CN114712400A - Probiotic fermented clam powder composition capable of reducing blood sugar and blood pressure and preparation method thereof - Google Patents
Probiotic fermented clam powder composition capable of reducing blood sugar and blood pressure and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
Abstract
The invention relates to a probiotic fermented clam powder composition capable of reducing blood sugar and blood pressure and a preparation method thereof, and solves the problem that the market is insufficient in probiotic drinks with the functions of reducing blood sugar, blood pressure and blood fat.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a probiotic fermented clam powder composition capable of reducing blood sugar and blood pressure and a preparation method thereof.
Background
The clams are common marine products distributed in China coastal areas, and also have medicinal value. How to further develop the clams and maximally utilize the medicinal value of the clams is still a problem at present. Meanwhile, although a lot of probiotic beverages are available in the market, few probiotic beverages with the functions of reducing blood sugar, blood pressure and blood fat are available.
Disclosure of Invention
Aiming at the technical problems, the invention provides a probiotic fermented clam powder composition capable of reducing blood sugar and blood pressure and a preparation method thereof, and solves the problem that the market is insufficient for probiotic drinks with the functions of reducing blood sugar, blood pressure and blood fat.
In order to achieve the aim, the invention provides a preparation method of a probiotic fermented clam powder composition, which comprises the following steps: enzymolysis of clam meat, adding prebiotics culture medium, inoculating probiotic bacteria, and culturing.
The method specifically comprises the following steps:
(1) raw material treatment: heating the clams, taking clam meat, drying, crushing and sieving to obtain clam dry powder;
(2) enzymolysis: adding water into the clam dry powder obtained in the step (1), adding trypsin and papain for enzymolysis, inactivating enzymes after the enzymolysis is finished to obtain an enzymolysis solution, filtering by using kieselguhr, and performing suction filtration to obtain a clear enzymolysis solution;
(3) blending and inactivating: adding glucose, peptone, yeast extract, inorganic salt, sweetener and prebiotics into the enzymolysis solution obtained in the step (2), adjusting the pH value, heating and sterilizing;
(4) fermentation: and (4) inoculating probiotics into the mixed liquor obtained in the step (3), fermenting to obtain fermentation liquor, and freeze-drying the fermentation liquor to obtain the probiotic fermented clam powder.
Further, in the step (1), heating the clams, taking and drying the clam meat, crushing the dried clam meat by a crusher, and sieving the dried clam meat by a sieve with 20-40 meshes to obtain the dried clam powder.
Further, in the step (2), the volume ratio of the clam dry powder to water is 1: 6.
further, in the step (2), the enzymolysis temperature is 40 ℃, and the enzymolysis time is 2 hours.
Further, in the step (2), the enzyme deactivation temperature is 100 ℃, and the enzyme deactivation time is 15 min.
Further, in the step (3), the pH value is 6.5-7.0.
Further, in the step (3), the sterilization temperature is 115 ℃ and the time is 30 min.
Further, the addition amount of the papain is 6500-8000U/g clam powder; the addition amount of the trypsin is 5000-.
Further, the inorganic salt is ammonium chloride, manganese sulfate and dipotassium hydrogen phosphate.
Further, the mass volume percentage (g/mL) of the clam powder in the probiotic fermented clam composition is 1-20%, the adding amount of glucose is 2-5%, the adding amount of peptone is 0.05-2%, the adding amount of yeast extract is 0.1-2%, the adding amount of ammonium chloride is 0.1-1%, the adding amount of manganese sulfate is 0.01-0.1%, and the adding amount of dipotassium hydrogen phosphate is 0.01-0.5%. Preferably, glucose is added in an amount of 2%, peptone is added in an amount of 1%, yeast extract is added in an amount of 0.5%, ammonium chloride is added in an amount of 0.6%, manganese sulfate is added in an amount of 0.02%, and dipotassium hydrogen phosphate is added in an amount of 0.1%.
The sweetening agent is tagatose with the mass volume percentage (g/mL) of 0.1-5% and trichlorosucrose with the mass volume percentage (g/mL) of 0.01-0.1%, and preferably, the adding amount of the tagatose is 0.2%, and the adding amount of the trichlorosucrose is 0.08%.
