CN115948296B - Lactobacillus plantarum for improving musculoskeletal health and/or improving exercise capacity, and composition and application thereof - Google Patents
Lactobacillus plantarum for improving musculoskeletal health and/or improving exercise capacity, and composition and application thereof Download PDFInfo
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- CN115948296B CN115948296B CN202211672215.9A CN202211672215A CN115948296B CN 115948296 B CN115948296 B CN 115948296B CN 202211672215 A CN202211672215 A CN 202211672215A CN 115948296 B CN115948296 B CN 115948296B
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Abstract
Lactobacillus plantarum (Lactobacillus plantarum) LP550 is preserved in China center for type culture collection of Wuhan China, with the preservation number of CCTCC NO: M2021437 in 4 months of 2021. The composition comprises the metazoan of the lactobacillus plantarum LP550, or the composition also comprises the live strain of the lactobacillus plantarum LP550 and the inactivated strain, the strain metabolite and the vitamin D thereof 2 Vitamin D 3 A mixture of one or more of lactobacillus rhamnosus and lactobacillus fermentum. The invention also discloses the application of the composition containing lactobacillus plantarum and its metacrylic acid in improving muscle and bone health and/or improving exercise capacity. Lactobacillus plantarum LP550 pair of the inventionD 2 And vitamin D 3 Has extremely strong metabolic capability, can obviously improve skeletal muscle mass index, limb skeletal mass index, whole body bone density, lumbar vertebra bone density, forelimb gripping power and exercise capability, and can improve musculoskeletal health and exercise capability through various ways.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to lactobacillus plantarum for improving muscle and bone health and improving exercise capacity, and a composition and application thereof.
Background
Vitamin D is a fat-soluble vitamin, and exists in two forms in nature, namely vitamin D 2 (ergosterol) and vitamin D 3 (cholecalciferol). Plant-derived vitamin D 2 And vitamin D 3 Cannot be synthesized in human body, and can be supplemented from diet, such as marine products, yeast, plant extracts, etc. With the diligent research in the field of vitamin D, it has been found so far that vitamin D metabolites play an important role in the human body, and the main function of vitamin D is to promote the absorption of calcium and phosphorus by small intestinal mucosal cells. Nature Communications, a cross-sectional study published in 567 elderly men, conducted an in-depth analysis of serum vitamin D metabolites, found that 8 specific intestinal bacterial taxa correlated with vitamin D active form-1, 25-dihydroxyvitamin D levels, and higher levels of 1, 25-dihydroxyvitamin D correlated with intestinal microbial diversity, suggesting that intestinal flora would alter intestinal vitamin D metabolism (Kado D, et al Vi tamin D Metabolites and the Gut Microbiome in Older Men[J].Innovation in Aging,2020.)。
In 2009, binkley et al proposed the concept of sarcopenia (OS) based on the pathophysiological basis and close correlation of similar sarcopenia and osteoporosis, mainly referring to patients who met the diagnostic criteria of osteoporosis and had a simultaneous decrease in muscle mass/function. OS patients show a decrease in bone density (BMD), an increase in bone fragility, a progressive and systemic decrease in muscle mass, muscle strength and muscle function, leading to an increased risk of weakness, falls, fractures, hospitalization, disability and mortality in the patient.
According to different diagnostic criteria for sarcopenia, the incidence of sarcopenia in people over 60 years old ranges from 5% to 37%, even with a tendency to younger. At present, no targeted medicine for preventing and treating the sarcopenia syndrome exists, the effective prevention and treatment means at present comprise traditional Chinese medicine treatment mainly for strengthening the spleen and tonifying the kidney, for example, CN202110982828.1 discloses a traditional Chinese medicine composition for preventing and treating sarcopenia osteoporosis, the composition comprises traditional Chinese medicines or extracts such as eucommia ulmoides, and the like, and in addition, early intervention measures such as resistance and balance training, nutrition intake (supplementing vitamin D and calcium) and the like can also effectively prevent and relieve the sarcopenia symptom. US15385786 discloses that lactobacillus plantarum compositions are capable of improving muscle mass and endurance, reducing blood lactic acid and improving exercise capacity.
