Method for fermenting ginseng by using compound strain, fermentation product and application
阅读说明：本技术 复合菌株发酵人参的方法及发酵产物和用途 (Method for fermenting ginseng by using compound strain, fermentation product and application ) 是由 林峰 马涛 沈晓勇 宋帅 于 2021-09-03 设计创作，主要内容包括：本发明提供了一种复合菌株发酵人参的方法及发酵产物和用途。其制备方法包括如下步骤：(1)将人参粉加水制备发酵匀浆,加入维生素C,调节发酵匀浆的pH为3.0～6.0；(2)向所述发酵匀浆中加入酶解剂,在50～60℃下酶解120～180min,调节pH到5.5～7.0,得到酶解液；(3)将所述酶解液在60～90℃下保温35～55min,得到灭菌液；(4)将所述灭菌液降温至10～30℃,向灭菌液中加入发酵菌粉溶液,以接入发酵菌粉溶液开始计时,在发酵温度20～30℃下发酵24～72h,得到发酵液。该方法可以提高发酵液中人参稀有皂苷Rg3的含量。(The invention provides a method for fermenting ginseng by using a compound strain, a fermentation product and application. The preparation method comprises the following steps: (1) adding water into ginseng powder to prepare fermented homogenate, adding vitamin C, and adjusting the pH value of the fermented homogenate to 3.0-6.0; (2) adding an enzymolysis agent into the fermented homogenate, carrying out enzymolysis for 120-180 min at 50-60 ℃, and adjusting the pH to 5.5-7.0 to obtain an enzymolysis liquid; (3) preserving the temperature of the enzymolysis liquid at 60-90 ℃ for 35-55 min to obtain a sterilized liquid; (4) and cooling the sterilized solution to 10-30 ℃, adding a zymophyte powder solution into the sterilized solution, starting timing by inoculating the zymophyte powder solution, and fermenting for 24-72 hours at the fermentation temperature of 20-30 ℃ to obtain fermentation liquor. The method can improve the content of rare ginsenoside Rg3 in the fermentation broth.)
1. A method for fermenting ginseng by using a compound strain is characterized by comprising the following steps:
adding water into ginseng powder to prepare fermented homogenate, adding vitamin C, and adjusting the pH value of the fermented homogenate to 3.0-6.0;
adding an enzymolysis agent into the fermented homogenate, carrying out enzymolysis for 120-180 min at 50-60 ℃, and adjusting the pH to 5.5-7.0 to obtain an enzymolysis liquid;
preserving the temperature of the enzymolysis liquid at 60-90 ℃ for 35-55 min to obtain a sterilized liquid;
cooling the sterilized solution to 10-30 ℃, adding a zymophyte powder solution into the sterilized solution, starting timing by inoculating the zymophyte powder solution, and fermenting for 24-72 hours at the fermentation temperature of 20-30 ℃ to obtain a fermentation liquid;
wherein the zymophyte powder solution is prepared from zymophyte powder and water according to the weight ratio of 1: 10; the lactobacillus casei powder comprises, by weight, 0.1-1 part of lactobacillus casei, 0.1-1 part of lactobacillus bulgaricus, 0.1-1 part of lactobacillus acidophilus, 0.1-1 part of lactobacillus delbrueckii, 0.1-1 part of leuconostoc mesenteroides intestinal membrane subspecies, 0.1-1 part of lactobacillus plantarum, 0.1-1 part of bifidobacterium lactis and 0.1-1 part of bifidobacterium adolescentis;
the enzymolysis agent consists of cellulase, pectinase, acid protease and medium-temperature amylase.
2. The method for fermenting ginseng by using composite bacterial strains according to claim 1, wherein the weight ratio of lactobacillus casei, lactobacillus bulgaricus, lactobacillus acidophilus, lactobacillus delbrueckii, leuconostoc mesenteroides, lactobacillus plantarum, bifidobacterium lactis and bifidobacterium adolescentis in the fermented bacterial powder is 1:1:1:1:1: 1;
the effective bacterium content of the lactobacillus casei is 1800-2500 hundred million CFU/g; the effective bacterium content of the lactobacillus bulgaricus is 1800-2500 hundred million CFU/g; the effective bacterium content of the lactobacillus acidophilus is 1800-2500 hundred million CFU/g; the effective bacterium content of the lactobacillus delbrueckii is 1800-2500 hundred million CFU/g; the effective bacteria content of leuconostoc mesenteroides subspecies mesenteroides is 1800-2500 hundred million CFU/g; the effective bacterium content of the lactobacillus plantarum is 1800-2500 hundred million CFU/g part; the effective bacterium content of the bifidobacterium lactis is 1800-2500 hundred million CFU/g; the effective bacteria content of the bifidobacterium adolescentis is 1800-2500 hundred million CFU/g.
3. The method for fermenting ginseng by using the composite fermentation strain according to claim 1, wherein the weight of the ginseng powder and the enzymolysis agent is (7-12): 1.
4. The method for fermenting ginseng by using composite fermentation strains according to claim 1, wherein the weight ratio of cellulase, pectinase, acid protease and medium-temperature amylase in the enzymolysis agent is 1:1:1: 1.
5. The method for fermenting ginseng by using composite bacterial strains according to claim 1, wherein the weight ratio of the ginseng powder to the zymophyte powder is (400-650): 1.
6. The method for fermenting ginseng by using composite strains according to claim 1, wherein the weight ratio (45-65) of the ginseng powder to the vitamin C in the step (1) is as follows: 1; the weight ratio of the ginseng powder to the water is 1: (6-9).
