Lactobacillus reuteri capable of promoting mineral absorption and improving bone metabolism, and its products and applications

Abstract

The present invention belongs to the field of microbial technology, and specifically relates to a Lactobacillus reuteri that can promote mineral absorption and improve bone metabolism, and its products and applications. Lactobacillus reuteri LR-01, deposited with the General Microbiology Center of the China National Committee for the Administration of Microbiological Culture Collection, under the accession number CGMCC No. 17754, was deposited on May 10, 2019. The Lactobacillus reuteri LR-01 provided by the present invention can balance height and weight, promote physical development in children, enhance mineral absorption, increase mineral bioavailability, improve bone metabolism, and promote healthy bone development. It has broad application prospects in the preparation of products that promote growth and development and nutrient absorption.

Description

Technical Field

[0001] The present invention belongs to the technical field of microorganisms, and in particular relates to a Lactobacillus reuteri capable of promoting mineral absorption and improving bone metabolism, and a product and application thereof. Background Art

[0002] Children's growth and development is a continuous, phased process, influenced by genetics and the environment, and exhibits significant individual variation. Therefore, children's growth and development are of great concern not only to parents but also to the medical community. Minerals are essential elements for the human body, closely linked to the functioning of various systems and organs, and significantly impact metabolism, intellectual function, immune regulation, and other aspects of life. Minerals are also necessary for maintaining the body's acid-base balance and normal osmotic pressure. For example, calcium, phosphorus, and magnesium are the primary building blocks of bones and teeth. Certain physiological substances in the human body, such as hemoglobin and thyroxine, require the presence of iron and iodine for their synthesis. Magnesium promotes growth and development; phosphorus participates in overall energy regulation; and zinc deficiency can lead to slowed growth, short stature, indigestion, and poor appetite in children.

[0003] 25-(OH)D (25-hydroxyvitamin D), OC (bone R-hydroxyglutamate protein), and PINP (amino-terminal propeptide of type I procollagen) are common indicators of bone metabolism. 25-(OH)D is the primary form of vitamin D in the body and is essential for promoting cell growth and development. OC is primarily synthesized by osteoblasts and odontoblasts and regulates bone metabolism. PINP is a marker of bone formation, primarily formed in bone tissue, and reflects the intensity of bone metabolism. In-depth research on bone health has revealed that late childhood and adolescence are critical periods for bone accumulation, reaching 95% of peak bone mass during this period. Failure to reach peak bone mass by the end of adolescence not only impacts the child's growth and development but also significantly increases the risk of pathological fractures and osteoporosis in adulthood. Therefore, maintaining bone health during childhood is crucial.

[0004] As a live bacterial preparation, probiotics can regulate the intestinal microecology and improve the balance of the intestinal environment. They have a significant inhibitory effect on the growth and reproduction of pathogenic microorganisms, can enhance the body's immunity, and fight inflammation. They can also penetrate deep into the intestines, accelerate the proliferation and maturation of intestinal epithelial cells, and thus promote the absorption of nutrients. In addition, probiotics can synthesize enzymes, B vitamins, and proteins, and the lactic acid they produce is also beneficial for promoting the absorption of trace elements such as calcium, iron, and phosphorus, which has a good promoting effect on children's neurological development. For children, since childhood is the golden period of growth and development, and it is also a critical period for establishing a normal flora, appropriate and reasonable supplementation of probiotics can not only effectively reduce a series of diseases caused by flora disorders, but also improve children's immune metabolic function, which is of great significance to children's growth and development.

[0005] Therefore, obtaining a probiotic preparation that can effectively promote children's physical development and mineral absorption is a very promising solution. Summary of the Invention

[0006] The present invention addresses the problems existing in the prior art and provides a Lactobacillus reuteri strain, a product thereof, and its application, which can promote mineral absorption and improve bone metabolism. The strain can effectively promote children's physical development, enhance mineral absorption, improve bone metabolism levels, promote healthy bone development, and have a positive effect on children's growth and development and nutrient absorption.

[0007] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0008] A strain of Lactobacillus reuteri that promotes mineral absorption and improves bone metabolism Lactobacillus reuteri ) LR-01, the preservation unit of the Lactobacillus reuteri LR-01 is the General Microbiology Center of the China Culture Collection Administration, the preservation number is CGMCC No.17754, and the preservation time is May 10, 2019.

