CN115067517B - Application of passion fruit and cordyceps sinensis complex in preparation of tablets and preparation method thereof - Google Patents
Application of passion fruit and cordyceps sinensis complex in preparation of tablets and preparation method thereof Download PDFInfo
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- CN115067517B CN115067517B CN202210362606.4A CN202210362606A CN115067517B CN 115067517 B CN115067517 B CN 115067517B CN 202210362606 A CN202210362606 A CN 202210362606A CN 115067517 B CN115067517 B CN 115067517B
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/20—Agglomerating; Granulating; Tabletting
- A23P10/28—Tabletting; Making food bars by compression of a dry powdered mixture
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention relates to the field of biotechnology, in particular to application of passion fruit and cordyceps sinensis complexes in preparing tablets and a preparation method thereof; the acid-resistant growth capacity of the cordyceps militaris strain can be remarkably improved through domestication of the lactic acid bacteria and the passion fruit juice, the acid environment of the passion fruit juice can be better adapted, the cordyceps militaris comprehensive body prepared by using the domesticated strain can remarkably improve the absorption of cordyceps militaris to the effective substances of the substrate, and the health-care performance of the comprehensive body is improved; the Chinese medicinal composition with the blood sugar reducing effect is added into the passion fruit shell culture medium, so that the passion fruit and cordyceps militaris complex has a good blood sugar reducing function; when the tablet is prepared, the auxiliary materials are used for improving the passion fruit and cordyceps militaris complex, so that the taste and the blood sugar reducing capability of the tablet can be improved, and more food forms are provided for the effective utilization of the passion fruit and cordyceps militaris complex.
Description
[ field of technology ]
The invention relates to the technical field of biology, in particular to application of passion fruit and cordyceps sinensis complexes in preparation of tablets and a preparation method thereof.
[ background Art ]
The passion fruit and cordyceps militaris complex is a food newly appeared in the culture process of cordyceps militaris strains on the market at present, and the processing method is to culture mycelia of cordyceps militaris by using passion fruits as a culture container, and related products are circulated on the market at present, for example, the passion fruit and cordyceps militaris complex disclosed in patent 202010902421.9 is mainly prepared by combining various food materials and passion fruit juice into a whole, and inoculating cordyceps militaris strains into the passion fruits, so that the whole passion fruit grows up to form cordyceps militaris mycelia, and cordyceps militaris complex is obtained. At present, the market has wide sales volume, and the subject group finds that in the research on passion fruit and cordyceps militaris complex: in the process of preparing the passion fruit complex, the passion fruit itself has higher acidity due to the need of pulp and fruit juice during cultivation, and is not suitable for growth of the cordyceps militaris plant, and finally the growth effect of the cordyceps militaris plant is not good. Therefore, it is necessary to domesticate the Cordyceps militaris strain to obtain Cordyceps militaris strain more suitable for growth in passion fruit, and develop health effects, and obtain more special health foods.
[ invention ]
In view of the above, in order to effectively domesticate the cordyceps militaris strain, a cordyceps militaris strain more suitable for growing in passion fruits is obtained, and the health care effect of the cordyceps militaris strain is developed, so that more special health care foods are obtained, and the research of the application is carried out.
To achieve the above object, the present application adopts the following method:
the passion fruit and cordyceps militaris complex is prepared by domesticating a cordyceps militaris strain by a domestication culture medium and inoculating the domesticated cordyceps militaris strain into a passion fruit shell culture medium for culture; the domestication culture medium contains lactobacillus; the passion fruit shell culture medium contains traditional Chinese medicine components.
Further, the lactobacillus described herein is strain Lactobacillus plantarum LDVS005, lactobacillus plantarum LDVS007 and/or Lactobacillus plantarum LDVS008; the strain Lactobacillus plantarum LDVS005 has a deposit number of: CGMCC NO:20027,Lactobacillus plantarum LDVS007 the deposit number is: CGMCC NO:20028,Lactobacillus plantarum LDVS008 the deposit number is: CGMCC NO: 20029. all the strains are preserved in the China general microbiological culture Collection center, address: the collection date is 6/8/2020 of the institute of microbiology, national academy of sciences, national institute of sciences, 1/3/north Chen West Lu, chaoyang, beijing.
In the application, the best choice is that the lactobacillus is prepared by mixing strains LDVS005, LDVS007 and LDVS008 according to the mass ratio of 1:1:1.
Furthermore, the domestication culture medium also contains passion fruit juice.
Further, the traditional Chinese medicine components in the passion fruit shell culture medium consist of the following components in parts by weight: 10 parts of ginseng, 10 parts of okra, 3 parts of astragalus, 10 parts of dwarf lilyturf tuber and 2 parts of medlar.
In addition, the passion fruit and cordyceps militaris complex can be used for preparing blood sugar reducing tablets.
The invention also comprises an application and a method for preparing tablets from the passion fruit and cordyceps militaris integrated preparation, wherein the method comprises the following steps: pulverizing the passion fruit and Cordyceps militaris complex, adding water to prepare paste, adding auxiliary materials according to 5% of the mass of the paste of the passion fruit and Cordyceps militaris complex, and tabletting to prepare passion fruit tablets.