In the prior art, vitamin D is directly compounded with probiotics to intervene related diseases, such as vitamin D published by Chen Dan 3 The combination of lactobacillus rhamnosus and P40 has the protective effect on enteritis and the mechanism research thereof, but no research report on the muscular-bone co-reduction syndrome and the improvement of the exercise capacity by the interaction of a specific strain with vitamin D and metabolites thereof is seen, so that the novel strain and the composition thereof are developed for preparing the novel strain which is convenient and efficient and has important social significance and market value for improving the muscular-bone health and the exercise capacity of organisms.
Disclosure of Invention
The invention aims to solve the technical problems that: overcomes the defects of the prior art, provides a preparation which has high biological activity, wide carbon source utilization and canHigh-efficiency metabolism of vitamin D 2 Or and vitamin D 3 The invention also provides a composition consisting of the lactobacillus plantarum and its metazoan, and provides a new application of the lactobacillus plantarum and the composition consisting of the lactobacillus plantarum in improving the muscle and bone health and the exercise ability.
One of the technical schemes adopted for solving the technical problems is as follows:
lactobacillus plantarum, designated Lactobacillus plantarum (Lactobacillus plantarum) LP550, was deposited at the China center for type culture collection of Wuhan, china, with a deposit number of CCTCCNO: M2021437, at month 23 of 2021.
Biological preservation description: lactobacillus plantarum (Lactobacillus plantarum) LP550, deposited at the chinese collection of typical cultures, accession number: eight-path 299 of Wuchang district in Wuhan, hubei province, the preservation organization is abbreviated as: cctccc, date of preservation: 2021, 4, 23 (2021, 4, 23 received registration book, 4, 30 detected as viable and preserved), biological preservation number cctcccno: m2021437, strain number: lactobacillus plantarum LP550.
Wherein the lactobacillus plantarum LP550 is isolated from a plant source, in particular, the lactobacillus plantarum LP550 is obtained by screening and separating from Pi county bean cotyledon of the farmer family of the Cheng Co.
The biological properties of lactobacillus plantarum LP550 of the invention are as follows:
1) Morphological features: the colony grows well on MRS agar culture medium, is round, medium in size, milky white, convex and glossy upwards, has tidy colony edges, is easy to pick, is rod-shaped under a microscope, and is positive after gram staining.
2) Biological identification: the lactobacillus plantarum LP550 is sent to China center for identification of 16SrRNA, the 16S rRNA gene sequence of the lactobacillus plantarum LP550 is shown as SEQ ID NO:1, NCBI BLAST comparison is carried out on the 16S rRNA gene sequence of the lactobacillus plantarum LP550, a Neighbor-Joining phylogenetic tree constructed by taking Holzapfelia floricola Ryu1-2 (AB 523780) as an outer branch is based on the 16S rRNA gene sequence comparison result, and the similarity with lactobacillus plantarum (Lactobacillus plantarum) in Genebank is larger than 99 percent as shown in figure 2, and the strain is identified as lactobacillus plantarum (Lactobacillus plantarum) LP550.
The culture medium for fermenting lactobacillus plantarum LP550 is MRS and vitamin D 2 +MRS Medium or vitamin D 3 +MRS Medium, the vitamin D 2 The +MRS culture medium comprises peptone 10.0g/L, beef powder 5.0g/L, yeast powder 4.0g/L, glucose 20.0g/L, tween 80 1.0g/L, K 2 HPO 4 ·7H 2 O2.0 g/L, anhydrous sodium acetate 5g/L, diammine citrate 2.0g/L, mgSO 4 ·7H 2 O 0.2g/L，MnSO 4 ·H 2 O0.038 g/L, 1000-4000 IU vitamin D 2 (agar powder 15g/L is added as a solid culture medium); the vitamin D 3 The +MRS culture medium comprises peptone 10.0g/L, beef powder 5.0g/L, yeast powder 4.0g/L, glucose 20.0g/L, tween 80 1.0g/L, K 2 HPO 4 ·7H 2 O2.0 g/L, anhydrous sodium acetate 5g/L, diammine citrate 2.0g/L, mgSO 4 ·7H 2 O 0.2g/L，MnSO 4 ·H 2 O0.038 g/L, 1000-4000 IU vitamin D 3 (agar powder 15g/L is added as solid culture medium), preferably vitamin D 3 Vitamin D in+MRS Medium 3 The growth of lactobacillus plantarum LP550 is obviously promoted, the acid production level is equivalent to that of a conventional MRS culture medium, and meanwhile, the increase of short-chain fatty acid can be promoted, and particularly, the butyric acid content playing an important role in intestinal myoaxis and intestinal bone axis is obviously improved; in addition, vitamin D 3 Can promote and improve the antibacterial capability of lactobacillus plantarum LP550 to pathogenic microorganisms, and has antibacterial effects on helicobacter pylori ATCC 26695, streptococcus mutans CGMCC 1.2499, staphylococcus aureus CMCC 26003, clostridium perfringens ATCC 13124 and candida albicans ATCC 10231.