The prebiotics are one or more of polydextrose, fructo-oligosaccharide, inulin, mannan, lactulose and galacto-oligosaccharide.
The addition amount of the prebiotics is 0.01-0.7% by mass volume percentage (g/mL), preferably, the addition amount of the polydextrose is 2-6% by mass volume percentage (g/mL); the addition amount of the fructo-oligosaccharide is 0.1-0.7% by mass volume (g/mL).
The probiotics are bifidobacteria and lactobacillus; the bifidobacterium is selected from bifidobacterium lactis, bifidobacterium longum or bifidobacterium bifidum; the lactobacillus is selected from lactobacillus gasseri, lactobacillus casei, lactobacillus paracasei, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus rhamnosus or lactobacillus helveticus. Preferably, the probiotics are bifidobacterium and lactobacillus casei, and the mass ratio of the bifidobacterium to the lactobacillus casei is 1: 1.
Further, in the step (4), the using amount of the probiotics is 1-5% of the volume percentage of the mixed liquor in the step (3).
Further, after inoculating probiotics, fermenting for 8 hours at 40 ℃, and fermenting for 2 days after placing at 4 ℃.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the probiotic fermented clam powder ensures the complete conversion of nutritional ingredients of the clam in the fermentation process, so that the fermented clam powder not only contains active ingredients such as amino acid, glycogen and the like in the nutritional ingredients of the clam, but also contains active probiotics, and the nutritional and health-care functions of the clam are ensured.
2. The clam polysaccharide is generally embedded or inlaid in protein, and different proteases are used, so that on one hand, the protein can be degraded into small-molecule polypeptide, and on the other hand, more clam polysaccharide can be released. Meanwhile, bitter peptides can be reduced by adding complex enzyme, so that enzymatic hydrolysate with good flavor can be obtained, and development and utilization of marine food are facilitated.
3. The compound protease primarily degrades protein in the clam powder, the molecular weight of the clam peptide after enzymolysis is reduced through probiotic fermentation, and the content of clam polysaccharide is increased, so that the substance with high activity and blood pressure reduction is prepared.
4. According to the probiotic fermented clam powder, the fermentation product contains clam polysaccharide, and the pharmacological effects of reducing blood sugar, reducing blood fat, improving immunity and the like are achieved. The clam peptide is rich in amino acids such as proline, phenylalanine, tyrosine and the like, has better ACE inhibitory activity and has the function of assisting in reducing blood pressure; prebiotics is added into the fermentation product, and the prebiotics and the clam polysaccharide in the fermentation product have the effect of reducing blood fat under the synergistic effect.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
Heating Concha Meretricis Seu Cyclinae, and collectingDrying clam meat, crushing by a crusher, sieving by a sieve of 20-40 meshes to obtain 500g of clam powder, adding 3L of purified water into the clam powder, adding papain according to 7000U/g of the clam powder, stirring uniformly, adding trypsin according to 8000U/g of the clam powder, keeping the temperature at 40 ℃, stirring for 2h, inactivating enzyme at 100 ℃ for 15min after enzymolysis is finished, filtering by diatomite, performing suction filtration to obtain clear enzymolysis liquid, adding 60g of glucose, 30g of peptone, 15g of yeast extract and 18g of NH into the enzymolysis liquid4CL，0.6g MnSO4·H2O，3g K2HPO4·3H2O, 6.0g of tagatose, 2.4g of trichlorosucrose and 3g of polydextrose, adjusting the pH value to 7.0, sterilizing at 115 ℃ for 15min to obtain a mixed culture medium, inoculating 5% of bifidobacterium and lactobacillus casei according to the volume percentage of enzymolysis liquid (the ratio of the two is 1:1), fermenting for 8 hours at 40 ℃, standing at 4 ℃ and fermenting for 2 days to obtain fermentation liquid, and freeze-drying the fermentation liquid to obtain the fermented clam powder.
