KR101489751B1 - High-yield, high-purity fucoidan manufacturing methods - Google Patents

Abstract

More particularly, the present invention relates to a method for producing a high-purity fucoidan having a high yield and a high purity by preventing the loss of the active ingredient in the tuna by using sea tangle, High-purity fucoidan, which improves the purity of lactose, lactose, lactose, lactose, lactose and lactose).
In the high yield and high purity fucoidan production method of the present invention, the kelp crushed by grinding is pyrolyzed using a tumbling machine and then subjected to low-temperature extraction using a digesting enzyme to obtain Lactobacillus brevis DL-25, Saccharomyces cerevisiae DS-7, Aspergillus oryaze DF-11 is cultured in seaweed extract for 10 to 30 hours, and then the fermentation broth is separated and purified.

Description

TECHNICAL FIELD [0001] The present invention relates to a high-yield fucoidan production method,

More particularly, the present invention relates to a method for producing a high-purity fucoidan having a high yield and a high purity by preventing the loss of the active ingredient in the tuna by using sea tangle, High-purity fucoidan, which improves the purity of lactose, lactose, lactose, lactose, lactose and lactose).

Seaweeds have a hard shell and cell wall as compared with land animals. When simple treatments such as general organic solvent, enzyme treatment, and ultrasonic treatment are used, it is difficult to obtain useful components, and it is required to produce offensive, already generated, low yield and high cost.

The existing seaweed extraction fucoidan is prepared by separation of fine powder and extract (centrifugation) by extraction of fine powder and organic solvent (acetone, isopropyl alcohol, ether, alcohol, etc.) without pretreatment and separation / purification of organic solvent from extract The process is necessary, so that it takes a lot of time and cost because of the large number of processes. Also, in the separation / purification part of the organic solvent, it leads to the safety problem of the food.

Conventional fucoidan extracts were extracted from marine algae such as seaweed and sea tangle. The conventional fucoidan extraction method is ① raw materials → ② raw material crushing (raw material improvement, crushing) → ③ washing (purified water agitation, washing) → ④ extraction → ⑤ filtration → ⑥ Fucoidan was extracted from seaweeds through 13 ~ 15 stages including separation / purification → condensation → ⑧ solidification → ⑨ removal of alcohol → ⑩ freeze drying → ⑪ crushing → ⑫ quality inspection (separation) → ⑬ packaging.

Korea has three seaweeds and has very favorable conditions for producing seaweeds. Since then, there have been active algae-related industries, among which more than 670,000 tons of brown algae are produced annually in Korea. Recently, domestic seaweeds The main varieties produce seaweeds of 243,000 tons, 210,000 tons of seaweed, 25,000 tons of kelp, 16,000 tons of tops, 9,000 tons of parasites, and about 4,000 tons of other seaweeds. Of the total seaweed production is 97.3%.

Kelp plants are native to the Pacific Ocean with over 20 species of carbohydrates, such as mannitol and laminarnin, and minerals such as iodine, potassium and calcium, and alginic acid, which are beneficial to humans.

In particular, kelp has 50 kinds of minerals and high dietary fiber content, and kelp extraction is difficult to extract and pretreatment process, so it is processed and sold in the form of powder processing or conversion in Korea, and high value-added products depend on imports have.

The development of high - function fucoidan using kelp extract is expected to increase the income of fishermen and the import substitution effect.

Here, 'fucoidan' has a variety of physiological activities, and especially when it is administered to cancer cells, 1 g of fucoidan separated and purified directly acts on 'various cancers'. After 72 hours, the cancer cells die and they are called 'apoptosis' Suicide induction by the suicide induction). In 1995, Takara Bio Research Institute published a paper on the function of fucoidan in the Japanese Cancer Society. The five major constituents of Fucoidan are Fucose, Glucose, Xylose, Callactose and Sulfate, which determine the quality of Fucoidan.

