Rapid fermentation method of pulse-activating decoction enzyme

Abstract

The invention discloses a rapid fermentation method of a pulse-activating beverage enzyme, and belongs to the field of food processing. The specific disclosed fermentation method comprises two common prescriptions, namely a pulse-activating decoction (ginseng prescription) and a pulse-activating decoction (codonopsis pilosula prescription). According to the method for fermenting the pulse-activating decoction (ginseng formula), the multi-strain lactic acid bacteria are used as a leavening agent, fresh products or decoction pieces of ginseng, radix ophiopogonis and schisandra chinensis are used as fermentation raw materials, enzymolysis and fermentation processes are combined for use, the whole processing period is shortened to 8-10 hours, the multi-strain lactic acid bacteria are also used as the leavening agent for the pulse-activating decoction (codonopsis pilosula formula), and the fermentation efficiency is improved. Fresh products or decoction pieces of codonopsis pilosula, radix ophiopogonis and schisandra chinensis are used as fermentation raw materials, and an enzymolysis process and a fermentation process are combined, so that the whole processing period is shortened to 10-12 hours, the production cost is greatly saved, and the utilization rate of the raw materials is increased. The traditional Chinese medicine composition can achieve the effects of tonifying qi, restoring pulse, nourishing yin and promoting the secretion of saliva or body fluid, has the effect of regulating intestinal probiotics, and provides health guarantee for diabetics and people with hyperglycemia.

Description

Technical Field

[0001] The invention relates to the field of food processing, and in particular to a rapid fermentation method of Shengmaiyin enzyme. Background Art

[0002] In clinical treatment of traditional Chinese medicine, Shengmaiyin is often used to treat symptoms such as fatigue, shortness of breath, thirst and sweating caused by deficiency of both qi and yin. It has the functions of replenishing qi and promoting body fluid, nourishing yin and benefiting the heart, and is suitable for daily health care. In the clinical treatment of diabetes, it can reduce blood sugar in patients with deficiency of both qi and yin, improve their pancreatic function, TCM syndrome and clinical symptoms. Among them, Shengmaiyin (Dangshen prescription) is more suitable for people with mild symptoms, weak constitution or mild qi deficiency.

[0003] In the processing regulations of Shengmaiyin oral liquid in the Chinese Pharmacopoeia, the raw materials are red ginseng: Ophiopogon japonicus: Schisandra chinensis = 1:2:1, which is quite different from the reference dosage of modern commonly used Shengmaiyin (ginseng prescription) given in the "Chinese Pharmacopoeia" (ISBN: 9787513268714), which is 9g ginseng, 9g Ophiopogon japonicus, and 6g Schisandra chinensis. The relevant content of the Codonopsis prescription Shengmaiyin oral liquid is recorded in the tenth volume of the Ministry of Health's drug standard Chinese medicine formula preparation (WS3-B-1911-95). Combined with the "Chinese Clinical Drug Dictionary·Chinese Medicine Formula Preparation Volume" (referred to as the Chinese Medicine Formula Preparation Volume), the raw drug dosage per serving is about 3g Codonopsis pilosula, 2g Ophiopogon japonicus, and 1g Schisandra chinensis, and the daily raw drug dosage is about 9g Codonopsis pilosula, 6g Ophiopogon japonicus, and 3g Schisandra chinensis. The oral liquid needs to be added with syrup to correct the taste, which is not friendly to diabetic patients. In addition, the intestinal flora of diabetic patients is usually in an unbalanced state, so it is necessary to optimize and innovate the conventional process of Shengmai Drink and develop Shengmai Drink products that are more suitable for diabetic patients.

[0004] At present, the fermentation technology of Shengmai Drink is to ferment the residue of the medicine, which is mainly used for animal feed. The processing technology is complicated and the processing time is long, which takes 5 days. The fermentation process of plant fermentation products alone takes several days or even longer, which increases the production cycle, resulting in high investment and low return, and severely restricts product development. Therefore, it is urgent to find a method that can ferment quickly without affecting the quality of Shengmai Drink enzyme products. Summary of the invention

[0005] In view of the above problems, the purpose of the present invention is to provide a rapid fermentation method for Shengmaiyin enzyme, by adding the treated raw materials to an enzyme preparation for enzymatic hydrolysis, adding a starter agent for fermentation after the enzyme is inactivated, and then adding a sweetener, with or without freeze-drying treatment, to make an enzyme product, and the entire production cycle only takes 8-12 hours (excluding freeze-drying).

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions.

[0007] The present invention discloses a rapid fermentation method of Shengmaiyin enzyme, characterized in that the specific steps of the method are as follows: (1) Weigh and crush the raw materials according to the following mass ratio: ginseng / codonopsis: ophiopogon: schisandra, add water and crush to obtain raw material liquid A; (2) Add saturated sodium bicarbonate aqueous solution to raw material solution A and adjust the pH to 5.5-6.5. Add 0.3-0.5% compound enzyme preparation for enzymatic hydrolysis to obtain raw material solution B; (3) Add 0.3-0.5% of the composite fermentation agent to the raw material liquid B for fermentation at a stirring speed of 60 rpm to obtain liquid C; (4) Centrifuge liquid C at 3000-4000 rpm for 10 min, take the supernatant, and obtain liquid D; (5) Add 0.2%-0.3% of compound sweetener to liquid D to adjust the taste and mix well to obtain product E; (6) Product E is freeze-dried to obtain product F.

[0008] Furthermore, in the method (1), the mass ratio of the raw materials of the ginseng prescription is: ginseng: Ophiopogon japonicus: Schisandra chinensis = 3:3:2; the raw materials of the ginseng prescription are fresh or decoction pieces of ginseng, Ophiopogon japonicus, and Schisandra chinensis; the mass ratio of the raw materials of the Codonopsis pilosula prescription is: Codonopsis pilosula: Ophiopogon japonicus: Schisandra chinensis = 3:2:1.

[0009] Furthermore, in the method (2), the composite enzyme preparation comprises cellulase, pectinase or hydrolyzing protease.

[0010] Furthermore, in the method (3), the fermentation agent includes Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum, Lactobacillus rhamnosus or Bifidobacterium lactis.

[0011] Furthermore, in the method (5), the composite sweetener comprises mannitol, sorbitol, stevioside or β-glucan.

[0012] The present invention also discloses a Shengmaiyin enzyme beverage, characterized in that the Shengmaiyin enzyme beverage is prepared by fermentation according to any of the above methods.

[0013] The present invention also discloses a rapid fermentation method of Shengmaiyin (ginseng prescription), characterized in that the specific steps of the method are as follows: (1) Weigh and crush the raw materials according to the following mass ratio: ginseng: Ophiopogon japonicus: Schisandra chinensis = 3:3:2, add 4-8 times water and crush to obtain raw material liquid A; (2) Add saturated sodium bicarbonate aqueous solution to raw material solution A and adjust the pH to 5.5-6.5; add 0.3-0.5% of a composite enzyme preparation and perform enzymolysis at 60±5°C for 1.5-2.5 hours to obtain raw material solution B; the composite enzyme preparation comprises 0.1%-0.3% cellulase, 0.1%-0.3% pectinase and 0.1%-0.3% hydrolyzing protease; (3) Add 0.3-0.5% of a composite fermentation agent to the raw material liquid B, ferment for 3-5 hours at a temperature of 33±3°C and a stirring speed of 60 rpm to obtain a raw material liquid C; the composite fermentation agent comprises: Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum, and Lactobacillus rhamnosus, and the mass ratio thereof is 1:1:1:1:1; (4) Centrifuge liquid C at 3000-4000 rpm for 10 min, take the supernatant, and obtain liquid D; (5) Add 0.2%-0.3% of a compound sweetener to the liquid D to adjust the taste, and mix well to obtain product E; the compound sweetener includes mannitol, sorbitol and stevioside, and the mass ratio of mannitol: sorbitol: stevioside is 2:2:1; (6) Product E is freeze-dried to obtain product F.