7. The method for fermenting ginseng with the complex fermentation strain according to claim 1, wherein sodium bicarbonate is further added to adjust the pH of the fermented homogenate in the step (1); and (3) adjusting the pH value by using sodium bicarbonate in the step (2).
8. A fermented ginseng product, which is prepared by the method of fermenting ginseng using the complex strain of any one of claims 1 to 7.
9. A fermented ginseng preparation characterized by being prepared by adding a suitable auxiliary material to the fermented ginseng product according to claim 8 and making into oral liquid, powder or paste by a conventional method.
10. Use of the ginseng fermentation product according to claim 8 or the fermented ginseng preparation according to claim 9 for the preparation of an anti-fatigue product.
Technical Field
The invention belongs to the field of fermentation, and particularly relates to a method for fermenting ginseng by using a composite strain, a fermentation product and application.
Background
Ginseng is the dry root of ginseng of the genus panax of the family araliaceae, listed as the top grade in Shen nong's herbal Jing, which is called to tonify five internal organs, calm spirit, stop palpitation, improve eyesight and benefit intelligence, has the efficacy of reducing weight and prolonging life after long-term administration, is widely applied to health care and medical treatment, and is determined as a new resource food in 2012. The modern pharmacological analysis shows that ginseng contains ginsenoside, ginseng polysaccharide, panaxynol, volatile oil, vitamins (B1, B2 and C), amino acids, various trace elements and the like. Wherein, the ginsenoside is the main active component of the ginseng. It has been found that there are more than 60 prototype ginsenosides in araliaceae plants, including major ginsenosides such as Rb1, Rd, Re and rare ginsenosides Rg3, Rh1, Rh2, the content of major ginsenosides being about 80% of the total ginsenoside content, the major ginsenosides being convertible into rare ginsenosides. The rare ginsenoside Rg3 is a panaxadiol tetracyclic triterpenoid saponin, and has a chemical formula as follows: c42H72O13The molecular weight is: 784.50. studies prove that the ginsenoside Rg3 has the effects of resisting tumors, relaxing blood vessels, enhancing immunity and resisting fatigue. The saponin in ginseng is extracted from plants such as pseudo-ginseng, ginseng and the like because of the complex structure and no feasible chemical synthesis.
Modern Chinese medicine fermentation is a new process, it combines modern processes such as fermentation process with traditional Chinese medicine fermentation, the fermentation process is mainly to add one or more beneficial flora in Chinese medicinal materials, utilize the enzyme produced in the course of microorganism's growth and metabolism to react with complicated active ingredient in Chinese medicine, so can not only improve the content of active ingredient, but also utilize the catalytic action of enzyme to accelerate the completion of reaction. Has been widely applied in the field of extracting ginsenoside.
For example, chinese patent CN111363774A discloses a method for fermenting ginseng with lactobacillus fermentum, and a ginseng fermentation product and use thereof, which comprises the following steps: (1) adding water into Ginseng radix powder to prepare Ginseng radix homogenate, and sterilizing to obtain Ginseng radix sterile solution; wherein the ginseng is added in the ginseng homogenate by 5-12 wt%; (2) after cooling, inoculating a Lactobacillus fermentum seed solution, and performing shaking table fermentation at a fermentation temperature of 24-36 ℃, wherein the rotation speed of the shaking table is 100-300 rmp, and the fermentation time is 20-50 h, so as to obtain a ginseng fermentation product; the lactobacillus fermentum seed liquid is prepared from an MRS liquid culture medium; the viable count of the lactobacillus fermentum seed liquid is 106-108 CFU/mL; the inoculation amount of the lactobacillus fermentum seed liquid is 2-6 wt% of the weight of the ginseng sterilization liquid. However, the fermentation product of the patent focuses on the preparation of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc and ginsenoside Rd, and the application of the product in antioxidant products is not the research on rare ginsenoside Rg 3.
Disclosure of Invention
Therefore, the invention aims to provide a method for fermenting ginseng by using a compound strain, which can improve the content of rare ginsenoside Rg3 in fermentation liquor.
The invention also aims to provide the ginseng fermentation product obtained by the method, wherein the content of rare ginsenoside Rg3 is obviously improved.
The invention also aims to provide a fermented ginseng preparation, which is prepared into oral liquid, powder or paste by adding proper auxiliary materials into the fermented ginseng product and adopting a conventional method, and the oral liquid, powder or paste can be used as a final product and can also be used as a raw material.
It is still another object of the present invention to provide use of the fermented ginseng product and the fermented ginseng preparation for preparing an anti-fatigue product.
The technical scheme of the invention is as follows:
a method for fermenting ginseng by using compound strains comprises the following steps: (1) adding water into ginseng powder to prepare fermented homogenate, adding vitamin C, and adjusting the pH value of the fermented homogenate to 3.0-6.0; (2) adding an enzymolysis agent into the fermented homogenate, carrying out enzymolysis for 120-180 min at 50-60 ℃, and adjusting the pH to 5.5-7.0 to obtain an enzymolysis liquid; (3) preserving the temperature of the enzymolysis liquid at 60-90 ℃ for 35-55 min to obtain a sterilized liquid; (4) cooling the sterilized solution to 10-30 ℃, adding a zymophyte powder solution into the sterilized solution, starting timing by inoculating the zymophyte powder solution, and fermenting for 24-72 hours at the fermentation temperature of 20-30 ℃ to obtain a fermentation liquid; wherein the zymophyte powder solution is prepared from zymophyte powder and water according to the weight ratio of 1: 10; the lactobacillus casei powder comprises, by weight, 0.1-1 part of lactobacillus casei, 0.1-1 part of lactobacillus bulgaricus, 0.1-1 part of lactobacillus acidophilus, 0.1-1 part of lactobacillus delbrueckii, 0.1-1 part of leuconostoc mesenteroides intestinal membrane subspecies, 0.1-1 part of lactobacillus plantarum, 0.1-1 part of bifidobacterium lactis and 0.1-1 part of bifidobacterium adolescentis; the enzymolysis agent consists of cellulase, pectinase, acid protease and medium-temperature amylase.
