Preparation method of strontium-enriched lactobacillus plantarum powder

Abstract

The invention discloses a preparation method of strontium-enriched lactobacillus plantarum powder, and relates to a preparation method of probiotic powder. The invention aims to solve the problems that at present, people can only supplement strontium through drinking water and cannot realize continuous intake by combining with daily diet, and free strontium is relatively low in absorption rate in human intestinal tracts and difficult to meet nutritional requirements. The method comprises the following steps: 1, preparing a strontium-rich culture medium; 2, fermenting the lactobacillus plantarum; and 3, preparing bacterial powder. The strontium-rich lactobacillus plantarum developed by the invention has definite market rationality and technical feasibility, lactobacillus plantarum can be protected from oxidative damage in the processing and intestinal transfer processes, and the survival rate of viable bacteria is improved; in addition, the bone protection function of strontium and the bone protection function of strontium are complementary; the strontium-enriched lactobacillus plantarum powder prepared by the method has the psychological expectations of'natural sources' and'no additive burden ', can be used as a daily diet supplement, can also be adapted to subdivided scenes such as mothers and infants, fitness and health care, and is adapted to special crowds with lactose intolerance and the like.

Description

Technical Field

[0001] This invention relates to a method for preparing probiotic powder. Background Technology

[0002] Strontium is an essential trace element for the human body, with its physiological functions mainly manifested in two aspects: first, maintaining bone health. As a key component of bone mineralization, strontium can promote osteoblast activity and inhibit osteoclast proliferation. Studies have shown that adequate daily strontium intake can reduce the risk of osteoporosis by 28%-35%. Second, cardiovascular protection. Strontium can regulate vascular smooth muscle contraction and reduce vascular calcification, thus having an auxiliary effect on improving hypertension and arteriosclerosis. According to the "Chinese Dietary Reference Intakes (2023 Edition)," the recommended daily intake of strontium for adults is 2 mg, while children, adolescents, pregnant women, and the elderly need to increase it to 2.5-3 mg. However, data from a 2024 survey by the National Center for Disease Control and Prevention shows that the average daily strontium intake of urban and rural residents in my country is only 1.2-1.5 mg, with a more significant deficiency among people over 60 years old and residents in northern regions. Approximately 68% of the population did not meet the recommended standard, forming a widespread "strontium nutrient deficit." Currently, the main way people obtain strontium through food is through "natural high-strontium drinking water." This means that strontium can only be obtained through drinking water and cannot be combined with a daily diet for sustained intake. Furthermore, the absorption rate of free strontium in the human intestine is low, making it difficult to meet nutritional needs. Alternatively, strontium can be obtained by consuming agricultural products grown in high-strontium areas, but this method is geographically limited and requires high-strontium water sources or high-strontium soil. Summary of the Invention

[0003] The purpose of this invention is to address the problems that people can only supplement strontium through drinking water, which cannot be combined with daily diet to achieve continuous intake, and that the absorption rate of free strontium in the human intestine is low, making it difficult to meet nutritional needs. Therefore, this invention provides a method for preparing strontium-enriched Lactobacillus plantarum powder.

[0004] A method for preparing strontium-enriched Lactobacillus plantarum powder is specifically carried out according to the following steps:

[0005] I. Preparation of Strontium-rich culture medium:

[0006] ① Dissolve strontium citrate in distilled water to obtain a strontium citrate solution;

[0007] ② Dissolve the dry powder of MRS broth culture medium in strontium citrate solution, sterilize at 121°C, and cool naturally to room temperature to obtain strontium-rich culture medium, which should be stored under cold storage.

[0008] II. Fermentation by Lactobacillus plantarum:

[0009] ① Dissolve the dry powder of MRS broth culture medium in deionized water, sterilize at 121℃, and cool naturally to room temperature to obtain MRS broth culture medium, which should be stored under cold storage.

[0010] ② Inoculate Lactobacillus plantarum from the slant culture medium into MRS broth medium for activation; obtain activated Lactobacillus plantarum;

[0011] ③ Inoculate the activated Lactobacillus plantarum into strontium-rich medium, then incubate in an incubator at 30-40℃, sterilize at 121℃, let stand at room temperature, centrifuge, and collect the lower wet cells; wash with sterile physiological saline to obtain purified wet cells;

[0012] III. Preparation of bacterial powder:

[0013] The purified wet bacterial cells were dried in an oven at 40℃~50℃ to obtain strontium-rich Lactobacillus plantarum powder, which was then sealed and stored.

[0014] The principles and advantages of this invention:

[0015] I. *Lactobacillus plantarum* can improve the nutrient absorption environment by regulating the intestinal flora structure and repairing the integrity of the intestinal barrier. More importantly, *Lactobacillus plantarum* has a synergistic effect with mineral absorption. It can enhance the bioavailability of minerals by activating related signaling pathways, providing a microbial carrier basis for the efficient absorption of strontium. The development of strontium-enriched *Lactobacillus plantarum* in this invention has clear market rationality and technical feasibility. It can protect *Lactobacillus plantarum* from oxidative damage during processing and intestinal transport, and improve the survival rate of live bacteria. In addition, it complements the skeletal protection function of strontium.