Further, the auxiliary materials are prepared by mixing xylitol, magnesium stearate, aloe colloid and passion fruit juice according to the mass ratio of 1:1:1:1.
The specific formula and the domestication method of the domestication culture medium of the cordyceps militaris strain are as follows:
tissue separation is carried out under the aseptic condition, the tissue with the size of mung bean grains is adopted on the fruiting body of the cordyceps militaris, inoculated on a 1-time domestication culture medium, placed in a constant temperature incubator for culture at 25 ℃, and the growth condition of hyphae is observed, and if white hyphae are fully distributed on a flat plate, transfer can be carried out; transferring corresponding mycelium onto 2 times of domestication culture medium, placing into a constant temperature incubator for culturing at 25deg.C, observing mycelium growth condition, and transferring if white mycelium is fully distributed on the plate; transferring corresponding mycelium to a 3-time domestication culture medium, placing the culture medium into a constant temperature incubator for culturing at 25 ℃, observing the growth condition of the mycelium, and finishing the domestication process if the white mycelium is fully distributed on a flat plate;
the preparation method of the composite strain of the domestication culture medium comprises the following steps: mixing an MRS liquid culture medium and filtered passion fruit juice according to a mass ratio of 1:1, respectively inoculating lactobacillus LDVS005, LDVS008 or LDVS007 into the mixed culture medium in an inoculum size of 0.5%, and carrying out anaerobic fermentation for 1h at 37 ℃ to obtain an MRS-passion fruit juice liquid mixture containing lactobacillus; mixing an MRS-passion fruit juice liquid mixture containing LDVS005, an MRS-passion fruit juice liquid mixture containing LDVS008 and an MRS-passion fruit juice liquid mixture containing LDVS007 according to a volume ratio of 1:1:1 to obtain a composite strain;
the preparation method of the 1-time domestication culture medium comprises the following steps: mixing a composite strain (prepared by the upper-stage preparation method) and a PDA culture medium according to a mass ratio of 1:5;
the preparation method of the 2-time domestication culture medium comprises the following steps: mixing a composite strain (prepared by the upper-stage preparation method) and a PDA culture medium according to a mass ratio of 2:5;
the preparation method of the 3-time domestication culture medium comprises the following steps: mixing the composite strain (prepared by the upper preparation method) and PDA culture medium according to the mass ratio of 3:5.
The method for preparing the cordyceps militaris complex by using the culture medium comprises the following steps:
(1) the traditional Chinese medicine comprises the following components: oven drying 10 parts of ginseng, 10 parts of okra, 3 parts of astragalus membranaceus, 10 parts of radix ophiopogonis and 2 parts of medlar, pulverizing, and mixing for later use;
(2) preparing a rice culture medium: weighing 50g of rice, putting into a 500mL can, adding nutrient solution into the can according to the requirement of 1.3mL of nutrient solution for each gram of rice, and soaking for 2-3h; wherein, the nutrient solution consists of the following components: according to the calculation of each nutrient substance required by 1000g of rice, 15g of peptone, 5g of glucose, 5g of yeast extract and KH 2 PO 4 1.5g、MgSO 4 0.75g, water was added to make up to 1L; sealing, sterilizing under high pressure, taking out, and cooling to room temperature.
(3) Collecting passion fruit: collecting fresh passion fruit with maturity of 8-9 as raw material, breaking passion fruit from the middle, digging out pulp and pulp juice in the interior to obtain pulp, and taking passion fruit shell with pulp removed as culture box;
(4) preparing a mixed culture medium: mixing the crushed traditional Chinese medicine components in the step (1) and the rice culture medium in the step (2) according to the mass ratio of 1:2, then adding the pulp in the step (3) to adjust the water content to be about 55-60%, subpackaging the pulp into a culture box made of passion fruit shells, and sterilizing the pulp according to a conventional method to obtain the passion fruit shell culture medium.
Inoculating and culturing: picking up domesticated Cordyceps militaris mycelia under aseptic condition, inoculating into passion fruit shell culture medium, culturing at 16-20deg.C with humidity of 70-75% and darkness for 3-5 days, adjusting room temperature of culture room to 20-25deg.C with humidity of 80-90%, allowing mycelia to see light, and culturing until Cordyceps militaris mycelia completely eat through the culture medium, and completing the culturing process of Cordyceps militaris mycelia after the mycelia surface is completely colored; taking out the cordyceps militaris mycelium at the moment to obtain the passion fruit cordyceps militaris complex.
The invention has the following beneficial effects:
1. according to the preparation method, the acid-resistant growth capacity of the cordyceps militaris strain can be remarkably improved through the domestication of the lactic acid bacteria and the passion fruit juice, during domestication, the lactic acid bacteria are added into the domestication culture medium in a subject group innovation mode, so that the lactic acid is slowly generated by the culture medium, the acid-resistant capacity of the cordyceps militaris strain is gradually improved, the acid environment of the passion fruit juice can be better adapted, the absorption of the cordyceps militaris to the substrate effective substances can be remarkably improved through the passion fruit cordyceps militaris comprehensive body prepared by using the domestication strain, and the health-care performance of the comprehensive body is improved.
