KR20050122289A - Manufacturing process of chitosan-selenium complex - Google Patents

Abstract

The present invention relates to a method for preparing an organic chitosan-selenium complex in the form of chitooligosaccharide by combining selenium as a mineral using chitosan and then decomposing it with chitosan degrading enzyme. It not only reduces the toxicity of inorganic selenium, but also increases the absorption in vivo, so it can be expected to boost the immune system of chitosan and anti-cancer and antioxidant activity of selenium, so it can be used as a health supplement, feed additive, and functional fertilizer. have.

Description

Manufacturing process of Chitosan-Selenium Complex

Selenium (Se) continued to be discovered in 1957 as it was essential for microbial and animal nutrition, and deficiencies in many animals were due to the lack of selenium, and in humans, as in animals, they are essential for metabolism. It became known. Selenium is an element belonging to group 6B of the chemical periodic table and is contained in large quantities in brewer's yeast, pig and beef kidneys, butter, malt, oyster, shrimp, milk, brown rice and garlic. In animals such as humans, they are mainly present in the form of seleno amino acids such as selenocysteine and selenomethionine, and they activate glutathione peroxidase, which acts to remove hydrogen peroxide, which causes damage to the cell membrane. Sies, H. et. Al, Oxidation in the NAKP system and release of GSSG from hemoglobin-free rat liver during peroxidatic acidation of glutathione by hydroperoxides, FFBS Letter , 27: 171-175, 1972). In addition, anemia caused by lead poisoning causes hematopoiesis by increasing the excretion of the urinary tract of aminolevulinic acid by inhibiting the activity of S-aminolevulinic acid dehydrase (ALAD) enzyme, which is involved in hemolysis of hemoglobin and hemoglobin synthesis In addition to inhibiting the action, it is known that lead is caused by inactivation of the SH group of ALAD and deteriorating hematopoietic function. It has been reported that lead poisoning can also be detoxified by selenium, known as an antioxidant in vivo (Hsu, PC, and Guo, YL, Antioxidant nutrients and lead toxicity, Toxicology , 180: 33-44, 2002). It is thought that glutathione peroxidase, a selenium-dependent antioxidant enzyme whose activity has been attenuated by selenium supplementation, significantly increases its activity. In addition, the antioxidant power of selenium is 1,975 times that of tocopherol, and the combination of tocopherol and selenium has a synergistic effect.It has a therapeutic effect on dandruff and psoriasis and increases the utilization of oxygen to improve heart and lung function. In addition, it inhibits the oxidation of unsaturated fatty acids such as vitamins C and E to suppress the production of lipid peroxide, and also decomposes waste materials, which are a combination of lipid peroxide and protein, and combines with heavy metals such as mercury, cadmium, and lead to burn them. It activates and prevents necrosis of hepatocytes, thereby preventing liver cirrhosis, improving immune function, improving sexual function, treating muscle nutrition and preventing cancer. Supplementation of selenium even in people who do not have enough intake of selenium may boost the immune system (Baum, MK, et al, Selenium and interleukins in persons infected with human immunodeficiency virus type 1., J. Infect. Dis. , 182: 69-73) and anti-cancer effects (Combs, GF, et al, Chemopreventive agents: selenium, Pharmacol. Ther. , 79 (3): 179-192, 1998). The recommended daily intake of selenium is 55 ㎍ in adult, 60 ㎍ in pregnant women and 70 ㎍ in lactation period. The insufficiency of selenium is inhabited by soils with low selenium content and those who have weakened selenium absorption due to gastrointestinal dysfunction. This has been observed in patients who have received nutritional supplements that do not contain selenium. Excessive intake of selenium has side effects such as gastrointestinal disorders and neurological disorders due to selenium toxicity, so the maximum daily intake is limited to 400 µg. Selenium, which is present in the form of an inorganic form, has a low absorption rate in the body after ingestion, and organic selenium combined with an amino acid has a high absorption rate in vivo. This suggests that selenium excretion and intracorporeal binding in milk are higher with the addition of organic selenium preparations than with the addition of selenite. Inorganic forms of selenium have low bioavailability and are therefore contained in livestock excretion and may cause environmental problems that accumulate in rivers and soils. Organic selenium combines with amino acids such as selenomethionine in grains. It is the ideal structure that has the best absorption rate, and the presence of selenium in selenium-containing yeast contains the constituents of the selenium protein, which is in the form of binding to amino acids. Recently, edible plants that have accumulated organic selenium by absorbing inorganic selenium to plants by utilizing selenium metabolism of plants have been developed as a source of selenium (see: Organic Selenium-containing PR Rice and Manufacturing Method thereof, Korean Patent Registration No. 10 -0404067), manufacturing method of yeast containing organic selenium (Reference: Republic of Korea Patent Registration No. 10-0434949, Republic of Korea Patent Registration No. 0142891) and Manufacturing Method of Selenium Organic Compound (Reference: Republic of Korea Patent Publication No. 1991-0002534) Studies have been conducted on.

