CN113559150B - Preparation method and application of silkworm chrysalis-mulberry juice composite fermentation preparation - Google Patents

Preparation method and application of silkworm chrysalis-mulberry juice composite fermentation preparation Download PDF

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CN113559150B
CN113559150B CN202110698201.3A CN202110698201A CN113559150B CN 113559150 B CN113559150 B CN 113559150B CN 202110698201 A CN202110698201 A CN 202110698201A CN 113559150 B CN113559150 B CN 113559150B
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silkworm chrysalis
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CN113559150A (en
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吴力克
罗赛
税朝毅
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Guangxi Kangkang Biotechnology Development Group Co ltd
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Abstract

The invention discloses a preparation method and application of a silkworm chrysalis-mulberry juice composite fermentation preparation, wherein the method comprises the following steps: putting fresh silkworm pupa into purified water, decocting in boiling water for a certain time, cooling to normal temperature, removing pupa skin, collecting pupa content, mixing uniformly, adding complex enzyme for enzymolysis for a certain time until the pupa content is decomposed into a small molecule pupa mixture; cleaning fresh folium Mori, mincing in a blender, adding small amount of purified water, decocting for a certain time, filtering, collecting supernatant, and concentrating to obtain folium Mori extract; and mixing the obtained small-molecule silkworm chrysalis mixture with the mulberry leaf juice extract, adding composite probiotics, and performing liquid submerged fermentation to obtain the silkworm chrysalis-mulberry juice composite fermentation preparation. The silkworm chrysalis-mulberry juice composite preparation prepared by the preparation method of the silkworm chrysalis-mulberry juice composite fermentation preparation is extracted from natural products, and can effectively treat diabetes.