2. The Chinese medicinal composition with the blood sugar reducing effect is added into the passion fruit shell culture medium, so that the passion fruit and cordyceps militaris complex has a good blood sugar reducing function.
When the tablet is prepared, the auxiliary materials are used for improving the passion fruit and cordyceps militaris complex, so that the taste and the blood sugar reducing capability of the tablet can be improved, and more food forms are provided for the effective utilization of the passion fruit and cordyceps militaris complex.
[ description of the drawings ]
FIG. 1 is a schematic diagram of the passion fruit and Cordyceps militaris complex culture of the invention.
[ detailed description ] of the invention
The invention will be further described with reference to the accompanying drawings and examples and experiments.
Example 1:
the embodiment is a domestication method for cordyceps militaris strains, which specifically comprises the following steps:
cordyceps militaris strain: is provided by Guangxi Zhuangren Tang Biotechnology Co.
1. Domestication research of cordyceps militaris:
acclimatization is carried out by adopting several different culture mediums:
CK: PDA medium: peeling 200g of potato, removing bud eyes, cutting, boiling for 10min, filtering with gauze, filtering with sucrose 20g, peptone 5g and agar 20g, adding water to 1000mL, and sterilizing under high pressure.
Culture medium (1): passion fruit juice medium: taking out pulp of passion fruit, squeezing juice, and filtering to obtain passion fruit juice; the passion fruit juice and PDA culture medium (CK) are then mixed in a mass ratio of 1:5.
Culture medium (2): lactic acid bacteria culture medium: lactic acid bacteria were inoculated into MRS liquid medium at an inoculum size of 0.5%, anaerobic fermentation was performed at 37℃for 1 hour, and then MRS liquid medium containing lactic acid bacteria strain and PDA medium (CK) were mixed at a mass ratio of 1:5.
Culture medium (3): lactic acid bacteria passion fruit juice medium: mixing MRS liquid culture medium and filtered passion fruit juice according to a mass ratio of 1:1, inoculating lactobacillus into the mixed culture medium with an inoculum size of 0.5%, anaerobic fermenting at 37 ℃ for 1h, and mixing passion fruit-MRS composite culture medium containing lactobacillus strain and PDA culture medium (CK) according to a mass ratio of 1:5.
The domestication method comprises the following steps: tissue separation is carried out under the aseptic condition, the tissue with the size of mung bean grains is adopted on the fruiting body of the cordyceps militaris, inoculated on the flat plates of the culture mediums (1) to (3) and the CK culture medium, placed in a constant temperature incubator for culture at 25 ℃, and observed for mycelium growth, and if white mycelium is fully distributed on the flat plates, transfer can be carried out; transferring corresponding mycelia to corresponding culture medium, transferring 2 times, namely performing 3 times of domestication; after the 3 rd domestication culture, the mycelium is inoculated into PDA culture medium containing 20% passion fruit juice by mass percent, and the culture is carried out under the conventional condition, the colony diameter is marked once a day, and the culture is carried out for 7d, so that the mycelium growth rate is calculated.
Wherein, the mass ratio of passion fruit juice of the culture medium (1) to the PDA culture medium (CK) is 2:5, the mass ratio of MRS liquid culture medium containing lactobacillus strains of the culture medium (2) to the PDA culture medium (CK) is 2:5, and the mass ratio of passion fruit-MRS composite culture medium containing lactobacillus strains of the culture medium (3) to the PDA culture medium (CK) is 2:5;
the passion fruit pulp and PDA culture medium (CK) of the culture medium (1) are mixed in a mass ratio of 3:5, the MRS liquid culture medium and PDA culture medium (CK) of the lactobacillus strains of the culture medium (2) are mixed in a mass ratio of 3:5, and the passion fruit-MRS compound culture medium and PDA culture medium (CK) of the lactobacillus strains of the culture medium (3) are mixed in a mass ratio of 3:5 at the time of transfer 2.
