CN113398151A - Probiotics and prebiotics combined fermented milk for treating hyperhomocysteinemia and preparation method and application thereof - Google Patents
Probiotics and prebiotics combined fermented milk for treating hyperhomocysteinemia and preparation method and application 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
- A61K35/66—Microorganisms or materials therefrom
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- 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
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
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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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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Abstract
The invention discloses probiotic and prebiotics combined fermented milk for treating hyperhomocysteinemia, which comprises lactobacillus plantarum KLDS 1.0317 and galacto-oligosaccharide; the concentration of the galacto-oligosaccharide is 2mg/100mL, and the polymerization degree is 5; the number of the lactobacillus plantarum KLDS 1.0317 is more than or equal to 1 multiplied by 108CFU/mL or more than or equal to 1X 108CFU/g. The invention also discloses a preparation method and application of the probiotic and prebiotics combined fermented milk for treating hyperhomocysteinemia. Lactobacillus plantarum KLDS 1 of the inventionThe 0317 and galactooligosaccharide complex can effectively treat hyperhomocysteinemia mice, increase folate and 5-MTHF levels in blood and liver of hyperhomocysteinemia mice, and reduce plasma cysteine levels of hyperhomocysteinemia mice.
Description
Technical Field
The invention relates to probiotic and prebiotic combined fermented milk for treating hyperhomocysteinemia and a preparation method and application thereof, belonging to the technical field of microorganisms.
Background
Hyperhomocysteinemia (HHcy) is a common public health problem, especially in developing countries, with serious health problems and socioeconomic burdens. When Folic Acid (FA) is deficient in the body, impaired homocysteine metabolism in the blood triggers HHcy, which can cause a number of diseases, such as: hypertension, cerebral apoplexy, depression, Alzheimer's disease, diabetes and the like. Therefore, folic acid supplementation has become an important means of treating HHcy.
Humans lack the genes for folic acid synthesis themselves and can only supplement folic acid by either external ingestion or metabolic synthesis by microorganisms in the intestinal tract. The two main forms of folic acid supplement are two, namely natural folic acid supplement from dietary sources and chemically synthesized folic acid supplement.
More and more researches find that the harm of excessive intake of the synthetic folic acid is covered, such as vitamin B12 deficiency covering, liver dihydrofolate reductase activity changing, fetal methylenetetrahydrofolate polymorphism (associated with debilitating diseases) and the like. Compared with the natural folic acid, the natural folic acid has higher safety and can be absorbed and utilized by the body under the condition of the defect of metabolic function of the body.
The traditional method for supplementing natural folic acid is to take the folic acid from plant tissues, but the folic acid is extremely unstable, and 50% -90% of the folic acid in the plant tissues is lost after long-time cooking. Therefore, it is necessary to supplement natural folic acid by other ways after three meals a day.
The folic acid synthesized by the lactic acid bacteria has good safety, and the fermented food prepared from the lactic acid bacteria capable of producing the folic acid is edible without processing, has more stable folic acid content, simple process and lower cost, and can be used as an important source of natural folic acid.
However, folic acid produced by the existing folate-producing bacteria is unstable, easy to decompose, and low in bioavailability, and cannot reach the treatment amount of hyperhomocysteinemia, so that hyperhomocysteinemia cannot be treated, and therefore, a probiotic and prebiotics combined fermented milk for treating hyperhomocysteinemia, and a preparation method and application thereof are urgently needed.
Disclosure of Invention
The invention aims to solve the technical problem of providing probiotic and prebiotics combined fermented milk for treating hyperhomocysteinemia and a preparation method and application thereof.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a probiotic and prebiotic-bound fermented milk for the treatment of hyperhomocysteinemia comprising Lactobacillus plantarum KLDS 1.0317 and galactooligosaccharides; the concentration of the galacto-oligosaccharide is 2mg/100mL, and the polymerization degree is 5; the number of the lactobacillus plantarum KLDS 1.0317 is more than or equal to 1 multiplied by 108CFU/mL or more than or equal to 1X 108CFU/g。
Use of probiotics and prebiotics combined fermented milk for treating hyperhomocysteinemia in preparation of medicines, health products or foods for preventing and/or treating and relieving hyperhomocysteinemia.
