KR20120113158A - Processing method of oligo -porphyran from porphyra sp - Google Patents

Abstract

The present invention red algae; A technique for the extraction, concentration, duck glycosylation and purification of porphyran with excellent physiological activity from laver ( Porphyra yezoensis ), specifically red algae; laver (100 kg) or dried (10 kg) Into the same volume of spirits (80%), extracted with karaoke tosyl-glycerol (Galactosyl-glycerol), pigments and fat overnight (12 hours) at room temperature, filtered and removed, and reverse osmosis distilled water 2 times the volume and heated After stirring for 4 hours at 90 ° C., it was filtered through a diatomaceous earth filter system to filter out the aggregated protein, and concentrated. Then, double the volume of alcohol (> 95%), and overnight at low temperature (4 ° C.) (12 hours). The porphyran was precipitated and centrifuged (1600 xg, 20 minutes) to obtain crude porpyran, washed with alcohol and dried or concentrated in 2.5 liters of 10 mM sodium phosphate buffer (pH6.8) and sterilized at 121 ° C. minute Then, add enzyme (Agarase EC 3.2.1.81) (940 ug, 3250units) at 40 ℃, mix well for 2 minutes, cool down the reaction temperature at 25 ℃ for 12 hours, heat treatment at 100 ℃ for 10 minutes to stop enzyme activity. Concentrate oligo porpyran to a small amount, and add twice the volume of alcohol (> 95%). At low temperature (4 ℃), stand overnight (12 hours) to precipitate protein (enzyme) and centrifuge (5,000 xg, 20 minutes). It is a research and development process for the production process of oligoporium, dried and healthy functional food ingredients, after 8 hours dialysis (4 ℃) in sterilized distilled water after washing with oligosaccharide (> 95%).

Description

Process method of oligo -porphyran from Porphyra sp

The present invention red algae; A technique for the extraction, concentration, duck glycosylation and purification of porphyran with excellent physiological activity from laver ( Porphyra yezoensis ), specifically red algae; laver (100 kg) or dried (10 kg) Into the same volume of spirits (80%), extracted with karaoke tosyl-glycerol (Galactosyl-glycerol), pigments and fat overnight (12 hours) at room temperature, filtered and removed, and reverse osmosis distilled water 2 times the volume and heated After stirring for 4 hours at 90 ° C., it was filtered through a diatomaceous earth filter system to filter out the aggregated protein, and concentrated. Then, double the volume of alcohol (> 95%), and overnight at low temperature (4 ° C.) (12 hours). The porphyran was precipitated and centrifuged (1600 xg, 20 minutes) to obtain crude porpyran, washed with alcohol and dried or concentrated in 2.5 liters of 10 mM sodium phosphate buffer (pH6.8) and sterilized (121 degrees Celsius). In 15 minutes), add enzyme (Agarase EC 3.2.1.81) (940 μg, 3250 units) at 40 ° C, mix well for 2 minutes, react for 12 hours at 25 ° C, and heat-process at 100 ° C for 10 minutes for enzymatic activity. Stop, raise oligo porphyran to a small amount, and add twice the volume of alcohol (> 95%). At low temperature (4 ℃), stand overnight (12 hours) to precipitate protein (enzyme) and centrifuge (5,000 xg, After 20 minutes), it is washed with alcohol (> 95%) and porpyran, and then dialyzed in sterilized distilled water (4 ℃) for 8 hours.

Red algae; Porphyra sp. Is harvested and washed in aquaculture farms, and is dried by light drying after acid treatment, as described in recently published Patent Publication No. 10-2009-0090237 (2009-08-25). It has reached the stage of developing with bar porphyran jam or seaweed jam, but it has not reached purity and concentration that can be functionalized through the extraction, concentration, oligomerization and purification process into porpyran, and is used as a raw material of health functional food. This is not. That is, the disclosed patent method of extracting using hydrochloric acid is a method that is not allowed to be used for food, and the extraction for use as a food in the food process should be improved because only water, alcohol and carbon dioxide can be used as the solvent. Porphyran is a polysaccharide, which cannot be broken down by the digestive enzymes that our body possesses, so it can only serve as a dietary fiber and must be absorbed in order to exhibit the following physiological activities: oligomers or monomers. It should be low molecular weight with (Monomer).

The following describes the biological activity and research literature of oligo porphyran.

1) lowering blood pressure and lowering serum cholesterol [D. Ren, et al. (1994). Study on antihypertensive and antihyperlipidemic effects of Marine algae. Fisheries Sci ., 60, 83-88 (1994)],

2) antitumor activity [H. Noda, et al. (1989) Antitumour activity of Polysaccharide and lipids from Marine Algae. Nippon Suisan Gakkaishi , 55, 1285-1271],

3) antiallergic action [K. Ishihara et al. (1998). Inhibitory Effect of Porphyran, prepared from Dried "Nori", on Contact Hypersensitivity in Mice. Biosci. Biotechnol. Biochem. 69 (10): 1824-1830],

4) Immune activity [Y. Yoshizawa et al. (1995) Macrophage Stimulation of the Polysaccharide fraction from Marine Algae ( Porphyra yezoensis ). Biosci. Biotechnol. Biochem., 59 (10): 1933-1937].

