the highest with 70 ~ 80% of sea urchin (Morocco, Jeju), and 41% of seaweed. Was the lowest. In addition, the content of non-carbohydrate (protein, lipid and other) was the highest in seaweed (59%) and the lowest in the woodworm (Moroccosan, Jeju), 20-28%, the red algae was most efficiently used as an ethanol production raw material. Can lose. Therefore, Moroccan loot with relatively high carbohydrate content was selected to produce ethanol from algae raw or algae polysaccharides.

3. Production of large scale ethanol from algae raw or algae grinds and polysaccharides

In general, there are two basic approaches to producing fuel grade ethanol from algae primitive or algae crushed and polysaccharide materials on a large scale, using Clostridium phytofermentans cells.

In the first method, the algae raw or algae polysaccharides including high molecular weight algae raw or algae pulverized and polysaccharides are first hydrolyzed into low molecular weight algae raw or algae polysaccharides, and then Clostridium phytofermentans cells Ethanol is produced by fermenting low molecular weight algae raw or algae polysaccharides.

In the second method, the milled products and the polysaccharides themselves are fermented without chemical and / or enzymatic treatment.

In the first method, hydrolysis is carried out by acid, for example Bronsted acid (e.g. sulfuric acid or hydrochloric acid), base (e.g. sodium hydroxide), hydrothermal process, ammonia fiber decay. (ammonia fiber explosion process: “EFEX”), lime process, enzyme, or a combination thereof.

Hydrolysis and / or steaming of the algae raw or algae polysaccharides, for example, can increase the porosity and / or surface area of the algae raw or algae polysaccharides, often where the ground and polysaccharides are Clostridium phytophers. By exposing more to mentans cells, efficiencies and yields can be increased. In addition, by increasing the porosity and / or surface area of the polysaccharide material, it is often possible to increase the efficiency and yield.

By one of the processes according to the invention it is possible to produce products other than ethanol. More generally, fermentation products include fuels such as alcohols and hydrogen, and other products such as organic acids. Clostridium phytofermentans may be used alone or in synergistic combination with one or more of the other microorganisms (eg, yeast or other bacteria) described herein.

Biofuel production method using the seaweed according to the present invention, by the stable supply of raw algae raw or algae raw material can significantly improve the supply and demand problems, and does not require the lignin removal process that is essential accompanying the use of conventional wood-based raw materials As a result, it is possible to lower the cost of the process, and in addition to glucose, most sugars contained in algae, such as galactose and 3,6-anhydrogalactose, can be converted into biofuels, which can drastically lower the production cost of fuels, thereby reducing energy resource problems. Not only can this be solved, but the algae have excellent CO ₂ absorption capacity, which contributes to the reduction of national greenhouse gas emissions and actively cope with international environmental regulations.

Hereinafter, with reference to the accompanying specific embodiments will be described in detail a biofuel manufacturing method using the seaweed according to the present invention. The following embodiments are merely illustrative of the present invention and are not intended to limit the scope of the present invention.

Example 1 Large-scale production of ground crushed seaweeds or algae

In this embodiment, Moroccan loot, Jeju loot, Kotani, kotoni were used as red algae, auditory was used as green alga, and kelp was used as brown algae.

After washing the loach with distilled water, it was dried at 40 ℃ and pulverized by a pulverizer (Universal Cutting Mill, Fritsch, Germany), and used to powder the pulverized matter less than 1mm with Vibratory Sieve Shaker (Fritsch, Germany).

Example 2. Large-scale production of polysaccharides from seaweeds or seaweeds

Daikon was immersed in a KOH aqueous solution for a certain time in Moroccan loot was washed with distilled water and semi-dried was extracted with distilled water or ethyl alcohol or methyl alcohol, dried at 40 ℃ and used to pulverize.

Fibrin is twice extracted with agar O ₃ (1h / 1 times) bleaching and twice at 60 ℃ to CIO ₂ (1.5h / 1 times) After bleaching, two times at 80 ℃ with H ₂ O ₂ (1h / 1) bleached to use.

1% aqueous sulfuric acid solution and 75 g of the substrate were added to a 4 L triangle flask and reacted at 121 ° C. for 15 minutes, and then cooled to room temperature to neutralize the solution with CaCO ₃ and centrifuge (VS-150FN, Vision Science Co., LTD.). , Korea) to remove CaSO ₄ by centrifugation at 8,000 rpm for 10 minutes.