[0014] The present invention also discloses a rapid fermentation method of Shengmaiyin (Dangshen prescription), characterized in that the specific steps of the method are as follows: (1) Weigh and crush the raw materials according to the following mass ratio: Codonopsis pilosula: Ophiopogon japonicus: Schisandra chinensis = 3:2:1, add 4-8 times water and crush to obtain raw material liquid A; (2) Add saturated sodium bicarbonate aqueous solution to raw material solution A and adjust the pH to 5.5-6.5; add 0.3-0.4% of a composite enzyme preparation and perform enzymolysis at 50-60° C. for 2-2.5 hours to obtain raw material solution B; the composite enzyme preparation comprises 0.2%-0.4% cellulase, 0.2%-0.4% pectinase, and 0.2%-0.4% hydrolyzing protease; (3) adding 0.3-0.5% of a composite fermentation agent to the raw material liquid B, fermenting for 5-6 hours at a temperature of 36-39° C. and a stirring speed of 60 rpm to obtain a raw material liquid C; the composite fermentation agent comprises: Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum, and Bifidobacterium lactis, and the mass ratio thereof is 1:1:1:1; (4) Centrifuge liquid C at 3000-4000 rpm for 10 min, take the supernatant, and obtain liquid D; (5) Add 0.2%-0.3% of a compound sweetener to the liquid D to adjust the taste, mix well, and obtain product E; the compound sweetener includes mannitol, β-glucan and stevioside, and the mass ratio of mannitol: stevioside: β-glucan is 2:1:1; (6) Product E is freeze-dried to obtain product F.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows.

[0016] (1) The method for fermenting Shengmai Yin enzyme provided by the present invention can shorten the processing cycle of ginseng prescription from several days to 8-10 hours, of which the fermentation time is only 3-5 hours, and shorten the entire processing cycle of Codonopsis prescription from several days to 10-12 hours, of which the fermentation time is only 4-6 hours, thereby greatly saving production costs and solving the current situation of high investment and low return of Shengmai Yin enzyme.

[0017] (2) The present invention uses lactic acid bacteria or composite bacteria that can improve intestinal flora instead of classic yeast as a fermentation agent to prepare Shengmaiyin enzyme, so that the product can achieve a good effect of regulating blood sugar in the body. Using composite sweeteners instead of syrup added to traditional enzymes or oral liquids greatly reduces the sugar content and calories of the product, and has a good intestinal probiotic effect. It is a rational choice for people who need to lose weight and maintain blood sugar levels.

[0018] (3) The present invention uses ginseng / codonopsis, ophiopogon, and schisandra as fermentation raw materials. The present invention has no strict requirements on the form of product raw materials, and can effectively utilize plant resources that are lost during the picking process, improve their utilization rate, solve the problem of waste in actual medicinal slice processing, and reduce production costs. Compared with medicinal slices, fresh schisandra has an advantage in anthocyanin content. The fermented product raw materials are shown in Figure 1 , red in color, and has a much better sensory experience than Chinese medicine oral liquid.

[0019] (4) The present invention uses ginseng, Ophiopogon japonicus, and Schisandra chinensis as fermentation raw materials, wherein ginseng can be fresh ginseng, sun-dried ginseng, or red ginseng; both dried and fresh Ophiopogon japonicus and Schisandra chinensis can be processed using the above process, the difference being the amount of water added. The present invention has no strict requirements on the form of the product raw materials, and can effectively utilize the plant resources that are lost during the picking process, improve their utilization rate, solve the waste problem in actual medicinal slice processing, and reduce production costs.

[0020] (5) Shengmai Drink used in the present invention has the effects of invigorating Qi and restoring pulse, nourishing Yin and promoting body fluid, and is often used in prescriptions for treating diabetes. This product has important significance in promoting traditional Chinese medicine culture and modernizing Chinese medicine products. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 They are Shengmaiyin enzyme products, of which A is a ginseng enzyme product and B is a codonopsis enzyme product.

[0022] Figure 2This is the enzyme selection diagram, where A is the ginseng enzyme and B is the codonopsis enzyme.

[0023] Figure 3 This is a curve chart of the single factor determination result of enzymatic hydrolysis pH, where A is the ginseng enzyme and B is the codonopsis enzyme.

[0024] Figure 4 This is a curve chart of the single factor determination result of enzymatic hydrolysis time, where A is the enzyme of ginseng formula and B is the enzyme of codonopsis formula.

[0025] Figure 5 This is a curve chart of the single factor determination result of enzymatic hydrolysis temperature, where A is the ginseng enzyme and B is the codonopsis enzyme.

[0026] Figure 6 This is a curve chart of the single factor determination results of enzyme addition, where A is the ginseng enzyme and B is the codonopsis enzyme.

[0027] Figure 7 This is the response surface diagram of the complex enzyme composition of Shengmaiyin enzyme (ginseng prescription). A is the contour line of the effect of hydrolyzing protease and hydrolyzing protease on taste evaluation; B is the response surface of the effect of hydrolyzing protease and hydrolyzing protease on taste evaluation; C is the contour line of the effect of hydrolyzing protease and cellulose on taste evaluation; D is the response surface of the effect of hydrolyzing protease and cellulose on taste evaluation; E is the contour line of the effect of cellulase and hydrolyzed protein on taste evaluation; F is the response surface of the effect of cellulase and hydrolyzed protein on taste evaluation.

[0028] Figure 8 This is the response surface diagram of the complex enzyme composition of Shengmaiyin enzyme (Dangshen Fang). A is the contour line of the effect of hydrolyzing protease and pectin lyase on taste evaluation; B is the response surface of the effect of hydrolyzing protease and pectin lyase on taste evaluation; C is the contour line of the effect of hydrolyzing protease and cellulase on taste evaluation; D is the response surface of the effect of hydrolyzing protease and cellulase on taste evaluation; E is the contour line of the effect of pectin lyase and cellulase on taste evaluation; F is the response surface of the effect of pectin lyase and cellulase on taste evaluation.

[0029] Fig. 9 This is a selection diagram of bacterial strains. A and B are ginseng enzymes, and C and D are codonopsis enzymes.

[0030] Fig.10 This is a curve chart of the single factor determination result of fermentation time, where A is the ginseng enzyme and B is the codonopsis enzyme.

[0031] Fig.11 This is a curve chart of the fermentation temperature single factor determination results. A is the ginseng enzyme and B is the codonopsis enzyme.

[0032] Fig.12This is a curve chart of the single factor determination result of stirring speed, where A is the ginseng enzyme and B is the codonopsis enzyme.

