CN114668144A - Hypoglycemic composite plant enzyme, preparation method and application - Google Patents
Hypoglycemic composite plant enzyme, preparation method and application Download PDFInfo
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- CN114668144A CN114668144A CN202210160311.9A CN202210160311A CN114668144A CN 114668144 A CN114668144 A CN 114668144A CN 202210160311 A CN202210160311 A CN 202210160311A CN 114668144 A CN114668144 A CN 114668144A
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- hypoglycemic
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- ferment
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Abstract
The invention provides a hypoglycemic compound plant enzyme, a preparation method and application thereof. The blood sugar reducing composite plant enzyme is prepared by composite fermentation of medicinal and edible traditional Chinese medicinal materials and plants, has the efficacy and edibility of reducing blood sugar, is not medicinal, is safe and reliable, and has low cost. The optimal effect of reducing blood sugar is achieved by optimizing the fermentation time, the fermentation temperature and the addition amount of the compound lactobacillus, the fermentation time is short, the activities of alpha-amylase and alpha-glucosidase can be effectively inhibited, the fermentation process is mature, simple and convenient, and the method is suitable for industrial large-scale production. The blood sugar-reducing composite plant enzyme provided by the application can enable the blood sugar reduction rate of a rat to be 55.70%, is higher than most enzyme products on the market, and has a relatively strong blood sugar-reducing effect. Therefore, the hypoglycemic composite plant enzyme can solve the problem of shortage of hypoglycemic functional foods in the current market, simultaneously reduce uncomfortable symptoms caused by medication to the body and delay the occurrence of complications.
Description
Technical Field
The invention relates to the technical field of food fermentation, in particular to a hypoglycemic compound plant enzyme, a preparation method and application thereof.
Background
Diabetes, one of the epidemic diseases in the world today, is second to malignant tumor and cardiovascular and cerebrovascular diseases, and becomes the third disease harmful to human health, wherein type 2 diabetes accounts for 85% -95% of the total number of diabetes. The modern medicine for treating type 2 diabetes mainly comprises oral hypoglycemic drugs and insulin, and is assisted by diet control and exercise therapy, but the ideal curative effect is not achieved, and certain limitations exist.
The traditional Chinese medicine therapy is characterized in that the individual difference and compound use of patients are emphasized, and the traditional Chinese medicine for treating diabetes not only reduces blood sugar, but also focuses on preventing and treating diabetic complications, thereby playing roles in improving life quality and prolonging life. However, the traditional Chinese medicine prescription has a series of relatively troublesome problems of grabbing, decocting, storing and the like, and brings inconvenience to patients, so that the functional food with the blood sugar reducing effect is developed, and the application prospect is wide.
The traditional Chinese medicine has the theory of homology of medicine and food since ancient times, namely, some medicines are food and medicines, and no absolute boundary exists between the two medicines. With the development of economy in China and the improvement of the living standard of the nation, the health-care consciousness of people is gradually enhanced, more and more traditional Chinese medicinal materials are listed as medicinal and edible substances, and the development of novel enzyme food with the blood sugar reducing function by utilizing medicinal and edible plant resources becomes a new trend. The edible ferment is a functional fermented food produced by fermenting fresh fruits, vegetables, edible fungi, Chinese herbal medicines and the like serving as raw materials through microorganisms. In recent years, the health care function of the edible ferment is keen, and the market demand is increasing. However, the edible ferment products with the blood sugar reducing effect imported at the present stage are generally expensive in price at home, and meanwhile, the edible ferment products in China cannot meet the market demand. Therefore, there is a need to develop functional food ferment with blood sugar lowering effect to meet the increasing domestic requirements of diabetes patients.
Disclosure of Invention
The invention provides a hypoglycemic compound plant enzyme, a preparation method and application thereof, and aims to solve the problems that an edible enzyme product with a hypoglycemic effect is expensive in price and poor in hypoglycemic effect.