Description

Preparation method and application of silkworm chrysalis-mulberry juice composite fermentation preparation
Technical Field
The invention relates to a preparation method and application of a silkworm chrysalis-mulberry juice composite fermentation preparation.
Background
Diabetes mellitus is a group of metabolic diseases characterized by hyperglycemia, and is a syndrome of disturbances of glucose, protein and lipid metabolism caused by insulin deficiency or defects in the cellular metabolism of insulin. In recent years, the number of diabetic patients has been increasing, and these patients are classified into type I diabetes and type II diabetes. Wherein the number of the type II diabetes patients accounts for 85-90% of the total number of the diabetes patients, and the health and the life of the patients are seriously threatened. If diabetes is not well controlled, many complications may arise, such as: diabetic nephropathy, retinopathy, hypertension, etc.
At present, most patients are treated with diabetes by injecting insulin or orally taking hypoglycemic drugs to help control blood sugar in vivo, or regulating insulin secretion and the like. However, these drugs have large side effects, cannot be clinically used for a long time, and cannot meet the clinical requirements of diabetics. If the medicine which can be extracted from natural products and is safe and effective for treating diabetes can be extracted, good news can be brought to diabetics.
The silkworm pupa is a high-nutrition delicious food and has medicinal value. The silkworm chrysalis is a natural resource which is rich in economic value and development value, has wide pharmacological action, has the effects of protecting the liver, promoting growth, improving immunity, reducing blood pressure and blood sugar, resisting cancer, treating cancer and the like, is a good medicinal material, has the oil content of 20-30% in dry silkworm chrysalis, contains 75% of unsaturated fatty acid and mainly consists of oleic acid, linoleic acid and linolenic acid. Wherein linoleic acid has protective effect on nerve cells. The silkworm pupa protein is a high-quality pure natural complete protein, the content of crude protein of dry silkworm pupas is up to 45-50%, the silkworm pupa protein contains 18 amino acids required by human bodies through determination, and the content of 8 essential amino acids contained in the silkworm pupas is about 2 times of that of pork and 4 times of that of milk, and the silkworm pupa protein is a high-quality protein source with proper proportion. The silkworm pupa also contains various vitamins (vitamin A, vitamin B, vitamin D, etc.), trace elements (iron, zinc, selenium, etc.) and a small amount of crude fiber, chitin, antibacterial skin, hormone, lysozyme, polysaccharide, etc., and has the effects of regulating the metabolic balance of the human body and health care.
According to the research, the dried mulberry leaves contain 25 to 45 percent of crude protein, 20 to 25 percent of carbohydrate, 5 percent of crude fat, abundant potassium, calcium, vitamin C and the like, and also contain various trace elements such as copper, zinc, boron, manganese and the like. The mulberry leaves contain ecdysterone, lupulone, various amino acids such as target glycoside and mulberrin, vitamins, taenigine acid, folic acid, fumaric acid, meso-creatine, phytoestrogen and other substances required by human bodies, and have the effects of resisting stress, resisting aging, enhancing the body endurance, regulating the epinephrine function and the like.
Since ancient times, the traditional Chinese medicine applies the mulberry leaves as the traditional Chinese medicine for treating diabetes (diabetes in modern medicine) to clinic, and Japanese ancient books (tea eating health preserving records) also record the function of the mulberry leaves for improving the 'drinking water disease' (diabetes in modern medicine). The research data at home and abroad prove that the alkaloid and the polysaccharide are main blood sugar reducing components in the mulberry leaves. The hypoglycemic effect of mulberry leaves is realized through two ways: firstly, alkaloid DNJ (1-deoxynojirimycin) has an inhibitory effect on the activity of disaccharide degrading enzymes, so that the absorption of disaccharide by small intestine is inhibited, and the high peak value of postprandial blood sugar is reduced (Kimura, 1995); secondly, the beta cell insulin is promoted by mulberry leaf alkaloid fagomine and mulberry leaf polysaccharide, and the insulin can promote the utilization of cells to sugar, the synthesis of hepatic glycogen and the improvement of glycometabolism, so that the effect of reducing blood sugar is achieved with the strongest effect of reducing blood sugar.
Disclosure of Invention
The invention provides a preparation method and application of a silkworm chrysalis-mulberry juice composite fermentation preparation, which adopts the following technical scheme:
a preparation method of a silkworm chrysalis-mulberry juice composite fermentation preparation comprises the following steps:
silkworm chrysalis treatment: putting fresh silkworm pupa into purified water, decocting in boiling water for a certain time, cooling to normal temperature, removing pupa skin, collecting pupa content, mixing uniformly, adding complex enzyme for enzymolysis for a certain time until the pupa content is decomposed into a small molecule pupa mixture;