[0033] Fig.13 This is a curve chart of the results of the single factor determination of the inoculation amount, where A is the ginseng enzyme and B is the codonopsis enzyme.

[0034] Fig.14 This is a comparison chart of the changes in polysaccharide content before and after processing. A is ginseng enzyme, and B is codonopsis enzyme.

[0035] Fig.15 This is a scoring chart for sweetener selection. A is ginseng enzyme and B is codonopsis enzyme.

[0036] Fig.16 This is a sweetener addition score chart. A is ginseng enzyme, and B is codonopsis enzyme.

[0037] Fig.17 This is a diagram showing the effect of the product on improving fasting blood sugar in diabetic rats. A is ginseng enzyme and B is codonopsis enzyme.

[0038] Fig.18 This is a diagram showing the effect of the product on improving glucose tolerance in diabetic rats. A is the enzyme from the Panax ginseng formula, and B is the enzyme from the Codonopsis pilosula formula. DETAILED DESCRIPTION

[0039] The present invention is further described in detail below with specific embodiments, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments, and all technologies realized based on the content of the present invention belong to the scope of the present invention.

[0040] Unless otherwise specified, all reagents and materials used in the present invention are commercially available.

[0041] Example 1 Shengmaiyin enzyme (ginseng prescription) fermentation method.

[0042] 1. Experimental materials.

[0043] Fermentation substrate raw materials: fresh and decoction pieces of ginseng, Ophiopogon japonicus, and Schisandra chinensis. Ginseng decoction pieces are raw sun-dried ginseng and red ginseng that are less than 5 years old, baking soda, and drinking water.

[0044] Enzyme preparations: cellulase, pectinase, hydrolyzing protease.

[0045] Fermentation agents: Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum, Lactobacillus rhamnosus.

[0046] Food additives: mannitol, sorbitol, stevioside, β-glucan, isomaltooligosaccharide, glucomannan, protein sugar, maltitol, fructooligosaccharide.

[0047] Experimental animals: Wistar male rats (weight 180-200 g ± 20 g).

[0048] Animal feed: high-sugar and high-fat feed.

[0049] Reagents: streptozotocin (STZ), glucose, and normal saline.

[0050] Consumables: Blood glucose test strips.

[0051] 2. Experimental method.

[0052] 2.1 Pretreatment of raw materials.

[0053] 2.1.1 Treatment of Chinese herbal medicines: Soak ginseng, Ophiopogon japonicus and Schisandra chinensis in 4-5 times distilled water for 30 minutes. Add 3-4 times water and crush them with a wall-breaking machine for later use.

[0054] 2.1.2 Processing method of fresh products: Mix ginseng (less than 5 years old, cut into pieces), Ophiopogon japonicus and Schisandra chinensis in a certain mass ratio, add 3-4 times water, use a wall-breaking machine to break them up, and set aside.

[0055] 2.2 Single factor experiment of enzymatic hydrolysis.

[0056] 2.2.1 Enzyme selection: Each enzyme preparation was added at 0.5% and hydrolyzed for 2 hours. The raw material solution of Shengmai Drink was selected based on the sensory evaluation of the enzymatic effect within the same time. The sensory evaluation standards are as follows.

[0057] Table 1. Sensory evaluation table of Shengmaiyin enzyme (ginseng formula) .

[0058] 2.2.2 Study on enzymatic hydrolysis pH: Add cellulase, pectinase and protease to the processed Shengmai Drink raw material liquid, adjust the pH of the substrate to 4.0, 4.5, 5.0, 5.5, 6.0, 6.5 and 7.0, keep other enzymatic hydrolysis conditions unchanged, and evaluate the taste of the enzymatic hydrolyzate.

[0059] 2.2.3 Study on enzymatic hydrolysis time: Add cellulase, pectinase and protease to the processed Shengmai Drink raw material liquid respectively, adjust the enzymatic hydrolysis time to 1h, 1.5h, 2h, 2.5h and 3h, keep other enzymatic hydrolysis conditions unchanged, and evaluate the taste of the enzymatic hydrolyzate.

[0060] 2.2.4 Study on enzymatic hydrolysis temperature: Add cellulase, pectinase and protease to the processed Shengmai Drink raw material liquid, and adjust the enzymatic hydrolysis temperature to 35℃, 40℃, 45℃, 50℃, 55℃, 60℃, 65℃, 70℃ and 75℃. Keep other enzymatic hydrolysis conditions unchanged and evaluate the taste of the enzymatic hydrolyzate.

[0061] 2.2.5 Study on the amount of enzyme added: Add cellulase, pectinase and protease to the processed Shengmai Drink raw material liquid, adjust the enzyme amount to 0.1%, 0.2%, 0.3%, 0.4% and 0.5%, keep other enzymatic hydrolysis conditions unchanged, and evaluate the taste of the enzymatic hydrolyzate.

[0062] 2.3 Optimization of enzymatic hydrolysis conditions: According to the results of the single-factor experiment, the proportion of enzymes that have a greater impact on the enzymatic hydrolysis effect of Shengmai Yin (ginseng prescription) (total addition amount 0.5%) was selected for response surface experiment, and a meaningful range of values ​​was selected. According to the Design-expert software, the experimental plan was designed, as shown in Table 2.

[0063] Table 2. Response surface experimental design table .

[0064] 2.4 Basic fermentation conditions of Shengmai Drink hydrolysate: The treated Shengmai Drink hydrolysate was inoculated with various lactic acid bacteria for food industry for fermentation. The polysaccharide content was used as an indicator to evaluate the effects of four process conditions, namely fermentation time, fermentation temperature, stirring speed, and inoculation amount, on the quality of Shengmai Drink fermented liquid. The value range of the single factor was set based on the preliminary experiments and literature review. The total polysaccharides in the fermentation liquid were extracted by water extraction and alcohol precipitation, and the total polysaccharide content in the product was measured by weighing. The basic fermentation conditions were fermentation temperature of 37°C, inoculation amount of 0.3%, rotation speed of 40rpm, and fermentation time of 10h.

[0065] 2.4.1 Selection of fermentation bacteria: Inoculate the treated Shengmaiyin hydrolysate with the following types of lactic acid bacteria: Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium lactis, Bifidobacterium bifidum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Bifidobacterium infantis. According to the experimental conditions of 2.4, take the fermentation liquid after fermentation to measure its polysaccharide content. At the same time, measure the time required for the pH of the fermentation liquid to drop to 4.5. Select lactic acid-producing bacteria and polysaccharide-producing bacteria to prepare a composite starter.

[0066] 2.4.2 Selection of fermentation temperature: Inoculate the processed Shengmai Yin (ginseng recipe) hydrolysate with compound lactic acid bacteria, and adjust the fermentation temperature to 31°C, 34°C, 37°C, 42°C, and 45°C according to the experimental conditions in 2.4. After fermentation, take 500 mL of the fermentation liquid to measure its polysaccharide content.

[0067] 2.4.3 Selection of fermentation time: inoculate the processed Shengmai Drink hydrolysate with composite lactic acid bacteria, and adjust the fermentation time to 1.5h, 2.5h, 3.5h, 4.5h, 5.5h, 6.5h, and 7.5h according to the experimental conditions in 2.4. After fermentation, take 500mL of the fermentation liquid to measure its polysaccharide content.