The invention provides a blood sugar-reducing composite plant enzyme, which comprises the following raw materials: red ginseng, fragrant solomonseal rhizome, purslane, medlar, pomelo, pumpkin, ginger, celery and Chinese yam. Wherein Ginseng radix Rubri is dry root of Ginseng radix, and has effects of invigorating spleen, benefiting lung, promoting fluid production, and quenching thirst, and ginsenoside has effects of lowering blood sugar; the polygonatum odoratum has the effects of nourishing yin and moistening dryness, and the polygonatum odoratum polysaccharide can reduce blood sugar and blood fat and effectively promote insulin secretion, and is particularly characterized in that the polygonatum odoratum polysaccharide effectively reduces the contents of FBG, HbA1C, TC and LDL-C and improves the contents of fasting insulin and C peptide; polysaccharides in the Chinese yam, the pumpkin and the medlar can improve the apoptosis of islet beta cells, repair damaged islet beta cells and restore the normal secretion function of the islet beta cells, so that the aim of reducing blood sugar is fulfilled; the purslane polysaccharide can increase insulin sensitivity, improve glycolipid metabolic capability, reduce lipid metabolic disorder and prevent vascular inflammation caused by diabetes; the pomelo has sweet and sour pulp, is cold in nature, can promote the circulation of qi, relieve the epigastric distention, stimulate the appetite and promote digestion, contains insulin-like components, has a certain blood sugar reducing effect, and has faint scent, sour and sweet taste which can neutralize the bitter taste of the traditional Chinese medicine; apigenin in celery is one of the main flavonoids in human diet, and the flavonoids are known to protect the function of islet beta cells and enhance the sensitivity of peripheral tissues to insulin.
Specifically, compound plant ferment's of hypoglycemic in this application preparation raw materials includes: 5-10g/100g of red ginseng, 5-15g/100g of polygonatum odoratum, 5-15g/100g of purslane, 5-10g/100g of medlar, 5-15g/100g of grapefruit, 5-15g/100g of pumpkin, 5-10g/100g of ginger, 5-15g/100g of celery and 5-15g/100g of Chinese yam.
The blood glucose reducing composite plant enzyme has an inhibiting effect on alpha-glucosidase and alpha-amylase, and can reduce fasting blood glucose value of rats and inhibit the absorption of the rats on sucrose and starch.
The application also provides a preparation method of the hypoglycemic composite plant enzyme, which comprises the following steps:
s01: pulverizing Ginseng radix Rubri, rhizoma Polygonati Odorati, herba Portulacae, fructus Lycii, fructus Citri Grandis, fructus Cucurbitae Moschatae, rhizoma Zingiberis recens, herba Apii Graveolentis, and rhizoma Dioscoreae to obtain solid pulverized material.
Pulverizing Ginseng radix Rubri, rhizoma Polygonati Odorati, herba Portulacae, fructus Lycii, fructus Citri Grandis, fructus Cucurbitae Moschatae, rhizoma Zingiberis recens, herba Apii Graveolentis, and rhizoma Dioscoreae to obtain solid pulverized material.
S02: adding the pulverized solid into distilled water, and homogenizing at 12000r for 5min to obtain homogeneous mixed solution.
Adding the solid pulverized material into distilled water, and homogenizing at 12000r for 5min to obtain homogeneous mixed solution. Wherein the mass ratio of the solid crushed material to the distilled water is 3: 10.
S03: and sterilizing the homogenized mixed solution, and cooling to room temperature.
Sterilizing the homogenized mixture at 115 deg.C for 30 min. And cooling the sterilized homogeneous mixed solution to room temperature.
S04: adding composite lactobacillus powder into the homogenized mixed solution cooled to room temperature, and fermenting at 28-40 deg.C for 0-84h to obtain ferment.
Adding composite lactobacillus powder into the homogeneous mixed solution cooled to room temperature, and fermenting at 28-40 deg.C for 0-84h to obtain ferment. Wherein the composite lactobacillus powder comprises bifidobacterium lactis, lactobacillus acidophilus, lactobacillus casei, lactobacillus rhamnosus and lactobacillus plantarum. The addition amount of the compound lactobacillus is 0-0.4% of the homogeneous mixed liquid.
S05: and centrifuging and filtering the fermented ferment under an aseptic condition to obtain the hypoglycemic composite plant ferment.
And (4) centrifuging and filtering the fermented ferment under an aseptic condition to obtain the hypoglycemic composite plant ferment. Wherein the centrifugal speed is 4500r, and the centrifugal time is 3 min.
The application also tests the preparation elements in the preparation method to determine the optimal preparation process so as to achieve the optimal blood sugar reducing effect. The following is a detailed description.
