(19)
(11)EP 2 963 003 B1

(12)EUROPEAN PATENT SPECIFICATION

(45)Mention of the grant of the patent:
11.12.2019 Bulletin 2019/50

(21)Application number: 13876093.9

(22)Date of filing:  26.12.2013
(51)International Patent Classification (IPC): 
C05D 5/00(2006.01)
C05F 11/10(2006.01)
C05D 9/02(2006.01)
(86)International application number:
PCT/CN2013/090593
(87)International publication number:
WO 2014/131309 (04.09.2014 Gazette  2014/36)

(54)

HIGH-ENERGY COMPOSITE PEPTIDE SELENOPROTEIN NUTRIENT SOLUTION, PREPARATION METHOD AND APPLICATION THEREOF

HOCHENERGETISCHE ZUSAMMENGESETZTE PEPTID-SELENOPROTEIN-NÄHRLÖSUNG, HERSTELLUNGSVERFAHREN UND ANWENDUNG DAVON

SOLUTION DE NUTRIMENTS COMPOSITE À HAUTE ÉNERGIE, À BASE D'UNE SÉLÉNOPROTÉINE PEPTIDIQUE, SON PROCÉDÉ DE PRÉPARATION ET APPLICATION


(84)Designated Contracting States:
AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

(30)Priority: 27.02.2013 CN 201310060994

(43)Date of publication of application:
06.01.2016 Bulletin 2016/01

(73)Proprietor: Xie, Chao
Beijing 100039 (CN)

(72)Inventors:
  • CUI, Jianjiu
    Beijing 100039 (CN)
  • FU, Huadiao
    Beijing 100039 (CN)

(74)Representative: Rhein, Alain 
Cabinet Bleger-Rhein-Poupon 4A, rue de l'Industrie
67450 Mundolsheim
67450 Mundolsheim (FR)


(56)References cited: : 
CN-A- 1 412 158
CN-A- 101 817 710
CN-A- 102 701 844
CN-A- 103 120 110
CN-A- 101 462 904
CN-A- 102 617 247
CN-A- 103 120 107
CN-A- 103 145 495
  
  • DATABASE WPI Week 200952 Thomson Scientific, London, GB; AN 2009-L05838 XP002762135, -& CN 101 462 904 A (LI G) 24 June 2009 (2009-06-24)
  • DATABASE WPI Week 201282 Thomson Scientific, London, GB; AN 2012-N83058 XP002762136, -& CN 102 617 247 A (CHANGSHU PINGYE MACHINERY CO LTD) 1 August 2012 (2012-08-01)
  • DATABASE WPI Week 201379 Thomson Scientific, London, GB; AN 2013-R11234 XP002762050, & CN 103 120 107 A (XIE C) 29 May 2013 (2013-05-29)
  
Note: Within nine months from the publication of the mention of the grant of the European patent, any person may give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention).


Description

Technical Field



[0001] The present invention relates to a crop selenium-rich solution, in particular, to a composite peptide selenoprotein nutrient solution, specifically a composite peptide selenoprotein nutrient solution containing peptide selenoprotein, rich minerals, vitamins and natural energy minerals, etc..

Background Art



[0002] Selenium deficiency is closely related to people's health. In 1973, the World Health Organization announced to the world, the trace element selenium is an important and indispensable element in human's life activities. In 1988, the trace element selenium was listed in one of the fifteen essential dietary nutrients that are taken up every day by Chinese Nutrition Society.

[0003] China is poor in selenium. According to measurement of relevant department, selenium is lack in 72% lands, and seriously lack in 29% lands. Seventy percent of populations (as high as one billion, covering 22 provinces) need to supplement selenium. The deficiency in selenium may directly lead to decreased immunity according to analysis of scientists. The clinical medical evidences also show that selenium deficiency is associated with over 40 kinds of diseases that threaten to human health and life, such as cancers, cardiovascular diseases, liver diseases, cataracts, pancreatic diseases, diabetes, Keshan disease, reproductive system diseases and so on.

[0004] The main functions of selenium are as follows: (1) anti-oxidation, anti-aging, "the first element to keep people young"; (2) to enhance immunity and prevent diseases. Selenium is called as "the king for anti-cancer" by the World Health Organization; (3) to detoxify and disperse blood stasis. Selenium can promote the tissues and organs to eliminate the heavy metals, pesticide residues and excessive blood sugar, blood fat and cholesterol from the body, and purify the circulatory system; (4) radiation protection; (5) to improve the quality of agricultural products, enhance the activity of plant cells, and more and adequate nutrition; (6) to resist pests and diseases. Selenium can partially replace pesticides, to reduce amount of pesticides and pesticide residues. Therefore, selenium plays an irreplaceable role in the health of human and plants.

[0005] The selenium deficiency in human body is associated with the health of millions of people. We shall do a good job of selenium supplement like iodine supplement, especially the selenium supplement in the elderly. At present, it is imperative to do two things:

First, it is necessary to publicize the knowledge of selenium related to human health to the residents, to enhance their awareness on how to prevent selenium deficiency.

Second, strengthen the development and production of selenium-rich products, and promote the selenium-rich products.



[0006] At present, the selenium can be supplemented through the following three channels:
  1. (1) Inorganic selenium: Sodium selenite-based, difficult to absorb, easy to be poisoned, unsafe. It should be used to patients under the guidance of physicians, which should not be popularized extensively. In some countries, it is prohibited to add inorganic selenium in foods by legislation.
  2. (2) High-selenium health-care product: It is easy to carry and convenient to take, but expensive.
  3. (3) Selenium-rich agricultural products: selenium is supplemented through normal diets. Besides, it is organic, nutritional selenium, with comprehensive nutrition. It is safe, reliable, affordable, and suitable for the masses. It is the main channels to supplement selenium for human.