The fermented milk is used for improving the folic acid level in the serum of homocysteinemia.
The fermented milk is used for improving the 5-MTHF level in the serum of homocysteinemia.
Use of the fermented milk for increasing folate levels in the liver of homocysteinemia.
Use of the fermented milk for increasing 5-MTHF levels in the liver of homocysteinemia.
Use of the fermented milk for reducing the level of homocysteine in the plasma of homocysteinaemia.
A preparation method of probiotic and prebiotic combined fermented milk for treating hyperhomocysteinemia comprises the following steps: preparing 12% w/v solution of skimmed milk with ultrapure water, adjusting pH to 7.2 with 1mol/L NaOH, pasteurizing at 95 deg.C for 5min, and cooling in ice water bath to below 42 deg.C; centrifuging the bacterial liquid cultured to logarithmic phase at 12000rpm for 10min at 4 ℃, discarding the supernatant, washing the thalli for 2 times by using 0.90% normal saline, then resuspending, adding the thalli into sterilized cow milk in an inoculation amount of 2%, uniformly mixing to obtain a cow milk culture medium, adding galacto-oligosaccharide into the cow milk culture medium, culturing at 37 ℃ for 24h, immediately performing ice bath, and cooling for later use to obtain fresh fermented milk; meanwhile, fresh fermented milk is taken and freeze-dried for later use.
The invention firstly evaluates the folic acid producing capability of lactobacillus plantarum KLDS 1.0317 in MRS and MRS containing galacto-oligosaccharide, and then researches the folic acid producing capability of lactobacillus plantarum KLDS 1.0317 and galacto-oligosaccharide in cow milk. The invention utilizes a BALB/c mouse model to carry out in vivo evaluation on the capability of the high-folate-producing strain combined fermented milk to relieve the folate deficiency state. The invention aims to screen a probiotic combination with high folic acid yield for treating hyperhomocysteinemia.
The invention provides probiotic and prebiotic combined fermented milk for treating hyperhomocysteinemia and a preparation method and application thereof, and the prebiotic and probiotic compound has the following beneficial effects: the lactobacillus plantarum KLDS 1.0317 and galactooligosaccharide compound can effectively treat mice with hyperhomocysteinemia, increase the levels of folic acid and 5-MTHF in blood and liver of the mice with hyperhomocysteinemia, and reduce the level of plasma cysteine of the mice with hyperhomocysteinemia.
The lactobacillus plantarum KLDS 1.0317 and galactooligosaccharide compound can reduce and improve the levels of folic acid and 5-methyltetrahydrofolic acid (5-MTHF) in serum and liver of a mouse, reduce the level of cysteine in blood and relieve hyperhomocysteinemia; galactooligosaccharides only slightly affect the above criteria to a certain extent. Compared with the galactooligosaccharide conjugate and the lactobacillus plantarum KLDS 1.0317, the lactobacillus plantarum KLDS 1.0317 can greatly improve the remission effect of single bacteria. Meanwhile, the research of orally taking the lactobacillus plantarum KLDS 1.0317 and the galactooligosaccharide compound by rats discovers that the concentration of homocysteine can be effectively reduced. The lactobacillus plantarum KLDS 1.0317 and galactooligosaccharide compound can be used for preparing functional food and medicines for relieving hyperhomocysteinemia and can be used for daily health care of hyperhomocysteinemia people.