Comparative example 1
Heating clams, taking clam meat, drying, crushing by a crusher, sieving by a sieve of 20-40 meshes to obtain 500g of clam powder, adding 3L of purified water into the clam powder, adding papain according to 7000U/g of the clam powder, stirring uniformly, adding trypsin according to 8000U/g of the clam powder, keeping the temperature at 40 ℃ and stirring for 2h, inactivating enzyme for 15min at 100 ℃ after enzymolysis is finished, filtering by diatomite, performing suction filtration to obtain clear enzymolysis liquid, adding 60g of glucose, 30g of peptone, 15g of yeast extract and 18g of NH into the enzymolysis liquid4CL，0.6g MnSO4·H2O，3g K2HPO4·3H2O, 6.0g of tagatose, 2.4g of sucralose and 3g of polydextrose, adjusting the pH value to 7.0, sterilizing at 115 ℃ for 15min, and freeze-drying the mixed solution to obtain the clam powder composition.
Comparative example 2
Heating clams, taking clam meat, drying, crushing by a crusher, sieving by a sieve of 20-40 meshes to obtain 500g of clam powder, adding 3L of purified water into the clam powder, and adding 60g of glucose, 30g of peptone, 15g of yeast extract and 18g of NH into the supernatant4CL，0.6g MnSO4·H2O，3g K2HPO4·3H2O, 6.0g tagatose, 2.4g sucraloseAnd 3g of polydextrose, adjusting the pH value to 7.0, sterilizing at 115 ℃ for 15min to obtain a mixed culture medium, inoculating 5% of bifidobacterium and lactobacillus casei (the ratio of the bifidobacterium to the lactobacillus casei is 1:1) according to the volume percentage of the mixed solution, fermenting for 8 hours at 40 ℃, fermenting for 2 days after standing at 4 ℃ to obtain a fermentation liquid, filtering with diatomite, performing suction filtration to obtain a supernatant, and freeze-drying the supernatant to obtain the fermented clam powder.
Comparative example 3
Heating clams, taking clam meat, drying, crushing by a crusher, sieving by a sieve of 20-40 meshes to obtain 500g of clam powder, adding 3L of purified water into the clam powder, adding papain according to 7000U/g of the clam powder, stirring uniformly, adding trypsin according to 8000U/g of the clam powder, keeping the temperature at 40 ℃ and stirring for 2h, inactivating enzyme for 15min at 100 ℃ after enzymolysis is finished, filtering by diatomite, performing suction filtration to obtain clear enzymolysis liquid, adding 60g of glucose, 30g of peptone, 15g of yeast extract and 18g of NH into the enzymolysis liquid4CL，0.6g MnSO4·H2O，3g K2HPO4·3H2O, 6.0g of tagatose and 2.4g of trichlorosucrose, adjusting the pH value to 7.0, sterilizing at 115 ℃ for 15min to obtain a mixed culture medium, inoculating 5% of bifidobacterium and lactobacillus casei (the ratio of the two is 1:1) according to the volume percentage of enzymolysis liquid, fermenting for 8 hours at 40 ℃, fermenting for 2 days after standing at 4 ℃ to obtain fermentation liquid, and freeze-drying the fermentation liquid to obtain the fermented clam powder.
The detection method comprises the following steps:
a detection method of clam polysaccharide in probiotic fermented clam powder comprises the following steps: and (3) determining the polysaccharide content of each sample according to a method specified in NY/T1676 determination of the crude polysaccharide content of the edible fungi. Treating a fermentation liquor sample: taking a proper amount of a composition of the fermented clam powder and a sample of an unfermented composition, adding 80% ethanol, and fixing the volume to 100ml to obtain precipitates, namely the clam polysaccharide, wherein the detection results are shown in table 1.