The other technical scheme adopted by the invention for solving the technical problems is as follows:
a composition comprises Lactobacillus plantarum LP550 metazoan obtained by culturing Lactobacillus plantarum LP550 metazoan in MRS medium or vitamin D 2 +MRS Medium or vitamin D 3 Concentrating fermentation broth obtained by fermenting and culturing lactobacillus plantarum in +MRS culture medium, spray drying to obtain thallus and its generationSolid powders of the product, optionally containing vitamin D 2 Vitamin D 3 。
Further, the composition also comprises the live strain of the lactobacillus plantarum LP550, an inactivated strain of the lactobacillus plantarum LP550, a strain metabolite and vitamin D 2 Vitamin D 3 A mixture of one or more of lactobacillus rhamnosus, preferably applicant-selected lactobacillus rhamnosus S24, and lactobacillus fermentum, preferably applicant-selected lactobacillus fermentum GF1800.
The composition includes but is not limited to biological agents, functional foods, health care products or medicines.
The functional food is any one of ferment, pickle, solid beverage, pill, tablet or microcapsule crystal ball.
The medicament also contains a pharmaceutically acceptable carrier.
The pharmaceutically acceptable carriers include, but are not limited to: one or more of a filler, binder, wetting agent, disintegrant, or lubricant.
The illustrated medicament is a medicament for improving musculoskeletal health and improving exercise capacity, preferably an oral medicament.
The filler is one or more of fucoidin, trehalose, lactose, chitosan, starch or dextrin; the adhesive is one or more of liquid glucose, starch paste or syrup; the wetting agent is one or more of glycerol or ethanol; the disintegrating agent is one or more of crospovidone, sodium carboxymethyl starch or sodium cross-linked carboxymethyl starch; the lubricant is one or more of silicon dioxide magnesium stearate or sodium stearate fumarate.
The lactobacillus plantarum LP550 and the metaplasia of the LP550 can obviously improve the whole bone density and lumbar bone density of an OS rat, thereby improving the muscle bone health or preventing the formation of muscle bone co-reduction (OS).
The lactobacillus plantarum LP550 and the metaplasia of the LP550 can obviously improve the forelimb grabbing force, the whole body skeletal muscle quantity and the limb skeletal muscle quantity of an OS rat, and realize the improvement of musculoskeletal health or the effective prevention and treatment of Sarcopenia (SP).
The lactobacillus plantarum LP550 can also obviously prolong the running time and running distance of the rats, further improve the movement capacity of the organism and further prevent and treat the sarcopenia OS.
Lactobacillus plantarum LP550 can also promote increasing of the butyric acid content in the feces of OS rats, thereby achieving the purpose of preventing and treating the sarcopenia OS.
Based on the above-mentioned characteristics of the lactobacillus plantarum LP550 of the invention, one of the applications of the lactobacillus plantarum of the invention and its metants is:
use of lactobacillus plantarum LP550 metazoan for the preparation of a functional food or medicament for improving musculoskeletal health and improving exercise capacity; alternatively, the use of a composition formed from one or more of Lactobacillus plantarum LP550 metazoan, lactobacillus plantarum LP550, an inactivated strain of Lactobacillus plantarum LP550 and a metabolite of the strain for the manufacture of a functional food or medicament for improving musculoskeletal health and improving exercise capacity.
In one exemplary embodiment, the amount of Lactobacillus plantarum LP550 is greater than or equal to 6X 10 8 CFU/per mouse/day, the amount of inactivated strain of Lactobacillus plantarum LP550 is 3-8X10 10 CFU/day, the amount of the metazoan of the Lactobacillus plantarum LP550 is more than or equal to 6 multiplied by 10 8 The number per mouse/day, free amino acid > 1mg/g.
