KR20110095646A - Preparation methods of sugar from sea algae by solution plasma treatment and preparation methods of bio-fuel using thereof - Google Patents

Abstract

PURPOSE: A method for manufacturing saccharide materials using marine algae is provided to produce bio fuel. CONSTITUTION: A method for manufacturing saccharide material comprises: a step of treating marine algae with an extraction solvent containing acidic drug for 1-600 minutes as acid treatment; a step of treating by plasma for 1-300 minutes. A method for manufacturing bio-fuel comprises a step of fermenting the saccharide materials using a microorganism.

Description

Preparation Methods of Sugar From Sea Algae by Solution Plasma Treatment and Preparation Methods of Bio-fuel Using Thereof}

The present invention relates to a method for preparing a saccharide material from seaweed and a method for producing a biofuel using the same, and more particularly, to prepare a reducing sugar of monosaccharides and / or disaccharides by sequentially performing acid treatment and plasma treatment of seaweeds, The present invention relates to a method for producing a saccharide material from algae for producing a biofuel by fermenting the prepared reducing sugar with a microorganism and a method for producing a biofuel using the same.

With high oil prices, energy security, and stricter greenhouse gas regulations, the development of alternative energy has been a hot topic, and biofuels are rapidly spreading as a next generation fuel worldwide.

The biofuel refers to energy obtained by using biomass as a raw material, and is obtained through direct combustion, alcohol fermentation, methane fermentation, and the like.

Biomass, 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 this case, it is possible to convert raw materials into biofuels immediately through a relatively simple pretreatment followed by a fermentation process.However, in the case of starch and wood based biofuels, the fermentation process using a saccharified solution that has undergone proper pretreatment and saccharification process is used. It can manufacture.

On the other hand, the algae growth is fast, depending on the depth of the sea green algae in the upper layer, brown algae in the middle layer, red algae grow in the lower layer because it can have a higher productivity than the land biomass through the complex culture. In addition, algae have the advantage of being easy to decompose because there is no hardly degradable lignin in woody biomass.

However, seaweed has a high water content of more than 80%, the salt concentration occupies 20-30% of the dry weight, and has a high content of proteins (10-15%) and fat (1-5%). In addition, the carbohydrate composition, which occupies 25-50% of the dry weight, is also significantly different from that of land plants. Therefore, studies on pretreatment methods, saccharification methods, and fermentation methods suitable for each seaweed are necessary.

In particular, in order to produce biofuels, three steps of glycosylation, fermentation and distillation of raw materials are largely performed. In the saccharification step, chemical treatment, heat treatment, and pretreatment with enzymes are used.

However, these pretreatments have a problem in that the yield of the saccharide material produced is low.

The present invention provides a method for preparing a sugar substance in high yield using seaweed.

The present invention also provides a method for producing biofuel using the prepared saccharide material.

The present invention

An acid treatment step of acid treating the algae with an extraction solvent consisting of an acidic drug; And

It provides a method of producing a saccharide material comprising a plasma treatment step of plasma treating the acid-treated seaweed and extractant mixture.

In addition,

It provides a method for producing a biofuel comprising fermenting a sugar material prepared by the method for producing a sugar material by a microorganism.

The present invention has the effect of producing a saccharide material in a high yield from the algae through sequential acid treatment and plasma treatment, to produce a biofuel using the same.

1 is a flow chart showing a method of manufacturing a biofuel according to the present invention.

In one aspect, the present invention is an acid treatment step of acid treating the algae with an extraction solvent consisting of an acidic drug; And a plasma treatment step of plasma treating the acid-treated seaweed and extractant mixture.

In another aspect, the present invention provides a method for producing a biofuel comprising fermenting a saccharide substance by a microorganism.

Herein, the saccharide material is preferably a reducing sugar, and the reducing sugar refers to a monosaccharide, a disaccharide, or both thereof.

The acid treatment step according to the present invention is to immerse algae in an extraction solvent made of an acidic drug, preferably for 1 to 600 minutes.

