Biological fermentation composition with anticancer effect and preparation method and application thereof
阅读说明：本技术 一种具有抗癌作用的生物发酵组合物及其制备方法和应用 (Biological fermentation composition with anticancer effect and preparation method and application thereof ) 是由 李宝恒 李倩 舒威 于 2021-03-22 设计创作，主要内容包括：本发明提出了一种具有抗癌作用的生物发酵组合物及其制备方法和应用,属于生物工程技术领域。该具有抗癌作用的生物发酵组合物由以下的原料按重量份制备而成：富硒纳豆发酵提取物50-100份、人参皂苷Rg2 2-5份、人参皂苷Rg3 1-3份、三七皂苷R1 2-5份、虫草多糖4-10份。本发明制备方法简单、原料来源广,制得的组合物具有很好的抗肿瘤作用,特别对甲状腺癌具有很好的抑制和治疗作用,且安全无副作用,可以作用食疗产品、保健食品以及药品的原料使用,具有广阔的应用前景。(The invention provides a biological fermentation composition with an anticancer effect, a preparation method and application thereof, and belongs to the technical field of biological engineering. The biological fermentation composition with the anticancer effect is prepared from the following raw materials in parts by weight: 50-100 parts of selenium-enriched natto fermentation extract, 5-5 parts of ginsenoside Rg 22, 3-3 parts of ginsenoside Rg 31, 26-5 parts of notoginsenoside R12 and 4-10 parts of cordyceps polysaccharide. The preparation method is simple, the raw materials are wide in source, the prepared composition has a good anti-tumor effect, particularly has good inhibition and treatment effects on thyroid cancer, is safe and free of side effects, can be used as raw materials of food therapy products, health-care foods and medicines, and has a wide application prospect.)
1. A preparation method of a selenium-rich natto fermentation extract is characterized by comprising the following steps: culturing monascus, strain Bacillus natto918 and selenium-enriched yeast into seed liquid, inoculating the seed liquid into a liquid culture medium for culturing to obtain a composite zymophyte liquid, curing detoxified soybeans, putting the cured soybeans into the composite zymophyte liquid for fermentation, filtering and drying to obtain the selenium-enriched natto fermentation extract.
2. The preparation method according to claim 1, comprising the following steps:
s1, preparing a liquid culture medium: dissolving carbon source, nitrogen source, vitamins and inorganic salt with sterile water, mixing well, adjusting pH of culture medium to 6.8-7.2 with PBS solution, and ultraviolet sterilizing;
s2, culturing the composite zymophyte liquid: respectively inoculating monascus, strain Bacillus natto918 and selenium-enriched yeast into a Gao' S culture medium for streaking, carrying out first aerobic culture, then respectively culturing into strain seed solutions, inoculating into the liquid culture medium prepared in the step S1, carrying out second aerobic culture, and mixing and diluting the three culture media in equal volumes to obtain a composite zymogen liquid;
s3, detoxification of soybeans: cleaning polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 2-4h, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 20-24h, decocting at 70-100 deg.C for 10-20min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing the selenium-rich natto fermentation extract: adding the soybean powder processed in the step S4 into the compound zymocyte liquid obtained in the step S2, fermenting for 24-48h at the constant temperature of 37 ℃, filtering, washing the solid with sterile water, combining the washing liquid and the filtrate, and freeze-drying to obtain the selenium-rich natto fermentation extract.
3. The preparation method according to claim 2, wherein the carbon source is selected from one or more of glucose, maltose, lactose, sucrose, fructose and starch; the nitrogen source is selected from ammonia water, urea, ammonium salt, nitrate and amino acid; the vitamins are selected from one or more of vitamin C, vitamin B1, vitamin B2, vitamin A, vitamin K, vitamin B12, vitamin D and vitamin E; the inorganic salt is selected from one or more of sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, ferric chloride, zinc sulfate, copper sulfate, manganese sulfate, zinc chloride, copper chloride and manganese chloride; the amino acid is selected from one or more of glycine, serine, threonine, valine, tryptophan, leucine, alanine, cysteine, methionine, lysine, isoleucine and phenylalanine; the mass ratio of the carbon source, the nitrogen source, the vitamins, the inorganic salt and the sterile water is (5-15): (3-7): (0.1-0.2): (0.5-1): 100.
4. the preparation method according to claim 2, wherein the mass ratio of the monascus to the strain bacillus natto918 to the selenium-enriched yeast is (2-5): (5-10): (1-3); the first aerobic culture condition is 35-40 ℃, the oxygen content is 20-30%, the humidity is 70-80%, and the culture time is 12-24 h; the second aerobic culture condition is 35-40 ℃, the oxygen content is 20-30%, the humidity is 75-85%, and the culture time is 24-36 h; the strain seed liquid has a bacteria content of 107-108cfu/mL; the inoculation amounts of the monascus, the strain Bacillus natto918 and the selenium-enriched yeast are respectively 1-3%, 3-5% and 0.5-1.5%; the dilution factor is 100-200 times.
5. The preparation method according to claim 2, wherein the mixed solution of acetic acid, ethanol and triethylamine contains 10 to 20 wt% of acetic acid, 25 to 40 wt% of ethanol, 5 to 10 wt% of triethylamine, and the balance of water; the solid-liquid ratio of the soybean meal to the compound zymocyte liquid is 1: (2-5).
6. A selenium enriched fermented natto extract obtained by the method according to any one of claims 1 to 5.
7. The biological fermentation composition with the anticancer effect is characterized by being prepared from the following raw materials in parts by weight: 50-100 parts of selenium-enriched natto fermented extract, Rg 22-5 parts of ginsenoside, Rg 31-3 parts of ginsenoside, R12-5 parts of notoginsenoside and 4-10 parts of cordyceps polysaccharide, which are disclosed by claim 6.