Food raw material extraction solvent applied to the food industry; In order to extract and obtain high-purity porpyran by using any or several kinds of water, ethanol and carbon dioxide gas, unnecessary materials other than porpyran (hereinafter referred to as "impurity") are removed by appropriate techniques before and after extraction and bioactive Small molecule sizes that can be absorbed from the digestive tract using enzymes while maintaining maximum sulfuric acid groups essential to the organs; Decomposes oligomers and removes impurities, including used enzymes, to produce porphyrans with high purity and sufficient physiological activity, especially when the microorganisms should not be contaminated, dried in a state where microorganisms cannot grow, and in high concentration liquids. It should be possible to supply sterilized seals filled in containers and supply them as functional food ingredients.

In order to obtain high purity oligo porphyran, the following four step process (work) is carried out in order.

First step (washing and desalting): Removes impurities and saltiness and provides clean raw materials. Drinking water is poured into an extraction tank with a double volume of seaweed ( porphyra sp), followed by agitation, followed by washing and dehydration.

Second step (degreasing and discoloring): The laver washed by the extraction and removal of impurities during grinding, with the same volume of alcohol (80%) in the blender, crushed and transferred to the extraction tank, and then the impurity garaxil griesol ( Stir galactosyl-glycerol), pigment and fat at room temperature for 12 hours, filter it, remove it, and transfer to porphyran extraction tank.

The third step (porpyran extraction): 2 times the volume of reverse osmosis treated water into the porphyran extraction tank, warmed and stirred for 4 hours while stirring at 90 ° C to filter and remove the aggregated protein by diatomaceous earth filter system. Next, a double volume of alcohol (> 95%) was added, and the mixture was allowed to stand overnight at low temperature (4 ° C) (12 hours), followed by centrifugation (5,000xg, 20 minutes) to recover the precipitated porpyran (> 95%). Wash and dry.

4th step (oligo poryranization): 2.5 liters of 10 mM sodium phosphate buffer (pH6.8) was added to the reaction tank, and concentrated or dried porpyran (50 g) was added and sterilized (15 minutes at 121 ° C.). Add enzyme (Agarase EC 3.2.1.81) (940 μg, 3250units) at 40 ℃, mix well for 2 minutes, react for 12 hours at 25 ℃, heat treatment at 100 ℃ for 10 minutes to stop enzymatic activity Reduce the concentration to a small amount and add twice the volume of alcohol (> 95%) to stand overnight (12 hours) at low temperature (4 ℃) to precipitate the protein (enzyme) to remove centrifugation (5,000 xg, 20 minutes) Oligo (> 95%), washed porpyran, then dialyzed in sterile distilled water for 8 hours (4 degrees Celsius), dried and packaged.

High purity porpyran (> 85%) can be obtained and oligopolpyrans can be absorbed by the human body without damaging or reducing sulfuric acid groups, which are essential for physiological activity, and can be supplied as raw materials for health functional foods.

Red algae laver fishermen harvested laver and harvested it from the cold winter sea to concentrate on producing corrosive laver. However, it is not enough to compensate for the efforts. You will expand your kite or seaweed production to produce more seaweed.

On the other hand, oligo porphyran-containing health functional foods will be more easily obtained, which will contribute to the improvement of national health by increasing the number of people who receive the four biological activities described above.

In order to commercialize the invention, it is necessary to first receive the advice of the Korea Food and Drug Administration, identify all the requirements necessary for obtaining porphyran as a raw material for dietary supplements, and to apply for and approve it as a dietary supplement. Apply for a patent for this manufacturing method, register it with a patent, apply for an international patent, and take advantage of existing facilities to produce and scale pilot scales and supply samples to health food manufacturers. In addition, we plan to encourage and support the commercialization of health functional foods, and to open the way of supplying raw materials to health functional food companies by expanding their promotional activities based on the results of market research and verification of their own prototypes. .

In addition, find health functional food manufacturers and health functional food raw material suppliers in the United States, Japan, Europe, Southeast Asia, etc. to promote raw materials (products) and send samples of raw materials to check properties and quality, and open trade. If possible, the company plans to exhibit at internationally renowned international fairs and expand the production to find demand.

Due to the above activities, demand will increase, and we plan to apply for occupancy in Ori Industrial Complex of Gijang, Busan. The demand generated before the completion of the facility will be rented out of the Marine Bioindustry Center. In addition, we are planning to utilize the facility [Kumilsu Hyup Fisheries Co., Ltd.] in Jukcheong Agricultural Industrial Complex in Wando, and if the patent is recognized and registered, we will make every effort to commercialize it and achieve the goal of exporting porphyran to the world market. .

M = moles / liter.

Claims (2)

A method of obtaining oligo porphyran, which is obtained by harvesting red algae ( Porphyra sp) and carrying out the operation of the dried or raw material based on the above four-stage process, that is, washing the impurities into the same amount of alcohol (80%). Extraction is removed by filtering, 4 hours of porphyran is extracted, concentrated and filtered through a diatomaceous earth filter system while stirring at 90 ° C with a double volume reverse osmosis treatment water to remove impurities (proteins) and concentrating. After standing for 12 hours at a low temperature (4 ℃) with a volume of alcohol (> 95%), porphyrane were precipitated and centrifuged to obtain centrifugation.The solution was sterilized with 10 mM sodium phosphate buffer solution (pH 6.8) and then 40 ℃. After cooling, the enzyme (beta agarase) was mixed well for 2 minutes, the temperature was kept at 25 ° C. for 12 hours, and heat treated at 100 ° C. for 20 minutes to stop enzymatic activity, followed by a double dose of alcohol (> 95%). Small way to remove by centrifugation the precipitate (protein) and concentrated to a low temperature in a sterile distilled water after 8 hours dialysis at (4 ° C), collected, dried up to prepare a formyl pyran. Oligo porphyran obtained by applying the claim 1