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), para-toluene sulfonic acid (PTSA) or commercial solid acid Or a mixture thereof.

In this case, the acidic chemicals are dissolved in a solvent to form an extraction solvent, and the solvent is not particularly limited as long as it is a substance capable of dissolving the acidic chemicals, but preferably water is used.

The acid treatment decomposes polysaccharides, such as cellulose, contained in seaweeds into reducing sugars such as monosaccharides and / or disaccharides.

In the plasma treatment step according to the present invention, the acid-treated algae and extractant mixtures are plasma-treated. Preferably, some of the disaccharides formed through acid treatment of the algae are decomposed into monosaccharides or polysaccharides are decomposed into monosaccharides and / or disaccharides. As the plasma treatment for this purpose, it is not particularly limited, but it is preferable to use a solution plasma treatment.

The plasma treatment, in particular solution plasma treatment, includes supporting the cathode in an acid-treated algae and extractant mixture, installing the anode apart from the mixture, and applying a current of 1 to 100 mA to each electrode.

In this case, the plasma treatment is preferably performed for 1 to 300 minutes.

The biofuel according to the present invention may be selected from the group consisting of C ₁ to C ₄ alcohols and C ₂ to C ₄ ketones, preferably methanol, ethanol, propanol, butanol or acetone, but not limited thereto. It doesn't happen.

On the other hand, in the step of fermenting a saccharide material, for example reducing sugars by a microorganism according to the present invention, the microorganism is not particularly limited as long as it can produce a biofuel by fermenting the reducing sugar, preferably Clostridium aceto Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium aurantibutylicum or Clostridium tetanomorphum, but is not limited thereto. They are more preferred for butanol and acetone fermentation. In addition, Saccharomyces cerevisiae, Sarcina ventriculi, Kluyveromyces fragilis, Zygomomonas mobilis or Kluyberomyces maximans (Kluyveromyces marxianus) IMB3, Bretanomyces custersii and the like can be used, which are more preferred for ethanol fermentation.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings. However, the following description is only for the purpose of specifically describing the present invention, and the scope of the present invention is not limited by the following description.

1 is a flow chart illustrating a method of manufacturing a biofuel according to the present invention.

As shown in FIG. 1, the method for producing a biofuel according to the present invention includes an acid treatment step of acid treating an algae with an extraction solvent made of an acidic drug; Plasma treatment of the acid-treated seaweed and extractant mixture to prepare sugars; And fermenting the saccharide produced through the plasma treatment by the microorganism.

The algae according to the present invention may be used without limitation for large algae or microalgae, the large algae include red algae, brown algae, green algae, and the like, chlorella, spirulina and the like.

The red algae include gamble, laver, kotoni, dog gambling, and gambling, gambling, round buckwheat, daikon radish, buckwheat, green grass, twine, jindubal, spiny radish, silk grass, vulgaris, stone starch, stone, jinari Etc. may be used, but the present invention is not limited thereto, and among them, gambling and / or loot may be used. The most diverse species of red algae are excellent in their growth ability, and on the basis of dry weight, about 15 to 25% of cellulose is composed of cellulose, and about 60 to 50% is made of galactan as a main ingredient. It is composed of less than% 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 the brown algae and green algae, brown algae contains 30-40% alginic acid, 5-6% fibrin, and green algae contains 40-50% starch, the main component of carbohydrate, and less than 5% fibrin. have.

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 in the case of lodge, it accounts for 15 to 25% of the total component. 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 composed of glucose. The starch may be converted into glucose, a monosaccharide, through a saccharification process using an appropriate enzyme or an acid catalyst.

The acid treatment step according to the present invention is to decompose polysaccharides contained in seaweeds and the like to decompose into reducing sugars such as monosaccharides and / or disaccharides, and is not particularly limited as long as it is a conventional acid treatment method in the art for this purpose.

In one embodiment, the acid treatment according to the present invention is to immerse algae in an extraction solvent consisting of an acidic drug.