Drawings
FIG. 1 is a graph showing the effect of galactooligosaccharides with different degrees of polymerization of the present invention on the high folate production of Lactobacillus plantarum KLDS 1.0317; wherein, the same letter indicates that the total folic acid content has no significant difference (P >0.05) among different groups, and different letters indicate that the total folic acid content has significant difference (P < 0.05);
FIG. 2 is a graph showing the effect of different concentrations of galactooligosaccharides on the folate production of Lactobacillus plantarum KLDS 1.0317 according to the present invention; wherein, the same letter indicates that the total folic acid content has no significant difference (P >0.05) among different groups, and different letters indicate that the total folic acid content has significant difference (P < 0.05);
FIG. 3 is a graph showing folate production of pure cow's milk, Lactobacillus plantarum KLDS 1.0317 and Lactobacillus plantarum KLDS 1.0317+ GOS complex according to the present invention; wherein, 1: pure cow milk; 2: lactobacillus plantarum KLDS 1.0317; 3: lactobacillus plantarum KLDS 1.0317+ GOS complex; the same letter indicates no significant difference of the total folic acid content among different groups (P >0.05), and different letters indicate significant difference (P < 0.05);
FIG. 4 is a graph showing the effect of Lactobacillus plantarum KLDS 1.0317 in combination with galacto-oligosaccharide on whole blood folate of hyperhomocysteinemia mice according to the present invention; wherein, 1: a folate-normal group; 2: a folate deficient group; 3: only GOS cow milk groups were added; 4: a lactobacillus plantarum ferment group for high folic acid yield; 5: lactobacillus plantarum and GOS fermentation group for high yield of folic acid; the same letter indicates no significant difference of the index among different groups (P >0.05), and different letters indicate significant difference (P < 0.05); a is a graph of FA content in whole mouse blood; b is a graph of the content of the mouse whole blood 5-MTHF; FIG. C is a graph showing the FA content in the liver of mice; FIG. D is a graph showing the 5-MTHF content in the liver of a mouse;
FIG. 5 is a graph showing the effect of Lactobacillus plantarum KLDS 1.0317 in combination with galactooligosaccharides on the plasma cysteine content in hyperhomocysteinemia mice according to the present invention; wherein, 1: a folate-normal group; 2: a folate deficient group; 3: only GOS cow milk groups were added; 4: a lactobacillus plantarum ferment group for high folic acid yield; 5: lactobacillus plantarum and GOS fermentation group for high yield of folic acid; the same letters indicate no significant difference in plasma Hcy concentration between different groups (P >0.05) and different letters indicate significant difference (P < 0.05).
Detailed Description
The present invention will be further described with reference to the accompanying drawings.
Example 1
In this example, Lactobacillus plantarum KLDS 1.0317, deposited at the university of agricultural culture Collection, northeast, was used. The galacto-oligosaccharides are derived from the company Felanthus. The prior art sources of lactobacillus plantarum KLDS 1.0317 in the present invention are: in vitro evaluation of the effect of ten strains of lactobacillus on immune cell activity [ J ] food industry science 2018, 01 (39): 312-317.
A probiotic and prebiotic-bound fermented milk for the treatment of hyperhomocysteinemia comprising Lactobacillus plantarum KLDS 1.0317 and galactooligosaccharides; the concentration of the galacto-oligosaccharide is 2mg/100mL, and the polymerization degree is 5; the number of the lactobacillus plantarum KLDS 1.0317 is more than or equal to 1 multiplied by 108CFU/mL。
The galacto-oligosaccharide GOS with different polymerization degrees (3, 4, 5, 6, 7 and 8) are respectively added into MRS liquid culture medium, then the lactobacillus plantarum KLDS 1.0317 is inoculated into different culture media according to the inoculation amount of 2 percent, the folic acid level is analyzed by using high performance liquid chromatography, and the influence of different oligosaccharide contents on the folic acid production level of the lactobacillus plantarum KLDS 1.0317 is observed, so that the optimal addition concentration is determined.
The content of folic acid in the fermentation liquid obtained by the lactobacillus plantarum KLDS 1.0317 is very high when the polymerization degree is 5, FA reaches 0.393 mu g/mL, and 5-MTHF reaches 0.223 mu g/mL, which is shown in figure 1. The results show that the folic acid in MRS fermented by lactobacillus plantarum KLDS 1.0317 is remarkably increased compared with oligosaccharides with other polymerization degrees, and the lactobacillus plantarum KLDS 1.0317 is more suitable for utilization with high folic acid yield when the polymerization degree is 5.