[0007] The selenium content in the existing grain, vegetables, fruits and other natural foods is low; therefore, it is necessary to develop the selenium-rich foods, to relieve the situation of selenium deficiency, which is the most effective channel to promote human health, and an important guarantee for enhancing the overall quality of Chinese nation and building a well-off society.

[0008] For example, it is known, from the document CN 101 462 904 a high-protein organic selenium-enriched fertilizer for producing selenium-enriched fruits and vegetables by spraying the fertilizer on the leave surface of the plant at different growth stages. The high-protein organic selenium-enriched fertilizer comprised (in weight part):
  • Selenium (2-9 - preferentially: sodium selenate or sodium selenite;
  • Zinc (1-5) - preferentially: zinc acetate or zinc nitrate or zinc sulfate;
  • Magnesium (1-5);
  • Calcium (1-5) - preferentially: calcium oxide or calcium nitrate or calcium dioxide;
  • Molybdenum (1-5);
  • Boron (1-5) - from boric acid or sodium tetraborate;
  • Manganese (1-5) - from potassium permanganate or manganese sulfate or manganese dioxide;
  • Surfactant (1-5) - for example washing powder;
  • Nitrogen (4-100) - preferentially urea or sodium bicarbonate or ammonium sulfate or monoammonium phosphate;
  • Phosphors (4-100);
  • Potassium (4-100) - preferentially monopotassium phosphate or potassium nitrate or potassium chloride or potassium sulfate;
  • Amino-acid (90-180) - preferentially produced by performing a zymo-hydrolysis of a high protein substance (soybean...);
All the components of the composition are mixed together with water (800-1100 weight parts) and then submitted to a complex reaction preferentially for 1 to 2.5 hours at a temperature between 40 and 90°C in order to obtain the fertilizer. The fertilizer is then sprayed on leaves at different growth stages of the plant.

[0009] It is also known in the prior art, from the document CN 102 617 247, a selenium - enriching nutrient solution for carambola, more particularly for preventing and treating the plant diseases. This selenium - enriching nutrient solution comprises, in part by mass:
  • Sodium selenite (25-28)
  • Ammonium sulfate (12-15)
  • Anhydrous boric acid (13-15)
  • Potassium sulfate (15-20)
  • Clay (5-10)
  • Diammonium phosphate (10-15)
  • Fulvic acid (2-5)
  • Zinc chloride (0.5-1)
  • Key acid sodium (0.5-1)
  • Manganese sulfate (0.5-1)
  • Ferrous sulfate (0.8-2)
  • Magnesium sulfate (0.3-0.8)
  • Copper sulfate (0.1-0.5)
  • Humic acid (5-10)
  • Bacteria liquid (5-10)
  • Sodium dodecyl benzene sulfonate (0.5-3)
  • Tiophanate methyl (15-30)
  • Salicylic acid (0.6-2)
  • Sodium nitrophenolate (1-6)
  • White carbon black (40-50)


[0010] This solution can also include ethylene glycol (20-30), EDTA (0.5-1), preservative (1-5), sodium carboxymethyl cellulose (5-30), emulsifier OP (20-30) and water (balance).

[0011] Thus, at present, a variety of selenium-rich plant growth regulators are available on the markets, but there are still some drawbacks. Over the past two decades, the present inventor(s) have focused on relevant studies and applications and made continuous improvements and innovations from the first generation of inorganic selenium agents, to the second generation of selenium complex agents, the third generation of selenoprotein agents and until the fourth generation of peptide selenoprotein agents. Their effects are described as follows:

[0012] The first generation of selenium-rich agents, mainly the inorganic selenium agents, could enhance the selenium content in the crops and Chinese herbal medicines. The second generation of selenium-rich agents, mainly the selenium complex agents, could enhance the selenium content in the crops and Chinese herbal medicines, and improve the quality of crop and Chinese herbal medicine products; the third generation of selenium-rich agents, mainly the selenoprotein agents, could enhance the selenium content in the crops and Chinese herbal medicines, and improve the quality of crops and Chinese herbal medicines, with good taste, and it can enhance the antiseptic ability with a long shelf life. The fourth generation of selenium-rich agents, mainly the peptide selenoprotein agents, could enhance the organic selenium content and the output of fruits and vegetables, crops, herbs, and improve resistance to pests and diseases, reduce the use of pesticides, and improve the antiseptic ability with a long shelf life.

Summary of the Invention



[0013] The present invention provides a composite peptide selenoprotein nutrient solution, also known as the fifth generation of selenium-rich agent, which is developed on the basis of the previous products. It can greatly increase the selenium contents, nutritional values, and improve the pest and disease resistance and antiseptic ability of crops.

[0014] The composite peptide selenoprotein nutrient solution is composed of selenium-containing material, hydrolysate of protein, natural mineral, acetic acid, EDTA, penetrating agent, mineral additives, vitamin supplements, organic silicone, soluble modified starch, emulsifier, and filtered water, according to the following ratios (in weight): (1) selenium-containing material, 1-3%, (2) natural mineral, 0.5‰, (3) protein material, 2-3%, (4) acetic acid, 1 ∼ 5 , (5) EDTA, 0.5∼1, (6) penetrating agent, 1 , (7) trace element additive 6-10 , (8) vitamin supplements, 0.2-0.3 , (9) organic silicone 1 , (10) soluble starch, 1%, (11) emulsifier, 1.5%-5%, (12) water, 87-93%.

[0015] Said water in the composite peptide selenoprotein nutrient solution is obtained after a filtration of drinking water through sand, carbon and membrane, and nano-tech electric ion exchange equipment.

[0016] The said selenium-containing material is one or more from the element selenium (SE), selenium oxide, sodium selenite, selenium salt, and organic selenium.

