KR101083608B1 - Preparation method of biofuel from sea algae using irradiation - Google Patents

Abstract

The present invention relates to a method for producing a biofuel from seaweed using irradiation.

The present invention is a pretreatment step of irradiating the seaweed or algae-derived polysaccharides with radiation; Fermentation by microorganisms after the saccharification step of saccharifying algae or algae-derived polysaccharides subjected to the irradiation with radiation, saccharification step by saccharification by hydrolysis catalyst, or saccharification step by saccharification by decomposition enzyme and hydrolysis catalyst. It relates to a method for producing a biofuel from seaweed using radiation to obtain a biofuel by the step;

The present invention provides a saccharification step of saccharifying algae or algae-derived polysaccharides by degrading enzyme under irradiation, a saccharification step of saccharifying by hydrolysis catalyst under irradiation, or a saccharification step of saccharifying by hydrolyzing enzyme and hydrolysis catalyst under irradiation. It relates to a method of producing a biofuel from seaweed using radiation to obtain a biofuel by a fermentation step by a microorganism.

 Irradiation, Seaweeds, Biofuels

Description

Preparation method of biofuel from sea algae using irradiation}

The present invention relates to a method for producing a biofuel from seaweed using irradiation.

The present invention is a pretreatment step of irradiating the seaweed or algae-derived polysaccharides with radiation; Fermentation by microorganisms after the saccharification step of saccharifying algae or algae-derived polysaccharides subjected to the irradiation with radiation, saccharification step by saccharification by hydrolysis catalyst, or saccharification step by saccharification by decomposition enzyme and hydrolysis catalyst. It relates to a method for producing a biofuel from seaweed using radiation to obtain a biofuel by the step;

The present invention provides a saccharification step for saccharifying algae or algae-derived polysaccharides with a degrading enzyme under irradiation, a saccharification step for saccharifying with a hydrolysis catalyst under irradiation, or a saccharification step for saccharifying by a degrading enzyme and a hydrolysis catalyst under irradiation. It relates to a method for producing a biofuel from seaweed using radiation to obtain a biofuel by a fermentation process by a microorganism.

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 largely a sugar-based raw material such as sugar cane and sugar beet; Starch-based raw materials such as corn, potatoes, and sweet potatoes; It is divided into wood-based raw materials such as wood, rice straw and waste paper.

Among the biomass, the saccharide-based biomass and the starch-based biomass have advantages in that they can be directly manufactured as biofuels through a subsequent fermentation process after relatively simple pretreatment of the raw materials. However, since these sugar-based biomass and starch-based biomass are mostly used as edible crops, the price of grain due to global grain shortages when these sugar-based biomass and starch-based biomass are used to produce biofuels. There may be a problem such as a hike or food shortage. In particular, as edible crops such as saccharide-based biomass and starch-based biomass are gradually reduced in production due to a decrease in cultivation, climate change, and environmental pollution, an increase in grain prices and food shortages are becoming visible. It is difficult to use the saccharide-based biomass, starch-based biomass for other purposes.

On the other hand, among biomass raw materials, wood-based biomass can use waste wood in urban waste form 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. However, in order to prepare biofuels from wood-based biomass, pretreatment of lignin removal contained in wood-based biomass is indispensable, which increases the process cost and characteristics of wood-based cellulose substrate. Due to the crystalline structure composed of hydrogen bonds has a low glycosylation yield has a low economical for producing biofuels.

Recently, research has been conducted on raw materials of new biofuels in addition to raw materials of saccharide biomass, starch biomass, and wood based biomass. In particular, the production of biofuels using sea resources such as seaweed, which occupies most of the earth, is being produced. Much attention has been gathered. Seaweeds have much higher growth potential than the above-mentioned sugar-based biomass, starch-based biomass, and wood-based biomass, and have a wider cultivation area because they are collected from a wide ocean, and have a higher cost of freshwater, land, and fertilizer. It has the advantage of using less resources. In addition, as in the case of wood-based biomass, there is no lignin component that must be removed, so the biofuel manufacturing process is simple and the total energy conversion yield is high. In addition, algae have the advantage of removing carbon dioxide, which is the cause of global warming, because the annual absorption of carbon dioxide is 5-7 times higher than that of wood. However, algae have various components and structures according to species, harvesting place, and harvesting time, and the yield of final biofuels is low because some of the enzymes and catalysts that can effectively decompose the polysaccharide constituting the algae are low. Only seaweed is used to produce biofuels. In addition, many enzymes and acid catalysts or base catalysts are used for the saccharification of algae, resulting in over-degradation or by-products, thereby inhibiting the growth of microorganisms and the production of biofuels in subsequent fermentation processes.