[0009] The present invention also provides a culture of Lactobacillus reuteri LR-01. The preservation number of the Lactobacillus reuteri LR-01 is CGMCC No.17754.

[0010] The present invention also provides a microbial preparation comprising the above-mentioned Lactobacillus reuteri LR-01 or the above-mentioned culture.

[0011] The present invention also provides a probiotic composition comprising the aforementioned Lactobacillus reuteri LR-01 or the aforementioned culture. The Lactobacillus reuteri LR-01 has good tolerance to gastric acid and bile salts, and thus can be used as a probiotic.

[0012] Preferably, the probiotic composition further comprises food-scientifically acceptable nutrients.

[0013] Preferably, the nutrients include at least one of dietary fiber, protein, lipids, minerals and vitamins.

[0014] The present invention also provides a medicine for promoting children's physical development, mineral absorption and improving bone metabolism, comprising the above-mentioned Lactobacillus reuteri LR-01 or the above-mentioned culture.

[0015] Preferably, the drug contains no less than 1×10 6 CFU / mL or 1×10 6 CFU / g of Lactobacillus reuteri LR-01.

[0016] Preferably, the medicine further comprises at least one of a drug carrier and a pharmaceutical excipient.

[0017] Preferably, the drug carrier comprises microcapsules, microspheres, nanoparticles or liposomes.

[0018] Preferably, the pharmaceutical excipients include erythritol, D-mannitol, pectin, sodium alginate, talc, sodium pyrophosphate, polydextrose, carrageenan, sodium ascorbate, L-malic acid, xylitol, citric acid, potassium citrate, sodium citrate, lactic acid, sodium lactate, sorbitol, sodium carbonate, vitamin C, vitamin E, ethanol, sodium acetate, stearic acid or dextrin.

[0019] Preferably, the dosage form of the drug is granules, capsules, tablets, pills or oral liquid.

[0020] The present invention also provides the use of the Lactobacillus reuteri LR-01 or the culture thereof in the preparation of medicines for promoting physical development, mineral absorption and improving bone metabolism.

[0021] Preferably, the promoting physical development, mineral absorption and improving bone metabolism includes balancing height and weight;

[0022] and / or, increase the levels of macronutrients calcium, phosphorus, magnesium, and trace minerals iron, copper, and zinc in children;

[0023] And / or, increasing the levels of bone metabolism markers 25-hydroxyvitamin D, bone R-hydroxyglutamate protein and type I procollagen amino-terminal propeptide in the blood.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The present invention provides Lactobacillus reuteri ( Lactobacillus reuteri LR-01 can balance height and weight, promote children's physical development, enhance mineral absorption, increase mineral bioavailability, and improve bone metabolism, thereby promoting healthy bone development, thereby achieving the purpose of promoting growth and development and nutrient absorption. It has broad application prospects in the preparation of products that promote growth and development and nutrient absorption.

[0026] Biological Deposit Description

[0027] A classification of Lactobacillus reuteri LR-01 is named: Lactobacillus reuteri ( Lactobacillus reuteri ); Depository unit: China General Microbiology Center of Culture Collection of Microorganisms, abbreviated as CGMCC; Deposit number: CGMCC No.17754; Deposit date: May 10, 2019; Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; The test result of this strain is survival. DETAILED DESCRIPTION

[0028] The present invention will be further described below in conjunction with specific examples, and the advantages and features of the present invention will become clearer as the description progresses. However, these examples are merely exemplary and do not constitute any limitation to the scope of the present invention. The methods, ingredients, and dosages involved in the following examples, unless otherwise specified, are conventional methods known to those skilled in the art. It is worth noting that the raw materials used in the present invention are all common commercially available products, among which MRS liquid culture medium was purchased from Beijing Luqiao Technology Co., Ltd.; Lactobacillus reuteri Tb8-1 was provided by the Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, Inner Mongolia Agricultural University.