[0017] The said natural mineral is one or more from tourmaline, volcanic rock or volcanic ash.

[0018] The said emulsifier is monostearin: glycerin monostearate (short as GMS), sodium stearyl lactate (ssl), calcium stearyl lactate (csl), diacetyl tartaric acid esters of mono-and diglycerides (datem), sugar esters (se) or distilled monoglycerides (dmg).

[0019] The said penetrating agent is methanol or ethylene glycol.

[0020] The said proteins are animal or plant proteins, microbial proteins or peptide compounds.

[0021] The compositions of said trace element additives are as follows: zinc 1-3 ; magnesium 1-2 ; manganese 1-2.5 ; molybdenum 0.1-0.15 ; boron 2-3 .

[0022] The compositions of said vitamin supplements are as follows: VC: 0.1; VB1 and VD: 0.1.-0.2.

[0023] The method for preparing the aforesaid composite peptide selenoprotein nutrient solution, comprising the following steps:
  1. 1) Protein enzymolysis: Protein enzymolysis and peptide treatment of protein materials, to get the purified protein digest, which is the amino-protein peptide;
  2. 2) Synthesis of peptide selenoprotein compound: add the organic silicone protective agent in the resulting substance in step 1), add selenium material, and evenly mix them for 1.5h at 55°C;
  3. 3) Mix natural mineral raw materials, the trace element additives, acetic acid, soluble modified starch together, and evenly stir them for 1h at 42∼70°C;
  4. 4) Complexation reaction: Add EDTA to the resulting mixture in step 2) to react 2.5h at 50∼85°C;
  5. 5) Fortification treatment: Add penetrating agent, vitamin supplements to the resulting mixture in step 3), and mix them 1h under room temperature;
  6. 6) Emulsification pretreatment: put emulsifier in container, heat it to melt into liquid; add water that has been obtained after a filtration of drinking water through sand, carbon and membrane and the nano-tech electric ion exchange equipment, the water being added at proximately 70°C with weight 4-5 times of the emulsifier to the high-speed mixer, start the mixer to violently stir the hot water. Slowly add the melted emulsifier to the hot water for mixing, to get a milky white pasty fluid, and then cool down to the room temperature for standby;
  7. 7) Emulsification treatment: mix the substances obtained in step 4), step 5) and step 6), and add appropriate amount of filtered water, stir them for 1h at room temperature;
  8. 8) Prepare the qualified composite peptide selenoprotein nutrient mother liquor for storage after passing inspection and sterilization.


[0024] The said protein enzymolysis adopts neutral protease, with the substrate concentration 3%, neutral protease concentration 4.5%, phosphate buffer solution at PH 7, temperature 50°C. The enzymolysis procedure lasts 1-2 hours, and the undigested proteins are centrifuged and separated, to get the purified protein digest, namely the amino protein peptide.

[0025] The said application of composite peptide selenoprotein nutrient solution is as follows: to spray the diluted composite peptide selenoprotein nutrient solution to the water, earth or plant leaves of crops during the growth process, to make crops rich in selenium.

[0026] The said crops are agricultural products or medicinal and edible herbs.

[0027] The said agricultural products are grains, vegetables, fruits, dried fruits or flowers.

[0028] In the present invention, the protein is digested to amino protein peptide, which reacts with selenium to synthesize peptide selenoprotein complex under the protection of organic silicone protective agent, and further enhances the product energy through the energy accelerant- natural energy mineral. Through adding vitamins, mineral supplements to make crops absorb more comprehensive nutrition. The penetrating agent and water, that has been obtained after a filtration of drinking water through sand, carbon and membrane and the nano-tech electric ion exchange equipment, can promote the absorption of various active ingredients more rapidly and effectively. The composite peptide selenoprotein nutrient solution can greatly increase the selenium contents, nutritional values, and improve the pest and disease resistance and antiseptic ability of crops.

[0029] The proteins, vitamins, minerals and energy accelerants added in the nutrient solution formulations can dramatically enhance the contents of essential mineral elements such as zinc, magnesium, manganese, molybdenum, boron and vitamins B, C, D in the crops, improve the smoothness and delicateness of selenium-rich fruits and vegetables with best taste, increase the antioxidant and bacteria resistance, to make selenium-rich agricultural products easy storage and transport.

[0030] The selenium and boron and other elements in the nutrient solution formulations can be converted to plant nutrients, and resist bacteria and pests. It also shows significant effect for nematodes, leek maggot and fruit rot disease and other serious diseases and pests, and can partially substitute the pesticides, to reduce the consumption of pesticides and the pesticide residues, and achieve the green standards for the selenium-rich agricultural products.

[0031] In the present invention, after sand filtration, carbon filtration and membrane filtration and the nano-tech electric ion exchange equipment, the harmful acidic substances in the ordinary drinking water can be separated, to retain the mineral ions in water, to get the water employed in the composite peptide selenoprotein nutrient solution. It is featured by alkalescence and small molecular clusters. Such water, being obtained after a filtration of drinking water through sand, carbon and membrane and the nano-tech electric ion exchange equipment, can facilitate the dissolution of above components and fast absorption by the crops, enhance the absorption rate of nutrition solution, to achieve comprehensive and balanced nutrition.

[0032] The composite peptide selenoprotein nutrient solution in the present invention can be directly applied to the soil in the roots after diluted as required, or can be sprayed to the plants or leaf after diluted with water. In the acid soil (PH 4.5-6.5), since absorption of selenium and sulfur exhibits antagonism and selenium is not easily absorbed and utilized by the crops, spraying method is applied. In alkaline soils (PH 7.5-8.5), selenium is easily absorbed and utilized by the crops, so both methods can be adopted. In the crop plantation process, it is recommended to use organic fertilizers, biological-based pesticides, and prohibited to use highly toxic pesticides with high residues, prohibited to irrigate with contaminated water, and restricted to use the use growth regulators, so that the selenium-rich crops can achieve the green standards.