Preferably, the weight ratio of lactobacillus casei, lactobacillus bulgaricus, lactobacillus acidophilus, lactobacillus delbrueckii, leuconostoc mesenteroides, lactobacillus plantarum, bifidobacterium lactis and bifidobacterium adolescentis in the zymocyte powder is 1:1:1:1:1:1: 1; the effective bacterium content of the lactobacillus casei is 1800-2500 hundred million CFU/g; the effective bacterium content of the lactobacillus bulgaricus is 1800-2500 hundred million CFU/g; the effective bacterium content of the lactobacillus acidophilus is 1800-2500 hundred million CFU/g; the effective bacterium content of the lactobacillus delbrueckii is 1800-2500 hundred million CFU/g; the effective bacteria content of leuconostoc mesenteroides subspecies mesenteroides is 1800-2500 hundred million CFU/g; the effective bacterium content of the lactobacillus plantarum is 1800-2500 hundred million CFU/g part; the effective bacterium content of the bifidobacterium lactis is 1800-2500 hundred million CFU/g; the effective bacteria content of the bifidobacterium adolescentis is 1800-2500 hundred million CFU/g.
Preferably, the weight ratio of the ginseng powder to the enzymolysis agent is (7-12): 1.
Further preferably, the weight ratio of the cellulase to the pectinase to the acid protease to the medium-temperature amylase in the enzymolysis agent is 1:1:1: 1.
Preferably, the weight ratio of the ginseng powder to the zymophyte powder is (400-650): 1.
Further, preferably, the weight ratio (45-65) of the ginseng powder to the vitamin C in the step (1) is as follows: 1; the weight ratio of the ginseng powder to the water is 1: (6-9).
Preferably, sodium bicarbonate can be added in the step (1) to adjust the pH of the fermentation homogenate; and (3) adjusting the pH value by using sodium bicarbonate in the step (2).
A fermented Ginseng radix product is prepared by the method for fermenting Ginseng radix with the above compound strain.
A fermented Ginseng radix preparation is prepared by adding appropriate adjuvants into fermented Ginseng radix product, and making into oral liquid, powder or paste by conventional method.
The use of the fermented ginseng product or the fermented ginseng preparation for preparing an anti-fatigue product.
The technical scheme of the invention has the following advantages:
1. the invention provides a method for fermenting ginseng by using a composite strain, which is characterized in that vitamin C is added into fermented homogenate to adjust the pH value of the fermented homogenate to be 3.0-6.0, so that the activity of enzyme is kept during enzymolysis; the cellulase, the pectinase, the acid protease and the moderate temperature amylase are added as enzymolysis agents, wherein the cellulase can decompose cellulose in the ginseng powder into protein of oligosaccharide or monosaccharide, the pectinase can hydrolyze pectin to decompose plant cell walls, the acid protease can effectively hydrolyze protein, the moderate temperature amylase can liquefy starch more thoroughly, the four enzymolysis agents are cooperatively matched to achieve the effect of quickly decomposing cellulose, pectin, protein and starch in ginseng, so that the cellulose, pectin and other components are converted into glucose under the action of the enzymes, the protein is decomposed into amino acid, and the enzymolysis is facilitatedThe subsequent strains can obtain nutrition more quickly, the rapid growth of the strains is promoted, more enzyme systems are generated, and energy is provided for the high yield of Rg 3; after enzymolysis, because the dissociation degree of carboxyl in the amino acid of the enzymolysis solution is far greater than that of amino, the solution of the amino acid can release a large amount of protons, so that the pH value is reduced, and the quick growth of probiotics is not facilitated, therefore, the pH value of the enzymolysis solution is adjusted to 5.5-7.0 after enzymolysis, namely weak acidity, and the quick growth and propagation of the probiotics are facilitated; sterilization before fermentation is beneficial to removing harmful bacteria in the fermentation liquor; when the zymocyte powder is composed of lactobacillus casei, lactobacillus bulgaricus, lactobacillus acidophilus, lactobacillus delbrueckii, leuconostoc mesenteroides, lactobacillus plantarum, bifidobacterium lactis and bifidobacterium adolescentis, the lactobacillus strains are cooperatively matched to generate beta-glucosidase in the fermentation process, and the beta-glucosidase has the characteristic of hydrolyzing ginsenoside glucoside, so that the ginsenoside Rg in the obtained fermentation liquid3The content is obviously improved.
2. According to the method for fermenting ginseng by using composite bacterial strains, when the weight of ginseng powder and fermentation bacteria powder is (400-650): 1, the weight ratio of each bacterial strain in the fermentation bacteria powder is 1:1:1:1:1:1, the weight ratio of lactobacillus casei, lactobacillus bulgaricus, lactobacillus acidophilus, lactobacillus delbrueckii, leuconostoc mesenteroides, lactobacillus plantarum, bifidobacterium lactis and bifidobacterium adolescentis is 1:1:1:1, and ginsenoside Rg in the obtained fermentation liquor3The content is higher.