Here, the time and temperature for dipping the algae in the extraction solvent can be changed according to the user's choice, but it is recommended to immerse in the temperature range of 50 to 300 ℃ for 1 to 600 minutes.

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), para-toluene sulfonic acid (PTSA) or commercial solid acid Or a mixture thereof, but is not limited thereto.

In this case, the acidic chemicals are dissolved in a solvent to form an extraction solvent, and the solvent is not particularly limited as long as it is a substance capable of dissolving the acidic chemicals, but preferably water is used.

Plasma treatment according to the present invention is to decompose partially formed disaccharides into monosaccharides through acid treatment of algae, or to decompose polysaccharides contained in algae into monosaccharides and / or disaccharides, and the like. Preferably, solution plasma treatment is used.

The plasma treatment may include applying a cathode to an acid-treated algae and extractant mixture, installing the anode to be spaced apart from the mixture, and applying a current of 1 to 100 mA to each electrode.

In this case, the plasma treatment is preferably carried out for 1 to 300 minutes, the separation distance of the anode and the mixture, specifically the seaweed and the extraction solvent mixture is not particularly limited, but is 0.1 to 3cm, more preferably about 0.5cm It is good.

In addition, in the plasma treatment, it is preferable to stir the algae and the extraction solvent mixture in order to produce more sugar material from the algae and the extraction solvent mixture.

Preferably, the sugar material according to the present invention is a reducing sugar, which refers to a monosaccharide, a disaccharide or a mixture thereof.

The monosaccharides are galactose, galactose derivatives, 3,6-anhydrogalactose, glucose, fucose, rhamnose, xylose, mannose or mixtures thereof.

In the step of fermenting the sugar material according to the present invention, that is, reducing sugar by the microorganism, the microorganism is not particularly limited as long as it can produce a biofuel by fermenting the reducing sugar, but preferably Clostridium acetobutylicum ( Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium aurantibutylicum or Clostridium tetanomorphum, but are not limited to these butanols And in acetone fermentation. Also, Saccharomyces cerevisiae, Sarcina ventriculi, Kluyveromyces fragilis, Zygomomonas mobilis or Kluyveromyces maxia (Kluyveromyces marxianus) IMB3, Bretanomyces custersii and the like can be used, which are more preferred for ethanol fermentation.

In addition, the fermentation conditions for fermenting the sugar substance by the microorganism is not particularly limited as long as the fermentation conditions are conventional in the art, preferably fermentation for a temperature range of 20 to 50 ℃ and / or 12 to 50 hours.

Hereinafter, the present invention will be described in detail through examples. However, the following examples are only for illustrating the present invention in detail and are not intended to limit the scope of the present invention by these examples.

First, prior to explaining the embodiment of the present invention, as a method for analyzing the saccharide material prepared according to the embodiment of the present invention, an analysis method using a Shimadzu HPLC apparatus will be described as follows.

HPLC apparatus for the analysis of saccharides includes pumps [Shimadzu LC-20AD, Shimadzu Corporation, Japan], ovens [CTO-20AC, Shimadzu Corporation, Japan], auto samplers [Sil-20AC auto-sampler, Shimadzu Corporation, Japan], Fluorescence detector [RF-10Axl fluorescence detector, Shimadzu Corporation, Japan], system controller [CBM-20A system controller, Shimadzu Corporation, Japan], CRB-6A reaction box LC Workstation software [Shimadzu Corporation, Kyoto, Japan].

In addition, saccharide substances, for example, reducing sugar separation, use an ion exchange shim-pack ISA-07 (4.0 × 250 mm) analytical column and Shim-pack ISA guard column (4.0 × 50.0 mm). It was.

The mobile phase used potassium borate (pH 8) as solution A and potassium borate (pH 9) as solution B, starting from 0 minutes to 0%, increasing the solvent to 50% in 30 minutes, and 100% in 50 minutes. It increased to and maintained at 100% for 15 minutes, then decreased to 0% after 65 minutes, and the total 90 minutes was the analysis time.