[0033] The large area of cultivation test in recent year showed that, after absorption, transmission and conversion of composite peptide selenoprotein in the invention, most of selenium can be gathered at the plant's roots, leaves and fruits in the form of organic form, and appropriate amount of selenium enters into the plant cells, which enhances the content of glutathione peroxidase, strengthen the in vivo antioxidant capacity of plants, improve the plant's resistance and anti-aging ability, to enhance the yields and nutrition of the crops. The grains can be of full seeds, the vegetables can be of original flavor and melons and fruits are large and sweet. The fruits will not change color after cutting and the fruit flesh will not be perishable. Therefore, the selenium-rich products developed by the nutrient solution are praised "delicious and good-looking, nutrition and healthcare, less disease and high yield".

[0034] The composite peptide selenoprotein nutrient solution in the present invention has the following advantages: after applying the solution in the plants, especially in crops and vegetables, the crops accelerate the absorption and enrichment of selenoprotein, which promote the in vivo absorption of various kinds of nutrients that can facilitate the growth and improvement of tastes. The tests have shown that, the selenium content in the fruits, vegetables, grain crops after applying the nutrient solution is in full compliance with Tolerance limit of selenium in foods(GB13105-91): ≤ 0.05mg/kg for fruits, 10-100 microgram/kg for fresh vegetables, 100-300 microgram/kg for grains, oils and their products, and 250-500 micrograms/kg for eggs.

[0035] The selenium-rich nutrient solution in the present invention can not only provide selenium for crops, but also provide a variety of trace elements and energy accelerants needed for the crop normal growth, promote the crop growth and disease resistance, and resistance to harmful substances, and improve crop quality and yield.

[0036] After the spray tests of the nutrient solution in Shandong Yiyuan apple base, Yinan vegetables, grapes, mulberry base, Henan wheat base, Shanxi wheat, lotus base, the selenium content can meet the national standard. The yield per mu is increased by around 5-10%, and the economic income can be increased by RMB 1,000 yuan per mu. After spraying in melons and fruits, the economic incomes per mu are far more than 2-5 times of that in grain crops.

[0037] Since the use of composite peptide selenoprotein nutrient solution is to produce selenium-rich rather than high-selenium agricultural products, it is necessary to implement the standardized management during the production and processing. The nutrient solution spraying frequency and concentration should vary for different crops.

[0038] The composite peptide selenoprotein nutrient solution in the present invention, different from previous selenium-rich agents, has advanced technology, greatly improves the stable and reliable selenium-enrich effect, the safety and nutritional values of selenium-rich foods, specifically as follows:
  1. 1. Improving the content of organic selenium in products safely;
  2. 2. Improving the quality of fruits and vegetables, farming, herbs, etc.;
  3. 3. Improving the taste of fruits and vegetables, and the fruits are sweet and crisp;
  4. 4. Enhancing the resistance to pests and diseases, and reducing the use of pesticides;
  5. 5. Enhancing the antiseptic ability, with a long shelf life;
  6. 6. Increasing the contents of glutathione, SOD, and enhancing the antioxidant capacity;
  7. 7. Enhancing the content of rubber flavonoids and smoke detoxification ability;
  8. 8. Enhancing human's resistance to virus and infections;
  9. 9. Improving body's ability against electromagnetic radiation;
  10. 10. Greatly enhancing the absorption and utilization of nutrients, and the healthcare effect by the energy treatment process.

Detailed Descriptions of the Preferred Embodiments



[0039] The invention is described herein in connection with certain specific embodiments.

[0040] The fifth generation of selenium agent composite peptide selenoprotein nutrient solution is composed of selenium-containing material, hydrolysate of protein, natural mineral, acetic acid, EDTA, penetrating agent, mineral additives, vitamin supplements, organic silicone, soluble modified starch, emulsifier, and water, said water being obtained after a filtration of drinking water through sand, carbon and membrane and nano-tech electric ion exchange equipment, the composite peptide selenoprotein nutrient solution comprising the aforesaid elements according to the following ratios (in weight): (1) selenium-containing material, 1-3%, (2) natural mineral, 0.5, (3) protein material, 2-3%, (4) acetic acid, 1 ∼ 5 , (5) EDTA, 0.5∼1, (6) penetrating agent, 1 , (7) trace element additives 6-10 , (8) vitamin supplements, 0.2-0.3 , (9) organic silicone 1 , (10) soluble starch, 1%, (11) emulsifier, 1.5%- 5%, (12) filtered water, 87-93%.

[0041] The selenium-rich agricultural products developed by the nutrient solution of the present invention can be classified into three categories:
The first category: Appropriate amount of nutrient solution is introduced to the plants timely, after the selenium element is absorbed by the plant, it will be converted to selenoprotein in vivo to form the agricultural products with selenium-rich leaves, stems and seeds.

[0042] The second category: The eggs and meat products produced with the feeds of the selenium-rich agricultural products converted in the first category.

[0043] The third category: the selenium-rich products deeply-processed by the above two categories of selenium-rich agricultural products.

[0044] The following embodiments merely serve to illustrate the invention, but the scope of the present invention is not limited to the following embodiments which shall include the following products developed with the nutrient solution in the invention: grains, vegetables, fruits, dried fruits, fishes, crustaceans (shrimp, crab, etc.), meat and eggs, flowers, medicinal and edible items announced by the Ministry of Health. The technicians of ordinary skill in the art can achieve the purposes in the invention according to the disclosed contents herein.