Secondly, bacteriostasis experiments show that: lactobacillus plantarum LP550 has a broad antibacterial activity against pathogenic bacteria. The specific expression is that the composition has good inhibition effect on helicobacter pylori ATCC 26695, streptococcus mutans CGMCC1.2499, staphylococcus aureus CMCC26003, clostridium perfringens ATCC13124 and candida albicans ATCC10231, wherein the inhibition effect on streptococcus mutans CGMCC1.2499 is strongest, and vitamin D is adopted 2 -MRS and vitamin D 3 Lactobacillus plantarum LP550 cultured in MRS medium has stronger bacteriostasis to pathogenic bacteria than Lactobacillus plantarum LP550 cultured in conventional MRS medium.
Based on the above-mentioned properties of lactobacillus plantarum LP550, another application of the lactobacillus plantarum of the invention and its metants:
use of lactobacillus plantarum LP550 and its progeny in the preparation of a functional food or medicament for inhibiting pathogenic bacteria.
The lactobacillus plantarum LP550 has very good tolerance to gastric acid and bile salts, and is suitable for oral administration.
Lactobacillus plantarum LP550 has a broad-spectrum carbon source utilization capacity and is resistant to vitamin D 2 And vitamin D 3 Has strong metabolic capability, strong acid production capability, rapid acid production in the initial stage of fermentation, large acid production amount, and stable acid production amount along with the increase of fermentation time. In particular, lactobacillus plantarum LP550 is found in vitamin D 2 The growth performance and the acid-producing capacity of the +MRS culture medium are greatly improved on the conventional MRS culture medium, and the vitamin D 3 The +MRS medium only has a promoting effect on the growth of the lactobacillus plantarum LP550, and has the acid generating capacity equivalent to that of the MRS medium, and based on the characteristics, the lactobacillus plantarum and the other application of the metazoan are:
Use of lactobacillus plantarum LP550 and its metazoan as a starter in the preparation of a fermented food, a health food or a dietary supplement.
The lactobacillus plantarum has the beneficial effects that:
the Lactobacillus plantarum LP550 is cultured on MRS agar and vitamin D 2 +MRS medium and vitamin D 3 The +MRS culture medium has good growth, high biological activity, wide carbon source utilization capacity, good acid production property and good tolerance to artificial gastric juice, intestinal juice and bile salt.
The lactobacillus plantarum LP550 is capable of metabolizing and utilizing vitamin D 2、 Vitamin D 3 Thereby improving the growth performance and the antibacterial effect and vitamin D 2 Also can promote the acidogenesis of lactobacillus plantarum LP 550.
The lactobacillus plantarum LP550 and the metazoan group can obviously improve the skeletal muscle mass index, the limb skeletal mass index, the whole body bone density, the lumbar vertebra bone density, the front limb holding power and the exercise capacity, increase the butyric acid content in excrement, improve the muscle bone health and the exercise capacity through various ways, and further delay or improve the muscle bone co-reduction syndrome.
The lactobacillus plantarum LP550 has good safety, and the lactobacillus plantarum LP550 has wide application, can be used for preparing functional foods or medicines for improving muscle and bone health and improving exercise capacity, or functional foods or medicines for inhibiting pathogenic bacteria, or can be used as a starter for preparing fermented foods and health-care foods.
Drawings
FIG. 1 shows colony morphology of Lactobacillus plantarum LP550 on MRS agar medium, wherein (A) plate colony morphology is shown and (B) microscopic strain morphology is shown;
FIG. 2 is a phylogenetic tree of Lactobacillus plantarum LP 550;
FIG. 3 shows Lactobacillus plantarum LP550 and other strains versus vitamin D 2 Vitamin D 3 A comparative analysis chart of the metabolic capacity of (a);
fig. 4 shows vitamin D 2 Vitamin D 3 Influence on the growth performance of lactobacillus plantarum LP 550;
fig. 5 shows vitamin D 2 Vitamin D 3 Influence on the acidogenic properties of lactobacillus plantarum LP 550;
FIG. 6 is a graph showing the comparison analysis of the whole body bone density and lumbar bone density of rats in different experimental groups; wherein, (A) the bone density of the whole body of the rat and (B) the lumbar bone density of the rat;
FIG. 7 is a graph of comparative analysis of forelimb grip of rats from different experimental groups;
FIG. 8 is a graph of comparative analysis of SMI (A) and RSMI (B) for rats from different experimental groups;
FIG. 9 is a graph of comparative analysis of the running time (A) and distance (B) of rats from different experimental groups;
FIG. 10 is a graph showing the comparison of butyric acid content in the feces of rats of different experimental groups.