8. A process for the preparation of a biofermentation composition with anticancer action according to claim 7, characterized by comprising the following steps: the biological fermentation composition with the anticancer function is obtained by uniformly mixing the selenium-enriched natto fermentation extract, the ginsenoside Rg2, the ginsenoside Rg3, the notoginsenoside R1 and the cordyceps polysaccharide.
9. Use of the biologically fermented composition with anticancer effect according to claim 7 in the preparation of anticancer food, health food and medicine.
10. The use of claim 9, wherein the cancer is thyroid cancer.
Technical Field
The invention relates to the technical field of bioengineering, in particular to a biological fermentation composition with an anticancer effect and a preparation method and application thereof.
Background
The technological progress brings convenience and rapidness to the life of people, greatly improves the physical living standard of people, and brings a series of negative problems: destruction and pollution of ecological environment, water pollution, damage of pesticide and fertilizer hormone, abuse of various additives in food, abuse of antibiotics and drugs, and the like. The ecological environment of people's life has been destroyed, simultaneously add the fast-paced life of modern people, the upper net clan radiation injury, the infringement of influenza virus, the improvement of working pressure, make most people's immunity reduce gradually, be in sub-health state for a long time, what is more worrying is that cancer diabetes AIDS and three high crowds rise year by year, human health faces unprecedented challenge.
Cancer generally refers to all malignant tumors, which are formed by abnormal proliferation and differentiation of cells of local tissues, which are caused by loss of normal regulation and control of growth of local tissues at gene level under the action of various tumorigenic factors, wherein once formed, the growth of the new organism is not stopped due to elimination of diseases, and the growth of the new organism is not regulated by normal organism physiology, but normal tissues and organs are damaged. At present, no medicine for preventing and treating cancer exists in the market, which brings harm to the health of people.
The natto is a bean product prepared by fermenting soybeans through bacillus natto, has viscosity, smelly smell and slightly sweet taste, not only retains the nutritive value of the soybeans, is rich in vitamin K2 and improves the digestibility of protein, but also generates various physiological active substances in the fermentation process, and has the health-care effects of dissolving fibrin in a body and regulating physiological functions.
The bacillus natto (Bacillus natto) is beneficial bacteria with acid resistance and heat resistance, the survival rate of the bacillus natto in gastric acid for four hours is 100 percent, and the bacillus natto has strong pathogenic bacteria inhibiting capability, is one of various beneficial bacteria, has best environmental tolerance and can directly reach the small intestine, can change the ecology of intestinal flora of human bodies after being taken orally, helps the normalization of the function of digestive tracts, enables defecation to be smooth, and maintains the physiological environment in vivo. Can produce acid, regulate intestinal flora, enhance animal cell immunity, and generate various proteases (especially alkaline protease), saccharifying enzyme, lipase, amylase, etc. The enzyme Nattokinase (NK) produced by the bacillus natto when the bacillus natto breeds by phagocytizing soybean protein has magical drug effect and can produce delicious amino acid.
Disclosure of Invention
The invention aims to provide a biological fermentation composition with an anticancer effect, a preparation method and application thereof, the preparation method is simple, the raw material sources are wide, the prepared composition has a good antitumor effect, particularly has good inhibition and treatment effects on thyroid cancer, is safe, does not have side effects, can be used as a raw material of a dietary therapy product, a health food and a medicine, and has a wide application prospect.
The technical scheme of the invention is realized as follows:
the invention provides a preparation method of a selenium-rich natto fermentation extract, which comprises the following steps: culturing monascus, strain Bacillus natto918 and selenium-enriched yeast into seed liquid, inoculating the seed liquid into a liquid culture medium for culturing to obtain a composite zymophyte liquid, curing detoxified soybeans, putting the cured soybeans into the composite zymophyte liquid for fermentation, filtering and drying to obtain the selenium-enriched natto fermentation extract.
As a further improvement of the invention, the method specifically comprises the following steps:
s1, preparing a liquid culture medium: dissolving carbon source, nitrogen source, vitamins and inorganic salt with sterile water, mixing well, adjusting pH of culture medium to 6.8-7.2 with PBS solution, and ultraviolet sterilizing;
s2, culturing the composite zymophyte liquid: respectively inoculating monascus, strain Bacillus natto918 and selenium-enriched yeast into a Gao' S culture medium for streaking, carrying out first aerobic culture, then respectively culturing into strain seed solutions, inoculating into the liquid culture medium prepared in the step S1, carrying out second aerobic culture, and mixing and diluting the three culture media in equal volumes to obtain a composite zymogen liquid;
s3, detoxification of soybeans: cleaning polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 2-4h, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 20-24h, decocting at 70-100 deg.C for 10-20min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing the selenium-rich natto fermentation extract: adding the soybean powder processed in the step S4 into the compound zymocyte liquid obtained in the step S2, fermenting for 24-48h at the constant temperature of 37 ℃, filtering, washing the solid with sterile water, combining the washing liquid and the filtrate, and freeze-drying to obtain the selenium-rich natto fermentation extract.
As a further improvement of the invention, the carbon source is selected from one or a mixture of more of glucose, maltose, lactose, sucrose, fructose and starch; the nitrogen source is selected from ammonia water, urea, ammonium salt, nitrate and amino acid; the vitamins are selected from one or more of vitamin C, vitamin B1, vitamin B2, vitamin A, vitamin K, vitamin B12, vitamin D and vitamin E; the inorganic salt is selected from one or more of sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, ferric chloride, zinc sulfate, copper sulfate, manganese sulfate, zinc chloride, copper chloride and manganese chloride; the amino acid is selected from one or more of glycine, serine, threonine, valine, tryptophan, leucine, alanine, cysteine, methionine, lysine, isoleucine and phenylalanine; the mass ratio of the carbon source, the nitrogen source, the vitamins, the inorganic salt and the sterile water is (5-15): (3-7): (0.1-0.2): (0.5-1): 100.
as a further improvement of the invention, the mass ratio of the monascus to the strain Bacillus natto918 to the selenium-enriched yeast is (2-5): (5-10): (1-3); the first aerobic culture condition is 35-40 ℃, the oxygen content is 20-30%, the humidity is 70-80%, and the culture time is 12-24 h; the second aerobic culture condition is 35-40 ℃, the oxygen content is 20-30%, the humidity is 75-85%, and the culture time is 24-36 h;the strain seed liquid has a bacteria content of 107-108cfu/mL; the inoculation amounts of the monascus, the strain Bacillus natto918 and the selenium-enriched yeast are respectively 1-3%, 3-5% and 0.5-1.5%; the dilution factor is 100-200 times.