Adding galactooligosaccharides with good determined effect into MRS liquid culture medium according to different concentrations respectively to enable the mass concentration to be 0, 1%, 2%, 3%, 4% and 5%, then inoculating lactobacillus plantarum KLDS 1.0317 into different culture media according to the inoculation amount of 2%, inoculating lactobacillus plantarum KLDS 1.0317 into MRS liquid culture medium without any additives according to the inoculation amount of 2%, analyzing the folic acid level by utilizing high performance liquid chromatography, and observing the influence of different oligosaccharide contents on the folic acid production level of lactobacillus plantarum KLDS 1.0317, thereby determining the optimal addition concentration.
When the mass concentration of galacto-oligosaccharide is 2%, the content of folic acid in the obtained fermentation liquid is very high, FA reaches 0.760 mu g/mL, 5-MTHF reaches 0.360 mu g/mL, and the specific figure is 2. The results show that the folic acid in MRS fermented by Lactobacillus plantarum KLDS 1.0317 is remarkably increased compared with other mass concentrations of galactooligosaccharides, and the folic acid is more suitable for the utilization of Lactobacillus plantarum KLDS 1.0317 to achieve high folic acid yield when the mass concentration is 2% (2mg/100 mL).
Centrifuging the bacterial liquid at 12000rpm for 10min at 4 ℃, discarding the supernatant, washing the thalli for 2 times by using 0.90% physiological saline, adding cow milk, uniformly mixing to prepare bacterial suspension (>8.00lg CFU/mL), inoculating the bacterial suspension into ultra-high temperature sterilized (UHT) cow milk by using the inoculation amount of 2%, adding GOS (GOS), culturing at 37 ℃ for 24h, immediately performing ice bath, and cooling for later use.
Meanwhile, fresh fermented milk is taken for animal experiments for standby after freeze-drying.
The combination of lactobacillus plantarum KLDS 1.0317 with galacto-oligosaccharide compound fermented milk produced a very high folate content, reaching 0.810 μ g/mL, as shown in fig. 3. The results show that the content of the fermented lactic acid prepared by the lactobacillus plantarum KLDS 1.0317 and the galacto-oligosaccharide is obviously increased, the produced folic acid is natural and non-toxic, the product form has reducibility and is easier to absorb, and the product can replace the folic acid supplement chemically synthesized by the traditional process.
Table 1 below is the grouping case of fig. 3:
TABLE 1 packet case
Lactobacillus plantarum KLDS 1.0317 and galactooligosaccharide complex fermented milk was used to treat homocysteinemia animal model.
Experimental animals and grouping design:
first 30, 3 week old female BALB/c mice were randomized into 2 groups: the folic acid-deficient control group was fed with a folic acid-deficient feed, and the folic acid-normal control group was fed with a normal feed, and the feed formulations were shown in table 2 and provided by australian corporation of beijing, japan. After 14d (modeling period), 24 mice fed folate-deficient diet were randomized into 4 groups, and normal mice were continued to be fed normal diet, with the groups shown in table 3.
TABLE 2 mouse feed formulation
| Composition (I) | Folic acid deficient feed (g/kg) | Normal feed (g/kg) |
| Casein protein | 200 | 200 |
| L- |
3 | 3 |
| Corn starch | 397.486 | 397.486 |
| Maltodextrin | 132 | 132 |
| Sucrose | 100 | 100 |
| Cellulose, process for producing the same, and process for producing the same | 50 | 50 |
| Soybean oil | 70 | 70 |
| Choline bitartrate | 2.5 | 2.5 |
| AIN minerals | 35 | 35 |
| AIN vitamin (without folic acid) | 10 | 10 |
| BHT | 0.014 | 0.014 |
| |
0 | 0.002 |
TABLE 3 animal experiment grouping and feeding method
At the end of the experiment, whole blood from the venous plexus of the eyeball of each group of mice was collected, then the mice were sacrificed by cervical dislocation, and liver tissues of the mice were harvested on an ice bench. Centrifuging the blood sample at 3500r/min for 15min, collecting the supernatant, and freezing at-80 deg.C for blood index analysis. The liver tissue is taken out and then quickly rinsed in pre-cooled normal saline to remove blood, is frozen at medium speed in liquid nitrogen and is transferred to be frozen at minus 80 ℃ for storage, and then is prepared into liver homogenate to measure related indexes.