[0029] Example 1

[0030] Lactobacillus reuteri ( Lactobacillus reuteri ) How to obtain LR-01:

[0031] Lactobacillus reuteri LR-01 was isolated from the intestine of healthy children in 2017. 0.5 mL of the sample was added to 5 mL of MRS liquid medium and anaerobically cultured at 37°C for 24 h to obtain the enriched sample. 0.5 mL of the enriched sample was added to 4.5 mL of sterile saline to obtain 10 -1 Then, 0.5 mL of the dilution was added to 4.5 mL of saline to obtain 10 -2 Dilution, follow this operation to obtain 10 -3 , 10 -4 , 10 -5 and 10 -6 Dilution: 100 μL of gradient dilution was applied to MRS solid medium, 10 -4 , 10 -5 and 10 -6 One plate per gradient was plated and anaerobically cultured at 37°C for 48 hours to obtain colonies. Colonies with typical characteristics of Lactobacillus reuteri on MRS solid medium were selected based on colony shape, size, edge, and transparency. Colonies were picked with an inoculating loop and streaked onto MRS solid medium. The plates were then anaerobically cultured at 37°C for 48 hours to obtain purified single colonies. The purified single colonies were inoculated into 5 mL of MRS liquid medium and cultured anaerobically at 37°C for 24 hours to obtain bacterial suspensions. The strains corresponding to the bacterial suspensions were numbered and Gram staining, bacterial species identification, physiological and biochemical experiments, and genomic analysis were performed according to the procedures described in the textbook Microbiology (edited by Shen Ping and Chen Xiangdong). A strain with typical characteristics of Lactobacillus reuteri was selected, resulting in strain LR-01.

[0032] Identification of bacterial species:

[0033] The strain LR-01 was taken and the genome of the strain LR-01 was extracted using a bacterial genome extraction kit. The universal 27F / 1492R primer pair was used to amplify the extracted genome of the strain LR-01 as a template to obtain the 16S rRNA of the strain LR-01. The 16S rDNA of the strain LR-01 was sequenced using the NCBI Blastn program. The results showed that this strain was Lactobacillus reuteri and was named Lactobacillus reuteri ( Lactobacillus reuteri ) LR-01, which was deposited in the General Microbiology Center of China Culture Collection Administration of Microorganisms with the deposit number CGMCC No.17754. The test result of the strain was survival.

[0034] Example 2

[0035] A Lactobacillus reuteri LR-01 bacterial powder is prepared using a conventional bacterial powder preparation method, comprising the following steps:

[0036] (1) Activation of bacteria

[0037] The strains stored at -40°C were inoculated into the corresponding liquid culture medium sterilized at 121°C for 15 minutes, and cultured anaerobically at 37°C for 18-24 hours. The activated strains were obtained by subculturing 1-2 times.

[0038] (2) Preparation of seed solution

[0039] The activated bacteria in step (1) were inoculated into MRS liquid culture medium and cultured anaerobically at 37°C until the pH value reached 4.5-4.8;

[0040] (3) Inoculation and fermentation

[0041] The seed liquid of step (2) was inoculated into MRS liquid culture medium at 1‰ and fermented for 18 h under controlled fermentation conditions; the fermentation conditions were controlled as follows: constant temperature culture at 30°C in the early stage of fermentation, and natural fermentation to a pH of 5.0; then the fermentation temperature was adjusted to constant temperature culture at 37°C, and the pH was controlled to be maintained at 6.0, maintaining anaerobic fermentation.

[0042] In the above process, pH is controlled by adding a neutralizing agent, and the neutralizing agent is NaOH.

[0043] The anaerobic condition was achieved by aerating nitrogen every two hours.

[0044] (4) Freeze drying

[0045] a. Bacteria concentration: The high-density fermentation broth was centrifuged at 12,000 g to concentrate the bacteria.

[0046] b. Add protective agent: Add 5 times the amount of protective agent solution to the bacterial concentrate; the protective agent solution composition is as follows: 14 kg of skim milk, 10 kg of lactose, 1.1 kg of vitamin C, 0.8 kg of sodium glutamate and 1000 L of distilled water.

[0047] c. Drying: The bacterial suspension after adding the protective agent is freeze-dried to obtain freeze-dried bacterial powder, and the total number of viable bacteria in the mixed bacterial powder is controlled to reach 1.0×10 11 CFU / g and above.

[0048] The total viable bacterial count of the Lactobacillus reuteri LR-01 powder is 1×10 11 CFU / g.

[0049] Example 3

[0050] A milk powder comprises milk powder and the probiotic powder of Lactobacillus reuteri LR-01 prepared in Example 2.

[0051] The preparation method comprises the following steps: first preparing milk powder by a conventional milk powder preparation process, adding probiotic powder of Lactobacillus reuteri LR-01 to the prepared milk powder, and uniformly mixing the two to obtain the milk powder.

[0052] The number of viable Lactobacillus reuteri LR-01 in the milk powder is not less than 1.0×10 6 CFU / g.