3. According to the method for fermenting ginseng by using the compound strain, the weight ratio of the ginseng powder to the enzymolysis agent is (7-12): 1, and the weight ratio of the cellulase to the pectinase to the acid protease to the medium-temperature amylase in the enzymolysis agent is 1:1:1:1, so that the strain can rapidly obtain nutrition, the rapid growth of the strain is promoted, more enzyme systems are generated, and energy is provided for the high yield of Rg 3.
4. The method for fermenting the ginseng by using the composite bacterial strain provided by the invention prepares the fermentation liquor into powder, is convenient to carry, can be used as a product raw material, has a wide application range, is not limited to a liquid preparation, and can be applied to solid beverages or tablets and the like.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in further detail below. All other embodiments, which can be derived by a person skilled in the art from the embodiments disclosed herein without making any creative effort, shall fall within the protection scope of the present disclosure.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional procedures or conditions described in the literature in the field. All reagents are not indicated by manufacturers, and are conventional reagent products which can be obtained commercially.
The Ginseng radix powder is prepared by selecting clean Ginseng radix, pulverizing, and sieving with 60 mesh sieve.
Example 1
The embodiment provides a method for preparing ginseng fermentation liquor by fermenting ginseng with a compound strain, which comprises the following steps:
(1) uniformly mixing 480g of ginseng powder with 3300mL of water to prepare fermented homogenate, adding 8g of vitamin C, uniformly stirring, and adding sodium bicarbonate to adjust the pH value of the fermented homogenate to 4.8;
(2) adding 12g of cellulase, 12g of pectinase, 12g of acid protease and 12g of medium-temperature amylase into the fermented homogenate, stirring at 60 ℃ for enzymolysis for 120min, and adding sodium bicarbonate to adjust the pH value to 5.6 to obtain an enzymatic hydrolysate; wherein the enzyme activity of the cellulase is 2 ten thousand U/g; the enzyme activity of the pectinase is 3 ten thousand U/g; the enzyme activity of the acid protease is 4000U/g; the enzyme activity of the medium-temperature amylase is 1 ten thousand U/g;
(3) preserving the temperature of the enzymolysis liquid at 90 ℃ for 35min to obtain a sterilized liquid;
(4) cooling the sterilized solution to room temperature of 30 ℃, adding a zymophyte powder solution, timing by inoculating the zymophyte powder solution, controlling the temperature to be 30 ℃, and fermenting for 72 hours to obtain a fermentation liquid;
wherein the zymophyte powder solution is prepared from 0.8g of zymophyte powder and 8g of water; the zymocyte powder consists of 0.1g of lactobacillus casei, 0.1g of lactobacillus bulgaricus, 0.1g of lactobacillus acidophilus, 0.1g of lactobacillus delbrueckii, 0.1g of leuconostoc mesenteroides intestinal membrane subspecies, 0.1g of lactobacillus plantarum, 0.1g of bifidobacterium lactis and 0.1g of bifidobacterium adolescentis. The effective bacteria content of the lactobacillus casei is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus bulgaricus is 2000 hundred million CFU/g; the effective bacteria content of the lactobacillus acidophilus is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus delbrueckii is 2000 hundred million CFU/g; the effective bacteria content of leuconostoc mesenteroides subspecies mesenteroides is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus plantarum is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium lactis is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium adolescentis is 2000 hundred million CFU/g.
Example 2
The embodiment provides a method for preparing ginseng fermentation liquor by fermenting ginseng with a compound strain, which comprises the following steps:
(1) uniformly mixing 540g of ginseng powder with 3240mL of water to prepare fermented homogenate, adding 12g of vitamin C, uniformly stirring, and adding sodium bicarbonate to adjust the pH value of the fermented homogenate to 3.0;
(2) adding 19g of cellulase, 19g of pectinase, 19g of acid protease and 19g of medium-temperature amylase into the fermented homogenate, stirring at 50 ℃ for enzymolysis for 180min, and adding sodium bicarbonate to adjust the pH value to 7.0 to obtain an enzymatic hydrolysate; wherein the enzyme activity of the cellulase is 2 ten thousand U/g; the enzyme activity of the pectinase is 3 ten thousand U/g; the enzyme activity of the acid protease is 4000U/g; the enzyme activity of the medium-temperature amylase is 1 ten thousand U/g;
(3) preserving the temperature of the enzymolysis liquid at 60 ℃ for 55min to obtain a sterilized liquid;
(4) cooling the sterilized solution to room temperature of 20 ℃, adding a zymophyte powder solution, timing by inoculating the zymophyte powder solution, controlling the temperature to be 20 ℃, and fermenting for 24 hours to obtain a fermentation liquid;
wherein the zymophyte powder solution is prepared from 1.2g of zymophyte powder and 12g of water; the zymocyte powder consists of 0.15g of lactobacillus casei, 0.15g of lactobacillus bulgaricus, 0.15g of lactobacillus acidophilus, 0.15g of lactobacillus delbrueckii, 0.15g of leuconostoc mesenteroides intestinal membrane subspecies, 0.15g of lactobacillus plantarum, 0.15g of bifidobacterium lactis and 0.15g of bifidobacterium adolescentis. The effective bacteria content of the lactobacillus casei is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus bulgaricus is 2000 hundred million CFU/g; the effective bacteria content of the lactobacillus acidophilus is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus delbrueckii is 2000 hundred million CFU/g; the effective bacteria content of leuconostoc mesenteroides subspecies mesenteroides is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus plantarum is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium lactis is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium adolescentis is 2000 hundred million CFU/g.