and (3) mulberry leaf treatment: cleaning fresh folium Mori, mincing in a blender, adding small amount of purified water, decocting for a certain time, filtering, collecting supernatant, and concentrating to obtain folium Mori extract;
mixing: and mixing the obtained small-molecule silkworm chrysalis mixture with the mulberry leaf juice extract, adding composite probiotics, and performing liquid submerged fermentation to obtain the silkworm chrysalis-mulberry juice composite fermentation preparation.
Further, in the silkworm pupa processing step, the selected fresh silkworm pupas are healthy silkworm pupas which are cocooned for 3-9 days.
Further, in the silkworm pupa treatment step, the fresh silkworm pupae are decocted in boiling water at 100 ℃ for 0.5-1.5 h.
Further, in the silkworm pupa treatment step, the appropriate mixing temperature of the silkworm pupa content is 20-30 ℃.
Further, the complex enzyme is protease, lipase and chitinase according to the ratio of 3: 2: 1, the complex is mixed, and the adding proportion of the complex enzyme in the silkworm chrysalis content is 2.5-5%.
Further, in the mulberry leaf treatment step, the crushed mulberry leaves are stirred to have the granularity of 2-8 meshes, purified water is added according to the proportion of 6-8 times of the weight of the mulberry leaves, and the mixture is stirred uniformly and decocted in boiling water for 4-6 hours.
Further, in the mulberry leaf treatment step, filtration was performed, and the supernatant was collected and concentrated to 1/3 of the total volume of the supernatant.
Further, in the mixing step, the obtained small molecule silkworm chrysalis mixture and the mulberry leaf concentrated solution are mixed according to the ratio of 1: 1, and uniformly mixing by a homogenizer under the condition that the rotation speed of the homogenizer is 4000-.
Further, in the mixing step, the silkworm chrysalis-mulberry leaf mixture is placed in a fermentation tank, activated bifidobacterium and lactobacillus productive bacteria liquid are inoculated according to the proportion of 2 percent of the total fermentation liquid, and mechanical stirring type liquid submerged fermentation is carried out, wherein the fermentation conditions are as follows: the initial pH value is 5.5-6.5, the stirring speed is 80-100r/min, the fermentation temperature is 30.0-35.0 ℃, and the fermentation termination condition is that the number of the probiotic live bacteria in the fermentation liquor reaches 2.0 multiplied by 10 9 The content of the/L, 1-deoxynojirimycin is more than or equal to 4000U/L.
The application of the silkworm chrysalis-mulberry juice compound fermentation preparation prepared by the preparation method of the silkworm chrysalis-mulberry juice compound fermentation preparation in treating diabetes is provided.
The silkworm chrysalis-mulberry juice composite fermentation preparation prepared by the preparation method has the advantages that the silkworm chrysalis-mulberry juice composite fermentation preparation is extracted from natural products and fermented, and diabetes can be effectively treated.
Detailed Description
The present invention will be described in detail with reference to the following embodiments.
The invention discloses a preparation method of a silkworm chrysalis-mulberry juice composite fermentation preparation, which mainly comprises the following steps:
silkworm chrysalis treatment: putting fresh silkworm pupa into purified water, decocting in boiling water for a certain time, cooling to normal temperature, removing pupa skin, collecting pupa content, mixing uniformly, adding complex enzyme for enzymolysis for a certain time until the pupa content is decomposed into a small molecule pupa mixture.
In a preferable embodiment, in the silkworm pupa treatment step, the selected fresh silkworm pupas are healthy silkworm pupas with cocoons for 3-9 days. Decocting fresh pupa Bombycis in boiling water at 100 deg.C for 0.5-1.5 h. The appropriate mixing temperature of the silkworm chrysalis content is 20-30 ℃. The compound enzyme is protease, lipase and chitinase according to the ratio of 3: 2: 1 (mass ratio), and the adding proportion of the complex enzyme in the silkworm pupa content is 2.5-5% (mass ratio).
And (3) mulberry leaf treatment: cleaning fresh folium Mori, mincing in a blender, adding small amount of purified water, decocting for a certain time, filtering, collecting supernatant, and concentrating to obtain folium Mori extract.
In the preferable embodiment, in the mulberry leaf treatment step, the crushed mulberry leaves are stirred to have the granularity of 2-8 meshes, purified water is added in the granularity of 6-8 times of the weight of the mulberry leaves, and the mixture is stirred uniformly and then decocted in boiling water for 4-6 hours. Filtering, collecting supernatant, and concentrating to 1/4-1/2 of the total volume of the supernatant.
Mixing: and mixing the obtained small-molecule silkworm chrysalis mixture with the mulberry leaf juice extract, adding composite probiotics, and performing liquid submerged fermentation to obtain the silkworm chrysalis-mulberry juice composite fermentation preparation.
In the mixing step, mixing the obtained small molecule silkworm chrysalis mixture with the mulberry leaf concentrated solution according to the ratio of 1: 1 (mass ratio), and mixing uniformly by a homogenizer under the condition that the rotation speed of the homogenizer is 4000-6000r/min, and continuously mixing for 20 min.