1. Influence of fermentation time on blood sugar reducing composite plant enzyme
The inhibition rate of alpha-amylase is taken as an optimization condition in the application to determine the influence of fermentation time on the hypoglycemic composite plant enzyme. Specifically, the hypoglycemic composite plant ferment is prepared by the preparation method according to different fermentation times, wherein the fermentation times are respectively 0h, 12h, 24h, 36h, 48h, 60h, 72h and 84 h. Meanwhile, except for the fermentation time, the other preparation process conditions are the same.
The inhibition of alpha-amylase by fermentation time is determined by the method for measuring the inhibition rate of alpha-amylase commonly used in the field. Specifically, 28. mu.L of the enzyme supernatant and 100. mu.L of an α -amylase solution were added to 1mL of a PBS (phosphate buffered saline; English full name: phosphate buffered saline; Chinese full name: phosphate buffered saline) solution at pH6.8, and the mixture was heated in a 37 ℃ water bath for 20 min. After the heating, 5mL of a 1% soluble starch solution was added, and the water bath heating was continued for 5 min. After the heating, 1mL of the sample solution was heated in a boiling water bath for 5 min. After the heating, 1mL of distilled water and 1.5mL of DNS (3, 5-Dinitrosalicylic acid, Chinese) were added and the mixture was heated in a boiling water bath for 5 minutes. After cooling, 6.5mL of distilled water was added while measuring absorbance at 540nm to determine the inhibition rate of alpha-amylase, and A was measured using PBS instead of enzyme as a blank Blank spaceSample control assay A with PBS instead of alpha-amylaseSample controlsAssay A with PBS instead of enzyme and alpha-amylase as blank controlBlank control. . Wherein, the alpha-starchThe inhibition rate of the enzyme is calculated by the absorbance, and the calculation formula is
A graph is drawn according to the fermentation time, pH and the inhibition rate of alpha-amylase to obtain FIG. 1. As can be seen from the figure 1, when the fermentation time is more than 48h, the inhibition rate and the pH of the alpha-amylase are basically stable, so that the optimal fermentation time of the hypoglycemic composite plant ferment is 48 h.
2. Influence of fermentation temperature on hypoglycemic composite plant enzyme
The inhibition rate of alpha-amylase is taken as an optimization condition in the application to determine the influence of the fermentation temperature on the hypoglycemic composite plant enzyme. Specifically, the hypoglycemic composite plant enzyme is prepared at different fermentation temperatures according to the preparation method, wherein the fermentation temperatures are respectively 28 ℃, 31 ℃, 34 ℃, 37 ℃ and 40 ℃. Meanwhile, the preparation process conditions are the same except for the fermentation temperature.
The inhibition of alpha-amylase by fermentation temperature is determined by the method for determining the inhibition rate of alpha-amylase commonly used in the field. The measurement method was the same as above. A curve chart is drawn according to the fermentation temperature, pH and the inhibition rate of alpha-amylase, and a graph of FIG. 2 is obtained. As can be seen from FIG. 2, when the fermentation temperature is 37 ℃, the inhibition rate of alpha-amylase is the highest, and thus, the optimal fermentation time of the hypoglycemic composite plant ferment is 37 ℃.
3. Influence of composite lactobacillus powder addition amount on blood sugar reducing composite plant enzyme
The inhibition rate of alpha-amylase is used as an optimization condition in the application to determine the influence of the addition amount of the composite lactobacillus powder on the hypoglycemic composite plant enzyme. Specifically, the hypoglycemic composite plant enzyme is prepared according to the preparation method by adding different composite lactic acid bacteria powder, wherein the composite lactic acid bacteria powder is added by 0%, 0.025%, 0.05%, 0.1%, 0.2%, 0.3% and 0.4%, respectively. Meanwhile, except the addition amount of the composite lactobacillus powder, the other preparation process conditions are the same.
The inhibition effect of the composite lactic acid bacteria powder addition amount on the alpha-amylase is measured by adopting an alpha-amylase inhibition rate measuring method commonly used in the field. The measurement method is the same as above. A curve chart is drawn according to the addition amount of the composite lactobacillus powder, pH and the inhibition rate of alpha-amylase, and a graph shown in figure 3 is obtained. As can be seen from FIG. 3, when the amount of the composite lactic acid bacteria powder is 0.1%, the inhibition rate of the alpha-amylase is the highest, and when the amount of the composite lactic acid bacteria powder is more than 0.1%, the inhibition rate of the alpha-amylase is basically unchanged. Therefore, the optimal compound lactobacillus powder adding amount of the blood sugar reducing compound plant ferment is 0.1 percent.