[0016] Second, the strontium-rich Lactobacillus plantarum powder prepared by this invention meets the psychological expectations of "natural source" and "no added burden". It can be used as a daily dietary supplement, and can also be adapted to specific scenarios such as mother and baby, fitness, and health care. It is suitable for special groups such as lactose intolerance, and solves the application limitations of traditional probiotics. It has extremely high market potential. Detailed Implementation

[0017] Specific Implementation Method 1: This implementation method is a preparation method of strontium-enriched Lactobacillus plantarum powder, specifically completed according to the following steps:

[0018] I. Preparation of Strontium-rich culture medium:

[0019] ① Dissolve strontium citrate in distilled water to obtain a strontium citrate solution;

[0020] ② Dissolve the dry powder of MRS broth culture medium in strontium citrate solution, sterilize at 121°C, and cool naturally to room temperature to obtain strontium-rich culture medium, which should be stored under cold storage.

[0021] II. Fermentation by Lactobacillus plantarum:

[0022] ① Dissolve the dry powder of MRS broth culture medium in deionized water, sterilize at 121℃, and cool naturally to room temperature to obtain MRS broth culture medium, which should be stored under cold storage.

[0023] ② Inoculate Lactobacillus plantarum from the slant culture medium into MRS broth medium for activation; obtain activated Lactobacillus plantarum;

[0024] ③ Inoculate the activated Lactobacillus plantarum into strontium-rich medium, then incubate in an incubator at 30-40℃, sterilize at 121℃, let stand at room temperature, centrifuge, and collect the lower wet cells; wash with sterile physiological saline to obtain purified wet cells;

[0025] III. Preparation of bacterial powder:

[0026] The purified wet bacterial cells were dried in an oven at 40℃~50℃ to obtain strontium-rich Lactobacillus plantarum powder, which was then sealed and stored.

[0027] Specific Implementation Method Two: This implementation method differs from Specific Implementation Method One in that the concentration of the strontium citrate solution mentioned in step one ① is 50 mg / L to 150 mg / L. The other steps are the same as in Specific Implementation Method One.

[0028] Specific Implementation Method 3: This implementation method differs from Specific Implementation Method 1 or 2 in that the mass ratio of the MRS broth culture medium powder to the volume ratio of the strontium citrate solution in step 1, ② is (40g~70g):(800mL~1500mL). The other steps are the same as in Specific Implementation Method 1 or 2.

[0029] Specific Implementation Method Four: This implementation method differs from Specific Implementation Methods One to Three in that: the sterilization time in step one ② is 30-40 minutes; the refrigeration temperature in step one ② is 4°C. Other steps are the same as in Specific Implementation Methods One to Three.

[0030] Specific Implementation Method Five: This implementation method differs from Specific Implementation Methods One to Four in that: the mass ratio of the dry powder of MRS broth culture medium to the volume of deionized water in step two① is (40g~70g):(800mL~1500mL); the sterilization time in step two① is 30min~40min. Other steps are the same as in Specific Implementation Methods One to Four.

[0031] Specific Implementation Method Six: This implementation method differs from Specific Implementation Methods One to Five in that: the *Lactobacillus plantarum* mentioned in step two ② was purchased from the Shanghai Preservation Microbiology Center (Shanghai Preservation Microbiology Co., Ltd.); the inoculation amount of *Lactobacillus plantarum* mentioned in step two ② is 2% to 5%. Other steps are the same as in Specific Implementation Methods One to Five.

[0032] Specific Implementation Method Seven: The difference between this implementation method and Specific Implementation Methods One to Six is ​​that the activation time in step two ② is 12h~48h. The other steps are the same as in Specific Implementation Methods One to Six.

[0033] Specific Implementation Method Eight: This implementation method differs from Specific Implementation Methods One to Seven in that: the inoculation amount of activated *Lactobacillus plantarum* in step two, step three is 5% to 10%; and the culture in step two, step three is carried out in an incubator at 30 to 40°C for 30 to 60 hours. Other steps are the same as in Specific Implementation Methods One to Seven.

[0034] Specific Implementation Method Nine: This implementation method differs from Specific Implementation Methods One to Eight in that: the settling time in step two ③ is 15 min to 30 min; the centrifugation speed is 4500 r / min to 5000 r / min, and the centrifugation time is 10 min to 20 min. Other steps are the same as in Specific Implementation Methods One to Eight.

[0035] Specific Implementation Method Ten: This implementation method differs from Specific Implementation Methods One to Nine in that: the sterilization time in step two ③ is 30-40 minutes; and the number of times sterile saline is used for rinsing in step two ③ is 2-3 times. Other steps are the same as in Specific Implementation Methods One to Nine.