Accordingly, the present inventors have been studying ways to efficiently produce biofuels from various seaweeds, and after performing pretreatment to irradiate seaweeds with radiation, found that the productivity and yield of biofuels increased by the saccharification process and the fermentation process. The invention was completed.

An object of the present invention is to provide a method for producing a biofuel in which the productivity and yield of the biofuel from seaweed is increased.

The present invention is a pretreatment step of irradiating the seaweed or algae-derived polysaccharides with radiation; Fermentation by microorganisms after the saccharification step of saccharifying algae or algae-derived polysaccharides subjected to the irradiation with radiation, saccharification step by saccharification by hydrolysis catalyst, or saccharification step by saccharification by decomposition enzyme and hydrolysis catalyst. It is intended to provide a method for producing a biofuel from seaweed using radiation to obtain a biofuel by the step;

The present invention provides a saccharification step of saccharifying algae or algae-derived polysaccharides by degrading enzyme under irradiation, a saccharification step of saccharifying by hydrolysis catalyst under irradiation, or a saccharification step of saccharifying by hydrolyzing enzyme and hydrolysis catalyst under irradiation. A fermentation process by microorganisms; to provide a biofuel production method from seaweed using radiation to obtain a biofuel.

The present invention can provide a method for producing a biofuel in which the productivity and yield of the biofuel from the algae is increased by the method for producing a biofuel from the seaweed using the radiation of the present invention.

The present invention mainly depends on petroleum since biofuel can be produced from algae using a pretreatment step of irradiating algae or algae-derived polysaccharides and / or a saccharification step of saccharifying algae or algae-derived polysaccharides under irradiation. It can help solve the problem of energy resources.

The present invention shows a method for producing a biofuel from seaweed using radiation.

The first invention of the present invention is a pre-treatment step of irradiating seaweed or algae-derived polysaccharides with radiation;

Fermentation by microorganisms after the saccharification step of saccharifying algae or algae-derived polysaccharides subjected to the irradiation with radiation, saccharification step by saccharification by hydrolysis catalyst, or saccharification step by saccharification by decomposition enzyme and hydrolysis catalyst. The manufacturing method of a biofuel from seaweed using the radiation which obtains a biofuel by a process is shown.

According to a second aspect of the present invention, a saccharification step of saccharifying algae or seaweed-derived polysaccharides with a degrading enzyme under irradiation, a saccharification step of saccharifying with a hydrolysis catalyst under irradiation, or a degrading enzyme and a hydrolysis catalyst under irradiation. A method for producing a biofuel from seaweed using irradiation to obtain a biofuel by a fermentation step with microorganisms after a saccharification step for saccharification.

In the first and second inventions, the algae may use any one or more selected from the group of macroalgae and microalga.

Large algae among the algae may use any one or more selected from the group of red algae, brown algae and green algae.

The red algae in the algae may be any one or more selected from the group of laver, boulders, loach, full starfish, buckwheat, ox radish, braids, jindubal, sesame gambling, sesame noir.

Brown algae in the seaweed may use any one or more selected from the group of seaweed, kelp, 톳, mabanban.

The green algae in the algae is blue, thorny blue, lattice blue, flat blue, leaf blue, intestinal blue, single blue, brown blue, hole blue blue, hearing, supine hearing, agglomerate hearing, broad hearing, falcon, prayer beads, You can use any one or more selected from the group consisting of a half moon, a brush cover, and Hakkham.

The algae among the algae may be any one or more selected from the group of chlorella, spirulina, euglena, Haslea ostrearia.

The algae-derived polysaccharide may be any one or more selected from the group of agar, carrageenan, alginic acid, laminarin, and fucoidan.

The algae-derived polysaccharides may be those currently available on the market or those obtained from seaweeds by known methods. At this time, a known method for preparing seaweed-derived polysaccharides from seaweed is sufficient if a person skilled in the art can use them appropriately, and thus the details thereof will be omitted.

In the first and second invention, the radiation may use any one or more selected from the group of gamma rays, electron beams, and X-rays.

In the first and second inventions, radiation may use gamma rays.

In the first and second invention, the radiation may be carried out at a radiation dose of 1 to 100,000 kGy / hr.

In the first and second invention, the radiation may be carried out at an irradiation dose of 10 to 10,000 kGy / hr.

In the first and second inventions, the radiation may be carried out at an irradiation dose of 10 to 1,000 kGy / hr.

In the first and second inventions, the radiation may be carried out at an irradiation dose of 10 to 500 kGy / hr.

Degrading enzymes in the glycosylation process in the first and second inventions are β-agarase, β-galactosidase, β-glucosidase, endo-1,4-β-glucanase, α-amylase , β-amylase, glucoamylase, cellulase, laminarinase, α-D-glucosidase, β-D-glucosidase, sukrase, maltase, isomaltase, lactase, trehalase You can use more than one.