Example 3
The embodiment provides a method for preparing ginseng fermentation liquor by fermenting ginseng with a compound strain, which comprises the following steps:
(1) uniformly mixing 540g of ginseng powder with 4800mL of water to prepare fermented homogenate, adding 8.5g of vitamin C, uniformly stirring, and adding sodium bicarbonate to adjust the pH value of the fermented homogenate to 6.0;
(2) adding 11g of cellulase, 11g of pectinase, 11g of acid protease and 12g of medium-temperature amylase into the fermented homogenate, stirring at 60 ℃ for enzymolysis for 120min, and adding sodium bicarbonate to adjust the pH value to 5.6 to obtain an enzymolysis solution; wherein the enzyme activity of the cellulase is 2 ten thousand U/g; the enzyme activity of the pectinase is 3 ten thousand U/g; the enzyme activity of the acid protease is 4000U/g; the enzyme activity of the medium-temperature amylase is 1 ten thousand U/g;
(3) preserving the temperature of the enzymolysis liquid at 90 ℃ for 35min to obtain a sterilized liquid;
(4) cooling the sterilized solution to room temperature of 30 ℃, adding a zymophyte powder solution, timing by inoculating the zymophyte powder solution, controlling the temperature to be 30 ℃, and fermenting for 72 hours to obtain a fermentation liquid;
wherein the zymophyte powder solution is prepared from 0.84g of zymophyte powder and 8.4g of water; the zymocyte powder consists of 0.105g of lactobacillus casei, 0.105g of lactobacillus bulgaricus, 0.105g of lactobacillus acidophilus, 0.105g of lactobacillus delbrueckii, 0.105g of leuconostoc mesenteroides intestinal membrane subspecies, 0.105g of lactobacillus plantarum, 0.105g of bifidobacterium lactis and 0.105g of bifidobacterium adolescentis. The effective bacteria content of the lactobacillus casei is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus bulgaricus is 2000 hundred million CFU/g; the effective bacteria content of the lactobacillus acidophilus is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus delbrueckii is 2000 hundred million CFU/g; the effective bacteria content of leuconostoc mesenteroides subspecies mesenteroides is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus plantarum is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium lactis is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium adolescentis is 2000 hundred million CFU/g.
Example 4
The embodiment provides a method for preparing ginseng fermentation liquor by fermenting ginseng with a compound strain, which comprises the following steps:
(1) uniformly mixing 480g of ginseng powder with 3300mL of water to prepare fermented homogenate, adding 8g of vitamin C, uniformly stirring, and adding sodium bicarbonate to adjust the pH value of the fermented homogenate to 4.8;
(2) adding 12g of cellulase, 12g of pectinase, 12g of acid protease and 12g of medium-temperature amylase into the fermented homogenate, stirring at 60 ℃ for enzymolysis for 120min, and adding sodium bicarbonate to adjust the pH value to 5.6 to obtain an enzymatic hydrolysate; wherein the enzyme activity of the cellulase is 2 ten thousand U/g; the enzyme activity of the pectinase is 3 ten thousand U/g; the enzyme activity of the acid protease is 4000U/g; the enzyme activity of the medium-temperature amylase is 1 ten thousand U/g;
(3) preserving the temperature of the enzymolysis liquid at 90 ℃ for 35min to obtain a sterilized liquid;
(4) cooling the sterilized solution to room temperature of 30 ℃, adding a zymophyte powder solution, timing by inoculating the zymophyte powder solution, controlling the temperature to be 30 ℃, and fermenting for 72 hours to obtain a fermentation liquid;
wherein the zymophyte powder solution is prepared from 0.84g of zymophyte powder and 8.4g of water; the zymocyte powder consists of 0.01g of lactobacillus casei, 0.2g of lactobacillus bulgaricus, 0.01g of lactobacillus acidophilus, 0.2g of lactobacillus delbrueckii, 0.01g of leuconostoc mesenteroides intestinal membrane subspecies, 0.2g of lactobacillus plantarum, 0.01g of bifidobacterium lactis and 0.2g of bifidobacterium adolescentis. . The effective bacteria content of the lactobacillus casei is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus bulgaricus is 2000 hundred million CFU/g; the effective bacteria content of the lactobacillus acidophilus is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus delbrueckii is 2000 hundred million CFU/g; the effective bacteria content of leuconostoc mesenteroides subspecies mesenteroides is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus plantarum is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium lactis is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium adolescentis is 2000 hundred million CFU/g.