Embodiment 1



[0045] Ingredients: (1) selenium-containing material 2KG, (2) natural mineral 0.05KG, (3) protein material 2KG, (4) acetate 0.3KG, (5) EDTA 0.1KG, (6) penetrating agent 0.1KG, (7) trace element additives, zinc: 0.1KG; magnesium: 0.1KG; manganese: 0.2KG; molybdenum: 0.01KG; boron: 0.21KG, (8) vitamin supplements, VC: 0.01KG; VB1 and VD: 0.02KG in total; (9) silicone 0.1KG, (10) soluble starch 1KG, (11) emulsifier 3KG, (12) water obtained after a filtration of drinking water through sand, carbon and membrane and nano-tech electric ion exchange equipment, around 90.7KG, total 100KG.

[0046] The composite peptide selenoprotein nutrient solution is prepared according to the above method.

Embodiment 2 The application of composite peptide selenoprotein nutrient solution in apples planting and growing



[0047] The composite peptide selenoprotein nutrient solution prepared in above embodiment 1 is a highly concentrated product. Prior to use, the nutrient solution is diluted by 100-fold to prepare the stock solution, which then diluted by 60 to 80-fold to prepare the nutrient solution that can be directly sprayed. The spray amount per mu each time is 30 ∼ 40kg diluted nutrient solution. Usually the nutrient solution is sprayed 5 times during the whole growing period of apples and the yield per mu is about 3,000 kg. In recent years, the inventor(s) has developed several hundred mu selenium-rich apple demonstration bases in Zibo Yiyuan, Yinan, Linqu, Shandong province. Before the germination of apple trees, the diluted nutrient solution is uniformly sprayed to the branches and trunks of apple trees mechanically or manually. In the fruit setting period, spray the nutrient solution to the branches and trunks and surfaces of leaves once again. Spraying is prohibited in the dew, rainy days and at noon in hot days, otherwise, the direct absorption of selenium on the leaf surface will be affected. In the beginning of May, mid-July middle fruit phase, and 20 days before fruit harvest, spray the nutrient solution to the surfaces of leaves three times, respectively.

[0048] After tested by Shandong Province Test Center (GB5009.93-2010), the selenium content of delivered samples is 20.6 microgrammes/kg, conforming to the limit specified in the national standard.

[0049] The selenium content in fruits should be ≤0.05mg/kg in accordance with Tolerance limit of selenium in foods (GB13105-91). Therefore, the selenium content in selenium-rich apples shall also meet this standard. The amount of composite peptide selenoprotein nutrient solution should be controlled. After 5-6 times of spraying selenium nutrient solution to branches and trunks and surfaces of leaves, the selenium content of selenium-rich apples can be up to 20 to 50 micrograms per kg, but if spraying too many times excessively, the selenium content of apples will exceed the limit specified in the standard.

[0050] As the composite peptide selenoprotein nutrient solution contains rich proteins, amino acids, energy accelerants, a variety of trace elements and other elements that can promote the plant growth, it can enhance the yield of apples. Moreover, the selenium-rich apples taste good, crisp and delicious, rich in glutathione and SOD, with high oxidation resistance and easy to storage. More importantly, the clinical applications have showed that, after a long-term consumption of this kind of apples, it would not increase the blood glucose of patients with diabetes, but rehabilated their pancreas islet and stabilize their blood glucose levels, so it is the safe selenium-rich fruits for patients with diabetes. For patients with liver diseases (including hepatitis B), after a long-term consumption, their liver functions increased, and resistance to infection also enhanced. Dozens of cases of cancer patients improved their physical fitness, prolonged life and reduced pains after a long term of consumption of the selenium-rich apples.

[0051] In the selenium-rich apple bases located in Zhongzhuang, Yiyuan, Shandong, by using the composite peptide selenoprotein nutrient solution, the apples produced are free of pesticides, fertilizer residue, and they have such unique features of good colors, thin peels, juicy and crisp fruit fleshes, contain unique, efficient selenium compounds, which prove to be selenium-rich foods with high activity. The base successfully established a safe, effective channel for human to supplement selenium through diets.

Embodiment 3: Fortified cultivation of selenium-rich rice using the nutrient solution prepared in Embodiment 1



[0052] In the present invention, the nutrient fortified selenium-rich rice that is rich in selenium and other trace elements is cultivated in Shandong Tancheng through the following methods:
  1. 1. Preparation of foliage spray nutrient solution: The composite peptide selenoprotein nutrient solution in the present invention is diluted to 100-fold using filtered water, to prepare the stock solution; take appropriate amount of stock solution, and dilute it by 60 to 80-fold with the active water, about 30 ∼ 50 kg diluted composite peptide selenoprotein nutrient solution for each mu of rice field. The water employed in the nutrient solution is a water obtained after a filtration of drinking water through sand, carbon and membrane and the nano-tech electric ion exchange equipment.
  2. 2. First time of spraying in the rice seedlings stage. Spray the composite peptide selenoprotein nutrient solution to the seedling foliage at a consumption of 10∼ 20kg/100 m2;
  3. 3. Second time of spraying in the heading to flowering stage. Spray the nutrient solution to the foliages of rice, 30-50 kg per mu. The spraying should be carried out in weak light days or in the afternoons, or early evenings.
  4. 4. In the grain filling stage, that is, 5-7 days after the second time of spraying, spray the nutrient solution for the third time; and harvest 15-20 days later.


[0053] After the above steps, the rice seedlings are harvested and processed to make the finished products. After tested in Shandong Province Test Center (GB5009.93-2010), the selenium content of delivered samples is 189 micro grammes/kg, conforming to the limit (100-300 microgrammes/kg) for grains and oil products specified in the Tolerance limit of selenium in foods (GB13105-91).


