KR100945684B1 - A Method of producing useful biomass using acid and heat or enzymatic pre-treatment - Google Patents

Abstract

The present invention relates to a method for extracting biological resources from algae, comprising: treating algae with water and treating 100 mL of 21% acid per 2 g of cellulose in the seaweed called water. A method for extracting a biological resource and a seaweed called water, preparing a mixture by mixing the algae soaked with water and a buffer solution of pH4.6 and Novozyme 188: Celluclast in the mixture. A method of extracting biological resources from seaweeds, comprising the steps of treating enzymes at a rate of 1.5 L = 1: 2, and the use of large-scale land biomass, the necessity of land, food A biomass extraction method that solves the problem of rising prices and large regional differences in resource consumption, and is capable of regenerating and converting marine algae, which are resources of global dispersion, into energy. It has a very good effect of providing the law.

Algae, biomass, biomass, enzymes, acids and heat

Description

A method of producing useful biomass using acid and heat or enzymatic pre-treatment

The present invention relates to a method for extracting biological resources from seaweed, and more specifically, to a step called seaweed and water and to 100% of 21% acid per 2g of cellulose (cellulose) to the seaweed called water. Biological resource extraction method characterized in that made and the step called algae in the water, the step of mixing the algae soaked in water and a buffer solution of pH 4.6 to prepare a mixture and Novozyme 188: cellulose The present invention relates to a method for extracting biological resources using acid, heat and enzyme from seaweeds, which comprises the step of treating enzyme at a rate of 1.5L = 1: 1.

Biomass is a renewable organic material derived from energy-only crops and trees, agricultural and feed crops, agricultural waste and debris, forest waste and debris, aquatic plants, animal waste, municipal waste, and other wastes. It refers to wood, plants, agricultural and forest by-products, municipal waste and organic components in industrial wastes.

Biomass is a huge energy warehouse that stores solar energy. If such biomass can be effectively used, it will be easy to solve the energy shortage caused by the overuse of fossil fuels. In addition, the biomass is used to cultivate the amount of energy required to prevent excessive accumulation of carbon dioxide in the atmosphere, which can solve the global warming problem that is currently a very serious problem.

Biomass is generally converted into energy through three methods: combustion, dry chemical process, and aqueous process.

First, combustion is the process of converting biomass into electrical energy through high pressure steam.

Second, depending on the nature of the biomass, the dry chemical process is divided into three processes, pyrolysis, gasification, and hydrogasification, into energy. Among them, pyrolysis converts biomass into oil, gas and char, while gasification converts biomass into methane, methanol, ammonia and electricity. To switch. Water gasification converts the biomass to methane, ethane and char.

Finally, the aqueous process converts biomass into energy through three methods: chemical reduction, alcohol fermentation, and anaerobic digestion. Here, chemical reduction converts biomass to oil, alcoholic fermentation converts biomass to ethanol, and anaerobic digestion converts biomass to methane (Slesser and Lewis, 1979).

There are many different types of biomass that can be converted into energy, but it is urgent to find resources that are easy to cultivate, do not affect diet, and are capable of mass production to meet energy demand. . In fact, one of the things that is currently in the spotlight as a biomass that satisfies the above conditions is corn stock.

In recent decades, techniques for converting cellulose to glucose using chemical and physical methods and enzymes have been actively studied to convert wood-based biomass such as grain into energy. Furthermore, studies on the production of ethanol through fermentation using microorganisms as a raw material of glucose obtained through the pretreatment of cellulose materials have been actively conducted.

All of these advances in pretreatment technology have used plants as cellulose materials to date. The most widely used method of decomposing cellulose in plants is pretreatment using enzymes. However, after discovering that cellulose in plants is protected by lignin and hemicellulose, a combination of methods to facilitate enzyme access to cellulose using chemical and physical methods such as acid treatment and heat treatment have been used. come. The principles of the preprocessing methods that have been developed in this way are as follows.

First, chemical hydrolysis is a method of weak acid treatment using acids such as H ₂ SO ₄ or HCl. After the acid treatment, most of the heat treatment for several hours at high temperature to maximize the glucose conversion of cellulose.

Next, the method using the enzyme is to adjust the cellulose to the optimum temperature and pH, the best activity of the enzyme, and then β-glucosidase, xylanase, β-xylosidase ( It is a method of adding enzymes involved in cellulose hydrolysis such as β-xylosidse and α-L-arabino-franosidase. This method of using enzymes may be combined with acid treatment or heat treatment to increase the glucose conversion of cellulose.

However, these wood-based biomass have a limitation in that a large area of land is required for cultivation. In addition, many of these wood-based biomass, if energized brings a rise in food prices.