It is an object of the present invention to solve the above-described problems of the prior art, and to provide a method for preventing the loss of effective ingredients in kelp by low temperature extraction, simplifying the process by reducing the extraction process, Purity, high-purity fucoidan, which improves the purity of the fucoidan, lactose, lactose, lactose, and sulfate groups.

In order to achieve the above object, the present invention provides a high-yield, high-purity fucoidan preparation method in which a kelp crushed with a crushing ratio is thermo-calcined using a tumbling machine and then treated with Cellulose, Viscozyme and Ultrazyme as degrading enzymes The low-temperature extract and the vacuum low-temperature extract are mixed, and the Lactobacillus brevis DL-25 strain is cultured for 10 to 30 hours in a seaweed extract, and then the fermentation broth is separated and purified.

According to the method of the present invention described above, it is possible to prevent the loss of the active ingredient in the tuna, to simplify the process by reducing the extraction process, and to improve the extraction yield and the fucoidan indicator substance (fucose, glucose, xylose, callactose, Can be improved.

1 is a flow chart of a high yield, high purity fucoidan manufacturing method according to a preferred embodiment of the present invention.

The high yield and high purity fucoidan production method of the present invention can be divided into a first step of pretreating kelp, a second step of enzymatic decomposition / low temperature extraction and vacuum low temperature extraction, and a third step of fermentation, separation and purification of microorganism.

First, the first step of pretreating kelp is a step of air washing to remove foreign matter from the dried kelp, a step of crushing the kelp into granules of a certain size (1 to 2 cm in size), the granular crushed kelp into a tortuosity A thermosetting step of applying heat, and a step of washing the thermally calcinated kelp with distilled water.

The granule crushing of the kelp facilitates the thermosetting and perfusion works and the thermosetting perfume works to weaken the rigid shell and cell wall of the kelp and increase the intercellular space to increase the extraction yield .

Next, the thermo-plasticization step may be referred to as a 'thermo-plasticization step' or a 'heating step' in the sense that heat is applied, and the term 'imbibed step' or 'thermo- plastic im- proved' Step ".

If you look at the meaning and effect of 'thermo-plastic impervious', all substances present have a unique unique flavor, and the flavor (odor) of the substance is the refreshment of the volatile substance contained in the substance itself, . The granular kelp, which is discussed in the present invention, has a flavor and cell structure that is completely different from the dry state in the thermo-plasticization (perfection) stage. The intrinsic flavor of the kelp, which occurs in the thermosetting stage, is contained in the kelp as the pores are closed during the drying process of the kelp. The intrinsic flavor of the kelp is dissipated by heat, It is sealed, and the coating of kelp fragrance is applied. It is important to weaken the rigid body and cell walls, to make it porous, and the term "thermoplastic coating" is not a dictionary term, and it is named.

Spraying an appropriate amount of alcohol (about 0.5 ~ 5 wt% with respect to kelp) into granular kelp (moisture content 5 ~ 10%) in dry state and closing the inside of the tulip and heating it, the pore of the kelp that has been closed starts slowly to open. Alcohol (alcohol) helps to open the pores of the kelp and helps to evaporate water. The reason for the slow rotation at the internal temperature of 70 ℃ ~ 80 ℃ is to accelerate the heat transfer and to vaporize the original fragrance quickly.

The effect of thermo-plasticization (incense-coated) is obviously different in olfactory and taste, even in the case of real products. Generally, a person feels a sense of smell (smell) is a feeling of intrinsic fragrance and intrinsic fragrance inherent to the human body. The present invention does not oxidize the granular seaweed, And even if the internal temperature of the tumbler is 100 ° C to 130 ° C, the inner granular kelp rotates due to a strong centrifugal force rather than rotating due to the whirlwind. Therefore, the temperature Is lower than the internal temperature of the enhancement, which is controlled by the tumbler itself.

 In this way, the inside and outside of the granular kelp are perfumed (coated) in the confined space, and the rigid shell and the cell wall are weakened and made porous. For example, if you smoke a cigarette, you smell nicotine on your fingers and you smell it on your fingers. The smell of it smells like a bass on your clothes, and when you heat the hard rice cake to soften it, And to make the cells porous by increasing the interval (gap) between the cells.