Example 3. Analysis of Polysaccharide Components by Seaweeds

0.3 g of seaweeds (Morocco root stump, Jeju root stump, skewer, kotoni, auditory, seaweed, kelp) and 3 ml of 72% sulfuric acid solution were added to a glass tube and reacted at 30 ° C. for 2 hours (first hydrolysis).

After the reaction, the reaction solution was placed in a 250 ml bottle and 84 ml of distilled water was added and hydrolyzed at 121 ° C. for 1 hour using a high pressure reactor (VS-150FN, Vision Science Co., LTD., Korea) (2 Secondary hydrolysis).

After the hydrolysis, take out the bottle when the internal temperature of the high-pressure reactor is 50 ℃, leave it at room temperature, cool it, take 1 ml of this, neutralize with CaCO ₃ and centrifuge (VS-150FN, Vision Science Co., LTD., Korea), centrifuged at 8,000 rpm for 10 minutes to remove CaSO ₄ and analyzed the fibrin and galactan components.

Example 4. Ethanol Production from Seaweed Herbs and Polysaccharide Materials of Seaweeds

Produced using a microbial fermenter (INNO 200603, Inno Bio, Korea). Clostridium phytofermentans were grown in a culture tube containing GS-2 medium containing 20 g / L or 40 g / L of algae primitive and polysaccharide material of seaweed, respectively. GS-2 medium contains yeast extract 6.0; Urea 2.1; K2HPO4 2.9; KH 2 PO 4 1.5; MOPS 10.0; Trisodium citrate dihydrate 3.0; Cysteine hydrochloride 2.0, each expressed in g / L. The initial pH of the medium was 7.5, and the initial Clostridium phytofermentans concentration was 0.8-1.1 x 10 ⁷ cells / mL.

Cultures were incubated under 30 ° C., N ₂ atmosphere. Polysaccharide mass degradation was monitored visually. In cultures containing algae primrose and algae polysaccharide material, algae primrose and algae polysaccharide material were degraded.

In this embodiment the algae raw and the polysaccharide material of the algae are fermented simultaneously by the culture of Clostridium phytofermentans, cellulose as the most abundant natural source of most algae raw or algae polysaccharide material and the second richest ingredient It may be advantageous if it contains a mixture of carbohydrates including hemicellulose, such as phosphorus xylan.

Example 5. Ethanol production from algae primitives and crushed algae

Produced using a microbial fermenter (INNO 200603, Inno Bio, Korea). Clostridium phytofermentans were grown in a culture tube containing GS-2 medium containing the indicated amounts of the algae primula and the algae crushed products, respectively. The initial pH of the medium was 7.5, and the initial Clostridium phytofermentans concentration was 0.8-1.1 x 10 ⁷ cells / mL.

Cultures were incubated under 30 ° C., N ₂ atmosphere. Algae raw or seaweed mill fermentation determined the concentration of ethanol upon completion of fermentation. Ethanol concentration was analyzed using HPLC equipped with RI detector (Breeze HPLC system, Waters Co., USA), and the column was Aminex HPX-87H (3007.8 mm, Bio-rad).

When the initial mill concentration was 10 g / L, the concentration of ethanol was 75-87 mM. When the initial algae raw or seaweed mill concentration was 20 g / L, the concentration of ethanol was 137-148 mM. When the initial mill concentration was 40 g / L, the concentration of ethanol was 146-162 mM.

From this example, it can be seen that since the concentration of ethanol increases as the initial concentration of the pulverization increases, a higher concentration of pulverization does not inhibit the action of Clostridium phytofermentans. It can also be seen from this example that Clostridium phytofermentans ferment these cellulose feedstocks with ethanol without chemical pretreatment of the seaweed feedstock and the addition of cellulase or other enzymes.

Claims (6)

Grinding algae raw or algae; And fermenting the ground product by microorganisms. According to claim 1, Between the grinding step and the microbial fermentation step further comprising the step of extracting a polysaccharide material from the algae raw or algae, Method of producing a biofuel using seaweeds. The method of claim 1 or 2, wherein the biofuel is selected from the group consisting of C1 to C4 alcohols and C2 to C4 ketones. The method of claim 2, wherein the polysaccharide material is selected from the group consisting of radish, starch, carrageenan, alginic acid and fibrin. The method according to claim 1 or 2, wherein the algae is a large algae selected from red algae, brown algae and green algae, or a microalgae selected from chlorella and spirulina. The method of claim 1 or 2, wherein the pulverized product comprises carbohydrates, the fermentation step, the fermentation by reacting the pulverized product containing Clostridium phytofermentans cells, the second microorganism and carbohydrate in the medium Method for producing a biofuel using seaweed, characterized in that.