Example 5
The embodiment provides a method for preparing ginseng fermentation liquor by fermenting ginseng with a compound strain, which comprises the following steps:
(1) uniformly mixing 500g of ginseng powder with 3400mL of water to prepare fermented homogenate, adding 10g of vitamin C, uniformly stirring, and adding sodium bicarbonate to adjust the pH value of the fermented homogenate to 4.8;
(2) adding 14g of cellulase, 14g of pectinase, 14g of acid protease and 14g of medium-temperature amylase into the fermented homogenate, stirring at 63 ℃ for enzymolysis for 140min, and adding sodium bicarbonate to adjust the pH value to 5.6 to obtain an enzymatic hydrolysate; wherein the enzyme activity of the cellulase is 3 ten thousand U/g; the enzyme activity of the pectinase is 3.5 ten thousand U/g; the enzyme activity of the acid protease is 3800U/g; the enzyme activity of the medium-temperature amylase is 1.2 ten thousand U/g;
(3) preserving the temperature of the enzymolysis liquid at 92 ℃ for 37min to obtain a sterilized liquid;
(4) cooling the sterilized solution to the room temperature of 28 ℃, adding a zymophyte powder solution, timing by inoculating the zymophyte powder solution, controlling the temperature to be 28 ℃, and fermenting for 68 hours to obtain a fermentation liquid;
wherein the zymophyte powder solution is prepared from 1.2g of zymophyte powder and 12g of water; the zymocyte powder consists of 0.15g of lactobacillus casei, 0.15g of lactobacillus bulgaricus, 0.15g of lactobacillus acidophilus, 0.15g of lactobacillus delbrueckii, 0.15g of leuconostoc mesenteroides intestinal membrane subspecies, 0.15g of lactobacillus plantarum, 0.15g of bifidobacterium lactis and 0.15g of bifidobacterium adolescentis. The effective bacteria content of the lactobacillus casei is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus bulgaricus is 2000 hundred million CFU/g; the effective bacteria content of the lactobacillus acidophilus is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus delbrueckii is 2000 hundred million CFU/g; the effective bacteria content of leuconostoc mesenteroides subspecies mesenteroides is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus plantarum is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium lactis is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium adolescentis is 2000 hundred million CFU/g.
Example 6
The embodiment provides fermented ginseng powder, which comprises the following steps:
(1) centrifuging the fermentation liquor prepared in the embodiment 1 at the centrifugal speed of 4000rpm/min for 10min, and taking supernatant;
(2) concentrating the supernatant to 25% of the original fermentation broth volume, and obtaining a concentrated product, wherein the solid content is 40%;
(3) adding 300g of maltodextrin into the concentrated product, mixing, and performing spray drying to obtain fermented ginseng powder, wherein the spray drying condition is that the inlet temperature is 115 ℃, the power of a blower is 20%, and the feeding speed is 8 mL/min.
Example 7
The embodiment provides a fermented ginseng extract, which comprises the following steps:
(1) centrifuging the fermentation liquor prepared in the embodiment 1 at the centrifugal speed of 4000rpm/min for 10min, and taking supernatant;
(2) concentrating the supernatant to 25% of original fermentation liquid volume, and making the solid content be 40% to obtain concentrated product, fermentation extract.
Comparative example 1
This comparative example differs from example 1 in that the strain type is different in the powder of the fermentation bacteria, which consists of 0.8g of Lactobacillus casei. The rest is the same as example 1.
Comparative example 2
This comparative example differs from example 1 in that the strain type in the fermented powder was different, and the fermented powder in this comparative example consisted of 0.8g of Lactobacillus bulgaricus. The rest is the same as example 1.
Comparative example 3
This comparative example differs from example 1 in that the strain type is different in the fermented powder, which consists of 0.8g of Lactobacillus delbrueckii. The rest is the same as example 1.
Comparative example 4
This comparative example differs from example 1 in that the strain type in the powder of the fermenting bacteria is different, the powder of the fermenting bacteria in this comparative example consisting of 0.8g of Lactobacillus acidophilus. The rest is the same as example 1.
Comparative example 5
The comparative example differs from example 1 in that the strain type in the powder of the fermented bacteria is different, and the powder of the fermented bacteria in this comparative example consists of 0.8g of leuconostoc mesenteroides subspecies mesenteroides. The rest is the same as example 1.
Comparative example 6
This comparative example differs from example 1 in that the strain type in the fermented powder was different, and the fermented powder in this comparative example consisted of 0.8g of Lactobacillus plantarum. The rest is the same as example 1.
Comparative example 7
The comparative example differs from example 1 in that the strain type in the fermented powder was different, and the fermented powder in this comparative example consisted of 0.8g of bifidobacterium lactis. The rest is the same as example 1.
Comparative example 8
The comparative example differs from example 1 in that the strain type in the fermented powder was different, and the fermented powder in this comparative example consisted of 0.8g of bifidobacterium adolescentis. The rest is the same as example 1.
Comparative example 9
The comparative example differs from example 1 in that the strain type is different in the fermented powder, which consists of 0.4g bifidobacterium adolescentis and 0.4g lactobacillus plantarum. The rest is the same as example 1.
Comparative example 10
This comparative example differs from example 1 in that the strain type is different in the fermented powder, which consists of 0.4g of bifidobacterium lactis and 0.4g of lactobacillus plantarum. The rest is the same as example 1.
Comparative example 11
This comparative example differs from example 1 in that the strain type is different in the fermented powder, which consists of 0.4g of Lactobacillus bulgaricus and 0.4g of Lactobacillus plantarum. The rest is the same as example 1.
Comparative example 12
This comparative example differs from example 1 in that the strain type differs in the powder of the fermented bacteria, which consists of 0.2g of Lactobacillus casei, 0.2g of Lactobacillus bulgaricus, 0.2g of Leuconostoc mesenteroides and 0.2g of Lactobacillus plantarum. The rest is the same as example 1.
Comparative example 13
This comparative example differs from example 1 in the different strain types in the fermented powder, which consisted of 0.16g of Lactobacillus casei, 0.16g of Lactobacillus bulgaricus, 0.16g of Lactobacillus acidophilus, 0.16g of Leuconostoc mesenteroides subspecies mesenteroides and 0.16g of Lactobacillus plantarum. The rest is the same as example 1.