In addition, injection volume is 20μL and using a post-column method, 1% arginine and 3% boric acid mixed solution as a reaction reagent to deduct the reducing sugar in the reaction box (150 ℃) reaction box (fluorescence detector (Ex = 320, Em = 430).

The reducing sugar standard was dissolved in deionized distilled water to prepare a standard solution in the range of 0.1 to 400 ppm and analyzed. A calibration curve was prepared and quantified from the peak area.

≪ Example 1 >

0.2M aqueous hydrochloric acid solution and 2 parts by weight of kelp in an Erlenmeyer flask were used as an extraction solvent for acid treatment, followed by reaction at 120 ° C. for 10 minutes using a high temperature autoclave [VS-1221, Vision Scientific Co., LTD., Korea]. Acid treatment.

Then, a solution loaded with kelp as an acid treated material was placed in a plasma reactor, and the negative electrode was supported in the solution, and the positive electrode was installed to be spaced about 0.5 cm from the solution.

Next, a 15 mA current was applied to a DC power supply device (KSL-500 / 200GC, Korea Switching, Korea) connected to each electrode, and the acid treated material was plasma treated for 3 minutes.

At this time, the acid treated material was stirred at a speed of 400rpm.

As a result, the produced reducing sugar is shown in Table 1.

<Example 2>

The same procedure as in Example 1 was carried out except that the acid treated material was plasma treated for 5 minutes instead of 3 minutes.

As a result, the produced reducing sugar is shown in Table 1.

<Example 3>

The same procedure as in Example 1 was carried out except that the acid treated material was plasma treated for 10 minutes instead of plasma treated for 3 minutes.

As a result, the produced reducing sugar is shown in Table 1.

Comparative Example 1

0.2M aqueous hydrochloric acid solution and 2% by weight kelp were placed in an Erlenmeyer flask, and then reacted at 120 ° C for 10 minutes using a high temperature autoclave [VS-1221, Vision Scientific Co., LTD., South Korea].

Subsequently, the material after the reaction was naturally cooled to room temperature and then centrifuged at 8,000 rpm for 10 minutes using a centrifuge [MICRO-12, HANIL, South Korea] to obtain only the supernatant.

Then, the obtained supernatant was subjected to a filtration process using a syringe filter [GD / X PVDF Filter, Whatman international LTD., UK].

As a result, the produced reducing sugar is shown in Table 1.

As shown in Table 1, the reducing sugar preparation method of Examples 1 to 3 subjected to the acid treatment of kelp as seaweed and plasma treatment may produce up to about 2 times more reducing sugar than Comparative Example 1, which was subjected only to acid treatment. It was found that the reduced sugar production amount of Example 3, which had the longest plasma treatment time, was higher than those of Examples 1 and 2.

<Example 4>

0.2M aqueous hydrochloric acid solution and 2% by weight gambling were added to the Erlenmeyer flask as extraction solvent for acid treatment, and then reacted at 120 ° C for 10 minutes using a high temperature autoclave [VS-1221, Vision Scientific Co., LTD., South Korea]. Acid treatment.

Then, after putting the solution with gambling as an acid treated material in the plasma reactor, the negative electrode was supported in the solution, and the positive electrode was installed to be spaced about 0.5 cm away from the solution.

Next, a 30 mA current was applied to a DC power supply device (KSL-500 / 200GC, Korea Switching, Korea) connected to each electrode, and the acid treated material was plasma treated for 5 minutes.

At this time, the acid treated material was stirred at a speed of 400rpm.

As a result, the prepared reducing sugar is shown in Table 2.

Example 5

The same procedure as in Example 4 was carried out except that the acid treated material was plasma treated for 10 minutes instead of 5 minutes.

As a result, the prepared reducing sugar is shown in Table 2.

<Example 6>

The process was performed in the same manner as in Example 4 except that the acid treated material was plasma treated for 30 minutes instead of 5 minutes.