In the first and second inventions, the hydrolysis catalyst in the saccharification process is H ₂ SO ₄ , HCl, HBr, HNO ₃ , CH ₃ COOH, HCOOH, HClO ₄ , H ₃ PO ₄ , PTSA (p-toluenesulfonic acid) And any one or more acid catalysts selected from the group of commercial solid acids.

As the hydrolysis catalyst in the saccharification step in the first and second inventions, any one or more alkali catalysts selected from the group of potassium hydroxide, sodium hydroxide, calcium hydroxide, and ammonia aqueous solution may be used.

In the first and second inventions, the hydrolysis catalyst in the saccharification process is H ₂ SO ₄ , HCl, HBr, HNO ₃ , CH ₃ COOH, HCOOH, HClO ₄ , H ₃ PO ₄ , PTSA (p-toluenesulfonic acid) And any one or more acid catalysts selected from the group of commercial solid acids, or any one or more alkali catalysts selected from the group of potassium hydroxide, sodium hydroxide, calcium hydroxide, aqueous ammonia solution.

A saccharification step of glycosylating algae or algae-derived polysaccharides subjected to pretreatment of radiation in the first invention with a degrading enzyme; Saccharification step of saccharification by a hydrolysis catalyst; Alternatively, the saccharification step of saccharification by a degrading enzyme and a hydrolysis catalyst may be carried out at 20 to 70 ° C. for 1 hour to 72 hours, preferably at 30 to 50 ° C. for 1 hour to 48 hours.

A saccharification step of glycosylating algae or algae-derived polysaccharides by a degrading enzyme under irradiation in the second invention; Saccharification step of saccharification by hydrolysis catalyst under irradiation; Alternatively, the saccharification step of saccharification by a decomposing enzyme and a hydrolysis catalyst under irradiation may be carried out at 20 to 70 ° C for 1 hour to 72 hours, preferably at 30 to 50 ° C for 1 hour to 48 hours.

Microorganisms for biofuel production in the first and second invention are Saccharomyces cerevisiae, Bretanomyces custersii, Clostridium acetobutylicum ), Clostridium beijerinckii, Clostridium aurantibutylicum, Clostridium tetanomorphum, Sarcina ventriculi, Cluj Veromai Any one or more selected from the group of Kluyveromyces fragilis, Zygomomonas mobilis or Kluyveromyces marxianus can be used.

In the first and second invention, the microorganism may use any one or more selected from the group of Saccharomyces cerevisiae and Bretanomyces custersii.

Fermentation process in the first and second invention can be carried out under optimal conditions during fermentation by the microorganisms. As an example of the fermentation conditions in the fermentation process, it can be carried out for 24 to 96 hours at 25 to 45 ℃, preferably for 24 to 72 hours at 30 to 37 ℃.

In the first and second inventions, the biofuel can obtain an alcohol having 1 to 4 carbon atoms. For example, the biofuel of methanol, ethanol, propanol and / or butanol may be obtained by the first and second inventions.

In the first and second invention, the biofuel can obtain ketones having 2 to 4 carbon atoms. For example, acetone biofuel may be obtained by the first and second inventions.

In the first and second inventions, the biofuel may be one or more selected from the group consisting of alcohols having 1 to 4 carbon atoms or ketones having 2 to 4 carbon atoms.

The method for producing a biofuel from seaweed using radiation according to the present invention was investigated under various conditions. In order to achieve the object of the present invention, a method for producing biofuel from seaweed using radiation under the above-mentioned conditions is provided. It is desirable to provide.

The present invention includes a biofuel produced by the above-mentioned method.

The present invention includes a biofuel produced by the above-mentioned method, wherein the bio-fuel is at least one selected from the group consisting of 1 to 4 carbon atoms alcohols, or 2 to 4 carbon atoms ketones. Indicates.

Hereinafter, the present invention will be described in detail with reference to Examples and Test Examples. However, these are intended to explain the present invention in more detail, and the scope of the present invention is not limited thereto.

Example 1 Results of Saccharification by Pretreatment of Seaweed Using Radiation

(1) Glycation yield measurement

The algae, a seaweed algae, was subjected to a pretreatment step of irradiating red seaweed, followed by a saccharification step at 45 ± 5 ° C. for 6 hours with a degrading enzyme, or a saccharification step at 45 ± 5 ° C. for a 6 hour period using a hydrolysis catalyst. It was set as an experimental group.

Seaweeds, seaweeds, were treated with saccharification for 6 hours at 45 ± 5 ° C. with a degrading enzyme or for 6 hours at 45 ± 5 ° C. with a hydrolysis catalyst as a control (electroless treatment).

In the experimental group, the irradiation in the pretreatment process was irradiated with gamma rays at irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively.