[0068] 2.4.4 Selection of stirring speed: inoculate the processed Shengmai Drink hydrolysate with compound lactic acid bacteria, and adjust the shaking speed to 0rpm, 60rpm, 120rpm, 180rpm, and 360rpm for fermentation according to the experimental conditions in 2.4. Take 500mL of the fermentation liquid to measure its polysaccharide content.

[0069] 2.4.5 Selection of inoculation amount: inoculate the processed Shengmai Drink hydrolysate with composite lactic acid bacteria. According to the experimental conditions of 2.4, adjust the lactic acid bacteria inoculation amount to 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%. Take 500 mL of the fermentation liquid to measure its polysaccharide content.

[0070] 2.4.6 Comparison of total polysaccharides before and after processing: The total polysaccharides in 500 mL of fermentation broth were extracted using the water extraction and alcohol precipitation method, and the weighing method was used for comparison.

[0071] 2.5 Adjustment of product taste: Use Table 3 for sensory evaluation. Organize 9 students to evaluate the texture, color, and taste of Shengmaiyin (ginseng prescription) fermented liquid or freeze-dried powder granules after using additives in the laboratory according to the sensory evaluation table.

[0072] Table 3. Evaluation table of sweetness adjustment of Shengmai Drink enzyme .

[0073] 2.5.1 Selection of sweeteners: According to the upper limit of sweeteners in GB2760-2018, each sweetener is added at 0.2% to examine its effect on the taste of Shengmai Yin (Ginseng Recipe) fermented liquid. The sweeteners involved in the examination are: mannitol, sorbitol, stevioside, β-glucan, isomaltooligosaccharide, glucomannan, protein sugar, maltitol oligosaccharide, and fructooligosaccharide.

[0074] 2.5.2 Study on the ratio of sweeteners: According to the results of 2.5.1, it was found that mannitol and sorbitol have a good effect on improving the taste of Shengmai Yin (ginseng prescription) fermented liquid, and can effectively cover up the bitterness of traditional Chinese medicine, but the sweetness of the two is limited and cannot balance the sourness, so stevia with high sweetness is added to make up for the shortcomings of the two. When stevia is used alone, the sweetness is felt quickly, but the aftertaste is slightly bitter, and the improvement effect is general, so the amount of stevia is not easy to add more. Therefore, the following ratios of mannitol, sorbitol, and stevia are examined respectively: mannitol: sorbitol: stevia = 1:1:1, mannitol: sorbitol: stevia = 1:2:1, mannitol: sorbitol: stevia = 1:3:1, mannitol: sorbitol: stevia = 2:1:1, mannitol: sorbitol: stevia = 3:1:1. Choose the right sweetener ratio.

[0075] 2.6 Animal experiments on Shengmai Yin (Ginseng Formula) product.

[0076] 2.6.1 Establishment of type 2 diabetes rat model: 24 SPF Wistar male rats weighing 180-220g were selected and weighed after one week of adaptive feeding with ordinary feed, and the rats were sorted and numbered according to the size of the body weight. At the same time, 6 rats were randomly selected by random number table and included in the blank control group (hereinafter referred to as the blank group) and fed with maintenance feed. The remaining 18 rats were fed with high-fat feed to induce model establishment. The above feeding method was maintained for 4 weeks. After fasting for 12 hours, the rats were weighed and injected with STZ intraperitoneally at a dose of 30 mg / kg to establish the type 2 diabetes rat model. The blank group rats were injected with the same volume of normal saline according to their body weight. 72 hours after injection, blood was collected from the tail tip to measure the fasting blood glucose (FBG) level. Rats with FBG ≥ 16.7 mmol / L were considered to have successfully established the model and were included in the experimental group for observation and control. If the blood sugar did not reach the standard, the above operation was repeated, STZ was injected intraperitoneally again after fasting, and the blood sugar level of the rats was tested 72 hours later. Rats with blood sugar values ​​that met the standard were included in the experimental group.

[0077] 2.6.2 Experimental animal grouping and treatment. The animals were grouped as follows: blank group: normal rats + ordinary feed + normal saline gavage; model group: model rats + high-fat feed + normal saline gavage; Shengmaiyin enzyme group: model rats + high-fat feed + Shengmaiyin enzyme gavage; Shengmaiyin decoction group: model rats + high-fat feed + Shengmaiyin decoction gavage. The drugs were administered continuously for 6 weeks.

[0078] 2.6.3 Determination of glucose metabolism indicators.

[0079] 2.6.3.1 Determination of fasting blood glucose. Use a blood glucose meter to measure the rat tail vein blood glucose (fasting for 12 hours) at a fixed time every week and keep records.

[0080] 2.6.3.2 OGTT determination After 6 weeks of continuous administration, the rats were fasted for 12 hours and then underwent an oral glucose tolerance test (OGTT). After measuring fasting blood glucose, 2 g / kg glucose solution was intraperitoneally injected, and blood glucose was measured at 30, 60, 90, and 120 minutes.

[0081] 3. Experimental results.

[0082] 3.1 Single factor experimental results of enzymatic hydrolysis.

[0083] 3.1.1 Enzyme selection: Figure 2 It can be seen that pectin lyase, cellulase and protease have better enzymatic hydrolysis effect on the raw material liquid. According to the composition of the raw material, the above three are selected for further study.

[0084] 3.1.2 Study results of enzymatic hydrolysis pH: Figure 3 As can be seen from A, a good enzymatic hydrolysis effect can be obtained at a pH of around 6.

[0085] 3.1.3 Research results of enzymatic hydrolysis time: Figure 4 A shows that the enzymatic hydrolysis effects of each enzyme are in a good state after 1.5 hours of enzymatic hydrolysis, and 2 hours can be used as a more appropriate enzymatic hydrolysis time. After 2.5 hours, no matter which enzyme is used, it is found that the stability and taste of the product are not improved, and the sourness and bitterness are increased.

[0086] 3.1.4 Research results of enzyme dosage: Figure 6 It can be seen that the three enzymes can play a certain hydrolysis effect after the addition amount reaches 0.3%. At the same time, the functions of enzymes are different, and the impact on product quality is also different. Cellulase and pectinase make the plant fiber in the product more delicate, and hydrolyzing protease can reduce the bitterness of the product. Therefore, in the subsequent process of process optimization, the ratio of the three enzymes will continue to be studied.

[0087] 3.1.5 Optimization results of enzymatic hydrolysis conditions: The Box-Benhnken program was used to regress the data results. After regression fitting of various factors, the regression equation obtained is as follows: (Where: A is pectin lyase, B is hydrolyzing protease, and C is the amount of cellulase used). As shown in Table 4, the model F value is 5.56, which means that the model is significant. Comparing the effects of the addition amount of the three enzymes on the taste of the fermentation broth, none of the three enzymes can have a significant effect on the quality of the enzymatic hydrolysate alone, but the interaction between pectin lyase and hydrolyzing protease is strong, and the combined use of the two will have a certain effect on the quality of the fermentation broth; pectin lyase and cellulase also have a certain interaction, which can be seen from the contour line and response surface diagram. Therefore, the optimal enzyme ratio after optimization is: cellulase: pectinase: hydrolyzing protease = 1:1:1.