As a further improvement of the invention, the mixed solution of acetic acid, ethanol and triethylamine contains 10-20 wt% of acetic acid, 25-40 wt% of ethanol, 5-10 wt% of triethylamine and the balance of water; the solid-liquid ratio of the soybean meal to the compound zymocyte liquid is 1: (2-5).
The invention further protects the selenium-rich natto fermentation extract prepared by the preparation method.
The invention further provides a biological fermentation composition with an anticancer effect, which is prepared from the following raw materials in parts by weight: 50-100 parts of the selenium-enriched natto fermented extract, 5-5 parts of ginsenoside Rg 22, 3-3 parts of ginsenoside Rg 31, 26-5 parts of notoginsenoside R12 and 4-10 parts of cordyceps polysaccharide.
The invention further provides a preparation method of the biological fermentation composition with the anticancer effect, which comprises the following steps: the biological fermentation composition with the anticancer function is obtained by uniformly mixing the selenium-enriched natto fermentation extract, the ginsenoside Rg2, the ginsenoside Rg3, the notoginsenoside R1 and the cordyceps polysaccharide.
The invention further protects the application of the biological fermentation composition with the anticancer effect in preparing anticancer food, health-care food and medicines.
As a further improvement of the present invention, the cancer is thyroid cancer.
The invention has the following beneficial effects: according to the invention, after mixed culture is carried out on strains Bacillus natto918 in monascus and Bacillus natto and selenium-enriched yeast, the obtained composite zymocyte liquid is used for fermenting and culturing soybeans after detoxification curing treatment, wherein monascus produces polyketone secondary metabolite monascus pigments in the fermentation process, and the polyketone secondary metabolite monascus pigments comprise monascus rubimine, monascus ruber amine, monascus rubicin, monascus anka monascus purpureus, monascus anka monascin, Monacolin K and other active substances, so that the composite zymocyte liquid has a good anti-tumor effect; meanwhile, due to the mixed culture of the three strains, the production of the citrinin by the monascus can be effectively inhibited, so that the biological toxicity is reduced; the Bacillus natto918 produces anti-cancer substances, namely soybean isoflavone such as genistein, genistin, infirabin and the like, in a high yield in the fermentation process, and the Bacillus natto can inhibit the proliferation of cancer cells, effectively destroy or kill the cancer cells, and can stimulate an immune system to induce and produce interferon to play a role in resisting cancer; the selenium-enriched yeast can generate a large amount of organic selenium nutrients in the fermentation process, the selenium element can influence the energy metabolism of cancer cells and interfere the protein synthesis of the cancer cells through the function of the phagocyte, so that the cancer is inhibited, the metabolism of chemical carcinogens can be influenced, the carcinogenic activity of the chemical carcinogens is lost, and the selenium-enriched natto fermentation extract can play a good anticancer effect.
The invention adds the notoginsenoside R1 and the cordyceps polysaccharide, has the synergistic effect, wherein, the notoginsenoside R1 achieves the anticancer effect by adjusting the hematopoietic function, the cordyceps polysaccharide has the functions of inhibiting the proliferation of tumor cells, enhancing the nonspecific immunity function of organisms and good immunoregulation, mixing with ginsenoside Rg2, ginsenoside Rg3 and prepared fermented extract of selenium-enriched natto, wherein ginsenoside Rg2 can inhibit tumor cell growth, induce tumor cell apoptosis, reverse abnormal differentiation of tumor cell, reverse drug resistance of tumor drug, and improve immunity to achieve anti-tumor effect, and ginsenoside Rg3 has good anti-tumor effect by enhancing immunity, inhibiting cancer cell growth, and resisting tumor metastasis, so as to obtain a biological fermented composition with anticancer effect, the composition has good anticancer effect, and has obvious therapeutic effect on thyroid cancer.
The preparation method is simple, the raw materials are wide in source, the prepared composition has a good anti-tumor effect, particularly has good inhibition and treatment effects on thyroid cancer, is safe and free of side effects, can be used as raw materials of food therapy products, health-care foods and medicines, and has a wide application prospect.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a graph of tumor volume over time for different groups in test example 5.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The Monascus purpureus is ACCC 30342 Monascus purpureus, provided by Shanghai Fuxiang Biotech limited; the strain Bacillus natto918 is provided by the strain Collection of the national academy of sciences of the Korean folk-leading people's republic of China; selenium-enriched yeast is provided by Angel Yeast GmbH.
Ginsenoside Rg2, CAS No. 52286-74-5, ginsenoside Rg3, CAS No. 14197-60-5, provided by Daliangfeng pharmaceutical Co., Ltd; notoginsenoside R1, CAS No. 88122-52-5, available from Kyoto Biotech Ltd; cordyceps sinensis polysaccharide, CAS number 73-03-0, was provided by Changdeli bioengineering, Inc.