When the glycation yield obtained in the control group was 100, the glycation yield of the experimental group pretreated with the irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively, was measured by gamma rays, and the results are shown in FIG. 1.

In Figure 1 it can be seen that the glycation yield in the experimental group (10kGy, 100kGy, 500kGy) is superior to the control group (electroless treatment), in particular the glycation yield obtained after a certain reaction time as the irradiation dose of radiation increases And it was found.

From the results of FIG. 1, it was found that the treatment of the experimental group with better saccharification yield result than that of the control group in order to obtain biofuel from seaweeds is preferable.

In the above, seaweed, seaweed, was used to remove water and then crushed. The radiation was irradiated with a high-level gamma irradiation facility located at the Korea Atomic Energy Research Institute.

The above-mentioned degrading enzyme used for glycosylation was used α-amylase, hydrolysis acid (HCl) was used.

The saccharification yield for the saccharification solution was analyzed using HPLC equipped with a current detector (ICS-3000, Dionex Co., USA), Carbopac PA1 (4250mm, Dionex Co., USA) and Carbopac PA 1 ( 450 mm, Dionex Co., USA) was used as a column. The mobile phase used a 16 mM NaOH solution, the flow rate was 1 ml / min and the column temperature was 30 ° C. Monosaccharide concentrations such as glucose and galactose were quantitatively analyzed using calibration curves of standards.

(2) Degrading enzyme and hydrolysis catalyst content

The algae, a seaweed algae, was subjected to a pretreatment step of irradiating red seaweed, followed by a saccharification step at 45 ± 5 ° C. for 6 hours with a degrading enzyme, or a saccharification step at 45 ± 5 ° C. for a 6 hour period using a hydrolysis catalyst. It was set as an experimental group.

Seaweeds, seaweeds, were treated with saccharification for 6 hours at 45 ± 5 ° C. with a degrading enzyme or for 6 hours at 45 ± 5 ° C. with a hydrolysis catalyst as a control (electroless treatment).

In the experimental group, the irradiation in the pretreatment process was irradiated with gamma rays at irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively.

In the experimental group pretreated with irradiation doses of 10 kGy, 100 kGy, and 500 kGy with gamma rays, respectively, when the content of the degrading enzyme and the hydrolysis catalyst required to obtain the same saccharification yield by the saccharification process of the control group and the experimental group was 100, respectively. The content of the degrading enzyme and the hydrolysis catalyst necessary to obtain the same saccharification yield is measured and the results are shown in FIG.

As shown in Figure 2 in the control group (electroless treatment) and the experimental group (10kGy, 100kGy, 500kGy) in the content of the enzyme and hydrolysis catalyst required to obtain the same glycation yield, the content of the enzyme and hydrolysis catalyst of the experimental group compared to the control group It can be seen that the decrease was very much, the pretreatment of irradiation can reduce the content of the degrading enzyme and hydrolysis catalyst required for saccharification, thereby improving the productivity during saccharification. On the other hand, as the radiation dose increased in the experimental group it was found that the content of the enzyme and hydrolysis catalyst required for saccharification can be further reduced.

In the above, seaweed, seaweed, was used to remove water and then crushed. The radiation was irradiated with a high-level gamma irradiation facility located at the Korea Atomic Energy Research Institute.

The above-mentioned degrading enzyme used for glycosylation was used α-amylase, hydrolysis acid (HCl) was used.

The saccharification yield for the saccharification solution was analyzed using HPLC equipped with a current detector (ICS-3000, Dionex Co., USA), Carbopac PA1 (4250mm, Dionex Co., USA) and Carbopac PA 1 ( 450 mm, Dionex Co., USA) was used as a column. The mobile phase used a 16 mM NaOH solution, the flow rate was 1 ml / min and the column temperature was 30 ° C. Monosaccharide concentrations such as glucose and galactose were quantitatively analyzed using calibration curves of standards.

<Example 2>

(1) Saccharification yield of algae-derived polysaccharides by radiation treatment pretreatment

The pretreatment of irradiation with laminarin, an algae-derived polysaccharide, was carried out, followed by a saccharification process at 45 ± 5 ° C. for 6 hours with a degrading enzyme.

Laminarine, an algae-derived polysaccharide, was glycosylated at 45 ± 5 ° C. for 6 hours with a degrading enzyme as a control.

Irradiation in the experimental group was irradiated with gamma rays at irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively.

The glycation yields obtained in the experimental and control groups were measured, respectively, and the results are shown in Table 1 below.

Table 1. Glycation yield of experimental and control groups

Item Control group Experimental group 10kGy 100kGy 500kGy Glycation yield 100 115 127 141

* In Table 1, the saccharification yield of the experimental group is equivalent to the conversion rate when the saccharification yield of the control group to 100 means that the higher the saccharification yield.