Comparative example 14
This comparative example differs from example 1 in the different strain types in the fermented powder, which consisted of 0.13g of Lactobacillus casei, 0.13g of Lactobacillus bulgaricus, 0.13g of Lactobacillus acidophilus, 0.13g of Leuconostoc mesenteroides, 0.13g of Bifidobacterium lactis and 0.15g of Lactobacillus plantarum. The rest is the same as example 1.
Comparative example 15
The comparative example provides a method for preparing ginseng fermentation liquor by fermenting ginseng with a composite strain, which comprises the following steps:
(1) uniformly mixing 480g of ginseng powder with 3300mL of water to prepare fermented homogenate, adding 8g of vitamin C, uniformly stirring, and adding sodium bicarbonate to adjust the pH value of the fermented homogenate to 4.8;
(2) preserving the temperature of the fermented homogenate for 35min at the temperature of 90 ℃ to obtain a sterilized solution;
(3) and cooling the sterilization liquid to the room temperature of 30 ℃, controlling the temperature to be 30 ℃, and preserving the temperature for 72 hours to obtain fermentation liquid.
Comparative example 16
The comparative example provides a method for preparing ginseng fermentation liquor by fermenting ginseng with a composite strain, which comprises the following steps:
(1) uniformly mixing 480g of ginseng powder with 3300mL of water to prepare fermented homogenate, adding 8g of vitamin C, uniformly stirring, and adding sodium bicarbonate to adjust the pH value of the fermented homogenate to 4.8;
(2) preserving the temperature of the fermented homogenate at 90 ℃ for 35min to obtain a sterilized solution;
(3) cooling the sterilized solution to room temperature of 30 ℃, adding a zymophyte powder solution, timing by inoculating the zymophyte powder solution, controlling the temperature to be 30 ℃, and fermenting for 72 hours to obtain a fermentation liquid;
wherein the zymophyte powder solution is prepared from 0.8g of zymophyte powder and 8g of water; the zymocyte powder consists of 0.1g of lactobacillus casei, 0.1g of lactobacillus bulgaricus, 0.1g of lactobacillus acidophilus, 0.1g of lactobacillus delbrueckii, 0.1g of leuconostoc mesenteroides intestinal membrane subspecies, 0.1g of lactobacillus plantarum, 0.1g of bifidobacterium lactis and 0.1g of bifidobacterium adolescentis. The effective bacteria content of the lactobacillus casei is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus bulgaricus is 2000 hundred million CFU/g; the effective bacteria content of the lactobacillus acidophilus is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus delbrueckii is 2000 hundred million CFU/g; the effective bacteria content of leuconostoc mesenteroides subspecies mesenteroides is 2000 hundred million CFU/g; the effective bacterium content of the lactobacillus plantarum is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium lactis is 2000 hundred million CFU/g; the effective bacteria content of the bifidobacterium adolescentis is 2000 hundred million CFU/g.
Experimental example 1
Determination of ginsenoside Rg3 content
Fermented ginseng powder as a sample was prepared according to the method of example 6 using the fermented liquids prepared in examples 1 to 5 and comparative examples 1 to 16. Determining the content of ginsenoside Rg3 in the fermented Ginseng radix powder by high performance liquid chromatography.
1 preparation of control solutions
Accurately weighing appropriate amount of ginsenoside standard Rg3, adding methanol to obtain standard solution with concentration of 0.1082mg/ml, and filtering with 0.45 μm filter membrane.
2 investigation of Linear relationship
Precisely sucking 2, 4, 6, 8 and 10 mu L of ginsenoside Rg3 standard solution, carrying out regression statistics by taking mass concentration as a horizontal coordinate (X) and chromatographic peak area as a vertical coordinate (Y), wherein a linear equation of Rg3 is Y =751288X +24057, r =0.9991, and the linear range is 0.32-1.70 mu g.
3 sample treatment
Precisely weighing 1g of each sample (parallel experiment for 2 times), performing Soxhlet extraction, degreasing and decoloring, removing impurities with water saturated n-butyl alcohol solution, collecting extract, and evaporating in water bath to obtain the extract. Dissolving the extract with methanol to obtain test solution with concentration of 0.5mg/ml, and filtering with 0.45 μm filter membrane.
4 chromatographic conditions
ZORBAX SB-C18Chromatography column (4.6 mm. times.250 mm, 5 μm); the column temperature is 35 ℃; the sample volume is 10 mu L; the flow rate is 1mL min-1; mobile phase water (A) -acetonitrile (B), gradient elution is carried out for 0-35 min, 19% of B, 35-55 min, 19-28% of B, 55-80 min, 28% of B, 80-105 min,28% -40% of B, 105-110 min, 40% of B, 110-113 min, 40% -19% of B, 113-120 min and 19% of B; the detection wavelength was 203 mm.