According to the experimental verification, the optimal process conditions for preparing the hypoglycemic composite plant enzyme are as follows: the fermentation time is 48h, the fermentation temperature is 37 ℃, and the addition amount of the composite lactobacillus powder is 0.1 percent.
The technical scheme provided by the embodiment of the invention can have the following beneficial effects:
the invention provides a hypoglycemic compound plant enzyme, a preparation method and application thereof. The blood sugar reducing composite plant enzyme is prepared by carrying out composite fermentation on medicinal and edible traditional Chinese medicinal materials and plants such as vegetables, fruits and the like, has the effect of reducing blood sugar, is edible, is not medicinal, safe and reliable, and has low cost. In the preparation process of the hypoglycemic composite plant enzyme, the optimal hypoglycemic effect is achieved by optimizing the fermentation time, the fermentation temperature and the addition amount of the composite lactic acid bacteria, the fermentation time is short, the activities of alpha-amylase and alpha-glucosidase can be effectively inhibited, the fermentation process is mature, simple and convenient, and the hypoglycemic composite plant enzyme is suitable for industrial large-scale production. The hypoglycemic compound plant enzyme provided by the application can enable the blood sugar reduction rate of rats to be 55.70%, is higher than most enzyme products on the market, and has a relatively strong hypoglycemic effect. Therefore, the hypoglycemic composite plant enzyme can solve the problem of shortage of hypoglycemic functional foods in the current market, simultaneously reduce uncomfortable symptoms caused by medication to the body and delay the occurrence of complications.
It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.
Drawings
In order to more clearly explain the technical solution of the present application, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious to those skilled in the art that other drawings can be obtained according to the drawings without any creative effort.
FIG. 1 is a graph showing the effect of fermentation time on the inhibition rate of alpha-amylase and pH, according to an embodiment of the present invention;
FIG. 2 is a graph showing the effect of fermentation temperature on α -amylase inhibition and pH according to an embodiment of the present invention;
FIG. 3 is a graph showing the effect of the addition of composite lactobacillus powder on the inhibition rate of alpha-amylase and pH value;
FIG. 4 is a graph comparing the α -amylase inhibition and α -glucosidase inhibition provided by the examples of the present invention;
FIG. 5 is a graph showing the results of the glucose tolerance of the enzyme in rats orally administered with glucose according to the present invention;
FIG. 6 is a graph showing the results of the tolerance of the enzyme to the oral starch of rats according to the present invention;
Detailed Description
The present application provides a hypoglycemic compound plant enzyme and a preparation method thereof, which are described in detail in the following with specific embodiments.
Example 1
The embodiment of the application provides compound plant ferment of hypoglycemic, and this compound plant ferment of hypoglycemic's preparation raw materials includes: 5g/100g of red ginseng, 5g/100g of polygonatum odoratum, 10g/100g of purslane, 10g/100g of medlar, 15g/100g of pomelo, 15g/100g of pumpkin, 10g/100g of ginger, 15g/100g of celery and 15g/100g of Chinese yam.
The embodiment of the application also provides a preparation method of the hypoglycemic composite plant ferment, which comprises the following steps:
s101: pulverizing Ginseng radix Rubri, rhizoma Polygonati Odorati, herba Portulacae, fructus Lycii, fructus Citri Grandis, fructus Cucurbitae Moschatae, rhizoma Zingiberis recens, herba Apii Graveolentis, and rhizoma Dioscoreae to obtain solid pulverized material.
S102: adding the solid pulverized material into distilled water, and homogenizing at 12000r for 5min to obtain homogeneous mixed solution. Wherein the mass ratio of the solid crushed material to the distilled water is 3: 10.
S103: sterilizing the homogenized mixture at 115 deg.C for 30 min. And cooling the sterilized homogeneous mixed solution to room temperature.
S104: adding composite lactobacillus powder into the homogeneous mixed solution cooled to room temperature, and fermenting at 28 deg.C for 0h to obtain ferment. Wherein the composite lactobacillus powder comprises bifidobacterium lactis, lactobacillus acidophilus, lactobacillus casei, lactobacillus rhamnosus and lactobacillus plantarum. The adding amount of the compound lactobacillus is 0 percent of the mass of the homogeneous mixed liquid.