The lactobacillus plantarum KLDS 1.0317 and galactooligosaccharide compound fermented milk improves the levels of folic acid and 5-MTHF of serum and liver of a mouse with hyperhomocysteinemia:
and (4) sucking the supernatant into a 1.5m L centrifugal tube to obtain a serum sample, and determining the contents of folic acid and 5-MTHF in the serum and the liver according to a determination method provided by a corresponding test kit.
The concentrations of FA in whole mouse blood and liver are shown in A and C in FIG. 4. The whole blood FA concentration in the experimental group was significantly increased compared to group 2 (P <0.05), but not as much as the control group (group 1 was 2.85 times higher than group 2). Group 5 folate concentrations were significantly higher than groups 3 and 4 (P <0.05) than groups 3, 4 by a factor of 1.39 and 1.22, respectively. Whereas the folate concentration in the liver was not significantly different between groups 3 and 4 (P >0.05), the highest folate concentration was in fermentation group 5, second only to the folate normal group.
The concentrations of 5-MTHF in mouse whole blood and liver are shown in B and D in FIG. 4. The whole blood 5-MTHF concentration in the experimental group was significantly increased (P <0.05) compared to the group 2, but significantly lower than the control group. Group 5 works best, with the highest concentration of 5-MTHF, significantly higher than groups 3 and 4. While the concentration of 5-MTHF in whole blood was not significantly different between groups 3 and 4 (P > 0.05).
Research shows that the supplement of galactooligosaccharides and lactobacillus plantarum can improve the concentrations of folic acid and 5-MTHF in blood and liver, but the galactooligosaccharides can obviously promote the lactobacillus plantarum to produce folic acid, so that the combination effect is optimal.
Lactobacillus plantarum KLDS 1.0317 fermented milk with galacto-oligosaccharide complex reduced the cysteine content in the plasma of hyperhomocysteinemia mice:
the collected plasma samples were measured using a mouse Hcy assay kit (beijing sincerine bio ltd) and the procedure was performed according to the instructions.
At the end of 42d, the plasma Hcy content of the mice in each group is shown in FIG. 5. Compared to group 1, Hcy concentration in plasma was significantly increased (2.29 fold) in group 2 mice (P < 0.05). Compared to the folate deficient group, the Hcy concentration in plasma was significantly reduced (P <0.05) after supplementation of the milk containing GOS, the lactobacillus plantarum fermented milk with high folate, and the lactobacillus plantarum + GOS fermented milk with high folate. However, compared to the single treatment group, the milk fermentation group containing GOS reduced Hcy concentration in plasma more effectively with significant difference (P < 0.05).
Example 2
A probiotic and prebiotic-bound fermented milk for the treatment of hyperhomocysteinemia comprising Lactobacillus plantarum KLDS 1.0317 and galactooligosaccharides; the concentration of the galacto-oligosaccharide is 2mg/100mL, and the polymerization degree is 5; the number of the lactobacillus plantarum KLDS 1.0317 is more than or equal to 1 multiplied by 108CFU/g。
Use of probiotics and prebiotics combined fermented milk for treating hyperhomocysteinemia in preparation of medicines, health products or foods for preventing and/or treating and relieving hyperhomocysteinemia.
The fermented milk is used for improving the folic acid level in the serum of homocysteinemia.
The fermented milk is used for improving the 5-MTHF level in the serum of homocysteinemia.
Use of the fermented milk for increasing folate levels in the liver of homocysteinemia.
Use of the fermented milk for increasing 5-MTHF levels in the liver of homocysteinemia.
Use of the fermented milk for reducing the level of homocysteine in the plasma of homocysteinaemia.