As a result, the prepared reducing sugar is shown in Table 2.

Comparative Example 2

0.2M aqueous hydrochloric acid solution and 2% by weight gambling were placed in an Erlenmeyer flask, and then reacted at 120 ° C for 10 minutes using a high temperature autoclave [VS-1221, Vision Scientific Co., LTD., South Korea].

Subsequently, the material after the reaction was naturally cooled to room temperature and then centrifuged at 8000 rpm for 10 minutes using a centrifuge [MICRO-12, HANIL, South Korea] to obtain only the supernatant.

Then, the obtained supernatant was subjected to a filtration process using a syringe filter [GD / X PVDF Filter, Whatman international LTD., UK].

As a result, the prepared reducing sugar is shown in Table 2.

As shown in Table 2, the reducing sugar preparation method of Example 4 to Example 6, which was subjected to an acid treatment of gambling as seaweed and then plasma treated, could produce a reducing sugar higher than that of Comparative Example 2 subjected to only acid treatment, and the plasma treatment time was The longer the concentration of reducing sugar was found to be lower.

This indicates that as the reaction time between plasma and gambling becomes longer, not only the bonds are decomposed from the polysaccharide to the monosaccharide, but also a part of the reducing sugar is changed by further decomposing the structure in which the bond is decomposed into the monosaccharide.

<Example 7>

In the same manner as in Example 5, 50 mA was used instead of 30 mA of current applied to a DC power supply device [KSL-500 / 200GC, Korea Switching, Korea] for plasma treatment.

The results are shown in Table 3.

<Example 8>

In the same manner as in Example 5, 70 mA was used instead of 30 mA of current applied to a DC power supply device [KSL-500 / 200GC, Korea Switching, Korea] for plasma treatment.

The results are shown in Table 3.

As shown in Table 3, the amount of reducing sugar produced showed the highest amount of reducing sugar produced in Example 7 to which 50 mA current was applied for plasma treatment.

As described above, those skilled in the art will understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. Therefore, the scope of the present invention should be construed as being included in the scope of the present invention all changes or modifications derived from the meaning and scope of the claims to be described later rather than the detailed description and equivalent concepts thereof.

Claims (8)

An acid treatment step of acid treating the algae with an extraction solvent consisting of an acidic drug; And
And a plasma treatment step of plasma treating the acid-treated seaweed and extractant mixture. The method of claim 1,
The acid treatment step is a method for producing a saccharide material, characterized in that the algae immersed for 1 to 600 minutes in the extraction solvent consisting of acidic chemicals. The method of claim 1,
In the plasma treatment, a cathode is supported on an acid-treated seaweed and extractant mixture, and a cathode is installed to be spaced apart from the mixture, and a current of 1 to 100 mA is applied to each electrode. The method of claim 1,
The plasma treatment is a method for producing a saccharide material, characterized in that for 1 to 300 minutes. A method for producing a biofuel comprising fermenting a saccharide substance prepared according to the method for producing a saccharide substance according to any one of claims 1 to 4 by microorganisms. The method of claim 5,
The fermentation microorganisms are Clostridium acetobutylicum, Clostridium Bayerky, Clostridium aurantibutyricum, Clostridium tetanomorphium, Saccharomyces cerevisiae, Sarcina ventriculum, Klui Veromaises Fragilis, Zaigomonas Mobilis, Cluj Veromaises Maxianus IMB3, Bretanomyces Custerssee, Clostridium Acetobutylicum, Clostridium Bayerki, Clostridium aurantibutyl Method for producing a biofuel, characterized in that selected from the group consisting of Qum and Clostridium tetanomorphium. The method of claim 5,
Wherein said biofuel is selected from the group consisting of C ₁ to C ₄ alcohols and C ₂ to C ₄ ketones. The method of claim 7, wherein
Wherein said biofuel is selected from the group consisting of methanol, ethanol, propanol, butanol and acetone.

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