S105: and (3) carrying out centrifugal filtration on the fermented ferment under the aseptic condition to obtain the blood sugar-reducing composite plant ferment. Wherein the centrifugation speed is 4500r, and the centrifugation time is 3 min.
Example 2
The embodiment of the application provides compound plant ferment of hypoglycemic, and this compound plant ferment of hypoglycemic's preparation raw materials includes: 10g/100g of red ginseng, 15g/100g of polygonatum odoratum, 15g/100g of purslane, 5g/100g of medlar, 10g/100g of pomelo, 10g/100g of pumpkin, 10g/100g of ginger, 10g/100g of celery and 15g/100g of Chinese yam.
The embodiment of the application also provides a preparation method of the hypoglycemic composite plant ferment, which comprises the following steps:
s201: pulverizing Ginseng radix Rubri, rhizoma Polygonati Odorati, herba Portulacae, fructus Lycii, fructus Citri Grandis, fructus Cucurbitae Moschatae, rhizoma Zingiberis recens, herba Apii Graveolentis, and rhizoma Dioscoreae to obtain solid pulverized material.
S202: adding the solid pulverized material into distilled water, and homogenizing at 12000r for 5min to obtain homogeneous mixed solution. Wherein the mass ratio of the solid crushed material to the distilled water is 3: 10.
S203: sterilizing the homogenized mixture at 115 deg.C for 30 min. And cooling the sterilized homogeneous mixed solution to room temperature.
S204: adding composite lactobacillus powder into the homogeneous mixed solution cooled to room temperature, and fermenting at 40 ℃ for 84h to obtain the ferment. Wherein the composite lactobacillus powder comprises bifidobacterium lactis, lactobacillus acidophilus, lactobacillus casei, lactobacillus rhamnosus and lactobacillus plantarum. The adding amount of the compound lactobacillus is 0.4 percent of the mass of the homogeneous mixed liquid.
S205: and (4) centrifuging and filtering the fermented ferment under an aseptic condition to obtain the hypoglycemic composite plant ferment. Wherein the centrifugal speed is 4500r, and the centrifugal time is 3 min.
Example 3
The embodiment of the application provides compound plant ferment of hypoglycemic, and this compound plant ferment of hypoglycemic's preparation raw materials includes: 10g/100g of red ginseng, 15g/100g of polygonatum odoratum, 15g/100g of purslane, 10g/100g of medlar, 15g/100g of pomelo, 15g/100g of pumpkin, 10g/100g of ginger, 5g/100g of celery and 5g/100g of Chinese yam.
The embodiment of the application also provides a preparation method of the hypoglycemic composite plant ferment, which comprises the following steps:
s301: pulverizing Ginseng radix Rubri, rhizoma Polygonati Odorati, herba Portulacae, fructus Lycii, fructus Citri Grandis, fructus Cucurbitae Moschatae, rhizoma Zingiberis recens, herba Apii Graveolentis, and rhizoma Dioscoreae to obtain solid pulverized material.
S302: adding the solid pulverized material into distilled water, and homogenizing at 12000r for 5min to obtain homogeneous mixed solution. Wherein the mass ratio of the solid crushed material to the distilled water is 3: 10.
S303: sterilizing the homogenized mixture at 115 deg.C for 30 min. And cooling the sterilized homogeneous mixed solution to room temperature.
S304: adding composite lactobacillus powder into the homogeneous mixed solution cooled to room temperature, and fermenting at 35 deg.C for 40h to obtain ferment. Wherein the composite lactobacillus powder comprises bifidobacterium lactis, lactobacillus acidophilus, lactobacillus casei, lactobacillus rhamnosus and lactobacillus plantarum. The adding amount of the compound lactobacillus is 0.2 percent of the mass of the homogeneous mixed liquid.
S305: and (3) carrying out centrifugal filtration on the fermented ferment under the aseptic condition to obtain the blood sugar-reducing composite plant ferment. Wherein the centrifugation speed is 4500r, and the centrifugation time is 3 min.
Example 4
The embodiment of the application provides compound plant ferment of hypoglycemic, and this compound plant ferment of hypoglycemic's preparation raw materials includes: 10g/100g of red ginseng, 15g/100g of polygonatum odoratum, 5g/100g of purslane, 10g/100g of medlar, 5g/100g of pomelo, 15g/100g of pumpkin, 10g/100g of ginger, 15g/100g of celery and 15g/100g of Chinese yam.