This thermosetting application maximizes the flavor of the kelp, weakens the shell and cell wall of the kelp, increases the intercellular space, and makes it porous, facilitating the extraction and improving the extraction yield.

Washing distilled water prevents the removal of other debris and deposits of kelp, and effectively dissolves contaminants and other debris to become neutralized.

2 process includes enzymatic digestion / low temperature extraction and vacuum low temperature extraction with granular crushed kelp with distilled water.

The distilled water acts as a strong solvent for removing the nutrients in the cells, which can enter the kelp cell without damaging the kelp, and the low temperature / vacuum low temperature extraction minimizes the loss of the active ingredient which can be lost at high temperature and high pressure, Enzymatic degradation breaks down the complex polysaccharide, finely cleaves it, and increases the content of sulfate groups.

3 process includes microbial fermentation and separation / purification process. Microorganism fermentation low molecular weight fucoidan indicator substance and removes protein, fat, ash and so on to increase the yield of fucoidan indicator material and facilitate separation and purification , The final product is highly purified and is effective in improving the quality.

With the above process, the yield and purity can be improved, and fucoidan having safety of food can be obtained.

Hereinafter, a method for extracting physiologically active fucoidan using the sea tangle according to the present invention will be described with reference to the accompanying drawings.

FIG. 1 is a process flow diagram of a high-yield, high-purity fucoidan production method for improving the extraction yield and the purity and safety of fucoidan indicator by preventing the loss of the active ingredient according to the preferred embodiment of the present invention.

The method shown in FIG. 1 is applied to kelp, which enables preservation of active ingredients during fucoidan production, improvement of purity and extraction yield, and safety-enhanced physiologically active fucoidan extraction.

The kelp is fixed on rocks and ropes of wild fish, aquaculture, and seabed, and foreign matter is much more than other parts. First, the foreign matter of kelp is cleaned with air cleaner. When washing the tide, the spray pressure is suitably adjusted to perform cleaning (S100).

The air-washed kelp is granulated (1-2 cm) (S200). The granule crushing of the sea tangle receives the indirect heat due to the strong rotational force during the subsequent thermosetting (incense impregnation) step, weakens the shell and cell wall of the kelp by thermo plasticity (improves) and increases the intercellular space, To facilitate washing and extraction.

Next, the thermosetting step is performed in the kneader (S300). The granular crushed kelp was put into a kneading machine and sprayed with an appropriate amount of alcohol (about 0.5 to 5 wt.% Relative to kelp) and the kneader was rotated at a low speed of 40 to 50 rpm at an internal temperature of 70 to 80 DEG C, When the intrinsic fragrance starts to evaporate, the internal temperature of the tumbling machine is sealed so that the internal temperature of the tumbling machine does not exceed 100 to 130 DEG C, and the high-speed rotation is performed at a speed of 200 to 250 rpm. To increase the extraction yield of the extraction process of the second step by doubling the fragrance of the kelp, weakening the shell and cell wall, and increasing the gap between the cells.

Next, distilled water is washed until the salinity is eliminated in the washing machine using rotation and vortex (S400). Dissolving and cleansing of distilled water effectively removes the foreign substances in the sea tangle, prevents the deposition and effectively dissolves the debris such as contaminants, and soaking the distilled water to neutralize the sea tangle and increase the antibacterial power.

The granular crushed kelp was washed with 10 times the weight of distilled water and 0.5 ~ 4 parts by weight of Celluclast, Viscozyme and Ultrazyme, which are complex polysaccharide degrading enzymes, were added at 100 ~ Dehydration).

Seaweed extracts are composed mainly of ash, lipids, carbohydrates, proteins and fucoidan indicators. Cellulose and Viscozyme, a complex polysaccharide degrading enzyme, decompose and break up complex polysaccharides. Fucoidan index materials with molecular weights of 300,000 ~ 1,500,000 And Ultrazyme decomposes the complex polysaccharide to increase the content of sulfate groups.