Detailed Description
The invention is further described below with reference to the drawings and examples.
The formula of the culture medium in each embodiment of the invention comprises the following components:
MRS medium (Lactobacillus plantarum LP 550): 10.0g/L peptone, 5.0g/L beef powder, 4.0g/L yeast powder, 20.0g/L glucose, 1.0g/L Tween 80, K 2 HPO 4 ·7H 2 O2.0 g/L, anhydrous sodium acetate 5g/L, diammine citrate 2.0g/L, mgSO 4 ·7H 2 O 0.2g/L，MnSO 4 ·H 2 O0.038 g/L, (15 g/L of agar powder is added as a solid culture medium).
Vitamin D 2 +MRS Medium (Lactobacillus plantarum LP 550) (abbreviated as VD) 2 +mrs): 10.0g/L peptone, 5.0g/L beef powder, 4.0g/L yeast powder, 20.0g/L glucose, 1.0g/L Tween 80, K 2 HPO 4 ·7H 2 O2.0 g/L, anhydrous sodium acetate 5g/L, diammine citrate 2.0g/L, mgSO 4 ·7H 2 O 0.2g/L，MnSO 4 ·H 2 O0.038 g/L,4000IU vitamin D 2 (agar powder 15g/L is added as solid culture medium).
Vitamin D 3 +MRS Medium (Lactobacillus plantarum LP 550) (abbreviated as VD) 3 +mrs): 10.0g/L peptone, 5.0g/L beef powder, 4.0g/L yeast powder, 20.0g/L glucose, 1.0g/L Tween 80, K 2 HPO 4 ·7H 2 O2.0 g/L, anhydrous sodium acetate 5g/L, diammine citrate 2.0g/L, mgSO 4 ·7H 2 O 0.2g/L，MnSO 4 ·H 2 O0.038 g/L,4000IU vitamin D 3 (agar powder 15g/L is added as solid culture medium).
Example 1
The lactobacillus plantarum LP550 (Lactobacillus plantarumLP) is preserved in China center for type culture Collection, with the preservation number of 2021, 4 months and 23 days: cctccc No. M2021437.
1) Isolation, screening and gastric acid and bile salt resistance determination of lactobacillus plantarum LP550
Microorganism screening samples are collected from Pi county bean cotyledon of farm in Sichuan Cheng, the collected samples are sheared, 1g of the samples are weighed, the samples are placed into 9mL of sterile physiological saline, and after full shaking and uniform mixing, 10-time gradient dilution is carried out, the samples are coated in MRS solid culture medium, and the samples are cultured for 48 hours at 37 ℃. Visually observing, picking single colonies with different forms and sizes in the culture medium, and repeatedly streaking, purifying and culturing; then the strain is preliminarily determined to be lactobacillus by gram staining and calcium dissolving method, and the purified strain is stored in a refrigerator at-80 ℃ for standby by 45% glycerol.
a) Morphological observation
Purified lactobacillus plantarum LP550 was streaked on MRS agar medium, and after culturing for 48 hours at 37 ℃ in an inverted manner, colony morphology of the strain was observed by electron microscopy, and the result is shown in FIG. 1: the strain grows well on MRS agar culture medium, and the bacterial colony is round, medium in size, milky white, upward convex in luster, relatively neat in edge of the bacterial colony, easy to pick, rod-shaped under a microscope and positive after gram staining.
b) Molecular biology identification of strains
The purified strain is sent to China center for 16S rRNA identification, NCBI BLAST comparison is carried out on the measured 16S rRNA sequence, the similarity with lactobacillus plantarum in Genebank is greater than 99%, and the strain can be primarily identified as lactobacillus plantarum (Lactobacillus plantarum). The 16S rRNA identification sequence of the strain is shown as SEQ ID NO. 1, and is named as lactobacillus plantarum LP550 (Lactobacillus plantarumLP), and the lactobacillus plantarum LP550 is a Neighbor-Joing phylogenetic tree constructed by taking Holzapfelia floricola Ryu1-2 (AB 523780) as an outer branch based on the 16S rRNA gene sequence comparison result, as shown in figure 2.