Table 1 shows that the glycation yield in the experimental group (10kGy, 100kGy, 500kGy) is superior to the control group (electroless treatment), as shown in the experimental results of Figure 1 of Example 1, in particular, the radiation dose of the experimental group also increased It was found that the yield of saccharification obtained after a certain reaction time increases.

In the above, seaweed, seaweed, was used to remove water and then crushed. The radiation was irradiated with a high-level gamma irradiation facility located at the Korea Atomic Energy Research Institute.

In the glycosylation process, α-amylase was used.

The saccharification yield for the saccharification solution was analyzed using HPLC equipped with a current detector (ICS-3000, Dionex Co., USA), Carbopac PA1 (4250mm, Dionex Co., USA) and Carbopac PA 1 ( 450 mm, Dionex Co., USA) was used as a column. The mobile phase used a 16 mM NaOH solution, the flow rate was 1 ml / min and the column temperature was 30 ° C. Monosaccharide concentrations such as glucose and galactose were quantitatively analyzed using calibration curves of standards.

(2) Saccharification yield of seaweed-derived polysaccharides by radiation treatment pretreatment

The experimental group was subjected to a pretreatment to irradiate laminarin, a seaweed-derived polysaccharide, with radiation, followed by a saccharification process at 45 ± 5 ° C. for 6 hours with a hydrolysis catalyst.

Laminarine, an algae-derived polysaccharide, was subjected to a saccharification process for 6 hours at 45 ± 5 ° C. with a hydrolysis catalyst as a control.

Irradiation in the experimental group was irradiated with gamma rays at irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively.

The glycation yields obtained in the experimental and control groups were measured, respectively, and the results are shown in Table 1 below.

Table 2. Glycation yield of experimental and control groups

Item
Control
Experimental group 10kGy 100kGy 500kGy Glycation yield 100 113 124 139

* In Table 2, the saccharification yield of the experimental group is equivalent to the conversion rate when the saccharification yield of the control group to 100 means that the higher the saccharification yield.

Table 2 shows that the glycation yield in the experimental group (10kGy, 100kGy, 500kGy) is superior to the control group (electroless treatment), as shown in the experimental results of Figure 1 of Example 1, in particular, the radiation dose of the experimental group also increased It was found that the yield of saccharification obtained after a certain reaction time increases.

In the above, seaweed, seaweed, was used to remove water and then crushed. The radiation was irradiated with a high-level gamma irradiation facility located at the Korea Atomic Energy Research Institute.

In the above saccharification process, sulfuric acid (H ₂ SO ₄ ) was used as the hydrolysis catalyst.

The saccharification yield for the saccharification solution was analyzed using HPLC equipped with a current detector (ICS-3000, Dionex Co., USA), Carbopac PA1 (4250mm, Dionex Co., USA) and Carbopac PA 1 ( 450 mm, Dionex Co., USA) was used as a column. The mobile phase used a 16 mM NaOH solution, the flow rate was 1 ml / min and the column temperature was 30 ° C. Monosaccharide concentrations such as glucose and galactose were quantitatively analyzed using calibration curves of standards.

<Example 3>

(1) Saccharification yield of seaweed-derived polysaccharides by radiation irradiation and saccharification process

The experimental group was subjected to a saccharification process for 6 hours at 45 ± 5 ° C. with a degrading enzyme under irradiation with laminarine, a seaweed-derived polysaccharide.

Laminarine, an algae-derived polysaccharide, was glycosylated at 45 ± 5 ° C. for 6 hours with a degrading enzyme as a control.

Irradiation in the experimental group was irradiated with gamma rays at irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively.

The glycation yields obtained in the experimental and control groups were measured, respectively, and the results are shown in Table 2 below.

Table 3. Glycation yield of experimental group and control group

Item
Control
Experimental group 10kGy 100kGy 500kGy Glycation yield 100 117 131 144

* In Table 3, the saccharification yield of the experimental group is equivalent to the conversion rate when the saccharification yield of the control group to 100 means that the higher the saccharification yield.

Table 3 shows that the glycation yield in the experimental group (10kGy, 100kGy, 500kGy) is superior to the control group (electroless treatment), as shown in the experimental results of Figure 1 of Example 1, in particular, the irradiation dose of radiation also increased in the experimental group It was found that the yield of saccharification obtained after a certain reaction time increases.

In the above, laminarin, a polysaccharide derived from seaweed, was used as a 10% suspension by crushing the pulverized water in purified water, and a high-level gamma irradiation facility located at Jeongeup Radiation Science Research Institute, Korea Atomic Energy Research Institute.

In the glycosylation process, α-amylase was used.