The results obtained from the tests are shown in the following table:
TABLE 1 influence of different culture media on growth of Cordyceps militaris strains
| Culture medium | Mycelium growth rate (mm/d) |
| Culture medium (1) | 3.97 |
| Culture medium (2) | 4.41 |
| Culture medium (3) | 5.01 |
| CK (PDA culture medium) | 4.33 |
As shown in the table, the culture medium (1)) added with passion fruit juice in the PDA culture medium has an inhibition effect on the growth of the cordyceps militaris, which is consistent with the conclusion of the prior research, and after the passion fruit juice is added in the culture medium, an environment with higher acidity can be formed, and the growth rate of the cordyceps militaris can be influenced;
the growth rate of mycelium of the culture medium (2)) of the lactobacillus strain is slightly higher than that of the CK group, so that the lactobacillus can form an acidic environment, but the acid production of the lactobacillus is a longer and progressive process, and after the lactobacillus reaches a fermentation peak, the cordyceps militaris strain is gradually adapted to the acidic environment, so that the acid tolerance of the cordyceps militaris strain is improved, and the purpose of acclimatizing and adapting to the acidic environment is achieved; the higher rate of mycelium growth in medium (2) than in medium (1) also illustrates: compared with the method that the cordyceps militaris strain is directly put into an acid solution (high acid environment) containing passion fruit juice, in the culture medium (2) (containing lactobacillus), the adaptability of the cordyceps militaris strain is better, and the domestication effect is better;
the addition of passion fruit juice and lactobacillus strain medium (3)) simultaneously in PDA medium, the mycelium growth rate of cordyceps militaris strain is optimal, and is significantly improved compared with CK group, thus indicating that this is the most suitable medium for domesticating cordyceps militaris strain, which we infer is because: the addition ratio of the passion fruit juice to the lactobacillus strain culture medium is 1:1, the acidity is not as high as that of the culture medium (1), but higher than that of the culture medium (2), under the condition, the growth of the cordyceps militaris strain is not affected, and meanwhile, the passion fruit juice culture medium can be slowly adapted to the growth environment with gradually increased acidity under the slow fermentation of the lactobacillus to generate stronger acid resistance, so that the passion fruit juice culture medium with 20 mass percent can well grow in the end, and preparation is provided for preparing passion fruit complexes in the next step.
(II) influence of different lactobacillus strains on growth of Cordyceps militaris:
strains selected by applicant per se were used: LDVS005 (preservation number: CGMCC NO: 20027), LDVS007 (preservation number: CGMCC NO: 20028), LDVS008 (preservation number: CGMCC NO: 20029), LDVS012 (preservation number: CGMCC NO: 20030) (the strains are preserved in China general microbiological culture center, address: national institute of sciences of China, no. 3, national academy of sciences of North Chen-West road 1, korea, beijing, and preservation date is 2020, 6 months and 8 days), respectively preparing corresponding lactobacillus strain culture mediums according to the formula of the point (one) according to the point (3), and then acclimating the same cordyceps militaris strain according to the method of the step (one), wherein the adopted culture mediums are culture mediums (3), but the lactobacillus strains are respectively LDVS005, LDVS007, LDVS008 and LDVS012; after acclimatization according to the above-mentioned acclimatization method, after the end of the 3 rd acclimatization culture, mycelia were inoculated into a PDA containing 20 mass% passion fruit juice, cultured under conventional conditions, colony diameters were marked once per day, cultured for 7d, and the hypha growth rate was calculated, with the following results:
TABLE 2
| Culture medium | Mycelium growth rate (mm/d) |
| LDVS005 | 5.01 |
| LDVS007 | 5.12 |
| LDVS008 | 5.07 |
| LDVS012 | 4.34 |
| CK (PDA culture medium) | 4.13 |
As can be seen from the table, the length and growth rate of mycelia of the domesticated Cordyceps militaris strain of LDVS012 are not very different from those of CK group of table 1, thus demonstrating that not all lactic acid bacteria strains can improve properties of Cordyceps militaris strain itself, which is likely to be different from lactic acid bacteria strains, and that fermentation products of different lactic acid bacteria strains may have an influence on growth of mycelia, so that Cordyceps militaris strain is more suitable for growth in passion fruit juice, and specific mechanism needs further study, while other lactic acid bacteria strains: the growth promotion effect of the LDVS005, the LDVS008 and the LDVS007 on the cordyceps militaris strain is particularly remarkable, and the fact that most of lactobacillus strains can be added into the passion fruit juice is indicated, so that the cordyceps militaris strain is promoted to adapt to the high acidity environment of the passion fruit juice, and a good domestication effect is achieved.
(III) combining the study of the first point and the second point in the embodiment, selecting an acclimation culture medium for preparing the cordyceps militaris strain by adopting the combination of LDVS005, LDVS008 and LDVS007 to prepare a composite strain, wherein the specific acclimation steps are as follows:
preparing a raw material culture medium: mixing an MRS liquid culture medium and filtered passion fruit juice according to a mass ratio of 1:1, respectively inoculating lactobacillus LDVS005, LDVS008 or LDVS007 into the mixed culture medium in an inoculum size of 0.5%, and carrying out anaerobic fermentation for 1h at 37 ℃ to obtain a passion fruit-MRS composite culture medium containing lactobacillus; mixing the passion fruit-MRS composite culture medium containing LDVS005, the passion fruit-MRS composite culture medium containing LDVS008 and the passion fruit-MRS composite culture medium containing LDVS007 according to the volume ratio of 1:1:1 to obtain composite strains;
then mixing the composite strain prepared by the method and the PDA culture medium according to the mass ratio of 1:5 to prepare a 1-time domestication culture medium, mixing the composite strain and the PDA culture medium according to the mass ratio of 2:5 to prepare a 2-time domestication culture medium, and mixing the composite strain and the PDA culture medium according to the mass ratio of 3:5 to prepare a 3-time domestication culture medium;
domestication culture:
tissue separation is carried out under the aseptic condition, the tissue with the size of mung bean grains is adopted on the cordyceps militaris fruiting body, inoculated on the 1-time domestication culture medium flat plate, placed in a constant temperature incubator for culture at 25 ℃, and the growth condition of hyphae is observed, if white hyphae are fully distributed on the flat plate, transfer can be carried out; transferring corresponding mycelia to a 2-time domestication culture medium plate, placing the plate in a constant temperature incubator for culturing at 25 ℃, observing the growth condition of mycelia, and transferring if white mycelia are fully distributed on the plate; transferring the corresponding mycelium onto a plate of a 3-time domestication culture medium, placing the plate into a constant temperature incubator for culturing at 25 ℃, observing the growth condition of the mycelium, and finishing the domestication process if the white mycelium is fully distributed on the plate.