In a preferred embodiment, in the mixing step, the silkworm pupa-mulberry leaf mixture is placed in a fermentation tank, activated bifidobacterium and lactobacillus productive bacteria liquid are inoculated according to the proportion of 2% of the total fermentation liquid, and mechanical stirring type liquid submerged fermentation is carried out, wherein the fermentation conditions are as follows: initial pH5.5-6.5, stirring speed of 80-100r/min, fermentation temperature of 30.0-35.0 deg.c, and final fermentation condition of 2.0 × 10 viable probiotic bacteria in the fermented liquid 9 The content of the/L, 1-deoxynojirimycin is more than or equal to 4000U/L.
One embodiment is described below:
putting healthy silkworm pupas which are cocooned for 3 days into purified water, decocting in boiling water at 100 ℃ for 1h, cooling to normal temperature, removing pupa skin, collecting silkworm pupa content, uniformly mixing at 25 ℃, adding protease, lipase and chitinase according to the ratio of 3: 2: 1 intoAnd carrying out enzymolysis on the mixed complex enzyme for 2 hours until the complex enzyme is decomposed into a small-molecule silkworm chrysalis mixture, wherein the adding proportion of the complex enzyme in the silkworm chrysalis content is 3%. Cleaning fresh folium Mori, mincing in a blender to 6 mesh, adding purified water 7 times of folium Mori, stirring, boiling for 5 hr, filtering, collecting supernatant, and concentrating to 1/3 of total volume of supernatant to obtain the final product. Mixing the obtained small molecule silkworm pupa mixture and the mulberry leaf juice extract according to the ratio of 1: 1, uniformly mixing by a refiner under the condition that the rotation speed of the refiner is 4000 plus 6000r/min, continuously mixing for 20min, putting the mixture into a fermentation tank, inoculating activated bifidobacterium and lactobacillus productive bacteria liquid according to the proportion of 2 percent of the total fermentation liquid, and performing mechanical stirring type liquid submerged fermentation under the fermentation condition that: the initial pH value of 6, the stirring speed of 80r/min, the fermentation temperature of 30.0 ℃, and the fermentation termination condition that the viable count of the probiotics in the fermentation liquor reaches 2.0 multiplied by 10 9 The content of the/L, 1-deoxynojirimycin is more than or equal to 4000U/L, and the silkworm chrysalis-mulberry juice composite fermentation preparation is obtained after the fermentation is finished.
In the invention, in order to improve the enzymolysis effect, in the preparation process of the above specific embodiment, on the basis of keeping other conditions unchanged, the ratio of the protease, the lipase and the chitinase is changed, and the enzymolysis effect of the protease, the lipase and the chitinase at different ratios is observed through multiple experiments. In the invention, the final core components after enzymolysis are flavone and ghost mycin, and the enzymolysis effect is measured by measuring the contents of the two substances, and the specific conditions are shown in the following table. As shown by the data in the table, the ratio of protease, lipase and chitinase to protease is 3: 2: 1, the enzymolysis effect is best.
TABLE 1 fruit test chart for enzymolysis effect of protease, lipase and chitinase in different proportions
Ratio of protease to Lipase Flavone content (ug/ml) Content of ghost mycin (ug/ml)
3:2：1 6097 5031
3.25:2:1 5725 4624
3.5:2:1 5321 4214
2.75:2:1 5625 4578
2.5:2:1 5231 4225
3:2:1.1 5135 4124
3:2:1.2 4754 3724
3:2:0.9 5115 4100
3:2:0.8 4687 3712
3：2.15:1 5521 4431
3:2.3:1 5145 4025
3:1.85:1 5510 4421
3:1.7:1 5089 4112
The silkworm chrysalis-mulberry juice compound fermentation preparation prepared by the preparation method of the silkworm chrysalis-mulberry juice compound fermentation preparation can be used for treating diabetes.
As shown in the following table, mice after being adaptively fed for one week are fasted for 24h, the mice are injected with physiological saline solution of alloxan (50mg/kg) in tail vein, after 72h, the mice are boosted once by the same dosage, after one week, blood is taken from tail vein after being fasted for 4-5h to measure blood sugar value, and the blood sugar concentration is more than or equal to 11.1mmol/L, so that the mice are diabetic until 40 mice are obtained. 40 diabetic mice were randomly divided into 4 groups of 10 mice each. Wherein 3 groups of the silkworm chrysalis-mulberry juice composite fermentation preparations are respectively given. The group 1 was administered intragastrically according to the administration standard of 0.035g/kg, based on body weight. Group 2 was dosed at 0.040g/kg/d, and group 3 was dosed at 0.045 g/kg. Group 4 served as a blank control group and was continuously gavaged with an equal volume of saline for 10 d. After 20d, the weight and blood sugar of the mice were recorded. The specific results are shown in the table below, and the silkworm chrysalis-mulberry juice composite fermentation preparation prepared by the invention has the effect of assisting in reducing blood sugar and improving the condition of a diabetic patient.
TABLE 2 experiment data table for diabetes treatment with silkworm pupa-mulberry juice composite fermentation preparation
Group of Initial blood glucose mean (mmol/L) Final blood glucose mean (mmol/L)
Group 1 12.2 5.9
Group 2 12.3 5.7
Group 3 12.4 5.4
Blank control group 12.3 12.1
The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It should be understood by those skilled in the art that the above embodiments do not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent variations fall within the scope of the present invention.