[0053] Example 4

[0054] A nut comprising nuts and the probiotic powder of Lactobacillus reuteri LR-01 prepared in Example 2.

[0055] The preparation method comprises the following steps: firstly preparing nuts by adopting a conventional nut preparation process, adding probiotic powder of Lactobacillus reuteri LR-01 to the prepared nuts, and uniformly mixing the two to obtain the nuts.

[0056] The number of viable Lactobacillus reuteri LR-01 in the nut is not less than 1.0×10 6 CFU / g.

[0057] Example 5

[0058] A crystal ball (Cruibobo), comprising a crystal ball and the probiotic powder of Lactobacillus reuteri LR-01 prepared in Example 2.

[0059] The preparation method comprises the following steps: preparing crystal balls of specific composition according to the composition requirements of crystal ball raw materials, adding probiotic powder of Lactobacillus reuteri LR-01 to the prepared crystal balls, and mixing the two evenly to obtain crystal balls.

[0060] The number of viable Lactobacillus reuteri LR-01 in the crystal sphere is not less than 1.0×10 6 CFU / g.

[0061] Example 6

[0062] A solid beverage powder comprises solid beverage powder and the probiotic powder of Lactobacillus reuteri LR-01 prepared in Example 2.

[0063] The preparation method comprises the following steps: preparing a solid beverage powder of a specific composition according to the composition requirements of the solid beverage powder raw material, adding probiotic powder of Lactobacillus reuteri LR-01 to the prepared solid beverage powder, and uniformly mixing the two to obtain the solid beverage powder.

[0064] The number of viable Lactobacillus reuteri LR-01 in the solid beverage powder is not less than 1.0×10 6 CFU / g.

[0065] Example 7

[0066] A meat floss comprises meat floss and the probiotic powder of Lactobacillus reuteri LR-01 prepared in Example 2.

[0067] The preparation method comprises the following steps: preparing meat floss according to conventional meat floss production process, adding probiotic powder of Lactobacillus reuteri LR-01 into the prepared meat floss, and uniformly mixing the two to obtain the meat floss.

[0068] The number of viable Lactobacillus reuteri LR-01 in the meat floss is not less than 1.0×10 6 CFU / g.

[0069] Example 8

[0070] A film candy comprises the film candy and the probiotic powder of Lactobacillus reuteri LR-01 prepared in Example 2.

[0071] The preparation method comprises the following steps: mixing raw materials of film candies, adding probiotic powder of Lactobacillus reuteri LR-01 during the raw material mixing process, uniformly mixing the raw materials and the probiotic powder of Lactobacillus reuteri LR-01, and then adopting a conventional film candy process to obtain the film candies.

[0072] The number of viable Lactobacillus reuteri LR-01 in the film candy is not less than 1.0×10 6 CFU / g.

[0073] Example 9

[0074] A compressed candy comprises the compressed candy and the probiotic powder of Lactobacillus reuteri LR-01 prepared in Example 2.

[0075] The preparation method comprises the following steps: mixing raw materials for tablet candies, adding probiotic powder of Lactobacillus reuteri LR-01 during the mixing process, uniformly mixing the raw materials and the probiotic powder of Lactobacillus reuteri LR-01, and then adopting a conventional tableting process to obtain the tablet candies.

[0076] The number of viable Lactobacillus reuteri LR-01 in the compressed candy is not less than 1.0×10 6 CFU / g.

[0077] Example 10

[0078] The invention discloses a fermented milk, which uses milk as a raw material and Lactobacillus reuteri LR-01 as a fermentation bacterium to ferment and obtain the fermented milk. The preparation process of the fermented milk is a conventional process.

[0079] The number of viable Lactobacillus reuteri LR-01 in the fermented milk is not less than 1.0×10 6 CFU / g.

[0080] Comparative Example 1

[0081] A Lactobacillus reuteri Tb8-1 bacterial powder is prepared in the same process as in Example 2, except that the bacterial strain used is Lactobacillus reuteri Tb8-1.

[0082] Experimental Example 1

[0083] 1. Experimental Subjects

[0084] A total of 150 children who underwent physical examinations at the Inner Mongolia People's Hospital between March 2023 and September 2023 were selected as research subjects. Inclusion criteria: (1) aged 3-12 years; (2) all were born at full term; (3) had no family history of genetic diseases.