Example 1 preparation method of selenium-enriched fermented natto extract
The method specifically comprises the following steps:
s1, preparing a liquid culture medium: dissolving 5g of fructose, 1g of ammonia water, 1g of threonine, 1g of valine, 0.1g of vitamin E, 0.2g of calcium chloride, 0.1g of magnesium sulfate and 0.2g of ferric chloride in 100mL of sterile water, uniformly mixing, adjusting the pH value of a culture medium to 6.8 by using a PBS solution, and sterilizing by ultraviolet rays for later use;
s2, culturing the composite zymophyte liquid: respectively inoculating 2g of Monascus, 5g of Bacillus natto918 and 1g of selenium-enriched yeast into a Gao's medium, streaking, carrying out aerobic culture for 12h under 35 ℃, 20% of oxygen content and 70% of humidity, and respectively culturing to obtain strain seed solutions with a bacterium content of 107cfu/mL, inoculating the mixture in the liquid culture medium prepared in the step S1, wherein the inoculation amounts of monascus, Bacillus natto918 and the selenium-enriched yeast are 1%, 3% and 0.5% respectively, carrying out aerobic culture for the second time under the culture condition of 35 ℃, the oxygen content of 20%, the humidity of 75% and the culture time of 24 hours, and mixing the three culture media in equal volumes and diluting the three culture media by 100 times to obtain a composite zymogen liquid;
s3, detoxification of soybeans: cleaning the polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine (the content of acetic acid is 10 wt%, the content of ethanol is 25 wt%, the content of triethylamine is 5 wt%, and the balance is water) in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 2-4h, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 20h, steaming at 70 deg.C for 1min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing the selenium-rich natto fermentation extract: adding 100g of the soybean powder processed in the step S4 into 200g of the compound zymocyte liquid obtained in the step S2, fermenting for 24h at the constant temperature of 37 ℃, filtering, washing the solid with 100mL of sterile water for 3 times, combining the washing liquid and the filtrate, and freeze-drying to obtain the selenium-rich natto fermentation extract.
Example 2 preparation method of selenium-rich fermented natto extract
The method specifically comprises the following steps:
s1, preparing a liquid culture medium: dissolving 15g of glucose, 5g of urea, 1g of ammonium nitrate, 1g of glutamic acid, 0.1g of vitamin K, 0.1g of vitamin B2, 0.2g of potassium chloride gg, 0.2g of calcium chloride, 0.2g of magnesium sulfate, 0.2g of ferric chloride and 0.2g of zinc sulfate in 100mL of sterile water, uniformly mixing, adjusting the pH value of a culture medium to 7.2 by using a PBS (phosphate buffer solution), and performing ultraviolet sterilization for later use;
s2, culturing the composite zymophyte liquid: respectively inoculating 5g of Monascus, 10g of strain Bacillus natto918 and 3g of selenium-enriched yeast into a Gao's medium, streaking, carrying out aerobic culture for 24h under the conditions of 40 ℃, 30% of oxygen content and 80% of humidity, and respectively culturing to obtain strain seed solutions with the strain content of 108cfu/mL, inoculating the mixture into the liquid culture medium prepared in the step S1, wherein the inoculation amounts of monascus, Bacillus natto918 and the selenium-enriched yeast are 3%, 5% and 1.5%, respectively, carrying out aerobic culture for the second time under the culture condition of 40 ℃, the oxygen content of 30%, the humidity of 85% and the culture time of 36h, and mixing the three culture media in equal volumes and diluting the three culture media by 200 times to obtain a composite zymogen liquid;
s3, detoxification of soybeans: cleaning the polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine (the content of acetic acid is 20 wt%, the content of ethanol is 40 wt%, the content of triethylamine is 10 wt%, and the balance is water) in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 4 hours, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 24h, steaming at 100 deg.C for 20min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing the selenium-rich natto fermentation extract: adding 100g of the soybean powder processed in the step S4 into 500g of the compound zymocyte liquid obtained in the step S2, fermenting for 48h at the constant temperature of 37 ℃, filtering, washing the solid with 100mL of sterile water for 3 times, combining the washing liquid and the filtrate, and freeze-drying to obtain the selenium-rich natto fermentation extract.
Example 3 preparation method of selenium-rich fermented natto extract
The method specifically comprises the following steps:
s1, preparing a liquid culture medium: dissolving 10g of maltose, 3g of urea, 1g of leucine, 1g of alanine, 0.1g of vitamin C, 0.05g of vitamin B1, 0.2g of zinc sulfate, 0.3g of copper sulfate and 0.2g of manganese sulfate in 100mL of sterile water, uniformly mixing, adjusting the pH value of a culture medium to 7 by using a PBS (phosphate buffer solution), and sterilizing by using ultraviolet rays for later use;
s2, culturing the composite zymophyte liquid: respectively inoculating 3.5g of monascus, 7g of strain Bacillus natto918 and 2g of selenium-enriched yeast into a Gao's medium, streaking, carrying out aerobic culture for 18h under the conditions of 37 ℃, 25% of oxygen content and 75% of humidity, and respectively culturing to obtain strain seed solutions with the strain content of 108cfu/mL, inoculating the mixture into the liquid culture medium prepared in the step S1, wherein the inoculation amounts of monascus, Bacillus natto918 and the selenium-enriched yeast are respectively 2%, 4% and 1%, carrying out aerobic culture for the second time under the culture condition of 37 ℃, the oxygen content of 25%, the humidity of 80% and the culture time of 30h, and mixing the three culture media in equal volumes and diluting the three culture media by 150 times to obtain a composite zymogen liquid;
s3, detoxification of soybeans: cleaning the polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine (the content of acetic acid is 15 wt%, the content of ethanol is 32 wt%, the content of triethylamine is 7 wt%, and the balance is water) in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 3 hours, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 22h, decocting at 85 deg.C for 15min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing the selenium-rich natto fermentation extract: adding 100g of the soybean flour processed in the step S4 into 350g of the compound zymocyte liquid obtained in the step S2, fermenting for 36h at the constant temperature of 37 ℃, filtering, washing the solid with 100mL of sterile water for 3 times, combining the washing liquid and the filtrate, and freeze-drying to obtain the selenium-rich natto fermentation extract.
Comparative example 1
Compared with the example 3, the selenium-enriched yeast is not added, and other conditions are not changed.