[0088] Table 4. Response surface results .

[0089] 3.2 Fermentation conditions of Shengmai Yin (ginseng prescription) enzymatic hydrolysate.

[0090] 3.2.1 Selection of fermentation bacteria: Fig. 9 A shows that the microorganisms that produce acid faster are Lactobacillus paracasei, Bifidobacterium bifidum and Lactobacillus acidophilus, which can quickly reduce the pH of the fermentation liquid to below 4.5, which is conducive to the growth and reproduction of other probiotics and improves the sour flavor of the product. Fig. 9B shows that the microorganisms that produce more polysaccharides are Lactobacillus plantarum, Bifidobacterium breve, Bifidobacterium bifidum, and Lactobacillus rhamnosus. In both cases, the same probiotics that are beneficial to the fermentation results are taken, Lactobacillus plantarum: Lactobacillus paracasei: Bifidobacterium breve: Bifidobacterium bifidum: Lactobacillus rhamnosus = 1:1:1:1:1 (mass ratio).

[0091] 3.2.2 Determination of fermentation time: Fig.10 A shows that the accumulation of polysaccharide components in the fermentation broth is beneficial at 3.5-5h of fermentation, indicating that the secondary metabolism of probiotics is active at this time, and the secretion of enzymes that can further hydrolyze the plant cell walls and membranes in the raw materials is beneficial to the release and accumulation of effective ingredients. As the fermentation time increases, the pH in the fermentation system decreases, which promotes the occurrence of acid hydrolysis of effective ingredients, but reduces the content of polysaccharides and other ingredients.

[0092] 3.2.3 Determination of fermentation temperature: Fig.11 AIt can be seen that probiotics are not very demanding on temperature. Good fermentation results can be obtained within the range of 33-42°C, and the difference is not obvious.

[0093] 3.2.4 Determination of stirring speed: Fig.12 A It can be seen that within the range of 0-60rpm, the stirring speed has a greater impact on the fermentation process. After exceeding 60rpm, the stirring speed cannot significantly affect the accumulation of polysaccharides in the system. The maximum rotation speed provided by the air shaker is 360rpm. Studies have found that the shear force generated by the high rotation speed will have a certain destructive effect on the accumulation of polysaccharides in the fermentation system.

[0094] 3.2.5 Determination of inoculation volume: Fig.13 A It can be seen that after the inoculation amount reaches 0.3%, the accumulation of polysaccharides in the fermentation system tends to be moderate, mainly because the substrate concentration limits the growth and development of microorganisms. Therefore, the inoculation amount can be controlled between 0.3-0.5%.

[0095] 3.2.6 Comparison of total polysaccharides before and after processing: Fig.14 A It can be seen that the use of optimized enzymatic hydrolysis and fermentation technology can achieve higher polysaccharide accumulation. After 30 minutes of decoction, Shengmai Yin (ginseng recipe) was left to stand at room temperature, and 500 mL of decoction was extracted with water and precipitated with alcohol. The polysaccharide obtained was 1.5 g, 3.1 g of polysaccharide was obtained from 500 mL of enzymatic hydrolysis solution, and 4.2 g of polysaccharide was accumulated in 500 mL of fermentation solution. This shows that enzymatic hydrolysis and fermentation technology is conducive to the release and accumulation of effective ingredients in Chinese medicinal materials, but due to the sensitivity of the fermentation process, it is necessary to control the fermentation conditions through metabolism to obtain a higher content of functional ingredients.

[0096] 3.3 Adjustment of product taste.

[0097] 3.3.1 Choice of sweeteners: Fig.15 A It can be seen that mannitol and sorbitol have a good effect on improving the taste of Shengmai Yin (ginseng prescription) fermented liquid, and can effectively cover up the bitterness of traditional Chinese medicine, but the sweetness of the two is limited and cannot balance the sourness, so stevia with high sweetness is added to make up for the shortcomings of the two. When stevia is used alone, the sweetness is felt quickly, but the aftertaste is slightly bitter, and the improvement effect is general, so it is not easy to add more stevia.

[0098] 3.3.2 Sweetener ratio: Fig.16 A It can be seen that when mannitol: sorbitol: stevia = 2:2:1, the taste of the fermentation liquid is moderately sweet and sour and slightly bitter, with the special flavor of ginseng and schisandra chinensis, which is a relatively ideal ratio.

[0099] 3.4 The blood sugar lowering effect of Shengmaiyin enzyme product.

[0100] 3.4.1 Fasting blood glucose test results: Fig.17 A It can be seen that the blood sugar level of the animals continued to decrease after administration, and the blood sugar of most of the animals in the administration group returned to the normal range. In the process of blood sugar decrease, the improvement effect of the Shengmai Drink fermented liquid group was slightly better than that of the Shengmai Drink decoction group.

[0101] 3.4.2 OGTT results: Fig.18 A It can be seen that the blood sugar levels of each experimental animal group increased 30 minutes after taking glucose and gradually decreased after 90 minutes. The area under the curve (AUC) of the fermentation liquid group was smaller than that of the water decoction group, and the change trend was more gentle, indicating that the fermentation liquid group had a better effect on improving the insulin resistance of the model animals. And the blood sugar curve of the fermentation liquid group returned to the normal range after 2 hours, indicating that Shengmai Yin (ginseng prescription) can effectively improve the blood sugar level of animals Example 2 Shengmaiyin enzyme (Codonopsis pilosula prescription) fermentation method.

[0102] 1. Experimental materials.

[0103] Fermentation substrate raw materials: fresh and decoction pieces of Codonopsis pilosula, Ophiopogon japonicus, Schisandra chinensis, baking soda, drinking water.

[0104] Enzyme preparations: cellulase, pectinase, hydrolyzing protease.

[0105] Fermentation agents: Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum, Lactobacillus rhamnosus.

[0106] Food additives: mannitol, sorbitol, stevioside, β-glucan, isomaltooligosaccharide, glucomannan, protein sugar, maltitol, fructooligosaccharide.

[0107] Experimental animals: Wistar male rats (weight 180-200 g ± 20 g).

[0108] Animal feed: high-sugar and high-fat feed.

[0109] Reagents: streptozotocin (STZ), glucose, and normal saline.

[0110] Consumables: Blood glucose test strips.

[0111] 2. Experimental method.

[0112] 2.1 Pretreatment of raw materials.

[0113] 2.1.1 Treatment of Chinese herbal medicines: Soak Codonopsis pilosula, Ophiopogon japonicus and Schisandra chinensis in 4-8 times distilled water for 30 minutes. Add 3-4 times water and crush with a wall breaker for later use.

[0114] 2.1.2 Processing method of fresh products: Mix Codonopsis pilosula, Ophiopogon japonicus and Schisandra chinensis in a certain mass ratio, add 3-4 times of water, use a wall-breaking machine to break them up, and set aside.

[0115] 2.2 Single factor experiment of enzymatic hydrolysis.

[0116] 2.2.1 Enzyme selection: Each enzyme preparation was added at 0.5% and hydrolyzed for 2 hours. The raw material solution of Shengmai Drink was selected based on the sensory evaluation of the enzymatic effect within the same time. The sensory evaluation criteria are as follows: Table 5. Sensory evaluation table of Shengmaiyin enzyme (Dangshen prescription) .