As described above, a technique for preparing organic selenium has been developed and commercialized by chelation of selenium with amino acids methionine and cysteine or by chelation of yeast intracellular protein with methionine and cysteine among constituents. Considering that no method has been developed to chelate selenium, while researching ways to improve the absorption of selenium and reduce the toxicity of inorganic selenium in living organisms, chelate inorganic selenium to chitosan, which is safe for human body and has excellent biological activity. The method was developed to complete the present invention.

Accordingly, an object of the present invention is to chelate inorganic selenium to chitosan, which is safe for human body and excellent in physiological activity, and not only reduces the toxicity of inorganic selenium, but also increases absorption in vivo, thereby enhancing the immune activity of chitosan and selenium. Anticancer activity can be expected to provide a method for producing a chitosan-selenium complex that can be utilized as a health supplement, feed additives, functional fertilizers, and the like.

As described above, the chitosan source for preparing the chitosan-selenium complex, which is an object of the present invention, may be a polymer chitosan having various molecular weights, but preferably, chitosan having a molecular weight of about 300,000, and sodium selenium as an inorganic selenium source. All compounds containing selenium such as Na 2 SeO 4), sodium selenite (Na 2 SeO 3), selenium dioxide (SeO 2), and the like can be used, but sodium selenite (Na 2 SeO 3) is preferably used.

The method for producing chitosan-selenium complex according to the present invention consists of seven steps.

Step (1): dissolving inorganic selenium in distilled water

Step (2): dissolving chitosan by adding an appropriate amount of acetic acid to step (1)

Step (3): organic passivation reaction with chelate of dissolved selenium and chitosan

Step (4): enzymatic oligoglycosylation reaction of chitosan-selenium complex of step (3)

Step (5): chitosan degrading enzyme inactivation step (4)

Step (6): ion exchange adsorption removal of unreacted inorganic selenium

Step (7): Microfiltration to remove insoluble matters and complete organic chitosan-selenium complex

Hereinafter, the present invention will be described in detail, but the scope of the present invention is not limited thereto.

Example Method for producing chitosan-selenium complex

1 liter of distilled water was added to a reactor controlled by temperature and stirring speed, and 4.386 grams of sodium selenite (Na2SeO3, Se content 45.6%, Shinko Chemicals Co., Japan) was dissolved as inorganic selenium, and the polymer chitosan (MW 300,000, Kluka Co. , Switzerland) was added and swelled chitosan for 1 hour with stirring at room temperature at 300 rpm. 15 milliliters of glacial acetic acid (Shinyo Pure Chemicals Co., Japan) was added to dissolve chitosan, and the reaction was stirred at 600 rpm while maintaining the organic passivation reaction of chitosan and selenium at 80 ° C for 3 hours, and then cooled to 50 ° C. To prepare a polymer chitosan and selenium complex. In order to convert the polymer chitosan and selenium complex into low molecular weight chitosan oligosaccharides that can be easily absorbed in vivo, 10 U chitosan degrading enzyme (Sigma Co., USA) was added and stirred at 600 rpm for 2 hours while maintaining 50 ° C, followed by 300 rpm for 8 hours. Enzymatic reaction was performed for 10 hours while stirring, and the enzyme reaction temperature was raised to 90 ° C. to inactivate chitosan degrading enzyme for 15 minutes, thereby preparing an oligosaccharide chitosan-selenium complex.

12 minutes per ion exchange column (300 × 20 mm) filled with an anion exchange resin (Amberlite S IRA-96, Sigma Co., USA) to remove unreacted inorganic selenium from the reaction solution containing chitosan-selenium complex After passing through a milliliter flow rate, the unreacted inorganic selenium was adsorbed and removed, and an insoluble substance was filtered through a Wattman filter paper to prepare a chitosan-selenium complex. A control was prepared in the same manner as above without adding only sodium selenite.

1 is a result of analyzing the selenium content in the chito and organic chitosan-selenium complex (se-chito) by the inductively coupled plasma analyzer (ICP) as requested by the Korea Testing and Research Institute, while the control was not detected selenium In the 2000 ppm organic chitosan-selenium complex of 1803ppm was detected it was confirmed that about 90% chelated.

The present invention prepared organic chitosan-selenium complex can not only reduce the toxicity of inorganic selenium and improve the absorption in vivo, but also can simultaneously expect physiological activities such as immune enhancement of chitosan and anticancer and antioxidant effects of selenium. It can be used as a health supplement, feed additives, and functional fertilizers.

1 is a selenium content of the chitosan oligosaccharide (denoted 'chito') and the organic chitosan-selenium complex (denoted 'se-chito') without selenium, commissioned by the Korea Research Institute of Chemical Testing (ICP). As a result of analysis, selenium-free chitosan oligosaccharides did not detect selenium, whereas the organic chitosan-selenium complex treated with 2000ppm detected 1803ppm.

Claims (1)

A method for producing a chitosan-selenium complex comprising combining inorganic selenium with polymer chitosan and organolating it and oligosaccharide polymerizing chitosan with chitosan degrading enzyme.