[0085] Exclusion criteria: (1) those with a history of dystocia or perinatal asphyxia at birth; (2) those with chronic diseases, neurological diseases, or infectious diseases; (3) those with intellectual disabilities; (4) those taking long-term medications that affect bone metabolism; (5) those who dropped out midway or were lost to follow-up. All guardians of the study subjects provided informed consent and signed the informed consent form.

[0086] 2. Experimental Methods

[0087] 2.1 Grouping

[0088] 150 children were randomly divided into three groups: probiotic group A (n=53), probiotic group B (n=50), and control group (n=47). In group A, there were 32 males and 21 females with an average age of (7.47±1.59) years; in group B, there were 30 males and 20 females with an average age of (7.61±1.40) years; in the control group, there were 25 males and 22 females with an average age of (7.56±1.62) years. There was no statistically significant difference in the baseline data of the three groups ( p >0.05), which is comparable.

[0089] 2.2 Dosage

[0090] Probiotic Group A consumed Lactobacillus reuteri LR-01 powder prepared in Example 2 (10 billion CFU / day); Probiotic Group B consumed Lactobacillus reuteri Tb8-1 powder prepared in Comparative Example 1 (10 billion CFU / day) daily for 60 consecutive days. The control group received no intervention and instead received an equal amount of placebo maltodextrin. All study groups maintained adequate sleep, increased longitudinal exercise, and a balanced diet.

[0091] 2.3 Index detection

[0092] Height, weight, and body mass index (BMI) levels.

[0093] Mineral content: 3 mL of fasting venous blood was drawn from children before and after the intervention, and the blood was centrifuged at 5000 r / min for 20 min. The mineral levels of calcium, phosphorus, magnesium, iron, copper, and zinc were measured using an automatic biochemical analyzer.

[0094] Bone metabolism indicators: 5 mL of fasting venous blood was drawn from the three groups, and the serum levels of bone metabolism markers 25-hydroxyvitamin D (25-(OH)D), bone R-hydroxyglutamate protein (OC), and type I procollagen amino-terminal propeptide (PINP) were measured using a CS-600A automatic biochemical analyzer.

[0095] 3. Experimental Results

[0096] 3.1 Analysis of subjects’ physical development indicators

[0097] Before and 60 days after the probiotic intervention, the subjects' physical development indicators of height, weight, and BMI were statistically analyzed. The results, shown in Table 1, show no significant differences in height and weight among the three groups of children before the probiotic intervention. After the probiotic intervention, the children's height and weight increased compared to before the intervention, with those in the Probiotic A group exceeding those in the Probiotic B group. This suggests that the probiotic Lactobacillus reuteri LR-01 can balance height and weight, effectively promoting physical development and healthy growth in children.

[0098] Table 1 Comparison of physical development indicators of subjects

[0099]

[0100] Note: Compared with the control group, * express p <0.05; compared with before intervention, # express p <0.05.

[0101] 3.2 Analysis of mineral content in subjects’ blood

[0102] Statistical analysis of blood mineral levels was performed before and 60 days after the probiotic intervention. The results, shown in Table 2, show that the probiotic intake increased the levels of macronutrients (calcium, phosphorus, and magnesium) and trace elements (iron, copper, and zinc) in the children compared with the control group. Furthermore, the effect of Probiotic A was significantly superior to that of Probiotic B. It is speculated that the probiotic Lactobacillus reuteri LR-01 produces short-chain fatty acids, which, through their solubilizing effect, enhance mineral absorption and increase mineral bioavailability, potentially promoting growth and nutrient absorption in children.

[0103] Table 2 Analysis of mineral content in blood

[0104]

[0105] Note: Compared with the control group, * and ** Respectively p <0.05, p <0.01; compared with before intervention, # and ## Respectively p <0.05, p <0.01.

[0106] 3.3 Statistical analysis of subjects’ bone metabolism indicators

[0107] Statistical analysis was performed on the levels of bone metabolism markers 25-(OH)D, OC, and PINP before and 60 days after probiotic intervention. The results, shown in Table 3, showed no significant differences in 25-(OH)D, OC, and PINP levels among the three groups of children before probiotic intervention. After 60 days of probiotic intervention, 25-(OH)D, OC, and PINP levels changed to varying degrees, with the probiotic group showing significantly greater changes than the control group, and Probiotic A significantly outperforming Probiotic B. 25-(OH)D, OC, and PINP are common bone metabolism markers, and the increased levels after ingestion of the probiotic Lactobacillus reuteri LR-01 suggest that this probiotic can improve bone metabolism and promote healthy bone development.