[0117] 2.2.2 Study on enzymatic hydrolysis pH: Add cellulase, pectinase and protease to the processed Shengmai Drink raw material liquid, adjust the pH of the substrate to 4.5, 5.0, 5.5, 6.0 and 6.5, keep other enzymatic hydrolysis conditions unchanged, and evaluate the taste of the enzymatic hydrolyzate.

[0118] 2.2.3 Study on enzymatic hydrolysis time: Add cellulase, pectinase and protease to the processed Shengmai Drink raw material liquid respectively, adjust the enzymatic hydrolysis time to 1h, 1.5h, 2h, 2.5h and 3h, keep other enzymatic hydrolysis conditions unchanged, and evaluate the taste of the enzymatic hydrolyzate.

[0119] 2.2.4 Study on enzymatic hydrolysis temperature: Add cellulase, pectinase and protease to the processed Shengmai Drink raw material liquid, adjust the enzymatic hydrolysis temperature to 40℃, 45℃, 50℃, 55℃ and 60℃, keep other enzymatic hydrolysis conditions unchanged, and evaluate the taste of the enzymatic hydrolyzate.

[0120] 2.2.5 Study on the amount of enzyme added: Add cellulase, pectinase and protease to the processed Shengmai Drink raw material liquid, adjust the enzyme amount to 0.1%, 0.2%, 0.3%, 0.4% and 0.5%, keep other enzymatic hydrolysis conditions unchanged, and evaluate the taste of the enzymatic hydrolyzate.

[0121] 2.3 Optimization of enzymatic hydrolysis conditions: According to the results of the single-factor experiment, the proportion of enzymes that have a greater impact on the enzymatic hydrolysis effect of Shengmai Yin (Dangshen prescription) was selected for response surface experiment (total addition amount 0.5%), and a meaningful range of values ​​was selected. According to the Design-expert software, the experimental plan was designed, as shown in Table 6.

[0122] Table 6. Response surface experimental design table .

[0123] 2.4 Basic fermentation conditions of Shengmai Drink hydrolysate: The treated Shengmai Drink hydrolysate was inoculated with various lactic acid bacteria for food industry for fermentation. The polysaccharide content was used as an indicator to evaluate the effects of four process conditions, namely fermentation time, fermentation temperature, stirring speed, and inoculation amount, on the quality of Shengmai Drink fermented liquid. The value range of the single factor was set based on the preliminary experiments and literature review. The total polysaccharides in the fermentation liquid were extracted by water extraction and alcohol precipitation, and the total polysaccharide content in the product was measured by weighing. The basic fermentation conditions were fermentation temperature of 36°C, inoculation amount of 0.3%, rotation speed of 40rpm, and fermentation time of 6h.

[0124] 2.4.1 Selection of fermentation bacteria: Inoculate the treated Shengmaiyin hydrolysate with the following types of lactic acid bacteria: Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium lactis, Bifidobacterium bifidum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Bifidobacterium infantis. According to the experimental conditions of 2.4, take the fermentation liquid after fermentation to measure its polysaccharide content. At the same time, measure the time required for the pH of the fermentation liquid to drop to 4.5. Select appropriate lactic acid-producing bacteria and polysaccharide-producing bacteria to prepare a composite starter.

[0125] 2.4.2 Selection of fermentation temperature: Inoculate the processed Shengmai Yin (Dangshen recipe) hydrolysate with compound lactic acid bacteria, and adjust the fermentation temperature to 30°C, 33°C, 36°C, 39°C, and 42°C according to the experimental conditions in 2.4. After fermentation, take 500 mL of the fermentation liquid to measure its polysaccharide content.

[0126] 2.4.3 Selection of fermentation time: inoculate the processed Shengmai Drink hydrolysate with composite lactic acid bacteria, and adjust the fermentation time to 2.5h, 3.5h, 4.5h, 5.5h, and 6.5h according to the experimental conditions in 2.4. After fermentation, take 500mL of the fermentation liquid to measure its polysaccharide content.

[0127] 2.4.4 Selection of stirring speed: inoculate the processed Shengmai Drink hydrolysate with compound lactic acid bacteria, and adjust the shaking speed to 0rpm, 60rpm, 120rpm, 180rpm, and 360rpm for fermentation according to the experimental conditions in 2.4. Take 500mL of the fermentation liquid to measure its polysaccharide content.

[0128] 2.4.5 Selection of inoculation amount: inoculate the processed Shengmai Drink hydrolysate with composite lactic acid bacteria. According to the experimental conditions of 2.4, adjust the lactic acid bacteria inoculation amount to 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%. Take 500 mL of the fermentation liquid to measure its polysaccharide content.

[0129] 2.4.6 Comparison of total polysaccharides before and after processing: The total polysaccharides in 500 mL of fermentation broth were extracted using the water extraction and alcohol precipitation method, and the weighing method was used for comparison.

[0130] 2.5 Adjustment of product taste: Use Table 3 for sensory evaluation. Organize 9 students to evaluate the texture, color, and taste of Shengmaiyin (Dangshen Fang) fermented liquid or freeze-dried powder granules after using additives in the laboratory according to the sensory evaluation table.

[0131] Table 7. Evaluation table of sweetness adjustment of Shengmaiyin enzyme (Dangshen prescription) .

[0132] 2.5.1 Selection of sweeteners: According to the upper limit of sweeteners in GB2760-2018, each sweetener is added at 0.2% to examine its effect on the taste of Shengmai Yin (Dangshen Fang) fermented liquid. The sweeteners involved in the examination are: mannitol, sorbitol, stevioside, β-glucan, isomaltooligosaccharide, glucomannan, proteose, maltitol, and fructooligosaccharide.

[0133] 2.5.2 Study on the ratio of sweeteners: According to the results of 2.5.1, it was found that mannitol has a good effect on improving the taste of Shengmai Yin (Dangshen Fang) fermented liquid, and can effectively cover up the bitterness of traditional Chinese medicine, but the sweetness of the two is limited and cannot balance the sourness, so stevia and β-glucan with higher sweetness are added to make up for the shortcomings of the two. However, adding too much stevia and β-glucan will lead to a quick sweetness sensation, a slightly bitter aftertaste, and a general improvement effect, so it is not easy to add more of the two. Therefore, the following ratios of mannitol, stevioside, and β-glucan were examined respectively: mannitol: stevioside: β-glucan = 1:1:1, mannitol: stevioside: β-glucan = 1:2:1, mannitol: stevioside: β-glucan = 1:3:1, mannitol: stevioside: β-glucan = 2:1:1, mannitol: stevioside: β-glucan = 3:1:1. Choose the appropriate ratio of sweeteners.

[0134] 2.6 Animal experiments on Shengmai Yin (Dangshen formula) product.