Acid resistance experiment of domesticated Cordyceps militaris strain:
culture medium: adding citric acid to the culture medium to adjust total acid of the culture medium (total acid of the culture medium is calculated by using a neutralization titration method) based on PDA culture medium; the total acid was adjusted to 10g/L, 20g/L, 30g/L, 40g/L, 50g/L, 60g/L and 70g/L.
Culturing the cordyceps militaris strains which are not domesticated and are domesticated by the method under the conventional conditions, marking the colony diameter once a day, culturing for 7 days, and calculating the hypha growth rate, wherein the obtained results are shown in the following table:
TABLE 3 acid resistance experiments of Cordyceps militaris strains
As can be seen from the table, after the cordyceps militaris strain is domesticated by the steps, the cordyceps militaris strain can grow in a culture medium with the total acid content of 50g/L (close to the acidity of passion fruit), and can keep a relatively fast growth rate in the culture medium with the total acid content of 10-20 g/L; the cordyceps militaris strain which is not domesticated can only grow in a culture medium with the total acid content of 20g/L at most, but the hypha growth rate is obviously reduced under the condition that the total acid content is 20 g/L.
Example 2:
the embodiment is the preparation of passion fruit and cordyceps militaris complex, and the blood sugar reducing effect of the passion fruit and cordyceps militaris complex prepared by the preparation is studied, and the specific steps are as follows:
(1) Selecting the cordyceps militaris mycelium domesticated in the third step of the embodiment 1 as an experimental group, and selecting the cordyceps militaris mycelium domesticated on a PDA culture medium as a control group;
(2) Preparing a culture material:
(1) the traditional Chinese medicine comprises the following components: oven drying 10 parts of ginseng, 10 parts of okra, 3 parts of astragalus membranaceus, 10 parts of radix ophiopogonis and 2 parts of medlar, pulverizing, and mixing for later use;
(2) preparing a rice culture medium: weighing 50g of rice, putting into a 500mL can, adding nutrient solution into the can according to the requirement of 1.3mL of nutrient solution for each gram of rice, and soaking for 2-3h; wherein, the nutrient solution consists of the following components: according to the calculation of each nutrient substance required by 1000g of rice, 15g of peptone, 5g of glucose, 5g of yeast extract and KH 2 PO 4 1.5g、MgSO 4 0.75g, water was added to make up to 1L; sealing, sterilizing under high pressure, taking out, and cooling to room temperature.
(3) Collecting passion fruit: collecting fresh passion fruit with maturity of 8-9 as raw material, breaking passion fruit from the middle, digging out pulp and pulp juice in the interior to obtain pulp, and taking passion fruit shell with pulp removed as culture box;
(4) preparing a mixed culture medium: mixing the crushed traditional Chinese medicine components in the step (1) and the rice culture medium in the step (2) according to the mass ratio of 1:2, then adding the pulp in the step (3) to adjust the water content to be about 55-60%, subpackaging the pulp into a culture box made of passion fruit shells, and sterilizing the pulp according to a conventional method to obtain the passion fruit shell culture medium.
(3) Inoculating and culturing: inoculating Cordyceps militaris strains of the experimental group and the control group in the culture medium of the step (2) under the aseptic condition, culturing for 3-5 days in the environment with the temperature of 16-20 ℃ and the humidity of 70-75% and the dark light, adjusting the room temperature environment of a culture room to 20-25 ℃ when mycelia grow to be full of the material surface, adjusting the humidity to 80-90%, letting the mycelia see light, and then continuously culturing until the Cordyceps militaris mycelia eat through the whole culture medium, and completing the whole culturing process of the Cordyceps militaris mycelia after the surfaces of the mycelia are completely colored; taking out the cordyceps militaris mycelium at the moment to obtain the passion fruit cordyceps militaris complex.
In addition, this embodiment further provides a traditional Chinese medicine group, and the traditional Chinese medicine group adopts: 10 parts of ginseng, 10 parts of okra, 3 parts of astragalus, 10 parts of dwarf lilyturf tuber and 2 parts of medlar are dried, crushed and mixed to prepare the mixed powder.
Drying and crushing passion fruit and cordyceps militaris complexes of the experimental group and the control group to prepare powder, and obtaining experimental group powder and control group powder.