The method comprises the following specific steps:
s1, preparing a liquid culture medium: dissolving 10g of maltose, 3g of urea, 1g of leucine, 1g of alanine, 0.1g of vitamin C, 0.05g of vitamin B1, 0.2g of zinc sulfate, 0.3g of copper sulfate and 0.2g of manganese sulfate in 100mL of sterile water, uniformly mixing, adjusting the pH value of a culture medium to 7 by using a PBS (phosphate buffer solution), and sterilizing by using ultraviolet rays for later use;
s2, culturing the composite zymophyte liquid: respectively inoculating 5.5g of Monascus and 7g of Bacillus natto918 into a Gauss culture medium, streaking, carrying out aerobic culture for 18h under the conditions of 37 ℃, 25% of oxygen content and 75% of humidity, and respectively culturing to obtain strain seed solutions with the strain content of 108cfu/mL, inoculating 3% and 4% of Monascus purpureus and Bacillus natto918 respectively in the liquid culture medium prepared in step S1, carrying out aerobic culture for the second time under the conditions of 37 ℃, 25% of oxygen content, 80% of humidity and 30h, mixing the two culture media in equal volume, and diluting by 150 times to obtain a composite zymogen liquid;
s3, detoxification of soybeans: cleaning the polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine (the content of acetic acid is 15 wt%, the content of ethanol is 32 wt%, the content of triethylamine is 7 wt%, and the balance is water) in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 3 hours, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 22h, decocting at 85 deg.C for 15min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing a natto fermentation extract: adding 100g of the soybean powder processed in the step S4 into 350g of the compound zymocyte liquid obtained in the step S2, fermenting for 36h at the constant temperature of 37 ℃, filtering, washing the solid with 100mL of sterile water for 3 times, combining the washing liquid and the filtrate, and freeze-drying to obtain the natto fermentation extract.
Comparative example 2
Compared with example 3, no monascus was added, and other conditions were not changed.
The method comprises the following specific steps:
s1, preparing a liquid culture medium: dissolving 10g of maltose, 3g of urea, 1g of leucine, 1g of alanine, 0.1g of vitamin C, 0.05g of vitamin B1, 0.2g of zinc sulfate, 0.3g of copper sulfate and 0.2g of manganese sulfate in 100mL of sterile water, uniformly mixing, adjusting the pH value of a culture medium to 7 by using a PBS (phosphate buffer solution), and sterilizing by using ultraviolet rays for later use;
s2, culturing the composite zymophyte liquid: respectively inoculating 7g of strain Bacillus natto918 and 5.5g of selenium-enriched yeast into a Gao's medium, streaking, carrying out first aerobic culture under the conditions of 37 ℃, 25% of oxygen content and 75% of humidity for 18h, and respectively culturing to obtain strain seed solutions with the strain content of 108cfu/mL, inoculating the cfu/mL into the liquid culture medium prepared in the step S1, wherein the inoculation amounts of the strain Bacillus natto918 and the selenium-enriched yeast are respectively 4% and 3%, carrying out aerobic culture for the second time under the culture conditions of 37 ℃, 25% of oxygen content, 80% of humidity and 30h, and mixing the two culture media in equal volumes and diluting the two culture media by 150 times to obtain a composite zymogen liquid;
s3, detoxification of soybeans: cleaning the polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine (the content of acetic acid is 15 wt%, the content of ethanol is 32 wt%, the content of triethylamine is 7 wt%, and the balance is water) in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 3 hours, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 22h, decocting at 85 deg.C for 15min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing the selenium-rich natto fermentation extract: adding 100g of the soybean flour processed in the step S4 into 350g of the compound zymocyte liquid obtained in the step S2, fermenting for 36h at the constant temperature of 37 ℃, filtering, washing the solid with 100mL of sterile water for 3 times, combining the washing liquid and the filtrate, and freeze-drying to obtain the selenium-rich natto fermentation extract.
Comparative example 3
Compared with example 3, only monascus was added, and other conditions were not changed.
The method comprises the following specific steps:
s1, preparing a liquid culture medium: dissolving 10g of maltose, 3g of urea, 1g of leucine, 1g of alanine, 0.1g of vitamin C, 0.05g of vitamin B1, 0.2g of zinc sulfate, 0.3g of copper sulfate and 0.2g of manganese sulfate in 100mL of sterile water, uniformly mixing, adjusting the pH value of a culture medium to 7 by using a PBS (phosphate buffer solution), and sterilizing by using ultraviolet rays for later use;
s2, culturing zymophyte liquid: inoculating 12.5g of Monascus into a Gauss culture medium, streaking, performing aerobic culture for 18h under 37 deg.C with oxygen content of 25% and humidity of 75%, and culturing to obtain strain seed solution with bacteria content of 108cfu/mL, inoculating the culture medium into the liquid culture medium prepared in the step S1, wherein the inoculation amount of monascus is 7%, carrying out aerobic culture for the second time under the culture condition of 37 ℃, the oxygen content of 25%, the humidity of 80% and the culture time of 30h, and diluting the culture medium by 150 times to obtain a zymogen liquid;
s3, detoxification of soybeans: cleaning the polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine (the content of acetic acid is 15 wt%, the content of ethanol is 32 wt%, the content of triethylamine is 7 wt%, and the balance is water) in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 3 hours, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 22h, decocting at 85 deg.C for 15min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing a natto fermentation extract: adding 100g of the soybean powder processed in the step S4 into 350g of the zymocyte liquid obtained in the step S2, fermenting at the constant temperature of 37 ℃ for 36h, filtering, washing the solid with 100mL of sterile water for 3 times, combining the washing liquid and the filtrate, and freeze-drying to obtain the natto fermentation extract.
Comparative example 4
Compared with example 3, only the strain Bacillus natto918 was added, and other conditions were not changed.
