CN1570093A

CN1570093A - Extraction method for SOD complex enzyme fine powder from corn

Info

Publication number: CN1570093A
Application number: CNA031502466A
Authority: CN
Inventors: 韩伯钧; 韩旺
Original assignee: Individual
Current assignee: Individual
Priority date: 2003-07-23
Filing date: 2003-07-23
Publication date: 2005-01-26
Anticipated expiration: 2023-07-23
Also published as: CN1313606C

Abstract

A SOD composite enzyme powder production method extracting from corn is disclosed. The technical scheme comprise: treating corn, crushing and beating it, at the same time adding natural antioxidant, obtaining homogenate, filtering, heating and removing protein, microfiltering, ultrafiltrating, nano filtering and concentrating it, column chromatography, nano filtering for concentrating chromatographic liquid, separating out SOD using organic solvent, separating SOD, low temperature vacuum drying and obtaining the products SOD composite fine powder. The advantages of the invention: adding natural antioxidant while crushing corn protection SOD activity to a large extent. The inventive SOD composite enzyme fine powder has high purity and activity.

Description

Method for extracting SOD complex enzyme fine powder from corn

The invention relates to an industrial production method for extracting superoxide dismutase (SOD) complex enzyme fine powder from corn.

Superoxide dismutase (SOD) is a great medical finding in the twentieth century. Since the first discovery of SOD from bovine blood cells by Muord in 1969, the international biochemical and medical communities have attracted great attention. SOD becomes the most popular topic of the international pharmaceutical industry in the twentieth century. A great deal of medical research proves that SOD is an important human superoxide anion free radical O₂ ^-Scavenger for clinically treating various diseases caused by superoxide anion free radical produced by human metabolism, such as various inflammations, rheumatism, and rheumatoid diseasesImmune diseases, alcoholic hepatotoxicity, atherosclerosis, coronary heart disease, acute myocarditis, pulmonary edema, emphysema, Parkinson's disease, acute and chronic renal failure, uremia and diabetes mellitus, and particularly has special effects on preventing cancer cell diffusion and metastasis, resisting radiation, enhancing immunity, resisting aging and beautifying.

SOD has been widely used as a pharmaceutical agent in developed countries such as the United states, Germany, Japan, Canada, Australia, France, etc. At home and abroad, there are some SOD-containing foods, beverages, cosmetics, skin care products and the like which have the effects of whitening, rejuvenating and delaying senility. Therefore, the SOD has wide market prospect in the fields of medicines, foods and cosmetics and health care products.

People firstly extract SOD from animal blood to prepare products, and the animal blood contains mad cow disease factors and morbid factors internationally, so people are unsafe after application. The international blood defence organization has stated that the extraction of SOD from animal blood is prohibited to produce various products. It is required to extract SOD from plants and microorganisms containing SOD to produce various products.

The plant contains many SOD plants, such as fructus Rosae Normalis, fructus Hippophae, semen glycines, horse radish, Aloe, etc. But the most promising development is the extraction of SOD from corn. China is the second big base of world high-yield corn, and annual output is billions of tons. The high-yield corn is partially eaten, and a part of the corn is subjected to deep processing except for producing feed. The deep processed product comprises starch, modified starch, and fructose-glucose syrup, and is prepared by fermenting corn flour to produce lysine, citric acid, nucleic acid, etc. However, the most promising and economic value in the deep processing of corn is to extract SOD from corn.

In the prior art, patents have been reported on the aspect of extracting SOD from corn, and finally, the prepared product is liquid; however, because SOD is extremely unstable, SOD is very easy to oxidize, degrade, denaturize and inactivate in the extraction process, and the measured activity unit of the finally obtained product is very low, even 0. Such SOD liquids are not well stored, transported and processed. Therefore, industrial implementation is difficult.

Aiming at the problems of SOD extraction from corn, the invention provides an industrial manufacturing method for extracting SOD complex enzyme fine powder from corn, which can well solve the problems of the prior method for extracting SOD from corn.

The invention is realized by the following process route:

grading and screening corn, sterilizing, washing, soaking in warm water, pulverizing, pulping while adding antioxidant for protection, stirring, plate-and-frame filter pressing, removing protein by heat treatment, micro-filtering, ultrafiltering, concentrating by nanofiltration, refining by column chromatography, concentrating by nanofiltration, and separating SOD and G with organic solvent₄Filtering and separating to obtain wet SOD product, and vacuum drying at low temperature to obtain SOD complex enzyme fine powder.

1. Grading and screening the fed corn, and removing the small, immature and brown corn kernels.

2. And (3) preparing 1% sodium hypochlorite to disinfect the corn kernels and kill bacteria on the surfaces of the corn kernels.

3. And washing the disinfected corn kernels with clear water until no disinfectant is left.

4. After cleaning, the corn kernels are soaked in water at 18-32 ℃ for 18-60 hr.

5. After the corn kernels swell, the corn kernels are crushed and pulped, natural antioxidant is added for protection, and the pH value is kept between 5.0 and 7.8 by using a phosphate buffer solution.