The saccharification yield for the saccharification solution was analyzed using HPLC equipped with a current detector (ICS-3000, Dionex Co., USA), Carbopac PA1 (4250mm, Dionex Co., USA) and Carbopac PA 1 ( 450 mm, Dionex Co., USA) was used as a column. The mobile phase used a 16 mM NaOH solution, the flow rate was 1 ml / min and the column temperature was 30 ° C. Monosaccharide concentrations such as glucose and galactose were quantitatively analyzed using calibration curves of standards.

(2) Saccharification yield of seaweed-derived polysaccharides by radiation irradiation and saccharification process

An experimental group was subjected to a saccharification process for 6 hours at 45 ± 5 ° C. with a hydrolysis catalyst under irradiation with laminarine, a seaweed-derived polysaccharide.

Laminarine, an algae-derived polysaccharide, was subjected to a saccharification process for 6 hours at 45 ± 5 ° C. under a hydrolysis catalyst as a control.

Irradiation in the experimental group was irradiated with gamma rays at irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively.

The glycation yields obtained in the experimental and control groups were measured, respectively, and the results are shown in Table 3 below.

Table 4. Glycation yield of experimental and control groups

Item
Control
Experimental group 10kGy 100kGy 500kGy Glycation yield 100 115 130 142

* In Table 4, the saccharification yield of the experimental group is equivalent to the conversion rate when the saccharification yield of the control group to 100 means that the higher the saccharification yield.

Table 4 shows that the glycation yield in the experimental group (10kGy, 100kGy, 500kGy) is superior to the control group (electroless treatment), as shown in the experimental results of Figure 1 of Example 1, in particular, the radiation dose of the experimental group also increased It was found that the yield of saccharification obtained after a certain reaction time increases.

The algae-derived polysaccharide laminarin was used to make a pulverized 10% suspension in purified water, and the irradiation of the radiation was used a high-level gamma irradiation facility located in the Jeongeup Radiation Science Research Institute of the Korea Atomic Energy Research Institute.

In the above saccharification process, a hydrolysis catalyst was used as a base catalyst of sodium hydroxide.

The saccharification yield for the saccharification solution was analyzed using HPLC equipped with a current detector (ICS-3000, Dionex Co., USA), Carbopac PA1 (4250 mm, Dionex Co., USA) and Carbopac PA 1 ( 450 mm, Dionex Co., USA) was used as the column. The mobile phase used a 16 mM NaOH solution, the flow rate was 1 ml / min and the column temperature was 30 ° C. Monosaccharide concentrations such as glucose and galactose were quantitatively analyzed using calibration curves of standards.

Example 4 Ethanol Production

Saccharomyces cerevisiae is used as a seaweed algae hydrolysed saccharification solution obtained by performing a pretreatment step of irradiating red seaweed seaweed as a seaweed and performing a saccharification step at 45 ± 5 ° C. under a degrading enzyme or a hydrolysis catalyst. The alcohol fermentation was carried out to the experimental group.

Algae fermentation of algae as a seaweed was performed by saccharomyces cerevisiae in the algae hydrolyzed saccharification solution obtained by performing saccharification process for 6 hours at 45 ± 5 ° C. under a degrading enzyme or a hydrolysis catalyst. It was made.

Irradiation in the experimental group was irradiated with gamma rays at irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively.

When the ethanol production amount obtained in the control group was 100, the ethanol production rate of the experimental group pretreated with irradiation doses of 10 kGy, 100 kGy, and 500 kGy, respectively, was measured by gamma rays, and the results are shown in FIG. 3.

In Figure 3 it can be seen that the ethanol production in the experimental group (10kGy, 100kGy, 500kGy) is superior to the control group (electroless treatment), in particular, the experimental group also showed that the ethanol production also increases with the radiation dose.

According to the results of FIG. 3, the ethanol production of the control group obtained by the saccharification process and the fermentation process without the irradiation pretreatment of the seaweeds as the present invention by the saccharification process and the fermentation process after the radiation treatment of the seaweed as shown in the present invention. It was found to be superior.

In the above, seaweed, seaweed, was used to remove water and then crushed. The radiation was irradiated with a high-level gamma irradiation facility located at the Korea Atomic Energy Research Institute.

The above-mentioned degrading enzyme used for glycosylation was used α-amylase, hydrolysis acid (HCl) was used.

The alcohol fermentation using Saccharomyces cerevisiae in the seaweed hydrolyzed saccharification solution was performed using the following method.

A 250 ml Erlenmeyer flask containing 100 ml of YEPD medium (yeast extract 10g / l, peptone 20g / l, dextrose 20g / l) was inoculated with platinum to Saccharomyces cerevisiae in a solid medium at 37 ℃. The strain was precultured at 150 rpm for 24 hours. The main culture was inoculated with the preculture solution 25% in 150ml medium containing 20% saccharified solution, 15% peptone, 15% yeast extract, and 0.5% magnesium sulfate obtained from the experimental group or control group, and then the initial pH was 5.25 ± 0.25. The culture was carried out for 48 hours at 37 ℃ alcohol fermentation was carried out.