Hypoglycemic experiment:
the hyperglycemia mouse animal model is adopted to analyze the hypoglycemic capability of the experiments, and the specific steps are as follows:
injecting tetraoxypyrimidine into abdominal cavity to prepare hyperglycemia mouse model; 50 diabetic model mice were randomly divided into 5 groups according to the weight and blood sugar balance principle, 10 mice in each group,
mixing the powders of the experimental group, the control group and the traditional Chinese medicine group with water according to the mass ratio of 1:20, and fully stirring to prepare suspension; then feeding the mice according to the dosage of 10 mg/kg.times, feeding the mice 2 times a day at intervals of 12 hours;
meanwhile, the experiment is also provided with a positive control group, wherein the positive control group adopts a hypoglycemic drug acarbose, and the hypoglycemic drug is used for the hyperglycemia mice according to the product specification;
furthermore, a blood glucose group was set: the blood sugar group is not taken with any hypoglycemic drugs, and is only infused with normal saline;
normal group: the normal group is normal mice.
The fasting blood glucose concentration was measured by administering the product continuously or for 30d in the above-described group administration mode, and fasting after the last administration. The specific test data are shown in Table 4.
TABLE 4 influence of different passion fruit and Cordyceps militaris complexes on glycemic effect
| Group of | Blood glucose level before feeding (mmol/L) | Blood glucose level after 30d of feeding (mmol/L) |
| Experimental group | 24.11 | 9.45 |
| Control group | 23.97 | 13.57 |
| Traditional Chinese medicine group | 24.57 | 9.04 |
| Positive control group | 25.14 | 8.13 |
| Normal group | 6.34 | 6.38 |
| Blood sugar group | 24.16 | 25.34 |
From the above table, the blood sugar content of the experimental group, the control group, the traditional Chinese medicine group and the positive control group is reduced after the experimental group, the control group, the traditional Chinese medicine group and the positive control group are fed, but the blood sugar value is still slightly higher than that of the normal group; the blood sugar value of the traditional Chinese medicine group is slightly higher than that of the positive control group, which shows that the blood sugar reducing effect of the traditional Chinese medicine group is close to that of acarbose; compared with the control group, the blood sugar reducing effect of the experimental group is obviously reduced, so that the growth speed of the cordyceps militaris strain is increased after domestication, and hyphae are robust, so that more effective components in the traditional Chinese medicine culture medium can be converted, and finally, the cordyceps militaris strain has better blood sugar reducing effect.
Example 3:
preparing passion fruit tablets:
from the experiment in example 2, after the lactic acid bacteria composite strain is domesticated, the blood sugar reducing effect of the passion fruit and cordyceps militaris complex is improved, and for convenient carrying, the applicant prepares the passion fruit and cordyceps militaris complex into tablets, and the specific preparation method is as follows:
because the tablet disclosed by the application is required to maintain the efficacy of reducing blood sugar, the passion fruit and cordyceps militaris complex has the characteristics of being relatively sour and relatively thick in traditional Chinese medicine taste compared with the conventional cordyceps militaris after being crushed, and is not good in taste, the auxiliary materials which can improve the blood sugar are required to be avoided in the selection of related forming agents and auxiliary materials, and meanwhile, some flavoring agents are required to be added to improve the taste, so that the applicant selects the following auxiliary materials to tablet and form the passion fruit and cordyceps militaris complex:
(1) the mass ratio of xylitol to magnesium stearate is 1:3;
(2) xylitol, magnesium stearate and aloe colloid have a mass ratio of 1:2:1;
(3) xylitol, magnesium stearate and passion fruit juice are mixed according to the mass ratio of 1:2:1;
(4) xylitol, magnesium stearate, aloe colloid and passion fruit juice are mixed according to the mass ratio of 1:1:1:1;
pulverizing the above Chinese medicinal materials, adding water, making into paste, adding adjuvants (1) - (4), and respectively pressing into passion fruit tablet; wherein the addition amount of the auxiliary materials is 5% of the mass of the passion fruit and cordyceps militaris complex paste.
After that, passion fruit tablets of the above several experimental groups were tasted, and finally found:
the passion fruit slices of group (1) are the hardest, next to group (2), next to group (4), and finally to group (3);
the best mouthfeel is group (4): the sweetness is moderate, no traditional Chinese medicine taste, delicate fragrance and thick fruit flavor are basically generated;
the group (3) has heavier sour taste, basically no traditional Chinese medicine taste and strong fruit flavor;
group (2) has moderate sweetness, basically no traditional Chinese medicine flavor and no fruit flavor.