[0108] Table 3 Comparison of bone metabolism indicators

[0109]

[0110] Note: Compared with the control group, ** and *** Respectively p <0.01 and p <0.001; compared with before intervention, # 、 ## and ### Respectively p <0.05, p <0.01 and p <0.001.

[0111] Experimental Example 2

[0112] 1. Experimental Animals

[0113] Forty Wistar rats, 5 weeks old, weighing 120–150 g, half male and half female, were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. They were housed in separate cages, two rats per cage, maintained at an ambient temperature of 22 ± 2°C, a humidity of 50–60%, a light / dark cycle of 12 h / 12 ​​h, and fed a standard rodent maintenance diet with regular bedding changes.

[0114] 2. Experimental Methods

[0115] 2.1 Grouping

[0116] After one week of adaptive feeding, the rats were randomly divided into a normal control group (P0), a low-dose group (P1), a medium-dose group (P2), and a high-dose group (P3), with 10 rats in each group.

[0117] 2.2 Dosage

[0118] Each rat in the P1 group was given 0.1 g of Lactobacillus reuteri LR-01 powder prepared in Example 2 (the total viable count of Lactobacillus reuteri LR-01 was 1×10 7 Each rat in the P2 group was given 0.1 g of Lactobacillus reuteri LR-01 powder prepared in Example 2 (the total viable count of Lactobacillus reuteri LR-01 was 2×10 8 CFU / day); each rat in the P3 group was given 0.1 g of Lactobacillus reuteri LR-01 powder prepared in Example 2 (the total viable count of Lactobacillus reuteri LR-01 was 1×10 9 CFU / day). 2 mL of sterile saline was used to fully suspend the bacterial powder to obtain bacterial solution. Rats in the P0 group were gavaged with an equal volume of sterile saline for 4 consecutive weeks, during which they were free to eat.

[0119] For three days prior to sampling, the rats' food intake was recorded daily, and feces were collected for 24 hours. The feces were placed in glass petri dishes pre-washed with deionized water and dried in a constant-temperature drying oven for 48 hours. Rats were fasted for 12 hours prior to sampling, but not water. For sampling, rats were anesthetized intraperitoneally with 2.5% sodium pentobarbital. Blood (3 mL) was collected from the abdominal aorta into a sterile glass tube. After standing at 4°C for 30 minutes, serum was separated by centrifugation at 3500 rpm for 10 minutes. The liver and femur were simultaneously collected, rinsed with deionized water, and blotted dry with filter paper. Serum and liver tissue were stored at -80°C, while the femur was oven-dried at 60°C until later use.

[0120] 2.3 Determination of mineral content

[0121] Approximately 0.3 g (accurate to 0.001 g) of liver, feed (wet weight), femur, and feces (dry weight) samples from each group of rats were weighed and placed in a 50-mL beaker. 20 mL of concentrated HNO3 was added overnight and heated and boiled the next day until yellow smoke disappeared. After cooling, 2 mL of perchloric acid was added and boiled until the solution was colorless. The ion concentrations in the nitrated solutions of each sample were detected using an inductively coupled plasma emission spectrometer, and the apparent absorbances of the minerals Ca, Fe, Zn, and Mg were calculated according to the following formulas.

[0122] Apparent absorption rate of minerals (%) = (mineral intake - mineral excretion) ÷ mineral intake × 100%.

[0123] 3. Experimental Results

[0124] 3.1 Changes in apparent absorption rate of minerals

[0125] The changes in the apparent absorption rate of minerals in each group of rats are shown in Table 4. It can be seen that the supplementation of probiotics Lactobacillus reuteri LR-01 can significantly improve the apparent absorption rate of Fe, Zn, Ca and Mg ( p<0.05 or p <0.01). Compared with the P0 group, the apparent absorption rates of Fe and Zn in the P1 group did not change significantly, while those of Ca and Mg increased significantly. The apparent absorption rates of Fe, Zn, Ca, and Mg in the P2 and P3 groups all increased significantly. This suggests that supplementation with the probiotic Lactobacillus reuteri LR-01 can increase the apparent absorption rates of Fe and Zn, although the differences were not significant. However, it has a significant effect on increasing the apparent absorption rates of Ca and Mg. Furthermore, the apparent absorption rates of Fe, Ca, and Mg gradually increased with increasing viable probiotic counts.