TABLE 1 content of clam polysaccharides in fermentation broth
Relative molecular mass GPC detection method of clam peptide: preparing standard solution by precisely weighing appropriate amount of cytochrome C, aprotinin, bacillus enzyme, alanine-tyrosine-arginine and alanine-alanine with relative molecular mass standard, respectively matching into peptide standard solution with concentration of 1.0g and different relative molecular mass with flowing phase, filtering with filter membrane with pore diameter of 0.2um, and respectively sampling to obtain chromatogram of series of standard products. Plotting the logarithm of the relative molecular mass (1gMW) against the retention time or performing linear regression to obtain a relative molecular mass calibration curve and an equation thereof; preparing a sample: weighing 125.0mg of sample, dissolving with mobile phase, diluting to 25mL, ultrasonically oscillating for 10min to fully dissolve and mix the sample, filtering with a polytetrafluoroethylene or nylon filter membrane with pore size of 0.2 μm, and measuring the filtrate; a chromatographic column: TSKgel G2000SWxL gel chromatography column (300 × 7.8mm, Tosoh), mobile phase: acetonitrile-water-trifluoroacetic acid (40:60: 0.05); detection wavelength: 220 nm; flow rate: 0.2 mL/min; column temperature: 35 ℃; sample introduction volume: 10 ul; and (3) carrying out isocratic elution. The results are shown in Table 2.
TABLE 2 relative molecular masses of clam peptides in fermentation broths
Effect example 1: effect of probiotic fermented clam powder on hyperglycemic mice
Taking 100 experimental mice, randomly taking 10 mice as a normal control group, fasting the rest mice for 16h, injecting a physiological saline solution of alloxan into the abdominal cavity according to the dose of 190mg/kg, establishing a diabetes animal model, injecting an equivalent amount of physiological saline into the normal control group, sampling blood from the tail vein after 72h (fasting for 5h) to measure the fasting blood glucose of the mice, randomly dividing 60 mice with the blood glucose value of 11-28mmol/L successfully molded into 6 groups according to the blood glucose value and the body weight, wherein the mice are respectively a model group and a metformin positive control group, an example 1 group, a comparative example 2 group, a comparative example 3 group and 10 mice each. Samples in examples and comparative examples are prepared into aqueous solutions, the animals are administrated in a gastric perfusion mode (1mL/100g), the model control group and the blank control group are respectively perfused with distilled water with the same volume, the probiotic fermented clam powder compositions of example 1 group, comparative example 2 group and comparative example 3 group are respectively perfused with 2g/kg bw, the metformin positive control group is perfused with 180mg/kg metformin hydrochloride aqueous solution with the same volume, the intragastric administration is carried out for 1 time every day, and the mice in each group freely eat and drink water during the experiment. The normal control group and the model group were administered with equal volume of distilled water for 6 weeks, and blood was collected from the tail vein at the end of 0, 2, 4 and 6 weeks of the experiment to determine fasting blood glucose. And (4) detecting the fasting blood glucose value by using a micro-glucometer.
The specific experimental results are shown in Table 3
TABLE 3 Effect of probiotic fermented clam powder compositions on hyperglycemic mice
※P<0.05, compared to a blank control group;#P<0.05, compared to the model set.
As can be seen from Table 3, there was no statistical difference in fasting blood glucose levels of rats of each group before molding, and the fasting blood glucose levels of rats of the diabetic model after molding were significantly higher (P <0.05) than those of the rats of the blank control group, indicating that the diabetic rat model was successfully constructed. After 4 weeks of gavage, the fasting plasma glucose decreased significantly in the group of example 1 (P < 0.05); compared with the diabetes model group, the fasting blood glucose of the group of the comparative example 1 and the group of the comparative example 2 has no obvious difference after the intragastric administration for 6 weeks; comparative example 3 group showed significant reduction in fasting glucose (P <0.05) after 6 weeks of gavage; the metformin control group had significantly lower fasting blood glucose after 2 weeks of gavage than the diabetes model group (P < 0.05). The composition of the probiotic fermented clam powder has the function of reducing blood sugar, and as can be seen from the table 1, the probiotic fermented clam powder in the group in the example 1 decomposes macromolecular proteins in the clam powder into oligopeptides, has high content of clam polysaccharide, and the group in the comparative example 1 does not add probiotics, so that the fasting blood sugar value of a mouse is reduced, but the time is longer, so that the blood sugar reducing function is slower, and the probiotic added in the group in the example 1 obviously reduces the fasting blood sugar value of the mouse after the stomach is filled for 4 weeks, so that the probiotic and the clam polysaccharide can synergistically act to help to maintain the healthy level of blood sugar. In the comparative example 2, no complex enzyme is added, the protein molecular weight in the clam powder is larger, the content of the clam polysaccharide is less, the blood sugar reducing effect is not obvious, no prebiotics are added in the comparative example 3, the fasting blood sugar of the mouse is obviously reduced after the gavage for 6 weeks, and the blood sugar reducing effect is smaller than that in the example 1.