Therefore, the (4) group is preferably used as the auxiliary material of the passion fruit tablet, however, the applicant finds that the auxiliary material contains passion fruit juice and has a certain fructose, so that the applicant performs research on the blood sugar reducing effect of the (3) group and the (4) group, and the specific experimental method is similar to the animal experimental method of the embodiment 2, namely:
the hyperglycemia mouse animal model is adopted to analyze the hypoglycemic capability of the experiments, and the specific steps are as follows:
injecting tetraoxypyrimidine into abdominal cavity to prepare hyperglycemia mouse model; the number of diabetic model mice is 40, and the diabetic model mice are randomly divided into 4 groups according to the weight and blood sugar balance principle, 10 mice in each group,
grinding the passion fruit slices prepared in the (3) and (4) into powder, mixing the powder with water according to a mass ratio of 1:20, and fully stirring to prepare suspension; then feeding the mice according to the dosage of 10 mg/kg.times, feeding the mice 2 times a day at intervals of 12 hours;
meanwhile, the experiment is also provided with a positive control group, wherein the positive control group adopts a hypoglycemic drug acarbose, and the hypoglycemic drug is used for the hyperglycemia mice according to the product specification;
furthermore, a blood glucose group was set: the blood sugar group is not taken with any hypoglycemic drugs, and is only infused with normal saline;
normal group: the normal group is normal mice.
The fasting blood glucose concentration was measured by administering the product continuously or for 30d in the above-described group administration mode, and fasting after the last administration. The specific test data are shown in Table 5.
TABLE 5 influence of different excipients on the hypoglycemic effect of tablets
| Group of | Blood glucose level before feeding (mmol/L) | Blood glucose level after 30d of feeding (mmol/L) |
| Group (3) | 25.34 | 12.14 |
| Group (4) | 25.11 | 8.47 |
| Positive control | 25.08 | 8.07 |
| Normal group | 6.24 | 6.29 |
| Blood sugar group | 24.87 | 25.04 |
From this, the (4) group adjuvant combination of the present application: xylitol, magnesium stearate, aloe colloid and passion fruit juice are in a mass ratio of 1:1:1:1, and the taste of the combination is optimal, so that the auxiliary materials of the group (4) are selected to be combined to prepare the passion fruit tablet most suitable.
In summary, the best preparation method of the passion fruit tablet is obtained as follows:
(1) Domesticating Cordyceps militaris:
tissue separation is carried out under the aseptic condition, the tissue with the size of mung bean grains is adopted on the fruiting body of the cordyceps militaris, inoculated on a 1-time domestication culture medium, placed in a constant temperature incubator for culture at 25 ℃, and the growth condition of hyphae is observed, and if white hyphae are fully distributed on a flat plate, transfer can be carried out; transferring corresponding mycelium onto 2 times of domestication culture medium, placing into a constant temperature incubator for culturing at 25deg.C, observing mycelium growth condition, and transferring if white mycelium is fully distributed on the plate; transferring corresponding mycelium to a 3-time domestication culture medium, placing the culture medium into a constant temperature incubator for culturing at 25 ℃, observing the growth condition of the mycelium, and finishing the domestication process if the white mycelium is fully distributed on a flat plate;
the preparation method of the composite strain of the domestication culture medium comprises the following steps: mixing an MRS liquid culture medium and filtered passion fruit juice according to a mass ratio of 1:1, respectively inoculating lactobacillus LDVS005, LDVS008 or LDVS007 into the mixed culture medium in an inoculum size of 0.5%, and carrying out anaerobic fermentation for 1h at 37 ℃ to obtain a passion fruit-MRS composite culture medium containing lactobacillus; mixing the passion fruit-MRS composite culture medium containing LDVS005, the passion fruit-MRS composite culture medium containing LDVS008 and the passion fruit-MRS composite culture medium containing LDVS007 according to the volume ratio of 1:1:1 to obtain composite strains;
the preparation method of the 1-time domestication culture medium comprises the following steps: mixing a composite strain (prepared by the upper-stage preparation method) and a PDA culture medium according to a mass ratio of 1:5;
the preparation method of the 2-time domestication culture medium comprises the following steps: mixing a composite strain (prepared by the upper-stage preparation method) and a PDA culture medium according to a mass ratio of 2:5;
the preparation method of the 3-time domestication culture medium comprises the following steps: mixing a composite strain (prepared by the upper-stage preparation method) and a PDA culture medium according to a mass ratio of 3:5;
(2) Preparing passion fruit and cordyceps militaris complex:
1) Preparing a culture material:
(1) the traditional Chinese medicine comprises the following components: oven drying 10 parts of ginseng, 10 parts of okra, 3 parts of astragalus membranaceus, 10 parts of radix ophiopogonis and 2 parts of medlar, pulverizing, and mixing for later use;
(2) preparing a rice culture medium: weighing 50g of rice, putting into a 500mL can, adding nutrient solution into the can according to the requirement of 1.3mL of nutrient solution for each gram of rice, and soaking for 2-3h; wherein, the nutrient solution consists of the following components: according to the calculation of each nutrient substance required by 1000g of rice, 15g of peptone, 5g of glucose, 5g of yeast extract and KH 2 PO 4 1.5g、MgSO 4 0.75g, water was added to make up to 1L; sealing and then high pressureSterilizing, taking out of the pot, and cooling to room temperature for standby.