The method comprises the following specific steps:
s1, preparing a liquid culture medium: dissolving 10g of maltose, 3g of urea, 1g of leucine, 1g of alanine, 0.1g of vitamin C, 0.05g of vitamin B1, 0.2g of zinc sulfate, 0.3g of copper sulfate and 0.2g of manganese sulfate in 100mL of sterile water, uniformly mixing, adjusting the pH value of a culture medium to 7 by using a PBS (phosphate buffer solution), and sterilizing by using ultraviolet rays for later use;
s2, culturing zymophyte liquid: inoculating 12.5g of Bacillus natto918 into a Gao's medium, streaking, performing aerobic culture for 18h under 37 deg.C with 25% oxygen content and 75% humidity, and culturing to obtain strain seed liquid with 10% bacteria content8cfu/mL, inoculating the cfu/mL into the liquid culture medium prepared in the step S1, wherein the inoculation amount of the strain Bacillus natto918 is 7%, carrying out aerobic culture for the second time under the culture condition of 37 ℃, the oxygen content of 25% and the humidity of 80%, and the culture time is 30h, and diluting the culture medium by 150 times to obtain a zymogen liquid;
s3, detoxification of soybeans: cleaning the polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine (the content of acetic acid is 15 wt%, the content of ethanol is 32 wt%, the content of triethylamine is 7 wt%, and the balance is water) in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 3 hours, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 22h, decocting at 85 deg.C for 15min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing a natto fermentation extract: adding 100g of the soybean powder processed in the step S4 into 350g of the zymocyte liquid obtained in the step S2, fermenting at the constant temperature of 37 ℃ for 36h, filtering, washing the solid with 100mL of sterile water for 3 times, combining the washing liquid and the filtrate, and freeze-drying to obtain the natto fermentation extract.
Comparative example 5
Compared with the example 3, only the selenium-enriched yeast is added, and other conditions are not changed.
The method comprises the following specific steps:
s1, preparing a liquid culture medium: dissolving 10g of maltose, 3g of urea, 1g of leucine, 1g of alanine, 0.1g of vitamin C, 0.05g of vitamin B1, 0.2g of zinc sulfate, 0.3g of copper sulfate and 0.2g of manganese sulfate in 100mL of sterile water, uniformly mixing, adjusting the pH value of a culture medium to 7 by using a PBS (phosphate buffer solution), and sterilizing by using ultraviolet rays for later use;
s2, culturing zymophyte liquid: inoculating 12.5g selenium-rich yeast into Gao's medium, streaking, performing aerobic culture for 18 hr at 37 deg.C under 25% oxygen content and 75% humidity, and culturing to obtain strain seed liquid with bacterium content of 108cfu/mL, inoculating the culture medium into the liquid culture medium prepared in the step S1, wherein the inoculation amount of the selenium-enriched yeast is 7%, carrying out aerobic culture for the second time under the culture condition of 37 ℃, the oxygen content of 25%, the humidity of 80% and the culture time of 30h, and diluting the culture medium by 150 times to obtain a zymogen liquid;
s3, detoxification of soybeans: cleaning the polished soybeans, placing the polished soybeans on an upper-layer screen, placing a mixed solution of acetic acid, ethanol and triethylamine (the content of acetic acid is 15 wt%, the content of ethanol is 32 wt%, the content of triethylamine is 7 wt%, and the balance is water) in a container, covering and installing a condenser pipe, heating to boil, performing steam treatment for 3 hours, and cooling to normal temperature to obtain detoxified soybeans;
s4, processing detoxified soybeans: cleaning the detoxified soybean obtained in step S3, soaking in sterile water for 22h, decocting at 85 deg.C for 15min, drying, pulverizing, and grinding to obtain soybean powder;
s5, preparing the selenium-rich natto fermentation extract: adding 100g of the soybean powder processed in the step S4 into 350g of the zymocyte liquid obtained in the step S2, fermenting for 36h at the constant temperature of 37 ℃, filtering, washing the solid with 100mL of sterile water for 3 times, combining the washing liquid and the filtrate, and freeze-drying to obtain the selenium-rich natto fermentation extract.
Example 4 biological fermentation composition with anticancer action
The raw materials comprise the following components in parts by weight: 70 parts of the selenium-enriched natto fermentation extract prepared in the embodiment 1, 23.5 parts of ginsenoside Rg, 32 parts of ginsenoside Rg, 13.5 parts of notoginsenoside R and 7 parts of cordyceps polysaccharide.
The preparation method comprises the following steps:
the biological fermentation composition with the anticancer function is obtained by uniformly mixing the selenium-enriched natto fermentation extract, the ginsenoside Rg2, the ginsenoside Rg3, the notoginsenoside R1 and the cordyceps polysaccharide.
Example 5
Compared with example 4, the selenium-enriched fermented natto extract was obtained in example 2 without changing other conditions.
Example 6
Compared with example 4, the selenium-enriched fermented natto extract is obtained in example 3 without changing other conditions.
Comparative example 6
Compared with example 6, the selenium-enriched fermented natto extract is prepared by comparative example 1, and other conditions are not changed.
Comparative example 7
Compared with example 6, the selenium-enriched fermented natto extract is prepared by comparative example 2, and other conditions are not changed.
Comparative example 8
Compared with example 6, the selenium-enriched fermented natto extract is prepared by comparative example 3, and other conditions are not changed.
Comparative example 9
Compared with example 6, the selenium-enriched fermented natto extract is prepared by comparative example 4, and other conditions are not changed.
Comparative example 10
Compared with example 6, the selenium-enriched fermented natto extract was obtained in comparative example 5, and other conditions were not changed.
Comparative example 11
Compared with example 6, the selenium-enriched fermented natto extract was not added, and other conditions were not changed.
The raw materials comprise the following components in parts by weight: ginsenoside Rg 23.5 parts, ginsenoside Rg 32 parts, notoginsenoside R13.5 parts, and Cordyceps polysaccharide 7 parts.
Comparative example 12
Compared with example 6, the ginsenoside Rg2 is not added, and other conditions are not changed.