[0135] 2.6.1 Establishment of type 2 diabetes rat model. 24 SPF Wistar male rats weighing 180-220g were selected and weighed after one week of adaptive feeding with ordinary feed. The rats were sorted and numbered according to the size of the weight value. At the same time, 6 rats were randomly selected by random number table and included in the blank control group (hereinafter referred to as the blank group) and fed with maintenance feed. The remaining 28 rats were fed with high-fat feed to induce model establishment. The above feeding method was maintained for 4 weeks. After fasting for 12h, the rats were weighed and injected with STZ intraperitoneally at a dose of 30mg / kg to establish the type 2 diabetes rat model. The blank group rats were injected with the same volume of normal saline according to their body weight. 72h after injection, blood was collected from the tail tip to measure the fasting blood glucose (FBG) level. Rats with FBG ≥ 16.7 mmol / L were considered to have successfully established the model and were included in the experimental group for observation and control. If the blood sugar did not reach the standard, the above operation was repeated, STZ was injected intraperitoneally again after fasting, and the blood sugar level of the rats was tested 72h later. Rats with blood sugar values ​​that met the standard were included in the experimental group.

[0136] 2.6.2 Experimental animal grouping and treatment. The animals were grouped as follows: blank group: normal rats + ordinary feed + normal saline gavage; model group: model rats + high-fat feed + normal saline gavage; Shengmaiyin enzyme group: model rats + high-fat feed + Shengmaiyin enzyme gavage; Shengmaiyin decoction group: model rats + high-fat feed + Shengmaiyin decoction gavage. The drugs were administered continuously for 6 weeks.

[0137] 2.6.3 Determination of sugar metabolism indicators.

[0138] 2.6.3.1 Determination of fasting blood glucose. Use a blood glucose meter to measure the rat tail vein blood glucose (fasting for 12 hours) at a fixed time every week and keep records.

[0139] 2.6.3.2 OGTT determination After 6 weeks of continuous administration, the rats were fasted for 12 hours and then underwent an oral glucose tolerance test (OGTT). After measuring fasting blood glucose, 2 g / kg glucose solution was intraperitoneally injected, and blood glucose was measured at 30, 60, 90, and 120 minutes.

[0140] 3. Experimental results.

[0141] 3.1 Single factor experimental results of enzymatic hydrolysis.

[0142] 3.1.1 Enzyme selection: Figure 2B It can be seen that pectin lyase, cellulase and protease have better enzymatic hydrolysis effect on the raw material liquid. According to the composition of the raw material, the above three are selected for further study.

[0143] 3.1.2 Study results of enzymatic hydrolysis pH: Figure 3 B shows that a good enzymatic hydrolysis effect can be obtained at a pH of about 5.75.

[0144] 3.1.3 Research results of enzymatic hydrolysis time: Figure 4 B shows that the enzymatic hydrolysis effects of each enzyme were in a good state after 2 hours of enzymatic hydrolysis, and 2.5 hours could be used as a more appropriate enzymatic hydrolysis time. After 2.5 hours, no matter which enzyme was used, it was found that the stability and taste of the product were not improved, and the sourness and bitterness increased.

[0145] 3.1.4 Research results of enzymatic hydrolysis temperature: Figure 5 B It can be seen that the enzymatic hydrolysis effect is relatively good in the range of about 55°C.

[0146] 3.1.5 Research results of enzyme dosage: Figure 6 B It can be seen that the three enzymes can play a certain hydrolysis effect after the addition amount reaches 0.3%. At the same time, the functions of enzymes are different, and the impact on product quality is also different. Cellulase and pectinase make the plant fiber in the product more delicate, and hydrolyzing protease can reduce the bitterness of the product. Therefore, in the process of subsequent process optimization, the ratio of the three enzymes will continue to be studied.

[0147] 3.1.6 Optimization results of enzymatic hydrolysis conditions: The Box-Benhnken program was used to regress the data results. After regression fitting of various factors, the regression equation obtained is as follows: (Where: A is the amount of hydrolyzing protease, B is pectinase, and C is the amount of cellulase used.) As shown in Table 8, the model F value is 8.04, which means that the model is significant. Comparing the effects of the addition of the three enzymes on the taste of the fermentation broth, none of the three enzymes can have a significant effect on the quality of the enzymatic hydrolysate alone, but pectin lyase and hydrolyzing protease have a strong interaction, and the combined use of the two will have a certain effect on the quality of the fermentation broth; pectin lyase and cellulase also have a certain interaction, which can be seen from the contour lines and response surface diagrams. Therefore, the optimal enzyme ratio after optimization is: 3:3:4.

[0148] Table 8. Response surface results .

[0149] 3.2 Fermentation conditions of Shengmai Yin (Dangshen prescription) enzymatic hydrolysate.

[0150] 3.2.1 Selection of fermentation bacteria: Fig. 9 C shows that the microorganisms that produce acid faster are Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum and Lactobacillus rhamnosus, which can quickly reduce the pH of the fermentation liquid to below 4.5, which is conducive to the growth and reproduction of other probiotics and improves the sour flavor of the product. Fig. 9 D shows that the microorganisms that produce more polysaccharides are Bifidobacterium breve, Bifidobacterium lactis, and Lactobacillus paracasei. In both cases, the same probiotics that are beneficial to the fermentation results are taken, Lactobacillus paracasei: Bifidobacterium breve: Bifidobacterium bifidum: Bifidobacterium lactis = 1:1:1:1 (mass ratio).

[0151] 3.2.2 Determination of fermentation time: Fig.10 B shows that the accumulation of polysaccharide components in the fermentation broth is beneficial during the fermentation period of 5-6.5h, indicating that the secondary metabolism of probiotics is active at this time, and the secretion of enzymes that can further hydrolyze the plant cell walls and membranes in the raw materials is beneficial to the release and accumulation of effective ingredients. As the fermentation time increases, the pH in the fermentation system decreases, which promotes the occurrence of acid hydrolysis of effective ingredients, but reduces the content of polysaccharides and other ingredients.

[0152] 3.2.3 Determination of fermentation temperature: Fig.11 B It can be seen that the temperature requirements of probiotics are not very strict. Good fermentation results can be obtained within the range of 33-39°C, and the difference is not obvious.

[0153] 3.2.4 Determination of stirring speed: Fig.12 B It can be seen that in the range of 0-60rpm, the stirring speed has a greater impact on the fermentation process. After exceeding 60rpm, the stirring speed cannot significantly affect the accumulation of polysaccharides in the system. The maximum rotation speed provided by the air shaker is 360rpm. The study found that the shear force generated by the high rotation speed will have a certain destructive effect on the accumulation of polysaccharides in the fermentation system.

[0154] 3.2.5 Determination of inoculation volume: Fig.13 B shows that after the inoculation amount reaches 0.3%, the accumulation of polysaccharides in the fermentation system tends to be moderate, mainly because the substrate concentration limits the growth and development of microorganisms. Therefore, the inoculation amount can be controlled between 0.3-0.5%.

[0155] 3.2.6 Comparison of total polysaccharides before and after processing: Fig.14B It can be seen that the use of optimized enzymatic hydrolysis and fermentation technology can achieve higher polysaccharide accumulation. After 30 minutes of decoction, Shengmai Yin (Dangshen Fang) was left to stand at room temperature, and 500 mL of decoction was extracted with water and precipitated with alcohol. The polysaccharide obtained was 4.5 g, the polysaccharide obtained from 500 mL of enzymatic hydrolysis solution was 6.5 g, and the polysaccharide accumulation from 500 mL of fermentation solution was 7.0 g. This shows that enzymatic hydrolysis and fermentation technology is conducive to the release and accumulation of effective ingredients in Chinese medicinal materials, but due to the sensitivity of the fermentation process, it is necessary to control the fermentation conditions through metabolism to obtain a higher content of functional ingredients.