HPLC results of 5 ginsenoside
TABLE 1
Rg3 content/mg g-1
Rg3 content/mg g-1
Rg3 content/mg g-1
Example 1
4.982
Comparative example 3
1.562
Comparative example 10
1.268
Example 2
4.875
Comparative example 4
1.320
Comparative example 11
1.789
Example 3
4.649
Comparative example 5
1.336
Comparative example 12
1.385
Example 4
4.213
Comparative example 6
0.989
Comparative example 13
1.826
Example 5
4.796
Comparative example 7
1.178
Comparative example 14
1.962
Comparative example 1
1.019
Comparative example 8
1.268
Comparative example 15
0.691
Comparative example 2
1.191
Comparative example 9
1.286
Comparative example 16
0.878
As can be seen from Table 1, the content of ginsenoside Rg3 in the fermentation broth of examples 1-5 is significantly higher than that of ginsenoside Rg3 in the fermentation broth of comparative example 15, which indicates that the enzymatic hydrolysis and fermentation of ginseng are helpful for improving the content of ginsenoside Rg 3; as can be seen from the content of the ginsenoside Rg3 in the example 1, the comparative example 15 and the comparative example 16, the enzymolysis is helpful for the release of the ginsenoside Rg3 in the fermentation process; the content of the ginsenoside Rg3 in the fermentation liquor of example 1 and comparative examples 1-14 shows that the strain combination provided by the invention is more beneficial to the release of the ginsenoside Rg3, and the strains are cooperated to ensure that the fermentation liquor contains high-content ginsenoside Rg 3.
Experimental example 2 anti-fatigue mouse experiment
Sample preparation: the fermented ginseng powder prepared in example 6 was selected as sample 1;
centrifuging the fermentation liquor obtained in the comparative example 15 at the centrifugal speed of 4000rpm/min for 10min, and taking supernatant; concentrating the supernatant to 25% of the original fermentation broth volume, and obtaining a concentrated product, wherein the solid content is 40%; the concentrated product was added to 300g of maltodextrin, and after mixing, spray-dried to obtain fermented ginseng powder as sample 2. The spray drying conditions were an inlet temperature of 115 ℃, a blower power of 20%, and a feed rate of 8 mL/min.
Procedure of experiment
50 mice in good condition were selected and randomly divided into 5 groups of 10 mice each with male and female halves. The groupings are shown in Table 2.
TABLE 2
Group of
Administering a medicament
Dosage to be administered
Blank group
Administration ofPhysiological saline
1.976g/kg
Comparative example group
Sample 1
1.976g/kg
Low dose group
Sample 1
0.657g/kg
Middle dose group
Sample 1
1.316g/kg
High dose group
Sample 2
1.976g/kg
The animals in each group were gavaged 1 time per day, measured as three times of low, medium and high. The administration is continued for 15 days. The room temperature is 23 +/-1 ℃, the mouse consciously takes food and drinks water, the mouse food is periodically replaced, the environment of the mouse is kept clean, and the condition of the mouse is observed and basically recorded in detail.
Weight bearing swimming test for mice
Animals of each dose group are placed in a deep water tank with the water temperature of 25 +/-0.5 ℃ for swimming according to the weight of a mouse load of 5% after the last dose for 1 hour, and an exhaustion load swimming test is carried out. After the mouse is put into the swimming tank, a stopwatch is used for timing immediately, the condition of the mouse is observed and recorded in detail, and when the condition is right, the mouse is stirred by a glass rod prepared in advance, so that the test mouse is always in a swimming activity and motion state. The time from swimming in the water tank to swimming in the water tank until the mouse sinks 10 seconds below the water surface without moving is taken as exhaustion weight swimming time/min. The results are shown in Table 3.
TABLE 3 results of the weight swimming test of mice
Group of
Animal number (only)
Dosage (g/kg)
Swimming time(s)
Blank group
10
1.976
496.3±92.3
Comparative example group
10
1.976
546.2±101.2
Low dose group
10
0.65
674.3±136.5
Middle dose group
10
1.316
723.5±157.2
High dose group
10
1.976
802.3±192.7
Through comparison of data, the fermented ginseng powder can obviously increase the load swimming time of a mouse and enhance the anti-fatigue capability of the mouse.
Measurement of blood lactic acid
Half an hour after the last administration of sample 1 and sample 2, the samples were placed in a water bath at a temperature of 30. + -. 1 ℃. The mouse bears a load of 4% and swims, and the stopwatch counts 10min to stop swimming. Then blood is taken from the canthus veins in the eyes which are not exercised in water, 0min after exercise, 15min after exercise and one hour after exercise respectively, and anticoagulant is added and mixed evenly. The content of the blood lactic acid is determined according to the blood lactic acid kit instruction provided by Jiangsu Nanjing institute of bioengineering. The results are shown in Table 4.
TABLE 4 measurement results (mmol/L) of blood lactic acid
Group of
Number of mice n
Before swimming
0min after swimming
15min after swimming
60min after swimming
Blank group
10
22.8±4.8
46.2±4.3
29.9±6.6
18.8±2.7
Comparative example group
10
23.7±6.0
44.8±4.2
29.2±6.1
18.7±3.7
Low dose group
10
22.3±4.6
40.9±5.5
24.6±3.9
17.3±5.2
Middle dose group
10
21.6±3.9
34.8±4.8
24.1±3.5
16.9±2.8
High dose group
10
22.3±4.9
30.3±4.9
21.7±2.8
16.3±3.3
After the mice move in a swimming manner for 0min and 15min, the mice of the high-dose stomach-perfused fermented ginseng powder group are compared with the mice of the stomach-perfused unfermented ginseng powder control group, the blood lactic acid content of the mice of the high-dose stomach-perfused fermented ginseng powder group is respectively reduced by 32.4% (P < 0.01) and 25.7% (P < 0.05), the influence of the stomach-perfused fermented ginseng powder on the blood lactic acid of the mice is shown, the blood lactic acid content of the mice is reduced within 15min after the mice swim, and the mice approach the stomach-perfused unfermented ginseng powder control group, so that the product has the effects of adjusting the metabolism of the organism and eliminating physical fatigue.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
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