In order to overcome the drawbacks of the woody system, research has recently been actively conducted to convert biomass in the ocean into energy. Biomass algae in the ocean grow rapidly and can be mass-produced, do not require separate fertilizer or agricultural water, and contain a large amount of various sugars and alginic acid, which is suitable for conversion to energy. Carbohydrate content is 1.5-2 times higher than that of wood-based raw materials, and there is no lignin component that must be removed from wood-based raw materials. The manufacturing process is simple. The methods for pretreatment of biomass in the ocean are mainly existing wood-based biomass. It is the application of pretreatment techniques used to energize them, and the technology to convert marine algae into energy is not yet clearly established.

Therefore, the present inventors have pretreated the biomass algae in the ocean to solve the problem of the need for a large area of land, the rise of food value and the regional difference in resource consumption in the use of the conventional wood-based biomass as described above. As a result of intensive research on the technology for the present invention, the present invention has been achieved.

Accordingly, an object of the present invention is to provide a method for producing a biomass through the enzymatic treatment using a marine algae, which is renewable and is a resource of a global dispersion type.

Another object of the present invention is to provide a biomass production method through heat treatment with acid using algae.

The object of the present invention was achieved by providing a pretreatment method using acid, heat and enzyme in brown algae to obtain glucose and mannitol as sugar components.

The present invention provides a method for extracting biomass from algae, which comprises the step of calling algae in water, and processing 100 mL of 21% acid per 2 g of cellulose and algae in water.

In the present invention, the seaweed is seaweed, and provides a biological resource extraction method characterized in that for 1 hour to 1 hour 30 minutes to react at 100 ℃ ~ 120 ℃.

The present invention also comprises the steps of the algae called water, mixing the algae soaked in water and a buffer solution of pH 4.6 to prepare a mixture and Novozyme 188: Celluclast 1.5L to the mixture It provides a method for extracting biological resources from algae, characterized in that the step of treating the enzyme at a ratio of 1: 1.

In the present invention, the seaweed is seaweed, and provides a biological resource extraction method characterized in that the reaction at 110 rpm.

In the present invention, the reaction time provides a biological resource extraction method, characterized in that 72 hours.

In the present invention, a method for producing a biomass from algae using acid and heat treatment or enzyme treatment requires a large area of land in the use of conventional wood-based biomass, a problem of increase in food value when energized, and a large regional difference in resource consumption. There is an excellent effect in eliminating disadvantages and providing a biomass extraction method for converting marine algae, which is renewable and renewable resources, into energy, which is very excellent in the biomass industry.

Hereinafter, a specific method of the present invention will be described in detail with reference to Examples, and the scope of the present invention is not limited to these Examples.

Example 1 : Algae pretreatment method

Naturally dried brown algae was prepared, and in the case of seaweed with a lot of alginic acid on the surface, it was washed with 3% NaCl, and then seaweed was cut into 4 mm size. The pH of the enzyme for use in the pretreatment is the most desirable pH, pH4.6, the citric acid (sodium citrate) is mixed with the buffer (buffer) and then the wakame and buffer solution by mixing the autoclave ) 4 mL of enzyme per 100 g of cellulose was added to break down cellulose in seaweed, such as β-glucosidase, xylanase and β-xylosidse. Then, the mixture was reacted for 72 hours by shaking at 45 ° C. and 110 rpm. The resulting reaction was analyzed by HPLC.

Experimental Example 1 : Brown algae brown algae Hydrolysis of

After cutting the dry seaweed into 4mm size, it is soaked in 20g water for 1 hour, and after draining the water in soaked seaweed, 25.5mL of 21.01g citric acid per 1L, 24.5mL of 29.41g sodium citrate and 50mL of water Buffer was mixed with seaweed. Thereafter, 4 mL of the enzyme was added at a ratio of Novozyme 188: Celluclast 1.5L = 1: 2 per 100 g of cellulose, followed by reacting for 72 hours while incubating at 100 ° C. and 110 rpm.

The resulting reaction was analyzed by HPLC. The HPLC analytical column used in the present invention was an Aminex ^® HPX-87H column (length 300 mm, inner diameter 7.8 mm), the mobile phase was 0.01 NH ₂ SO ₄ , and the temperature was 50 ° C. . 50 μL of the reactant was flowed to the column at a flow rate of 0.6 mL per minute, and glucose and mannitol were eluted with retention times of 9.8067 and 10.84 minutes, respectively.