[0156] 3.3 Adjustment of product taste.

[0157] 3.3.1 Choice of sweeteners: Fig.15 B It can be seen that mannitol has a good effect on improving the taste of Shengmai Yin (Dangshen Fang) fermented liquid, and can effectively cover up the bitterness of traditional Chinese medicine, but the sweetness of the two is limited and cannot balance the sourness, so stevioside and β-glucan with higher sweetness are added to make up for the shortcomings of the two. However, adding too much stevioside and β-glucan will lead to a quick sweetness sensation and a slightly bitter aftertaste, and the improvement effect is general, so it is not easy to add more of the two.

[0158] 3.3.2 Sweetener ratio: Fig.16 B It can be seen that when mannitol: stevia: β-glucan = 2:1:1, the taste of the fermentation liquid is moderately sweet and sour and slightly bitter, with the special flavor of dangshen and schisandra, which is a relatively ideal ratio.

[0159] 3.4 The blood sugar lowering effect of Shengmaiyin enzyme product.

[0160] 3.4.1 Fasting blood glucose test results: Fig.17 B shows that the blood sugar level of animals continued to decrease after administration, and the blood sugar level of most of the animals in the administration group returned to the normal range. In the process of blood sugar decrease, the improvement effect of Shengmaiyin fermented liquid group was slightly better than that of Shengmaiyin decoction liquid group.

[0161] 3.4.2 OGTT results: Fig.18 B shows that the blood glucose levels of each experimental animal group increased 30 minutes after taking glucose and gradually decreased after 90 minutes. The area under the curve (AUC) of the fermented liquid group was smaller than that of the decoction group, and the change trend was more gentle, indicating that the fermented liquid group had a better effect on improving the insulin resistance of the model animals. The blood glucose curve of the fermented liquid group returned to the normal range after 2 hours, indicating that Shengmai Yin (Dangshen Fang) can effectively improve the blood glucose level of animals.

[0162] Embodiment 3: .

[0163] The above description is only a preferred embodiment of the present invention and is not intended to limit the scope of the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A rapid fermentation method of Shengmaiyin enzyme, characterized in that: The specific steps of the method are as follows: (1) Weigh and crush the raw materials according to the following mass ratio: ginseng / codonopsis: ophiopogon: schisandra, add water and crush to obtain raw material liquid A; (2) Add saturated sodium bicarbonate aqueous solution to raw material solution A and adjust the pH to 5.5-6.5; add 0.3-0.5% compound enzyme preparation for enzymatic hydrolysis to obtain raw material solution B; (3) Add 0.3-0.5% of the composite fermentation agent to the raw material liquid B for fermentation at a stirring speed of 60 rpm to obtain liquid C; (4) Centrifuge liquid C at 3000-4000 rpm for 10 min, take the supernatant, and obtain liquid D; (5) Add 0.2%-0.3% of compound sweetener to liquid D to adjust the taste and mix well to obtain product E; (6) Product E is freeze-dried to obtain product F.

2. The method according to claim 1, characterized in that: The mass ratio of the raw materials of the ginseng prescription in the method (1) is: ginseng: Ophiopogon japonicus: Schisandra chinensis = 3:3:2; the raw materials of the ginseng prescription are fresh or decoction pieces of ginseng, Ophiopogon japonicus and Schisandra chinensis; the mass ratio of the raw materials of the Codonopsis pilosula prescription is: Codonopsis pilosula: Ophiopogon japonicus: Schisandra chinensis = 3:2:

1.

3. The method according to claim 1, characterized in that In the method (2), the composite enzyme preparation comprises cellulase, pectinase or hydrolyzing protease.

4. The method according to claim 1, characterized in that In the method (3), the fermentation agent includes Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum, Lactobacillus rhamnosus or Bifidobacterium lactis.

5. The method according to claim 1, characterized in that The compound sweetener in the method (5) includes mannitol, sorbitol, stevioside or β-glucan.

6. A Shengmaiyin enzyme drink, characterized in that: The Shengmaiyin enzyme beverage is prepared by fermentation according to the method described in any one of claims 1 to 5.

7. A rapid fermentation method for Shengmaiyin (ginseng prescription), characterized in that: The specific steps of the method are as follows: (1) Weigh and crush the raw materials according to the following mass ratio: ginseng: Ophiopogon japonicus: Schisandra chinensis = 3:3:2, add 4-8 times water and crush to obtain raw material liquid A; (2) Add saturated sodium bicarbonate aqueous solution to raw material solution A and adjust the pH to 5.5-6.5; add 0.3-0.5% of a composite enzyme preparation and perform enzymolysis at 60±5°C for 1.5-2.5 hours to obtain raw material solution B; the composite enzyme preparation comprises 0.1%-0.3% cellulase, 0.1%-0.3% pectinase and 0.1%-0.3% hydrolyzing protease; (3) Add 0.3-0.5% of a composite fermentation agent to the raw material liquid B, ferment for 3-5 hours at a temperature of 33±3°C and a stirring speed of 60 rpm to obtain a raw material liquid C; the composite fermentation agent comprises: Lactobacillus plantarum, Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum, and Lactobacillus rhamnosus, and the mass ratio thereof is 1:1:1:1:1; (4) Centrifuge liquid C at 3000-4000 rpm for 10 min, take the supernatant, and obtain liquid D; (5) Add 0.2%-0.3% of a compound sweetener to the liquid D to adjust the taste, and mix well to obtain product E; the compound sweetener includes mannitol, sorbitol and stevioside, and the mass ratio of mannitol: sorbitol: stevioside is 2:2:1; (6) Product E is freeze-dried to obtain product F.

8. A rapid fermentation method for Shengmaiyin (Dangshen prescription), characterized in that: The specific steps of the method are as follows: (1) Weigh and crush the raw materials according to the following mass ratio: Codonopsis pilosula: Ophiopogon japonicus: Schisandra chinensis = 3:2:1, add 4-8 times water and crush to obtain raw material liquid A; (2) Add saturated sodium bicarbonate aqueous solution to raw material solution A and adjust the pH to 5.5-6.5; add 0.3-0.4% of a composite enzyme preparation and perform enzymolysis at 50-60° C. for 2-2.5 hours to obtain raw material solution B; the composite enzyme preparation comprises 0.2%-0.4% cellulase, 0.2%-0.4% pectinase, and 0.2%-0.4% hydrolyzing protease; (3) adding 0.3-0.5% of a composite fermentation agent to the raw material liquid B, fermenting for 5-6 hours at a temperature of 36-39° C. and a stirring speed of 60 rpm to obtain a raw material liquid C; the composite fermentation agent comprises: Lactobacillus paracasei, Bifidobacterium breve, Bifidobacterium bifidum, and Bifidobacterium lactis, and the mass ratio thereof is 1:1:1:1; (4) Centrifuge liquid C at 3000-4000 rpm for 10 min, take the supernatant, and obtain liquid D; (5) Add 0.2%-0.3% of a compound sweetener to the liquid D to adjust the taste, mix well, and obtain product E; the compound sweetener includes mannitol, β-glucan and stevioside, and the mass ratio of mannitol: stevioside: β-glucan is 2:1:1; (6) Product E is freeze-dried to obtain product F.

Images