Claims (7)

1. The preparation method of the silkworm chrysalis-mulberry juice composite fermentation preparation is characterized by comprising the following steps:
silkworm chrysalis treatment: putting fresh silkworm pupa into purified water, decocting in boiling water for a certain time, cooling to normal temperature, removing pupa skin, collecting pupa content, mixing uniformly, adding complex enzyme for enzymolysis for a certain time until the pupa content is decomposed into a small molecule pupa mixture;
and (3) mulberry leaf treatment: cleaning fresh folium Mori, mincing in a blender, adding small amount of purified water, decocting for a certain time, filtering, collecting supernatant, and concentrating to obtain folium Mori extract;
mixing: mixing the obtained small-molecule silkworm chrysalis mixture with the mulberry leaf concentrated solution according to the weight ratio of 1: 1, uniformly mixing by a homogenizer under the condition that the rotation speed of the homogenizer is 4000-;
the compound enzyme is protease, lipase and chitinase according to the ratio of 3: 2: 1, mixing the complex, wherein the adding proportion of the complex enzyme in the silkworm pupa content is 2.5-5%;
in the mixing step, the silkworm chrysalis-mulberry leaf mixture is placed in a fermentation tank, activated bifidobacterium and lactobacillus producing bacterial liquid are inoculated according to the proportion of 2 percent of the total fermentation liquid, and mechanical stirring type liquid submerged fermentation is carried out, wherein the fermentation conditions are as follows: the initial pH value is 5.5-6.5, the stirring speed is 80-100r/min, the fermentation temperature is 30.0-35.0 ℃, and the fermentation termination condition is that the number of the probiotic live bacteria in the fermentation liquor reaches 2.0 multiplied by 10 9 The content of the/L, 1-deoxynojirimycin is more than or equal to 4000U/L.
2. The method for preparing a silkworm chrysalis-mulberry juice composite fermentation preparation according to claim 1, wherein the silkworm chrysalis-mulberry juice composite fermentation preparation,
in the silkworm pupa treatment step, the selected fresh silkworm pupas are healthy silkworm pupas with cocoons for 3-9 days.
3. The method for preparing a silkworm chrysalis-mulberry juice composite fermentation preparation according to claim 1, wherein the silkworm chrysalis-mulberry juice composite fermentation preparation,
in the silkworm pupa treatment step, the fresh silkworm pupas are decocted in boiling water at 100 ℃ for 0.5-1.5 h.
4. The method for preparing a silkworm chrysalis-mulberry juice composite fermentation preparation according to claim 1, wherein the silkworm chrysalis-mulberry juice composite fermentation preparation,
in the silkworm pupa treatment step, the appropriate mixing temperature of the silkworm pupa content is 20-30 ℃.
5. The method for preparing the silkworm chrysalis-mulberry juice composite fermentation preparation according to claim 1, wherein,
in the mulberry leaf treatment step, the crushed mulberry leaves are stirred to have the granularity of 2-8 meshes, purified water is added according to the proportion of 6-8 times of the weight of the mulberry leaves, and the mixture is stirred uniformly and decocted in boiling water for 4-6 hours.
6. The method for preparing a silkworm chrysalis-mulberry juice composite fermentation preparation according to claim 1, wherein the silkworm chrysalis-mulberry juice composite fermentation preparation,
in the mulberry leaf treatment step, filtration is carried out, and the supernatant is collected and concentrated to 1/3 of the total volume of the supernatant.
7. The use of the silkworm chrysalis-mulberry juice composite fermentation preparation prepared by the method for preparing the silkworm chrysalis-mulberry juice composite fermentation preparation according to any one of claims 1 to 6 in preparing a medicament for treating diabetes.
CN202110698201.3A 2021-06-23 2021-06-23 Preparation method and application of silkworm chrysalis-mulberry juice composite fermentation preparation Active CN113559150B (en)

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CN105661547A (en) * 2016-01-22 2016-06-15 湖南营养树科技有限公司 Probiotics and Bombyx mori compounded diabetic patient nutritional supplement
CN107670002A (en) * 2017-10-12 2018-02-09 广州普罗艾森健康科技有限公司 A kind of hypoglycemic composition and its preparation method and application
CN110862897A (en) * 2019-10-29 2020-03-06 陕西理工大学 Mulberry leaf and silkworm chrysalis yellow wine and its making process

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105661547A (en) * 2016-01-22 2016-06-15 湖南营养树科技有限公司 Probiotics and Bombyx mori compounded diabetic patient nutritional supplement
CN107670002A (en) * 2017-10-12 2018-02-09 广州普罗艾森健康科技有限公司 A kind of hypoglycemic composition and its preparation method and application
CN110862897A (en) * 2019-10-29 2020-03-06 陕西理工大学 Mulberry leaf and silkworm chrysalis yellow wine and its making process

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