Example 5

As a seaweed, a pretreatment step of irradiation with red seaweed seaweed was carried out, and saccharomyces cerevisiae was used for saccharified saccharification liquid obtained by performing a saccharification process at 45 ± 5 ° C. for 6 hours with a degrading enzyme and a hydrolysis catalyst. Except that the alcohol fermentation was performed, the rest of the conditions were carried out alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, the irradiation was carried out at a 100kGy irradiation dose.

<Example 6>

Seaweed saccharification liquid obtained by performing saccharification process for 6 hours at 45 ± 5 ° C. with degrading enzyme under irradiation with red seaweed seaweed as seaweed except alcohol fermentation using Saccharomyces cerevisiae. The remaining conditions of the alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 7>

Algae algae were red algae, and the algae saccharification solution obtained by performing a saccharification process at 45 ± 5 ° C. for 6 hours with a hydrolysis catalyst under irradiation, except that alcohol fermentation was performed using Saccharomyces cerevisiae. The rest of the conditions were subjected to alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 8>

As a seaweed, Saccharomyces cerevisiae was used for the algae saccharification solution obtained by performing a pretreatment step of irradiating the algae, a brown algae, with a saccharification step at 45 ± 5 ° C. for 6 hours using a degrading enzyme and a hydrolysis catalyst. Except that the alcohol fermentation was performed, the rest of the conditions were carried out alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

Example 9

Algae algae, algae algae, were obtained by carrying out saccharification solution obtained by performing saccharification process for 6 hours at 45 ± 5 ° C. with degrading enzyme under irradiation, except that alcohol fermentation was performed using Saccharomyces cerevisiae. The remaining conditions of the alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 10>

Algae algae, algae algae, were obtained by carrying out a saccharification solution obtained by performing a saccharification process at 45 ± 5 ° C. for 6 hours with a hydrolysis catalyst under irradiation, except that alcohol fermentation was performed using Saccharomyces cerevisiae. The rest of the conditions were subjected to alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 11>

Saccharomyces cerevisiae was added to the algae saccharification solution obtained by performing a pretreatment step of irradiating the green sea algae as a seaweed and performing a saccharification step at 45 ± 5 ° C. for 6 hours with a degrading enzyme and a hydrolysis catalyst. Except that the alcohol fermentation was carried out using the rest of the conditions in the same manner as in Example 4 was subjected to alcohol fermentation to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 12>

Seaweed saccharification liquid obtained by performing saccharification process for 5 hours at 45 ± 5 ° C. with degrading enzyme under irradiation with algae thorny seaweed as a seaweed, except alcohol fermentation using Saccharomyces cerevisiae. The rest of the conditions were subjected to alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

Example 13

Seaweed saccharification liquid obtained by performing saccharification process for 6 hours at 45 ± 5 ° C. by hydrolysis catalyst under irradiation with algae spiny seaweed as algae except for alcohol fermentation using Saccharomyces cerevisiae And the rest of the conditions were carried out alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 14>

The algae saccharified liquid obtained by performing a pretreatment step of irradiating chlorella to the algae and then saccharifying the algae solution at 45 ± 5 ° C. for 6 hours by a degrading enzyme and a hydrolysis catalyst was used as alcohol using Saccharomyces cerevisiae. Except that the fermentation was performed, the rest of the conditions were carried out alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 15>

Except for ethanol fermentation using Saccharomyces cerevisiae with algae saccharification liquid obtained by performing the saccharification process of chlorella as a seaweed for 6 hours at 45 ± 5 ° C. with degrading enzyme under irradiation. Ethanol was prepared by alcohol fermentation in the same manner as in Example 4.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 16>

Algae was obtained by subjecting chlorella to saccharification liquid obtained by performing saccharification process at 45 ± 5 ° C. for 6 hours by hydrolysis catalyst under irradiation, except for alcohol fermentation using Saccharomyces cerevisiae. Under the same conditions as in Example 4, the alcohol was fermented to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 17>

Saccharomyces cerevisiae was used for the algae saccharification solution obtained by performing a pretreatment step of irradiating spirulina with seaweed as a seaweed, followed by a saccharification step at 45 ± 5 ° C. for 6 hours with a degrading enzyme and a hydrolysis catalyst. Except that the fermentation was performed, the rest of the conditions were carried out alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

&Lt; Example 18 >

Except for ethanol fermentation using Saccharomyces cerevisiae with algae saccharification liquid obtained by saccharification of spirulina as a seaweed for 6 hours at 45 ± 5 ° C. with degrading enzyme under irradiation. Ethanol was prepared by alcohol fermentation in the same manner as in Example 4.

In the above, irradiation was carried out at a 500kGy irradiation dose.