Effect example 2 Effect of probiotic fermented clam powder on hyperlipidemic rats
Male rats 70 were acclimatized to normal diet for 7 days, and then 60 rats were fed with high-ester diet for 30 days, and then randomized into 6 groups (n-10) each of which was: the samples in the example 1 and the comparative example 1 are prepared into aqueous solutions and are fed to animals in a gastric perfusion mode (1mL/100g), the blank control group and the model control group are respectively perfused with distilled water with the same volume every day, the probiotic fermented clam powder compositions in the example 1 group, the comparative example 2 group and the comparative example 3 group are separately perfused with 2g/kg bw, the simvastatin positive control group is perfused with 5mg/kg of simvastatin aqueous solution with the same volume every day, the simvastatin positive control group is perfused once every day for a fixed time, and the rats in each group freely eat and drink water during the experiment. All rats were subjected to submandibular blood sampling on day 30 after gavage, serum was collected by centrifugation, and the specific test results of the contents of TC, TG and LDL-C in the serum were determined according to the kit shown in table 4.
TABLE 4 Effect of probiotic fermented clam powder composition on hyperlipidemic rats
※P<0.05, compared to a blank control group;#P<0.05, compared to the model set.
As shown in table 4, when rats were fed with the high-fat diet for 30 days, the contents of Total Cholesterol (TC), Triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in the serum of the diabetes model group were significantly increased (P <0.05) compared with the blank control group, which indicates that the mice fed with the high-fat diet for a long time cause lipid metabolism disorder and hyperlipidemia. After administration of the probiotic clam powder composition for gavage to mice fed with high-fat feed, the TC content in the serum of rats in the group of example 1 and the simvastatin positive group is obviously reduced (P is less than 0.05) compared with that of the model control group after 30 days; the TG in the serum of rats in the example 1 group, the simvastatin positive group and the comparative example 3 group is obviously reduced (P < 0.05); the serum LDL-C (P <0.05) content of rats in the group of example 1, the simvastatin positive group, the comparative example 1 and the comparative example 3 is obviously reduced. Example 1 the TC (P >0.05), TG (P >0.05) and LDL-C (P >0.05) were not significantly different in rat sera in the high dose group compared to the simvastatin positive group.
Effect example 3 Effect of probiotic fermented clam powder on ACE inhibitory Activity
Taking a proper amount of probiotic fermented clam powder of the groups of example 1, comparative example 2 and comparative example 3, dissolving the probiotic fermented clam powder in a 0.1mol/L boric acid solution, preparing 100mU/mL ACE and 5mmol/L HHL solutions by using the boric acid solution, uniformly mixing 30L HHL and 10L samples, preserving heat at 37 ℃ for 5min, adding 20L ACE to start reaction, uniformly mixing, reacting at 37 ℃ for 1h, and rapidly adding 70L HCL (1mol/L) to stop the reaction. And (3) measuring the ACE inhibitory activity by using high performance liquid chromatography, wherein a boric acid solution is used as a blank control. A chromatographic column: Xbridge-C18 column, mobile phase: acetonitrile-0.05% aqueous trifluoroacetic acid; detection wavelength: 228 nm; flow rate: 0.8 mL/min; column temperature: 30 ℃; sample introduction volume: 10 ul; gradient conditions: acetonitrile 10-60% (0-40 min). The results are shown in Table 5.