[0126] Table 4 Changes in apparent absorption rate of minerals in rats of each group

[0127]

[0128] Note: Compared with P0 group, * and ** Respectively p <0.05 and p <0.01.

[0129] 3.2 Changes in serum mineral content

[0130] The changes in serum mineral content in each group of rats are shown in Table 5. Supplementation of probiotic Lactobacillus reuteri LR-01 had a significant effect on increasing serum Fe, Zn and Ca contents ( p <0.05 or p <0.01), and there was a certain dosage effect, while the Mg content did not change significantly.

[0131] Table 5 Changes in serum mineral content in rats of each group

[0132]

[0133] Note: Compared with P0 group, * and ** Respectively p <0.05 and p <0.01.

[0134] 3.3 Changes in Mineral Content in the Liver

[0135] The changes in mineral content in the livers of rats in each group are shown in Table 6. Compared with the P0 group, the Fe and Zn contents in the livers of the P2 and P3 groups were significantly increased, while the Ca and Mg contents were significantly decreased. This indicates that supplementation with the probiotic Lactobacillus reuteri LR-01 significantly increased the Fe and Zn contents and decreased the Ca and Mg contents in the liver, promoting the transfer of Ca and Mg from the liver.

[0136] Table 6 Changes in mineral content in the liver of rats in each group

[0137]

[0138] Note: Compared with P0 group, * and ** Respectively p <0.05 and p <0.01.

[0139] 3.4 Changes in mineral content in femur

[0140] The changes in mineral content in the femur of rats in each group are shown in Table 7. Probiotic Lactobacillus reuteri LR-01 had a significant effect on increasing the Fe and Ca content in the femur ( p <0.05 or p <0.01), and has a certain dose effect, increasing the deposition of Fe, Ca and Mg in bones.

[0141] Table 7 Changes in mineral content in femur of rats in each group

[0142]

[0143] Note: Compared with P0 group, * and ** Respectively p <0.05 and p <0.01.

[0144] In summary, the intake of probiotic Lactobacillus reuteri LR-01 can balance height and weight, promote children's physical development, improve mineral absorption, increase mineral bioavailability, improve bone metabolism, promote healthy bone development, and have a positive effect on children's growth and development and nutrient absorption.

[0145] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A Lactobacillus reuteri that promotes mineral absorption ( Lactobacillus reuteri ) LR-01, characterized by, The preservation unit of the Lactobacillus reuteri LR-01 is the General Microbiology Center of the China Culture Collection Administration, the preservation number is CGMCC No.17754, and the preservation time is May 10, 2019.

2. A culture of Lactobacillus reuteri LR-01, characterized in that The preservation number of the Lactobacillus reuteri LR-01 is CGMCC No.17754.

3. A microbial preparation, characterized in that The method comprises the Lactobacillus reuteri LR-01 according to claim 1 or the culture according to claim 2.

4. A probiotic composition, characterized in that The method comprises the Lactobacillus reuteri LR-01 according to claim 1 or the culture according to claim 2.

5. The probiotic composition according to claim 4, characterized in that The probiotic composition further comprises food-scientifically acceptable nutrients, which include at least one of dietary fiber, protein, lipid, minerals and vitamins.

6. A drug for promoting physical development, mineral absorption and bone metabolism in children, characterized in that: The method comprises the Lactobacillus reuteri LR-01 according to claim 1 or the culture according to claim 2.

7. The medicine according to claim 6, characterized in that The number of viable bacteria in the drug is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g of Lactobacillus reuteri LR-01.

8. The medicine according to claim 7, characterized in that The medicine further comprises at least one of a drug carrier and a pharmaceutical excipient, and the dosage form of the medicine is granules, capsules, tablets, pills or oral liquid.

9. Use of the Lactobacillus reuteri LR-01 according to claim 1 or the culture according to claim 2 in the preparation of a drug for promoting physical development, mineral absorption and improving bone metabolism, characterized in that: The minerals are major elements calcium, phosphorus, magnesium, and trace elements iron, copper and zinc.

10. The use according to claim 9, characterized in that The promotion of physical development, mineral absorption and improvement of bone metabolism include balancing height and weight; and / or increasing the content of macroelements calcium, phosphorus, magnesium, and trace elements iron, copper and zinc in children's bodies; and / or increasing the levels of bone metabolism markers 25-hydroxyvitamin D, bone R-hydroxyglutamate protein and type I procollagen amino-terminal propeptide in the blood.

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