Claims

1. A composite peptide selenoprotein nutrient solution, comprising selenium-containing material, hydrolysate of protein, natural mineral, acetic acid, EDTA, penetrating agent, trace element additives, vitamin supplements, organic silicone, soluble modified starch, emulsifier, and water, said water being obtained after a filtration of drinking water through sand, carbon and membrane and the nano-tech electric ion exchange equipment, the composite peptide selenoprotein nutrient solution comprising the aforesaid elements according to the following ratios (in weight): (1) selenium-containing material, 1-3%, (2) natural mineral, 0.5‰ which is one or more from tourmaline, volcanic rock or volcanic ash , (3) protein material, 2-3%, (4) acetic acid, 1-5 ‰, (5) EDTA, 0.5∼1‰, (6) penetrating agent, 1 ‰ which is methanol or ethylene glycol, (7) trace element additives, 6-10 consisting in zinc 1-3 ‰; magnesium 1-2 ‰; manganese 1-2.5 ‰; molybdenum 0.1-0.15 ‰; boron 2-3 ‰, (8) vitamin supplements, 0.2-0.3 ‰, (9) organic silicone 1 , (10) soluble starch, 1%, (11) emulsifier, 1.5%- 5% which is glycerin monostearate (GMS), sodium stearyl lactate (ssl), calcium stearyl lactate (csl), diacetyl tartaric acid esters of mono-and diglycerides (datem), sugar esters (se) or distilled monoglycerides (dmg), and (12) filtered water, 87-93%.
 
2. The composite peptide selenoprotein nutrient solution according to claim 1, wherein the said selenium-containing material is one or more from the element selenium (SE), selenium oxide, sodium selenite, selenium salt, and organic selenium.
 
3. The composite peptide selenoprotein nutrient solution according to claim 1, wherein said proteins are animal or plant proteins, microbial proteins or peptide compounds.
 
4. The composite peptide selenoprotein nutrient solution according to claim 1, wherein the compositions of said vitamin supplements are as follows: VC: 0.1‰; VB1 and VD: 0.1.-0.2.
 
5. A method for preparing the aforesaid composite peptide selenoprotein nutrient solution according to claims 1-4, comprising the following steps:

1) Protein enzymolysis: Protein enzymolysis and peptide treatment of protein materials, to get the purified protein digest, which is the amino-protein peptide, wherein the said protein enzymolysis adopts neutral protease, with the substrate concentration 3%, neutral protease concentration 4.5%, phosphate buffer solution at PH 7.0, temperature 50 °C, said enzymolysis procedure lasts 1-2 hours, and the undigested proteins are centrifuged and separated, to get the purified protein digest, namely the amino protein peptide.;

2) Synthesis of peptide selenoprotein compound: add the organic silicone protective agent in the resulting substance in step 1), add selenium material, and evenly mix them for 1.5h at 55°C;

3) Mix natural mineral raw materials, trace element additives, acetic acid, soluble modified starch together, and evenly stir them for 1h at 42∼70°C;

4) Complexation reaction: Add EDTA to the resulting mixture in step 2) to react 2.5h at 50∼85°C;

5) Fortification treatment: Add penetrating agent, vitamin supplements to the resulting mixture in step 3), and mix them 1h under room temperature;

6) Emulsification pretreatment: put emulsifier in container, heat it to melt into liquid; add water that has been obtained after a filtration of drinking water through sand, carbon and membrane and the nano-tech electric ion exchange equipment, the water being added at proximately 70 °C with weight 4-5 times of the emulsifier to the high-speed mixer, start the mixer to violently stir the hot water. Slowly add the melted emulsifier to the hot water for mixing, to get a milky white pasty fluid, and then cool down to the room temperature for standby;

7) Emulsification treatment: mix the substances obtained in step 4), step 5) and step 6), and add appropriate amount of filtered water, stir them for 1h at room temperature;

8) Prepare the qualified composite peptide selenoprotein nutrient mother liquor for storage after passing inspection and sterilization.


 
6. An application of composite peptide selenoprotein nutrient solution according to claims 1-4 is as follows:

- Dilution of said solution with water;

- Direct application of the diluted solution to the soil, in the roots of plants, or spraying of the diluted solution to the plants or leaves, in crops and vegetables, during the growth process, to make crops rich in selenium.


 
7. The application according to claim 6, wherein the said crops are agricultural products or medicinal and edible herbs.
 
8. The application according to claim 6, wherein said agricultural products are grains, vegetables, fruits, or flowers.
 


Ansprüche

1. Komposit-Peptid-Selenoprotein-Nährlösung, umfassend selenhaltiges Material, Proteinhydrolysat, natürliches Mineral, Essigsäure, EDTA, Penetrationsmittel, Spurenelement-Zusatzstoffe, Vitaminzusätze, organisches Silikon, lösliche modifizierte Stärke, Emulgator und Wasser, wobei das besagte Wasser nach einer Filtration von Trinkwasser durch Sand, Kohlenstoff und Membran und elektrische Ionenaustausch-Nano-Tech-Ausrüstung erhalten wird, wobei die Komposit-Peptid-Selenoprotein-Nährlösung die oben genannten Elemente in den folgenden Gewichtsverhältnissen umfasst: (1) selenhaltiges Material, 1 bis 3 %, (2) natürliches Mineral, 0,5 %o, das ein oder mehrere Elemente unter Turmalin, Vulkangestein oder Vulkanasche ist, (3) Proteinmaterial, 2 bis 3 %, (4) Essigsäure, 1 ∼ 5 ‰, (5) EDTA, 0,5 ∼ 1 ‰, (6) Penetrationsmittel, 1 ‰, das Methanol oder Äthylenglykol ist, (7) Spurenelement-Zusatzstoffe, 6 bis 10 %o bestehend aus Zink 1 bis 3 %o; Magnesium 1 bis 2 %o; Mangan 1 bis 2,5 %o; Molybdän 0,1 bis 0,15 %o; Bor 2 bis 3 ‰, (8) Vitaminzusätze, 0,2 bis 0,3 ‰, (9) organisches Silikon 1 ‰, (10) lösliche Stärke, 1 %, (11) Emulgator, 1,5 % bis 5 %, der Glycerinmonostearat (GMS), Natriumstearyllactat (ssl), Calciumstearyllactat (csl), Diacetylweinsäureester von Mono- und Diglyceriden (datem), Zuckerester (se) oder destillierten Monoglyceriden (dmg) ist, und (12) gefiltertes Wasser, 87 bis 93 %.
 