For the extraction at low temperature, the fluidity is given to the distilled water using an air brower (1.7㎥ / min) and a stirrer (100 ~ 120 rpm / 0.5hp) .

After the low temperature extraction, kelp extract is vacuum low temperature extraction at 35 ~ 50 ℃ by adding distilled water of 5 times of the individual quantity to the vacuum extractor and maintaining vacuum of 0.06 Torr (0.079 mbar, 0.0079 kPa).

Vacuum cold extraction is to extract the active ingredients which are not completely extracted by low temperature extraction. The low temperature extract and the vacuum low temperature extract are mixed at a ratio of 2: 1 to prepare the next step.

Next, a step of fermenting a mixture (2: 1 ratio) of the low-temperature extraction and the vacuum low-temperature extract after the enzymatic degradation with the microorganism fermentation, the microorganism strain used is Generally Recognized As Safe (USS FDA) And fermented with a strain having excellent fermentation ability for seaweed extract. Traditional fermented foods Lactobacillus , a strain isolated from kimchi, miso, leaven, and bread brevis DL-25, Saccharomyces cerevisiae DS-7, Aspergillus oryaze DF-11 in a culture for 10 to 30 hours in a seaweed extract. Microbial fermentation low molecular weight of fucoidan index substance with molecular weight of 200,000 ~ 1,000,000 and removes protein, fat, ash and so on to increase the yield of fucoidan indicator material, facilitate separation and purification, It is effective for improvement.

Next, to carry out the separation and purification step, the filtrate obtained by separating the microbial cells from the culture broth using a filter (Membrane filtration: MF) is obtained, and the obtained product is purified by ion chromatography (Ion-exchange Chromatography) .

Next, a vacuum state of 0.06 Torr (0.079 mbar, 0.0079 kPa) was maintained in the vacuum extractor, and the solidified fucoidan was obtained by distillation at 35 to 50 ° C or less at 2,500 to 3,000 cc per hour. The obtained fucoidan can be formulated and commercialized.

Hereinafter, embodiments of the present invention will be described

Example

The extracted kelp is air-washed in the first step of the present invention, granulated, and then subjected to thermo-plastic impregnation, followed by washing with distilled water, followed by low-temperature extraction and enzymatic decomposition / vacuum low-temperature extraction, followed by fermentation and separation of the extract. A physiologically active fucoidan having improved purity and extraction yield was prepared by preserving the active ingredient by the concentration method.

First, kelp (3 kg) was put in an automatic air washer (Rice Korea, Korea, standard: W1000xL4500xH1450) and cleaned. Then, granules were crushed to a size of (1 to 2 cm) using a granule crusher (Washing Tech, Korea, standard: W800xL800xH1400).

The shredded kelp was put into a kneader (THA-18, Taihwa Automated Industry Co., Ltd.). When 30 g of fermented seaweed (alcoholic beverage: 14 degrees) is injected evenly into the kelp, the low temperature is rotated at a rate of 50 rpm while the internal temperature of the sonicator is 70 ° C. When the original flavor of the kelp begins to evaporate, The internal temperature of the machine was set at 120 ° C and rotated at a high speed of 250rpm to induce an indirect heat of high speed rotation to double the flavor of the kelp, weakening the shell and cell wall of the kelp, and increasing the intercellular space to promote porosity .

Thermosetting (impermeable) kelp was put into a washing machine (Hyosung, Korea, standard: 1000xH1200) and distilled water was washed / dehydrated until the salinity became '0' by generating vortex in distilled water. When washing, dry seaweed (3kg) absorbs moisture and becomes 5 ~ 6 times (15 ~ 18kg) of initial modified weight.