6. Stirring for 1-2hr, filtering with plate frame, and collecting filtrate.

7. Heating the filtrate to 40-65 ℃, stirring for 10-60 minutes, cooling the filtrate to-4 ℃, performing centrifugal separation to precipitate protein, and collecting the filtrate.

8. Microfiltering with inorganic membrane, and collecting filtrate.

9. And (4) carrying out ultrafiltration on the filtrate to remove macromolecular protein and miscellaneous enzymes.

10. After ultrafiltration, the filtrate is concentrated to the original volume of 1/2 by nanofiltration, and the molecular weight cut-off is above 15000 daltons.

11. Column chromatography on nanofiltration solution (DEAE-Sephardex G)₁₅₀Column), active fractions were collected.

12. And (4) carrying out nanofiltration concentration on the column chromatography liquid until the original volume is 1/5-1/10.

13. Adding organic solvent into the nanofiltration concentrated solution to separate out SOD.

13、G₄Filtering and separating to obtain wet SOD product.

13. Vacuum drying wet SOD product at low temperature to obtain SOD complex enzyme fine powder.

15. The SOD complex enzyme fine powder is packed in a nontoxic plastic container in vacuum according to a certain weight specification.

The invention has the following characteristics:

1. the SOD complex enzyme fine powder prepared by the method takes SOD as a main enzyme and contains one complex enzyme of auxiliary enzymes catalase CAT and peroxidase POD, and the process of eliminating superoxide anion free radicals in vivo by the SOD comprises the following steps:

SOD enzyme making superoxide anion free radical O in vivo₂ ^-Hydrogen peroxide H after disproportionation₂O₂If the hydrogen peroxide is not cleared in time, it is still toxic to human cells.Because the SOD extracted from the corn is the compound enzyme containing catalase CAT and peroxidase POD, the compound enzyme can completely eliminate the free radicals in the body under the catalytic action, and protect the cells of the human body from being damaged by the superoxide anion free radicals.

2. The SOD complex enzyme fine powder prepared by the method adopts the measures of protecting SOD in the extraction and processing, so that the SOD is not oxidized, degraded and denatured and inactivated in the processing, and the SOD activity is kept to the maximum extent.

3. The invention prepares SOD complex enzyme fine powder; has the characteristics of high purity and strong vitality unit, and can be used as food, health food, daily chemical skin care and beauty products and various medical products such as: tablet, capsule, oral liquid, injection and infusion solution, and additives.

China is a world-high corn-producing big country, the annual output is hundreds of millions of tons, and how to digest the high-yield corn becomes a big matter concerned by governments at all levels, which is related to the problems that vast farmers are off poverty and rich and the structures of agricultural products are adjusted. Because some provinces are in the corn deep processing and business recruitment project. After the industrial implementation of the invention, the invention has profound influence on the upgrading and updating of corn deep-processing products in China and the production and application of SOD medical products.

Claims

1. A method for extracting SOD complex enzyme fine powder from corn is technically characterized by comprising the following steps:

1) after the corn is pulped and pretreated (the pretreatment process comprises corn grading screening, disinfection, washing and soaking), the expanded corn is crushed and pulped, meanwhile, a natural antioxidant is added for protection, and a phosphate buffer solution is used for enabling the pH value to reach 5.0-7.8.

2) Stirring for 1-2hr after homogenizing, and filtering with plate-and-frame filter to collect filtrate.

3) Heating the filtrate to 40-65 ℃, and stirring for 10-60 minutes. Cooling the filtrate to-4 deg.c, centrifugally separating to precipitate protein and collecting the filtrate.

4) Filtering the filtrate with inorganic membrane, and collecting the filtrate.

5) And (4) carrying out ultrafiltration on the filtrate to remove macromolecular protein and miscellaneous enzymes.

6) After ultrafiltration, the filtrate is concentrated by nanofiltration to the original volume of 1/2, and the molecular weight cut-off is above 15000 daltons.

7) Column chromatography on nanofiltration solution (DEAE-Sephardex G)₁₅₀Column) the active fraction was collected.

8) And performing nanofiltration on the column chromatography liquid to concentrate the original volume of 1/5-1/10.

9) Adding organic solvent into the nanofiltration concentrated solution to separate out SOD. G₄Filtering and separating to obtain wet SOD product.

10) Vacuum drying wet SOD product at low temperature to obtain SOD complex enzyme fine powder.

11) The SOD complex enzyme fine powder is packed in a nontoxic plastic container in vacuum according to a certain weight specification.

2. SOD complex prepared by the method of claim 1The enzyme fine powder contains not only superoxide dismutase SOD main enzyme, but also auxiliary enzymes catalase CAT and peroxidase POD, and the compound enzyme can make the superoxide anion free radical in vivo

Completely eliminates the toxicity of superoxide anion free radicals to cells fundamentally.

3. The SOD complex enzyme fine powder prepared by the method of claim 1 can be used as food, healthfood, daily chemical skin care and beauty products and various medical products, such as: tablet, capsule, oral liquid, injection and medicinal raw material and additive for infusion.