[0036] The beneficial effects of the present invention are verified using the following embodiments:

[0037] Example 1: A method for preparing strontium-enriched Lactobacillus plantarum powder, specifically completed according to the following steps:

[0038] I. Preparation of Strontium-rich culture medium:

[0039] ① Dissolve strontium citrate in distilled water to obtain a strontium citrate solution;

[0040] The concentration of the strontium citrate solution mentioned in step 1① is 100 mg / L;

[0041] ② Dissolve the dry powder of MRS broth culture medium in strontium citrate solution, sterilize at 121℃ for 30 min, cool naturally to room temperature to obtain strontium-rich culture medium, and store at 4℃.

[0042] The mass ratio of the dry powder of MRS broth culture medium mentioned in step 1② to the volume ratio of the strontium citrate solution is 50g:1000mL;

[0043] II. Fermentation by Lactobacillus plantarum:

[0044] ① Dissolve the dry powder of MRS broth culture medium in deionized water, sterilize at 121℃ for 30 min, and cool naturally to room temperature to obtain MRS broth culture medium, which should be stored under cold storage.

[0045] The mass ratio of the dry powder of MRS broth culture medium to the volume ratio of deionized water in step 2① is 50g:1000mL.

[0046] ② Inoculate *Lactobacillus plantarum* from the slant culture medium into MRS broth medium and activate for 12 hours; thus obtaining activated *Lactobacillus plantarum*.

[0047] The *Lactobacillus plantarum* mentioned in step 2② was purchased from the Shanghai Center for the Preservation of Microbial Cultures.

[0048] The inoculum size of *Lactobacillus plantarum* mentioned in step 2② is 5%;

[0049] ③ Inoculate the activated Lactobacillus plantarum into strontium-rich medium, then incubate at 35°C for 48 hours, sterilize at 121°C for 30 minutes, let stand at room temperature, centrifuge, and collect the lower wet cells; wash three times with sterile physiological saline to obtain purified wet cells;

[0050] The inoculation amount of activated Lactobacillus plantarum mentioned in step 2③ is 10%;

[0051] The settling time mentioned in step 2③ is 15 minutes; the centrifugation speed is 5000 r / min, and the centrifugation time is 10 minutes;

[0052] III. Preparation of bacterial powder:

[0053] The purified wet bacterial cells were dried in an oven at 40°C to obtain strontium-rich Lactobacillus plantarum powder, which was then sealed and stored.

[0054] The strontium-rich *Lactobacillus plantarum* powder prepared in Example 1 was nitrated with nitric acid, and the content of other ions was measured by atomic absorption spectrometry. The strontium content in each powder is shown in Table 1 below.

[0055] Table 1

[0056]

Claims

1. A method for preparing strontium-enriched Lactobacillus plantarum powder, characterized in that... The preparation method is specifically carried out according to the following steps: I. Preparation of Strontium-rich culture medium: ① Dissolve strontium citrate in distilled water to obtain a strontium citrate solution; ② Dissolve the dry powder of MRS broth culture medium in strontium citrate solution, sterilize at 121°C, and cool naturally to room temperature to obtain strontium-rich culture medium, which should be stored under cold storage. II. Fermentation by Lactobacillus plantarum: ① Dissolve the dry powder of MRS broth culture medium in deionized water, sterilize at 121℃, and cool naturally to room temperature to obtain MRS broth culture medium, which should be stored under cold storage. ② Inoculate Lactobacillus plantarum from the slant culture medium into MRS broth medium for activation; Activated Lactobacillus plantarum was obtained; ③ Inoculate the activated Lactobacillus plantarum into strontium-rich medium, then incubate in an incubator at 30-40℃, sterilize at 121℃, let stand at room temperature, centrifuge, and collect the lower wet cells; wash with sterile physiological saline to obtain purified wet cells; III. Preparation of bacterial powder: The purified wet bacterial cells were dried in an oven at 40℃~50℃ to obtain strontium-rich Lactobacillus plantarum powder, which was then sealed and stored.

2. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The concentration of the strontium citrate solution mentioned in step 1① is 50 mg / L to 150 mg / L.

3. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The mass ratio of the dry powder of MRS broth culture medium to the volume ratio of the strontium citrate solution in step 1② is (40g~70g):(800mL~1500mL).

4. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The sterilization time mentioned in step 1② is 30 min to 40 min; the refrigeration temperature mentioned in step 1② is 4℃.

5. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The mass ratio of the dry powder of MRS broth culture medium to the volume ratio of deionized water in step 2① is (40g~70g):(800mL~1500mL); the sterilization time in step 2① is 30min~40min.

6. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The inoculation amount of *Lactobacillus plantarum* mentioned in step 2② is 2%~5%.

7. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The activation time mentioned in step 2② is 12h~48h.

8. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The inoculation amount of activated Lactobacillus plantarum in step 2③ is 5%~10%; in step 2③, it is cultured in an incubator at 30~40℃ for 30h~60h.

9. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The settling time mentioned in step 2③ is 15min~30min; the centrifugation speed is 4500r / min~5000r / min, and the centrifugation time is 10min~20min.

10. The method for preparing a strontium-enriched Lactobacillus plantarum powder according to claim 1, characterized in that... The sterilization time mentioned in step 2③ is 30 min to 40 min; the number of times sterile saline is used for rinsing in step 2③ is 2 to 3 times.

Images