The raw materials comprise the following components in parts by weight: 70 parts of the selenium-enriched natto fermentation extract prepared in the embodiment 3, 35.5 parts of ginsenoside Rg, 13.5 parts of notoginsenoside R and 7 parts of cordyceps polysaccharide.
Comparative example 13
Compared with example 6, the ginsenoside Rg3 is not added, and other conditions are not changed.
The raw materials comprise the following components in parts by weight: 70 parts of the selenium-enriched natto fermentation extract prepared in the embodiment 3, 25.5 parts of ginsenoside Rg, 13.5 parts of notoginsenoside R and 7 parts of cordyceps polysaccharide.
Comparative example 14
Compared with example 6, without adding notoginsenoside R1, the other conditions were unchanged.
The raw materials comprise the following components in parts by weight: 70 parts of the selenium-enriched natto fermentation extract prepared in the embodiment 3, 23.5 parts of ginsenoside Rg, 32 parts of ginsenoside Rg and 10.5 parts of cordyceps polysaccharide.
Comparative example 15
Compared with example 6, no cordyceps polysaccharide was added, and other conditions were not changed.
The raw materials comprise the following components in parts by weight: 70 parts of the selenium-enriched natto fermentation extract prepared in the embodiment 3, 23.5 parts of ginsenoside Rg, 32 parts of ginsenoside Rg and 110.5 parts of notoginsenoside R.
Test example 1
The selenium-enriched fermented natto extracts prepared in examples 1 to 3 and comparative examples 1 to 5 of the present invention were subjected to the ingredient measurement, and the results are shown in table 1.
TABLE 1
As can be seen from the above table, the selenium-enriched fermented natto extract prepared by the embodiment of the invention contains rich nutrient elements.
Test example 2 test for enhancing immunity:
650 volunteers with certain symptoms of insomnia, lassitude, physical fatigue and the like were selected, 50 volunteers were administered with the products prepared in examples 1 to 3 and comparative examples 6 to 15, respectively, once a day, every 5 days, and 1 month was observed, and the observation results are as follows in table 1 (wherein a indicates slight relief, B indicates relief, C indicates recovery, and the number after the alphabet indicates the number of volunteers):
table 2: immunity index evaluation table
As can be seen from the data in the above table, the products prepared in examples 4-6 have significant improvement effects on insomnia, lassitude and physical fatigue, with example 6 being the most effective. The volunteer symptoms of the products of examples 4-6 were all somewhat alleviated at day 10, and at day 30, the volunteer taking the products of examples 4-6 had a complete recovery ratio of over 94%; it can be seen that the product of the present invention has a significant improvement effect on the above symptoms. The volunteers taking the product have the advantages of improving sleep quality, completely relieving physical fatigue, improving spirit and obviously improving immunity.
Test example 3 test for auxiliary anticancer Effect
140 tumor sections were taken and 10 tumor sections were divided into one group, wherein groups 1 to 13 were cultured with the compositions prepared in examples 1 to 3 and comparative examples 6 to 15, respectively, and group 14 was cultured using LB medium, and the proliferation rate of tumor cells was observed and calculated, and the results are shown in Table 3 below.
Table 3: auxiliary anticancer action evaluation table
Group of
Day 1
Day 2
Day 5
Day 10
Example 4
75％
62％
42％
27％
Example 5
74％
60％
38％
24％
Example 6
72％
57％
35％
21％
Comparative example 6
78％
72％
56％
42％
Comparative example 7
77％
70％
54％
40％
Comparative example 8
79％
75％
61％
51％
Comparative example 9
79％
74％
63％
52％
Comparative example 10
78％
73％
60％
48％
Comparative example 11
80％
78％
75％
72％
Comparative example 12
76％
65％
46％
33％
Comparative example 13
75％
64％
47％
32％
Comparative example 14
75％
63％
44％
29％
Comparative example 15
76％
63％
45％
30％
LB Medium
80％
80％
80％
80％
As can be seen from the data in the above table, the product of the present invention has a certain inhibitory effect on the proliferation of cancer cells, of which the effect of example 6 is the most excellent.
Test example 4 inhibition of human thyroid cancer cells
1. Cell line and culture method
Human papillary thyroid carcinoma cells (K1), human medullary thyroid carcinoma cells (TT), and human thyroid squamous carcinoma cells (SW579) were obtained from Sainbur Hippocastanea Biotech, and human thyroid follicular carcinoma cells (FTC-133) were obtained from North Nay Biotech, Guangzhou. K1 cells were cultured in the culture medium containing 10% Australia fetal bovine serum RPMI-1640; TT cells were cultured in culture medium containing 10% Australian fetal bovine serum F-12K; FTC-133 cells were cultured in DMEM medium containing 10% fetal bovine serum from Australia, and the 3 cells were cultured at a relative humidity of 95% and a temperature of 37 ℃ in the presence of 5% CO2The environment of (2) is grown as a monolayer. SW579 cells were cultured in a medium containing 10% Australian fetal bovine serum L-15 under conditions of 95% relative humidity and 100% air at 37 ℃. Cells in logarithmic growth phase were taken for all experiments.
2. Cell proliferation assay
Taking cells in logarithmic growth phase by MTT method, digesting with pancreatin, counting, adjusting cell density to 1 × 105mL, 100 μ L per well plated in 96-well plates; adding into each well after cell adherenceCulture solution preparation 100. mu.L of solutions (0.001, 0.01, 0.1, 1, 10, 100, 500, 1000. mu.g/mL) for sterile water culture of products prepared in examples 4-6 and comparative examples 6-15 at various concentrations, and control group to which only 100. mu.L of complete culture solution was added, 6 parallel wells per concentration group. According to the results of preliminary experiments, 24h was selected as the drug duration. After the cells were cultured for 24 hours, MTT was added to each well in a concentration of 5mg/mL in 10. mu.L PBS, and 5% CO was added at 37 ℃ to each well2Culturing in 100% air environment, discarding supernatant after 4 hr, adding 100 μ L DMSO into each well to dissolve crystal particles, shaking table for 10min, measuring absorbance at 490nm wavelength of microplate reader, calculating cell inhibition rate, and calculating IC50。
3. Results
See table 4.