The embodiment of the application also provides a preparation method of the hypoglycemic composite plant ferment, which comprises the following steps:
s401: pulverizing Ginseng radix Rubri, rhizoma Polygonati Odorati, herba Portulacae, fructus Lycii, fructus Citri Grandis, fructus Cucurbitae Moschatae, rhizoma Zingiberis recens, herba Apii Graveolentis, and rhizoma Dioscoreae to obtain solid pulverized material.
S402: adding the solid pulverized material into distilled water, and homogenizing at 12000r for 5min to obtain homogeneous mixed solution. Wherein the mass ratio of the solid crushed material to the distilled water is 3: 10.
S403: sterilizing the homogenized mixture at 115 deg.C for 30 min. And cooling the sterilized homogeneous mixed solution to room temperature.
S404: adding composite lactobacillus powder into the homogeneous mixed solution cooled to room temperature, and fermenting at 30 ℃ for 72h to obtain the ferment. Wherein the composite lactobacillus powder comprises bifidobacterium lactis, lactobacillus acidophilus, lactobacillus casei, lactobacillus rhamnosus and lactobacillus plantarum. The adding amount of the compound lactobacillus is 0.3 percent of the mass of the homogeneous mixed liquid.
S405: and (4) centrifuging and filtering the fermented ferment under an aseptic condition to obtain the hypoglycemic composite plant ferment. Wherein the centrifugal speed is 4500r, and the centrifugal time is 3 min.
Example 5
The embodiment of the application provides compound plant ferment of hypoglycemic, and this compound plant ferment of hypoglycemic's preparation raw materials includes: 10g/100g of red ginseng, 15g/100g of polygonatum odoratum, 15g/100g of purslane, 10g/100g of medlar, 10g/100g of pomelo, 5g/100g of pumpkin, 5g/100g of ginger, 15g/100g of celery and 15g/100g of Chinese yam.
The embodiment of the application also provides a preparation method of the hypoglycemic composite plant ferment, which comprises the following steps:
s501: pulverizing Ginseng radix Rubri, rhizoma Polygonati Odorati, herba Portulacae, fructus Lycii, fructus Citri Grandis, fructus Cucurbitae Moschatae, rhizoma Zingiberis recens, herba Apii Graveolentis, and rhizoma Dioscoreae to obtain solid pulverized material.
S502: adding the solid pulverized material into distilled water, and homogenizing at 12000r for 5min to obtain homogeneous mixed solution. Wherein the mass ratio of the solid crushed material to the distilled water is 3: 10.
S503: sterilizing the homogenized mixture at 115 deg.C for 30 min. And cooling the sterilized homogeneous mixed solution to room temperature.
S504: adding composite lactobacillus powder into the homogeneous mixed solution cooled to room temperature, and fermenting at 37 ℃ for 48h to obtain the ferment. Wherein the composite lactobacillus powder comprises bifidobacterium lactis, lactobacillus acidophilus, lactobacillus casei, lactobacillus rhamnosus and lactobacillus plantarum. The addition amount of the compound lactobacillus is 0.1 percent of the mass of the homogeneous mixed liquid.
S505: and (3) carrying out centrifugal filtration on the fermented ferment under the aseptic condition to obtain the blood sugar-reducing composite plant ferment. Wherein the centrifugal speed is 4500r, and the centrifugal time is 3 min.
In order to verify that the hypoglycemic composite plant enzyme provided by the application has an inhibition effect on both alpha-glucosidase and alpha-amylase, the application takes the hypoglycemic composite plant enzyme prepared in example 5 as an example to perform an experimental test, wherein the method for determining the inhibition rate of alpha-amylase is the same as above, and is not repeated herein.