2. Komposit-Peptid-Selenoprotein-Nährlösung nach Anspruch 1, bei der das selenhaltige Material ein oder mehrere Elemente unter Spurenelementenselen (SE), Selenoxid, Natriumselenit, Selensalz und organisches Selen ist.
 
3. Komposit-Peptid-Selenoprotein-Nährlösung nach Anspruch 1, bei der die Proteine tierische oder pflanzliche Proteine, Mikrobenproteine oder Peptidverbindungen sind.
 
4. Komposit-Peptid-Selenoprotein-Nährlösung nach Anspruch 1, bei der die Zusammensetzungen der Vitaminzusätze wie folgt sind: VC: 0,1 ‰; VB1 und VD: 0,1 bis 0,2 ‰.
 
5. Verfahren zur Herstellung der vorgenannten Komposit-Peptid-Selenoprotein-Nährlösung nach den Ansprüchen 1 bis 4, umfassend die folgenden Schritte:

1) Proteinenzymolyse: Proteinenzymolyse und Peptidbehandlung von Proteinmaterialien, um den gereinigten Proteinverdau zu erhalten, der das Amino-Protein-Peptid ist, wobei die besagte Proteinenzymolyse eine neutrale Protease mit einer Substratkonzentration von 3 %, einer neutralen Proteasekonzentration von 4,5 %, einer Phosphatpufferlösung bei pH 7,0, einer Temperatur von 50°C annimmt, wobei das besagte Enzymolyseverfahren 1 bis 2 Stunden dauert und die unverdauten Proteine zentrifugiert und getrennt werden, um den gereinigten Proteinverdau, nämlich das Amino-Protein-Peptid, zu erhalten;

2) Synthese der Peptid-Selenoproteinverbindung: Zugeben des organischen Siliconschutzmittels zu der in Schritt 1) resultierenden Substanz, Selenmaterial zugeben und 1,5 Stunden lang gleichmäßig mischen bei 55°C;

3) Mischen der natürlichen mineralischen Rohstoffe, Spurenelement-Zugabestoffe, Essigsäure und lösliche modifizierte Stärke und diese 1 Stunde lang bei 42∼70°C gleichmäßig durchrühren.

4) Komplexierungsreaktion: Zugeben von EDTA zu der in Schritt 2) resultierenden Mischung, um 2,5 Stunden lang bei 50 bis 85° C zu reagieren;

5) Verstärkungsbehandlung: Zugeben des Penetrationsmittels und der Vitaminzusätze zu der in Schritt 3) resultierenden Mischung, und diese 1 Stunde lang bei Raumtemperatur mischen;

6) Emulgierungsvorbehandlung: Emulgator in einen Behälter giessen, erhitzen, um ihn in der Flüssigkeit zu schmelzen; Wasser, das nach Filtration von Trinkwasser durch Sand, Kohlenstoff und Membran und elektrische Ionenaustausch-Nano-Tech-Ausrüstung erhalten wurde zugeben, wobei das Wasser bei einer Temperatur von etwa 70°C mit einem Gewicht gleich das 4- bis 5-fache Gewicht des Emulgators zu dem Hochgeschwindigkeitsmischer zugegeben wird, den Mischer starten, um das heiße Wasser heftig zu rühren. Den geschmolzenen Emulgator langsam zu dem heißen Wasser für Mischung zugeben, um eine milchig weiße pastöse Flüssigkeit zu erhalten, und dann bis auf Raumtemperatur abkühlen lassen für Bereitstellung;

7) Emulgierungsbehandlung: Mischen der in Schritt 4), Schritt 5) und Schritt 6) erhaltenen Substanzen und Zugeben einer geeigneten Menge an gefiltertem Wasser, sie 1 Stunde lang bei Raumtemperatur durchrühren;

8) Aufbereiten der qualifizierten Komposit-Peptid-Selenoprotein-Nährungsmutterlauge zur Lagerung nach Inspektion und Sterilisation.


 
6. Anwendung der Komposit-Peptid-Selenoprotein-Nährlösung nach Ansprüchen 1 bis 4, wie folgt:

- Verdünnen der Lösung mit Wasser;

- Direktes Auftragen der verdünnten Lösung auf den Boden, in die Wurzeln von Pflanzen oder Aufsprühen der verdünnten Lösung auf die Pflanzen oder Blätter, bei Pflanzen und Gemüsen, während des Wachstumsprozesses, um selenreiche Pflanzen zu erhalten.


 
7. Anwendung nach Anspruch 6, bei der die besagten Pflanzen landwirtschaftliche Produkte oder medizinische und essbare Kräuter sind.
 
8. Anwendung nach Anspruch 6, bei der die besagten landwirtschaftlichen Produkte Körner, Gemüse, Früchte oder Blumen sind.
 