TABLE 4 proliferation inhibition IC for different tumor cell lines50
Test example 5 Effect on thyroid cancer-bearing mice
1. Principal materials and reagents
Thyroid cancer TT cell line was purchased from pituitarious biotechnology; 140 SPF-grade BALB/c mice, female and male half of which are purchased from Beijing Witonglihua laboratory animal technology Limited company, and the license numbers are as follows: cxk (Jing) 2019-; fetal bovine serum was purchased from hyelene; DMEM medium was purchased from Beijing ancient cooking country; RIPA lysate was purchased from Beijing Dingguo; the fluorescent quantitative PCR instrument was purchased from ABI (model 970).
2. Animal grouping and handling
SPF grade BALB/c mice, 140, were randomly divided into 14 groups of 10 mice each, and the experimental groups were: in the tumor-bearing control group, examples 4 to 6 and comparative examples 6 to 15, the mice were injected with 0.1mL (about 2X 10) of thyroid cancer TT cell suspension on the back6One), 4w later, the medicine is started to be taken after the tumor formation. The tumor-bearing control group is subjected to intragastric administration with physiological saline, 0.5 mL/tumor, 2 times a day; EXAMPLES 4-6 AND COMPARATIVE EXAMPLES 6-15 compositions prepared by EXAMPLES 4-6 and COMPARATIVE EXAMPLES 6-15Prepared into 1g/mL, and respectively administered to the group of mice for intragastric administration, wherein each time is 0.5mL, 2 times a day; after the tumor formation of the mice, the growth of the inoculated tumor was observed 2 times 1 week and the longest and shortest diameters a and b of the tumor nodules were measured with a vernier caliper, and the tumor volume was calculated according to the formula: v (mm)3)＝1/6π(ab2). After 3 weeks of administration, mice were sacrificed, solid tumors were detached, wet weights were weighed, and tumor inhibition rates were calculated according to the formula: tumor inhibition (%) rate [1- (mean tumor weight of experimental group/mean tumor weight of control group) ]]×100％。
3. Results
The tumor volume changes over time for each group of mice are shown in FIG. 1, and the tumor inhibition results are shown in Table 5.
TABLE 5
Group of
Tumor suppression ratio (%)
Example 4
34.7
Example 5
38.2
Example 6
40.9
Comparative example 6
20
Comparative example 7
20.4
Comparative example 8
17.4
Comparative example 9
16.9
Comparative example 10
17.8
Comparative example 11
2.6
Comparative example 12
31.7
Comparative example 13
31.0
Comparative example 14
32.2
Comparative example 15
32.2
The mice of the groups of examples 4-6 exhibited a tumor growth reduction compared to the tumor-bearing control group of mice that had not been treated at the same time (see FIG. 1). After the materials are obtained, the tumor weights of mice in the tumor-bearing control group are (2.25 +/-0.57) g, the tumor weights of mice in the examples 4-6 groups are (1.47 +/-0.47) g, (1.39 +/-0.59) g and (1.33 +/-0.62) g respectively, and the tumor inhibition rates are 34.7%, 38.2% and 40.9% respectively.
Compared with the prior art, after mixed culture is carried out on strains Bacillus natto918 in monascus and Bacillus natto and selenium-enriched yeast, the obtained composite zymocyte liquid is used for fermenting and culturing soybeans after detoxification and curing treatment, wherein monascus produces polyketone secondary metabolite monascus pigments in the fermentation process, and the monascus secondary metabolite monascus pigments comprise monascus rubimine, monascus ruber amine, monascus rubicin, monascin, ankaflavin and the like, and active substances such as Monacolin K and the like, so that the composite zymocyte liquid has a good anti-tumor effect; meanwhile, due to the mixed culture of the three strains, the production of the citrinin by the monascus can be effectively inhibited, so that the biological toxicity is reduced; the Bacillus natto918 produces anti-cancer substances, namely soybean isoflavone such as genistein, genistin, infirabin and the like, in a high yield in the fermentation process, and the Bacillus natto can inhibit the proliferation of cancer cells, effectively destroy or kill the cancer cells, and can stimulate an immune system to induce and produce interferon to play a role in resisting cancer; the selenium-enriched yeast can generate a large amount of organic selenium nutrients in the fermentation process, the selenium element can influence the energy metabolism of cancer cells and interfere the protein synthesis of the cancer cells through the function of the phagocyte, so that the cancer is inhibited, the metabolism of chemical carcinogens can be influenced, the carcinogenic activity of the chemical carcinogens is lost, and the selenium-enriched natto fermentation extract can play a good anticancer effect.
The invention adds the notoginsenoside R1 and the cordyceps polysaccharide, has the synergistic effect, wherein, the notoginsenoside R1 achieves the anticancer effect by adjusting the hematopoietic function, the cordyceps polysaccharide has the functions of inhibiting the proliferation of tumor cells, enhancing the nonspecific immunity function of organisms and good immunoregulation, mixing with ginsenoside Rg2, ginsenoside Rg3 and prepared fermented extract of selenium-enriched natto, wherein ginsenoside Rg2 can inhibit tumor cell growth, induce tumor cell apoptosis, reverse abnormal differentiation of tumor cell, reverse drug resistance of tumor drug, and improve immunity to achieve anti-tumor effect, and ginsenoside Rg3 has good anti-tumor effect by enhancing immunity, inhibiting cancer cell growth, and resisting tumor metastasis, so as to obtain a biological fermented composition with anticancer effect, the composition has good anticancer effect, and has obvious therapeutic effect on thyroid cancer.
The preparation method is simple, the raw materials are wide in source, the prepared composition has a good anti-tumor effect, particularly has good inhibition and treatment effects on thyroid cancer, is safe and free of side effects, can be used as raw materials of food therapy products, health-care foods and medicines, and has a wide application prospect.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
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