The method for measuring the alpha-glucosidase inhibition rate specifically comprises the following steps: heating 40 μ L of ferment and 40 μ L of alpha-glucosidase at 37 deg.C for 10min, adding 20 μ L of NPG solution (English name: 4-Nitrophenyl-alpha-D-glucopyranoside; Chinese name: 4-nitrobenzene-alpha-D-glucopyranoside), and heating in water bath for 10 min. After heating, 100. mu.L of NaCO was added at a concentration of 0.1moL/L3Solution, then absorbance was measured at 405nm to determine the inhibition rate of α -glucosidase, and PBS was used as a blank instead of enzyme to determine aBlank spaceSample control assay A with PBS instead of alpha-glucosidaseSample controls,. Wherein the inhibition rate of the alpha-glucosidase is calculated by absorbance, and the calculation formula is
Drawing a relational graph according to the alpha-amylase inhibition rate and the alpha-glucosidase inhibition rate to obtain the attached figure 4. As can be seen from fig. 4, the inhibition rate of the hypoglycemic composite plant enzyme provided in the embodiment of the present application to α -amylase is 96.3%, and the inhibition rate to α -glucosidase is 90.4%. Therefore, the hypoglycemic composite plant enzyme provided by the embodiment of the application has obvious inhibition effect on alpha-amylase and alpha-glucosidase. Meanwhile, compared with the products sold in the market, the sugar-reducing composite plant enzyme provided by the embodiment of the application has a far higher inhibition rate on alpha-amylase and alpha-glucosidase than similar enzyme products.
In addition, the examples of the present application also perform the relevant tests on the influence of fasting blood glucose, oral glucose tolerance and oral starch tolerance of rats, and the specific test procedures are as follows:
male Wister rats of age class 6 are bred in an SPF laboratory, are fed with common feed adaptively for one week, and are weighed and numbered randomly. And (3) randomly selecting 10 experimental rats to be included into a normal control group, feeding the rats with normal feed, feeding the rest rats with high-fat diet, and inducing model establishment. After feeding for 4 weeks, streptozotocin STZ with the dose of 30mg/kg is injected into the abdominal cavity, and a type 2 diabetes rat model is established. Normal control rats were injected with the same amount of buffer i.p. And taking blood from the tail tip after 72 hours of injection to measure the fasting blood glucose level, regarding the rat with the final fasting blood glucose value being more than or equal to 16.7mmol/L as successful model building, and randomly dividing the rat into a model group, a traditional Chinese medicine group, a western medicine group, a raw material group and a ferment group. After grouping is finished, different groups are subjected to intervention with the same dosage and different formulations. Specifically, the model group: the concentration is 10mL/kg d -1The normal saline is used for gastric perfusion; the traditional Chinese medicine comprises the following components: the concentration is 10mL/kg d-1The Chinese herbal compound Yitangkang enema is prepared by mixing the raw materials; western medicine group: gavage with acarbose at a concentration of 10 mg/Kg; raw material group: the concentration is 10mL/kg d-1The raw materials of the ferment are perfused into the stomach; enzyme group: the concentration is 10mL/kg d-1The hypoglycemic composite plant enzyme is used for intragastric administration. Fasting blood glucose values of blood taken from the tail tips of rats of each group before intervention, after 2 weeks after intervention, after 4 weeks after intervention, after 6 weeks after intervention and after 8 weeks after intervention are respectively recorded, and fasting is carried out for 12 hours before measurement, so as to obtain the fasting blood glucose values of the rats shown in the table 1.
In addition, at the 6 th week of intervention, after fasting for 16h and gavage for 30min, the rats in each group were gavaged with a sucrose solution at a weight of 2 g/Kg; then, blood was collected from the tail tip at 0, 30, 60, and 120min, and blood glucose was measured by the glucose peroxidase method, thereby obtaining a time-blood glucose value graph shown in fig. 5. At 7 weeks of intervention, fasting each group of rats for 16h, and after intragastric administration for 30min, intragastric administration of starch solution at a weight of 2 g/Kg; then, blood was collected from the tail tip at 0, 30, 60, and 120min, and blood glucose was measured by the glucose peroxidase method, thereby obtaining a time-blood glucose value graph shown in fig. 6.
Table 1: fasting blood glucose/mmol/L of rats in each group
Note: p < 0.05, P < 0.01, compared to normal controls; in comparison with the set of models, #P＜0.05，##P＜0.01
As can be seen from table 1, the blood glucose of the rats in the enzyme group was significantly decreased after 8 weeks of intervention, compared to before the intervention, and the decrease rate was 55.7%. Because the hypoglycemic compound plant enzyme provided by the application is not a therapeutic drug, the hypoglycemic effect is not as good as that of traditional Chinese medicines and western medicines, but the effect is obviously better than that of like enzyme products.