Revendications

1. Solution nutritive composite de peptides et sélénoprotéines, comprenant une matière contenant du sélénium, un hydrolysat de protéine, un minéral naturel, de l'acide acétique, de l'EDTA, un agent pénétrant, des additifs à base d'oligo-éléments, des suppléments vitaminiques, du silicone organique, de l'amidon modifié soluble, un émulsifiant et de l'eau, ladite eau étant obtenue après une filtration d'eau potable à travers du sable, du carbone et une membrane et l'équipement d'échange d'ions électrique nano-tech, la solution nutritive composite de peptides et sélénoprotéines comprenant les éléments précités selon les rapports suivants (en poids): (1) matière contenant du sélénium, 1 à 3 %, (2) minéral naturel, 0.5 ‰, lequel est l'un ou plusieurs parmi la tourmaline, une roche volcanique ou une cendre volcanique, (3) matière protéique, 2 à 3 %, (4) acide acétique, 1 ∼ 5 ‰, (5) EDTA, 0,5 ∼ 1 ‰, (6) agent pénétrant, 1 ‰, qui est du méthanol ou de l'éthylène glycol, (7) additifs à base d'oligo-éléments, 6 à 10 ‰, consistant en zinc, 1 à 3 ‰; magnésium, 1 à 2 ‰; manganèse, 1 à 2,5 ‰; molybdène, 0,1 à 0,15 ‰; bore, 2 à 3 ‰, (8) suppléments vitaminiques, 0,2 à 0,3 ‰, (9) silicone organique, 1 ‰, (10) amidon soluble, 1%, (11) émulsifiant, 1,5% à 5%, qui est du monostéarate de glycérine (GMS), du stéaryl-lactate de sodium (ssl), du stéaryl-lactate de calcium (csl), des esters de l'acide diacétyltartrique des mono- et diglycérides (datem), des esters de sucre (se) ou des monoglycérides distillés (dmg) et (12) eau filtrée, 87 à 93 %.
 
2. Solution nutritive composite de peptides et sélénoprotéines selon la revendication 1, dans laquelle ladite matière contenant du sélénium est un ou plusieurs éléments choisis parmi le sélénium élémentaire (SE), l'oxyde de sélénium, le sélénite de sodium, le sel de sélénium et le sélénium organique.
 
3. Solution nutritive composite de peptides et sélénoprotéines selon la revendication 1, dans laquelle lesdites protéines sont des protéines animales ou végétales, des protéines microbiennes ou des composés peptidiques.
 
4. Solution nutritive composite de peptides et sélénoprotéines selon la revendication 1, dans laquelle les compositions desdits suppléments vitaminiques sont les suivants: VC: 0,1 ‰; VB1 et VD: 0,1 à 0,2 ‰.
 
5. Procédé de préparation de la solution nutritive composite de peptides et sélénoprotéines susmentionnée selon les revendications 1 à 4, comprenant les étapes suivantes:

1) Enzymolyse de protéines: Enzymolyse des protéines et traitement des peptides de matières protéiques, pour obtenir un lysat de protéines purifié, qui est le peptide d'amino-protéine, dans lequel ladite enzymolyse de protéines adopte une protéase neutre, avec une concentration de substrat de 3 %, une concentration de protéase neutre de 4,5 %, une solution tampon de phosphate à pH 7,0, une température de 50°C, ladite procédure d'enzymolyse dure de 1 à 2 heures et les protéines non digérées sont centrifugées et séparées pour obtenir le lysat de protéines purifié, à savoir le peptide de l'amino-protéine;

2) Synthèse du composé de peptides et sélénoprotéines: ajouter l'agent de protection de silicone organique à la substance résultante à l'étape 1), ajouter de la matière de sélénium et les mélanger uniformément pendant 1,5 heure à 55°C;

3) Mélanger les matières premières minérales naturelles, les additifs à base d'oligo-éléments, l'acide acétique, l'amidon modifié soluble, et les agiter uniformément pendant 1 heure à une température de 42 ∼ 70°C;

4) Réaction de complexation: ajouter de l'EDTA au mélange résultant à l'étape 2) pour réagir pendant 2,5 h à une température de 50 ∼ 85°C;

5) Traitement de fortification: Ajouter l'agent pénétrant, les suppléments vitaminiques au mélange résultant à l'étape 3) et les mélanger pendant 1 heure à température ambiante;

6) Prétraitement d'émulsification: placer l'émulsifiant dans le récipient, le chauffer pour le faire fondre dans le liquide; ajouter l'eau qui a été obtenue après filtration d'eau potable à travers du sable, du carbone et la membrane et le matériel d'échange d'ions électrique nano-tech; l'eau étant ajoutée à une température d'environ 70°C à un poids de 4 à 5 fois celui de l'émulsifiant au mélangeur à grande vitesse, mettre en marche le mélangeur pour agiter violemment l'eau chaude. Ajouter lentement l'émulsifiant fondu à l'eau chaude pour mélange, pour obtenir un fluide pâteux blanc laiteux, et ensuite refroidir à la température ambiante pour rester en attente;

7) Traitement d'émulsification: mélanger les substances obtenues aux étapes 4), 5) et 6), et ajouter une quantité appropriée d'eau filtrée, les agiter pendant 1 heure à température ambiante;

8) Préparer la liqueur mère nutritive composite de peptides et sélénoprotéines qualifiée en vue de son entreposage après inspection et stérilisation.


 
6. Application d'une solution nutritive composite de peptides et sélénoprotéines selon les revendications 1 à 4 comme suit:

- dilution de ladite solution avec de l'eau;

- application directe de la solution diluée au sol, dans les racines de plantes, ou pulvérisation de la solution diluée sur les plantes ou les feuilles, dans les cultures et les légumes, pendant le processus de croissance, pour rendre les cultures riches en sélénium.


 
7. Application selon la revendication 6, dans laquelle lesdites cultures sont des produits agricoles ou des herbes médicinales et comestibles.
 
8. Application selon la revendication 6, dans laquelle lesdits produits agricoles sont des céréales, des légumes, des fruits ou des fleurs.
 






Cited references

REFERENCES CITED IN THE DESCRIPTION



This list of references cited by the applicant is for the reader's convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

Patent documents cited in the description