As shown in FIG. 1A, it was confirmed that glucose was eluted at 9.74 minutes. FIG. 1B is a peak of glucose taken at the time of analyzing the reaction product of FIG. 1A by HPLC, and FIG. 1C shows the molar concentration of glucose by time. The figure shown.

Experimental Example 2 : Brown algae brown algae Hydrolysis of

After cutting the dry seaweed into 4mm size, it was soaked in 20g water for 1 hour, drained from the soaked seaweed, washed the surface of the seaweed with 3% NaCl, 25.5mL per 1L citric acid, 29.41g per 1L A buffer solution of 24.5 mL of sodium citrate and 50 mL of water was mixed with seaweed. Thereafter, 4 mL of the enzyme was added at a ratio of Novozyme 188: Celluclast 1.5L = 1: 2 per 100 g of cellulose, followed by reacting for 72 hours while incubating at 100 ° C. and 110 rpm.

The resulting reaction was analyzed by HPLC, HPLC column used in the present invention is Aminex ^® HPX-87H column (length 300mm, inner diameter 7.8mm), mobile phase is 0.01NH ₂ SO ₄ , temperature is 50 ℃. . When 50 μL of the reactant was flowed to the column at a flow rate of 0.6 mL per minute, as shown in FIG. 2, it was confirmed that glucose was eluted maximum at 20 hr. After 20 hours, it was estimated that glucose was broken down again and gradually disappeared. In addition, it was confirmed that mannitol was eluted from seaweed exposed for a long time in an acidic buffer.

Example  2: pretreatment of algae through acid and heat treatment

Prepare seaweed dried naturally and wash it with 3% NaCl if seaweed with a lot of alginic acid on the surface. Thereafter, the seaweed was cut into 4 mm size, and 100 mL of 21% H ₂ SO ₄ per 2 g of cellulose was added to the cut seaweed. At this time, the optimal concentration of H ₂ SO ₄ may vary depending on the type of algae.

Thereafter, heat treatment was performed at 100 ° C. for 1 hour. Then, the sample taken for a certain time was analyzed by HPLC.

Experimental Example  1: brown algae through acid and heat treatment ( brown algae Hydrolysis of

After cutting the dry seaweed into 4 mm size, 20 g of dry seaweed was soaked in water for 1 hour, and the water was drained from the soaked seaweed. The surface of the seaweed was then washed with NaCl 3% and 21% H ₂ SO ₄ per 2g cellulose. 100 mL was mixed with seaweed and heated at 100 ° C. for 1 h 30 min.

As a result, the obtained reaction product was analyzed by HPLC, and the column for HPLC analysis Aminex ^® HPX-87H column (length 300mm, internal diameter 7.8mm) used in the present invention, the mobile phase is a 0.01NH ₂ SO _4, the temperature was 50 ℃ . When 50 μL of the reactant was flowed to the column at a flow rate of 0.6 mL per minute, as shown in FIG. 3A, a substance, glucose, and mannitol, which were estimated as cellobiose or lamminaran but could not be confirmed by HPLC, were retained for 8.1533 minutes, 9.9150 minutes, and 11.8017 minutes, respectively. Eluted with. 0.05 M of mannitol and 0.1 M of mannitol were added to the reactants and analyzed by HPLC under the same conditions as above. As a result, conclusions as shown in FIGS.

As is clear from the above examples, the present invention provides glucose and mannitol as sugar components by providing a pretreatment method using acid, heat and enzymes in seaweed, which are used as raw materials for daily life through ethanol fermentation using microorganisms as raw materials. It can be used to produce the necessary biological products and energy.

1 is a diagram illustrating (a) HPLC analysis of biomass extraction results using enzymes from wakame.

(b) Fig. 1a shows the peak of glucose taken together when the reaction of HPLC was analyzed.

(c) FIG. 1A is a diagram showing the glucose molar concentration for each time zone.

FIG. 2 is a diagram analyzing the result of extracting biological resources using enzyme from wakame seaweed,

Figure 3 (a) is a diagram analyzing the biological resource extraction results using acid and heat from seaweed by HPLC.

(b) 0.05M mannitol was added to the reaction product of FIG. 3A and analyzed by HPLC. It can be seen that the reactant contains mannitol.

(c) is 0.1 M mannitol (mannitol) in the reaction of Figure 3a is a diagram analyzed by HPLC.

Claims (5)

delete delete Calling algae into water; Preparing a mixture by autoclaving the algae soaked in water with citric acid and sodium citrate mixed buffer at pH4.6; And Adding cellulose hydrolase to the mixture and culturing at 45 ° C. and 110 rpm for 72 hours to convert cellulose in algae to glucose, and converting algae cellulose into glucose. delete delete

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