(3) Collecting passion fruit: collecting fresh passion fruit with maturity of 8-9 as raw material, breaking passion fruit from the middle, digging out pulp and pulp juice in the interior to obtain pulp, and taking passion fruit shell with pulp removed as culture box;
(4) preparing a mixed culture medium: mixing the crushed traditional Chinese medicine components in the step (1) and the rice culture medium in the step (2) according to the mass ratio of 1:2, then adding the pulp in the step (3) to adjust the water content to be about 55-60%, subpackaging the pulp into a culture box made of passion fruit shells, and sterilizing the pulp according to a conventional method to obtain the passion fruit shell culture medium.
2) Inoculating and culturing: selecting the cordyceps militaris mycelium domesticated in the step (1) under the aseptic condition, inoculating the cordyceps militaris mycelium as a strain to the passion fruit shell culture medium in the step (1), culturing for 3-5 days in an environment with the temperature of 16-20 ℃ and the humidity of 70-75% and dark light, adjusting the room temperature environment of a culture chamber to 20-25 ℃ and the humidity to 80-90% when the mycelium grows to a full level, allowing the mycelium to see light, continuing culturing until the cordyceps militaris mycelium is eaten through the whole culture medium, and completing the whole cordyceps militaris mycelium culturing process after the surface of the mycelium is completely colored; taking out the cordyceps militaris mycelium at the moment to obtain the passion fruit cordyceps militaris complex.
(3) Preparing tablets:
pulverizing the passion fruit and cordyceps militaris complex in the step (2), adding water to prepare paste, adding auxiliary materials according to 5% of the mass of the paste of the passion fruit and cordyceps militaris complex, and tabletting to prepare passion fruit tablets; wherein, the auxiliary materials are as follows: xylitol, magnesium stearate, aloe colloid and passion fruit juice are mixed according to the mass ratio of 1:1:1:1.
In summary, the problem that cordyceps militaris cannot effectively grow due to too high acidity in preparing the passion fruit cordyceps militaris complex is solved by domesticating the cordyceps militaris strain, meanwhile, the prepared passion fruit cordyceps militaris complex has the effect of reducing blood sugar by improving the culture medium for preparing the passion fruit cordyceps militaris complex, and in order to conveniently utilize the passion fruit cordyceps militaris complex, the passion fruit tablet with good taste and good blood sugar reducing effect can be obtained by improving auxiliary materials, and finally, the passion fruit cordyceps militaris complex tablet which is convenient to carry and has good blood sugar reducing effect is prepared, and is worthy of popularization and wide application.
The foregoing description is directed to the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the invention, and all equivalent changes or modifications made under the technical spirit of the present invention should be construed to fall within the scope of the present invention.
Claims (5)
1. The passion fruit and cordyceps militaris complex is characterized in that the passion fruit and cordyceps militaris complex is prepared by domesticating a cordyceps militaris strain by a domestication culture medium and then inoculating the domesticated cordyceps militaris strain into a passion fruit shell culture medium for culture; the domestication culture medium contains lactobacillus composite strains; the passion fruit shell culture medium contains traditional Chinese medicine components;
the lactobacillus composite strain is strain LDVS005, LDVS007 and LDVS008; the strain LDVS005 has a deposit number of: CGMCC NO:20027, ldvs007 accession number: CGMCC NO:20028, accession number of ldvs008 is: CGMCC NO:20029;
the preparation method of the lactic acid bacteria composite strain of the domestication culture medium comprises the following steps: mixing an MRS liquid culture medium and filtered passion fruit juice according to a mass ratio of 1:1, respectively inoculating lactobacillus LDVS005, LDVS008 or LDVS007 into the mixed culture medium in an inoculum size of 0.5%, and carrying out anaerobic fermentation for 1h at 37 ℃ to obtain an MRS-passion fruit juice liquid mixture containing lactobacillus; and mixing the MRS-passion fruit juice liquid mixture containing LDVS005, the MRS-passion fruit juice liquid mixture containing LDVS008 and the MRS-passion fruit juice liquid mixture containing LDVS007 according to a volume ratio of 1:1:1 to obtain the composite strain.
2. The passion fruit and cordyceps militaris complex as claimed in claim 1, wherein the traditional Chinese medicine components consist of the following components in parts by weight: 10 parts of ginseng, 10 parts of okra, 3 parts of astragalus, 10 parts of dwarf lilyturf tuber and 2 parts of medlar.
3. Use of a passion fruit and Cordyceps militaris complex as claimed in any one of claims 1 or 2 in the preparation of a hypoglycemic tablet.
4. A method for preparing a tablet by using the passion fruit and cordyceps militaris complex as claimed in any one of claims 1 or 2, wherein the method is as follows: pulverizing the passion fruit and Cordyceps militaris complex, adding water to prepare paste, adding auxiliary materials according to 5% of the mass of the paste of the passion fruit and Cordyceps militaris complex, and tabletting to prepare passion fruit tablets.
5. The method according to claim 4, wherein the auxiliary materials are prepared by mixing xylitol, magnesium stearate, aloe vera gel and passion fruit juice according to a mass ratio of 1:1:1:1.
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| CN112358988A (en) * | 2020-11-10 | 2021-02-12 | 广西壮族自治区农业科学院 | Lactobacillus plantarum LDVS008 strain and application thereof |
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