(30kg) of distilled water of the initial individual (3kg of dried sea tangle) was added to the washed / dehydrated granule crushed kelp, 30g of Celluclast, Viscozyme and Ultrazyme were added and the temperature was maintained at 35-50 ° C. The extracts were subjected to low temperature extraction and enzymatic decomposition for 2 to 4 hours and compressed / dehydrated using an air brower (1.7 m 3 / min) and a stirrer (120 rpm / 0.5 hp) Respectively.

Next, 5 times (15 kg) of distilled water of the initial amount (dried seaweed-3 kg) was added to a vacuum extruder (Kyeongseo Machinery Co., Ltd., Korea / COSMOS 660, specification: W1000xL500xH1800, capacity: 80,000 cc) The vacuum state of Torr (0.079 mbar, 0.0079 kPa) was maintained and the vacuum low-temperature extraction was carried out at 35 to 50 ° C for 2 to 4 hours. Then, 12 kg of kelp extract was obtained by compression / dehydration.

10 kg of the extracted 26 kg of low-temperature extract and 5 kg of 12 kg of vacuum low-temperature extraction were mixed (2: 1), and 15 kg of the extract was put into a fermenter (Kobiotec Korea, 50 L) Lactobacillus brevis DL-25 was added and cultured for 9 to 12 hours.

14 kg of the filtrate obtained by separating the microbial cells from the culture broth by using a filter (Membrane filtration: MF) was obtained and the obtained product was purified by ion chromatography (Ion-exchange chromatography) to obtain 12 kg of purified liquid.

The vacuum extractor was maintained at 0.06 Torr (0.079 mbar, 0.0079 kPa) in vacuum and distilled at 45 ° C for 4 hours (2,500 to 3,000 cc / h) to obtain 120 g of solid component and 11.8 kg of distillate.

The solid components thus obtained were analyzed for general components, monosaccharide, and sulfate groups. The solid content obtained by the above method was subjected to monosaccharide analysis and sulfate measurement by a laboratory of the Marine Resources Utilization Engineering Laboratory of the Marine Science University of Cheju National University.

General components: According to AOAC (1990), moisture was measured by atmospheric pressure 105 heat drying method, fat was measured by Soxhlet method, crude protein was measured by semi-micro Kjeldahl method and ash was measured by dry painting method. Carbohydrate quantification was performed by using Dubois et al. Respectively.

Monosaccharide analysis: Samples were prepared at a concentration of 10 mg / ml and mixed with 4M TFA in a 1: 1 ratio (sample concentration: 5 mg / ml) and then hydrolyzed in a 105 ° C oven for 4 hours. 200 μl of the hydrolyzate was transferred to an e-tube and then removed by using a vacuum filter to remove TFA. 1 ml of D. w was added to prepare a sample at a concentration of 1 mg / ml.

Sulfate group measurement: Ammonium sulfate was used as a standard and the content was measured using BaCl2 / gelation method.

The measured results are shown in Table 1. That is, the weight ratio of fucoidan indicator substance contained in seaweed (solid content) was shown.

Fucose 3.24 g / 100 g Galactose 0.46 g / 100 g Glucuronic acid 0.7g / 100g Xylose 0.35 g / 100 g Sulfate content 1.52 g / 100 g system 6.27 / 100g

As shown in Table 1, the distribution of fucose, galactose, clucuronic acid, xylose, and sulfate groups that determine the quality of fucoidan was evenly distributed. The yield was also about 50% at about 6.27% And the yield was increased.

Therefore, according to the method of the present invention, it is confirmed that although the process is reduced compared to the conventional method, the yield is high, the loss of the effective ingredient is small, and the purity of the indicator substance of fucoidan is improved.

The above method is not limited to sea tangle, but can be applied to all seaweeds such as seaweed, marigold, and the like.

Claims (1)

The low-temperature extract and the vacuum low-temperature extract were mixed with Celluclast, Viscozyme, and Ultrazyme as degrading enzymes, and Lactobacillus brevis DL-25 strain was inoculated for 10 to 30 hours with sea tangle extract , And then separating and purifying the fermentation broth. The method for producing high purity fucoidan according to claim 1,

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