A preparation method of probiotic and prebiotic combined fermented milk for treating hyperhomocysteinemia comprises the following steps: preparing 12% w/v solution of skimmed milk with ultrapure water, adjusting pH to 7.2 with 1mol/L NaOH, pasteurizing at 95 deg.C for 5min, and cooling in ice water bath to below 42 deg.C; centrifuging the bacterial liquid cultured to logarithmic phase at 12000rpm for 10min at 4 ℃, discarding the supernatant, washing the thalli for 2 times by using 0.90% normal saline, then resuspending, adding the thalli into sterilized cow milk in an inoculation amount of 2%, uniformly mixing to obtain a cow milk culture medium, adding galacto-oligosaccharide into the cow milk culture medium, culturing at 37 ℃ for 24h, immediately performing ice bath, and cooling for later use to obtain fresh fermented milk; meanwhile, fresh fermented milk is taken and freeze-dried for later use.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
The above description is only of the preferred embodiments of the present invention, and it should be noted that: it will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the principles of the invention and these are intended to be within the scope of the invention.
Claims (8)
1. Probiotic and prebiotic-bound fermented milk for the treatment of hyperhomocysteinemia comprising Lactobacillus plantarum KLDS 1.0317 and galactooligosaccharides; the concentration of the galacto-oligosaccharide is 2mg/100mL, and the polymerization degree is 5; the number of the lactobacillus plantarum KLDS 1.0317 is more than or equal to 1 multiplied by 108CFU/mL or more than or equal to 1X 108CFU/g。
2. Use of probiotic and prebiotic combined fermented milk for treating hyperhomocysteinemia according to claim 1 in the preparation of medicaments, health products or food for preventing and/or treating and relieving hyperhomocysteinemia.
3. Use according to claim 2, wherein the fermented milk is for increasing the serum folate level of homocysteinemia.
4. Use according to claim 2, wherein the fermented milk is for increasing the level of 5-MTHF in the serum of homocysteinemia.
5. Use according to claim 2, wherein the fermented milk is for increasing folate levels in the liver of homocysteinemia.
6. Use according to claim 2, wherein the fermented milk is for increasing 5-MTHF levels in the liver of homocysteinemia.
7. Use according to claim 2, wherein the fermented milk is for reducing the level of homocysteine in the plasma of homocysteinemia.
8. The method for preparing probiotic and prebiotic-bound fermented milk for the treatment of hyperhomocysteinemia according to claim 1, comprising the following steps: preparing 12% w/v solution of skimmed milk with ultrapure water, adjusting pH to 7.2 with 1mol/L NaOH, pasteurizing at 95 deg.C for 5min, and cooling in ice water bath to below 42 deg.C; centrifuging the bacterial liquid cultured to logarithmic phase at 12000rpm for 10min at 4 ℃, discarding the supernatant, washing the thalli for 2 times by using 0.90% normal saline, then resuspending, adding the thalli into sterilized cow milk in an inoculation amount of 2%, uniformly mixing to obtain a cow milk culture medium, adding galacto-oligosaccharide into the cow milk culture medium, culturing at 37 ℃ for 24h, immediately performing ice bath, and cooling for later use to obtain fresh fermented milk; meanwhile, fresh fermented milk is taken and freeze-dried for later use.
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Citations (2)
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| CN102099044A (en) * | 2008-05-16 | 2011-06-15 | 益生菌股份公司 | Use of probiotic bacteria for the treatment of hyperhomocysteinaemia |
| CN111529553A (en) * | 2020-05-28 | 2020-08-14 | 东北农业大学 | Application of plant lactobacillus capable of degrading tryptophan and tryptophan mixture |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102099044A (en) * | 2008-05-16 | 2011-06-15 | 益生菌股份公司 | Use of probiotic bacteria for the treatment of hyperhomocysteinaemia |
| CN111529553A (en) * | 2020-05-28 | 2020-08-14 | 东北农业大学 | Application of plant lactobacillus capable of degrading tryptophan and tryptophan mixture |
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| JIAN ZHANG等: "Screening of folate-producing lactic acid bacteria and modulatory effects of folate-biofortified yogurt on gut dysbacteriosis of folate-deficient rats", 《FOOD & FUNCTION》 * |
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