TABLE 5 inhibition assay results
As can be seen from table 5, the ACE inhibition rate of the group in example 1 is the best, and the ACE inhibition rate of the group in comparative example 3 is better compared with that of the group in comparative example 1 and the group in comparative example 3, because no probiotics are added in the group in comparative example 1, and the small-molecule polypeptide in the group in comparative example 1 is less; the comparative example 2 group has no enzymolysis, the protein is not basically degraded into small molecules, the content of the clam polysaccharide is low, and the ACE inhibition rate is only 7.49 percent, which proves that the example 1 has good ACE inhibition activity and is helpful for maintaining healthy blood pressure level after long-term use.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A preparation method of a probiotic fermented clam powder composition is characterized by comprising the following steps: enzymolysis of clam meat, adding prebiotics culture medium, inoculating probiotic bacteria, and culturing.
2. The method according to claim 1,
(1) treating raw materials: heating the clams, taking clam meat, drying, crushing and sieving to obtain clam dry powder;
(2) enzymolysis: adding water into the clam dry powder obtained in the step (1), adding trypsin and papain for enzymolysis, inactivating enzymes after the enzymolysis is finished to obtain an enzymolysis solution, filtering by using kieselguhr, and performing suction filtration to obtain a clear enzymolysis solution;
(3) blending and inactivating: adding glucose, peptone, yeast extract, inorganic salt, sweetener and prebiotics into the enzymolysis solution obtained in the step (2), adjusting the pH value, heating and sterilizing;
(4) fermentation: and (4) inoculating probiotics into the mixed liquor obtained in the step (3), fermenting to obtain fermentation liquor, and freeze-drying the fermentation liquor to obtain the probiotic fermented clam powder.
3. The preparation method according to claim 2, wherein in the step (1), the clam is heated, the clam meat is taken and dried, the clam meat is crushed by a crusher and is sieved by a sieve with 20-40 meshes to obtain the clam dry powder.
4. The preparation method according to claim 2, wherein in the step (2), the volume ratio of the clam dry powder to water is 1: 6; the enzymolysis temperature is 40 ℃, and the enzymolysis time is 2 hours; the enzyme deactivation temperature is 100 deg.C, and the enzyme deactivation time is 15 min.
5. The method according to claim 2, wherein in the step (3), the pH value is 6.5 to 7.0; sterilizing at 115 deg.C for 30 min.
6. The preparation method according to claim 2, wherein the addition amount of the papain is 6500-8000U/g clam powder; the addition amount of the trypsin is 5000-.
7. The preparation method of claim 2, wherein the mass volume percentage (g/mL) of the clam powder in the probiotic fermented clam composition is 1-10%, the adding amount of glucose is 2-5%, the adding amount of peptone is 0.05-2%, the adding amount of yeast extract is 0.1-2%, the adding amount of ammonium chloride is 0.1-1%, the adding amount of manganese sulfate is 0.01-0.1%, and the adding amount of dipotassium hydrogen phosphate is 0.01-0.5%; preferably, glucose is added in an amount of 2%, peptone is added in an amount of 1%, yeast extract is added in an amount of 0.5%, ammonium chloride is added in an amount of 0.6%, manganese sulfate is added in an amount of 0.02%, and dipotassium hydrogen phosphate is added in an amount of 0.1%.
8. The production method according to claim 2, wherein the inorganic salts are ammonium chloride, manganese sulfate and dipotassium hydrogen phosphate; the prebiotics are one or more of polydextrose, fructo-oligosaccharide, inulin, mannan, lactulose and galacto-oligosaccharide; the probiotics are bifidobacteria and lactobacillus; the bifidobacterium is selected from bifidobacterium lactis, bifidobacterium longum or bifidobacterium bifidum; the lactobacillus is selected from lactobacillus gasseri, lactobacillus casei, lactobacillus paracasei, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus rhamnosus or lactobacillus helveticus; preferably, the probiotics are bifidobacterium and lactobacillus casei, and the mass ratio of the bifidobacterium to the lactobacillus casei is 1: 1.
9. The preparation method according to claim 2, wherein in the step (4), the amount of the probiotics is 1-5% of the volume percentage of the mixed solution in the step (3); inoculating probiotic bacteria, fermenting at 40 deg.C for 8 hr, and fermenting at 4 deg.C for 2 days.
10. The probiotic fermented clam product with the efficacy of reducing blood sugar and blood pressure, which is prepared by the preparation method according to any one of the preceding claims.
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