&Lt; Example 19 >

Algae obtained from the saccharification liquid obtained by performing the saccharification process of spirulina as a seaweed for 6 hours at 45 ± 5 ° C. with a hydrolysis catalyst under irradiation, except that alcohol fermentation was performed using Saccharomyces cerevisiae. Under the same conditions as in Example 4, the alcohol was fermented to prepare ethanol.

In the above, irradiation was carried out at a 500kGy irradiation dose.

Example 20

Saccharomyces cerevisiae is used as a fucoidan saccharification solution obtained by performing a pretreatment step of irradiating fucoidan, a seaweed-derived polysaccharide, with a saccharification step at 45 ± 5 ° C. for 6 hours with a degrading enzyme and a hydrolysis catalyst. Except that the alcohol fermentation was performed, the rest of the conditions were carried out alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, fucoidan was used as a crushed product in purified water to make a 10% suspension.

In the above, irradiation was carried out at a 500kGy irradiation dose.

&Lt; Example 21 >

Fucoidan, a seaweed-derived polysaccharide, was obtained by performing a saccharification process for 6 hours at 45 ± 5 ° C. with a degrading enzyme under irradiation, except that alcohol fermentation was performed using Saccharomyces cerevisiae. The remaining conditions of the alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, fucoidan was used as a crushed product in purified water to make a 10% suspension.

In the above, irradiation was carried out at a 500kGy irradiation dose.

<Example 22>

Fucoidan, an algae-derived polysaccharide, was obtained by performing a saccharification process for 6 hours at 45 ± 5 ° C. with a hydrolysis catalyst under irradiation, except that alcohol fermentation was performed using Saccharomyces cerevisiae. The rest of the conditions were subjected to alcohol fermentation in the same manner as in Example 4 to prepare ethanol.

In the above, fucoidan was used as a crushed product in purified water to make a 10% suspension.

In the above, irradiation was carried out at a 500kGy irradiation dose.

As described above, although described with reference to a preferred embodiment of the present invention, those skilled in the art will be variously modified and modified within the scope of the present invention without departing from the spirit and scope of the invention described in the claims below. It will be appreciated that it can be changed.

In the present invention, a high yield of biofuel can be obtained by a pretreatment step of irradiating seaweed, which is a marine resource, which can be obtained from the sea corresponding to 70% of the earth's area.

In the present invention, the pre-treatment of seaweed using radiation and / or the glycosylation process of simultaneously irradiating the algae when the algae are glycosylated can be developed to produce biofuels from algae, thereby solving the energy resource problem, It can be used to replace fossil fuels and contribute to greenhouse gas reduction by fossil fuels.

Figure 1 shows the relative yield of saccharification according to the pretreatment using the radiation of seaweed.

Figure 2 shows the relative amount of catalyst required to obtain a constant saccharification yield according to the pre-treatment with seaweed radiation.

Figure 3 shows the ethanol production yield of the hydrolyzate obtained by saccharification according to the pretreatment using the radiation of seaweed.

Claims (11)

Pretreatment process to irradiate gamma-rays with irradiation dose of 10-500 kGy to any one or more seaweed-derived polysaccharides selected from the group of agar, carrageenan, alginic acid, laminarin and fucoidan. , Glycosylated polysaccharides obtained by pretreatment of radiation were glycated by any one or more degrading enzymes selected from the group of β-agarase, β-galactosidase, sucrase, maltase, isomaltase, lactase, trehalase A saccharification step to make; The algae-derived polysaccharides subjected to the radiation treatment were treated with H ₂ SO ₄ , HCl, HBr, HNO ₃ , CH ₃ COOH, HCOOH, HClO ₄ , H ₃ PO ₄ , PT-toluenesulfonic acid (PTSA), potassium hydroxide, sodium hydroxide, Saccharification step of saccharification by hydrolysis catalyst of calcium hydroxide or aqueous ammonia solution; Or a saccharification step of saccharifying the algae-derived polysaccharide subjected to the pre-treatment of the radiation for 1 to 72 hours at 20 to 70 ° C. with a mixed component of the degrading enzyme and the hydrolysis catalyst, and Saccharomyces cerevisiae, Bretanomyces custersii, Sarcina ventriculi, Kluyberomai after the saccharification process of the algae-derived polysaccharides subjected to the irradiation. At least one microorganism selected from the group of Kluyveromyces fragilis or Kluyveromyces marxianus obtains biofuel by a fermentation process at 25-45 ° C. for 24 to 96 hours. Method for producing biofuel from seaweed using radiation. delete delete delete delete delete delete delete delete delete The method of claim 1, wherein the biofuel is at least one selected from the group consisting of alcohols having 1 to 4 carbon atoms or ketones having 2 to 4 carbon atoms. Way.

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