As shown in figure 5, compared with the rats in the normal group, all intervention groups had obvious inhibition after taking sucrose for 30min, and the blood sugar lowering effect of the enzyme group was also superior to that of the raw material group. As shown in the attached figure 6, the intervention effect of the enzyme group on blood sugar can reach the result of the traditional Chinese medicine group.
Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the disclosure herein. This application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims.
It is to be understood that relational terms such as "first" and "second," and the like, may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. The invention is not limited to the precise arrangements described above and shown in the drawings, and various modifications and changes may be made without departing from the scope thereof. The scope of the invention is limited only by the appended claims.
Claims (9)
1. The blood sugar-reducing composite plant enzyme is characterized in that the blood sugar-reducing composite plant enzyme is prepared from the following raw materials: red ginseng, fragrant solomonseal rhizome, purslane, medlar, pomelo, pumpkin, ginger, celery and Chinese yam.
2. The hypoglycemic composite plant ferment of claim 1, wherein the hypoglycemic composite plant ferment is prepared from the following raw materials: 5-10g/100g of red ginseng, 5-15g/100g of polygonatum odoratum, 5-15g/100g of purslane, 5-10g/100g of medlar, 5-15g/100g of grapefruit, 5-15g/100g of pumpkin, 5-10g/100g of ginger, 5-15g/100g of celery and 5-15g/100g of Chinese yam.
3. The preparation method of the hypoglycemic compound plant enzyme is characterized by comprising the following steps:
pulverizing Ginseng radix Rubri, rhizoma Polygonati Odorati, herba Portulacae, fructus Lycii, fructus Citri Grandis, fructus Cucurbitae Moschatae, rhizoma Zingiberis recens, herba Apii Graveolentis, and rhizoma Dioscoreae to obtain solid pulverized material;
adding the solid crushed material into distilled water, and homogenizing for 5min under the condition of 12000r to obtain a homogeneous mixed solution;
sterilizing the homogenized mixed solution and cooling to room temperature;
adding composite lactobacillus powder into the homogenized mixed solution cooled to room temperature, and fermenting at 28-40 deg.C for 0-84h to obtain ferment;
and centrifuging and filtering the fermented ferment under an aseptic condition to obtain the hypoglycemic composite plant ferment.
4. The method for preparing the hypoglycemic composite plant ferment of claim 3, wherein the mass ratio of the solid crushed material to the distilled water is 3: 10.
5. The method for preparing the hypoglycemic composite plant ferment of claim 3, wherein the homogenized mixture is sterilized at 115 ℃ for 30 min.
6. The method for preparing the hypoglycemic composite plant ferment of claim 3, wherein the composite lactobacillus powder comprises bifidobacterium lactis, lactobacillus acidophilus, lactobacillus casei, lactobacillus rhamnosus and lactobacillus plantarum.
7. The preparation method of the hypoglycemic composite plant ferment of claim 3, wherein the addition amount of the composite lactic acid bacteria is 0-0.4% of the mass of the homogeneous mixed solution.
8. The hypoglycemic composite plant ferment of claim 1 or 2 or the hypoglycemic composite plant ferment prepared by the preparation method of any one of claims 3 to 7 has an inhibitory effect on both alpha-glucosidase and alpha-amylase.
9. The hypoglycemic composite plant ferment of claim 1 or 2 or the hypoglycemic composite plant ferment prepared by the preparation method of any one of claims 3 to 7 can reduce the fasting blood glucose value of mice and inhibit the absorption of sucrose and starch by the mice.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103550754A (en) * | 2013-11-25 | 2014-02-05 | 三株福尔制药有限公司 | Probiotic fermented traditional Chinese medicinal compound composition for assisting in reducing blood sugar as well as preparation method and application of composition |
| CN110710677A (en) * | 2019-11-18 | 2020-01-21 | 李桂丽 | Composite enzyme and preparation method thereof |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103550754A (en) * | 2013-11-25 | 2014-02-05 | 三株福尔制药有限公司 | Probiotic fermented traditional Chinese medicinal compound composition for assisting in reducing blood sugar as well as preparation method and application of composition |
| CN110710677A (en) * | 2019-11-18 | 2020-01-21 | 李桂丽 | Composite enzyme and preparation method thereof |
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| Title |
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| 于婷等: "发酵技术改善中药降血糖作用的研究进展", 《延边大学农学学报》, vol. 37, no. 1, pages 80 - 84 * |
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