2) Comparative test of vitamin D metabolism by Strain
Activating candidate strains of lactobacillus plantarum LP550, lactobacillus plantarum 360, lactobacillus plantarum 220, lactobacillus plantarum M71, M72, lactobacillus fermentum GF1800 and lactobacillus paracasei S6 in MRS culture medium twice, inoculating the activated lactobacillus plantarum LP550 and bacterial liquids of other strains into vitamin D according to an inoculum size of 5 percent 2 +MRS medium, vitamin D 3 In +MRS culture medium, mixing, and packaging into sterile test tubes (18 mm×180 mm) at a ratio of 8 ml/branch; placing the separated candidate strain bacterial liquid in a constant temperature incubator at 37 ℃ for static culture for 24, and measuring the vitamin D by HPLC 2 With vitamin D 3 Content (counting M) 1 ) Vitamin D is added into the culture medium 2 With vitamin D 3 Content as initial content (calculated as M 0 ) Wherein the medium was not inoculated as a control, the strain metabolized vitamin D calculation formula (%) = (M) 0 -M 1 )/M 0 *100％。
The experimental results show thatFIG. 3 Lactobacillus plantarum LP550 vs. vitamin D in candidate strains 2 With vitamin D 3 The metabolic capacities of (a) were 32.15%, 42.57%, respectively, and the vitamin D was reacted with Lactobacillus plantarum LP550 2 With vitamin D 3 Is significantly more metabolizable than other candidate strains.
The inventor further analyzes the influence of vitamin D on the growth and metabolism of the lactobacillus plantarum LP550 based on the strong metabolic capacity of the lactobacillus plantarum LP550 on the vitamin D, and further explores new application of the lactobacillus plantarum LP 550.
3) Physiological and biochemical characteristics of vitamin D on lactobacillus plantarum LP550
(1) Growth performance of lactobacillus plantarum LP550
Lactobacillus plantarum LP550 strain is inoculated into MRS culture medium for 24 hours according to the inoculation amount of 5 percent, and is activated twice continuously. Inoculating activated lactobacillus plantarum LP550 bacterial liquid into MRS and vitamin D according to an inoculum size of 5 percent 2 +MRS, vitamin D 3 In +MRS liquid culture medium, mixing, and packaging into sterile test tubes (18 mm×180 mm) at a ratio of 8 ml/branch; placing the subpackaged lactobacillus plantarum LP550 bacterial liquid in a constant-temperature incubator at 37 ℃ for standing culture for 24 hours, taking 3 test tubes to measure absorbance values OD of the bacterial liquid for 15 hours and 24 hours 600 Calculating an average value; absorbance value OD with time on the abscissa 600 The growth curve is plotted on the ordinate.
The experimental results of growth of the lactobacillus plantarum LP550 are shown in FIG. 4, and the lactobacillus plantarum LP550 is used for vitamin D 2 +MRS, vitamin D 3 The growth in the +MRS liquid culture medium is respectively improved by 22.99 percent and 33.70 percent, and the growth is obviously superior to that of the MRS culture medium, which shows that the vitamin D 2 With vitamin D 3 And the metabolites thereof not only do not influence the growth of the lactobacillus plantarum LP550, but also promote the growth of the lactobacillus plantarum LP 550.
(2) Acid production test of lactobacillus plantarum LP550
Lactobacillus plantarum LP550 strain is inoculated into MRS culture medium for 24 hours according to the inoculation amount of 5 percent, and is activated twice continuously. Inoculating activated lactobacillus plantarum LP550 bacterial liquid into MRS and vitamin D according to an inoculum size of 5 percent 2 +MRS, vitamin D 3 +MRS liquid culture mediumAfter mixing, the mixture was dispensed into sterile test tubes (18 mm. Times.180 mm) at 8 ml/serving. Placing the subpackaged lactobacillus plantarum LP550 bacterial liquid in a constant temperature incubator at 37 ℃ for static culture, taking 3 test tubes to measure the total acid of the bacterial liquid, and calculating the average value; at regular intervals, 3 test tubes are taken to measure total acid of bacterial liquid, and the time is taken as an abscissa, and the acid yield is taken as an ordinate, so that an acid production curve is drawn, as shown in fig. 5.
Referring to FIG. 5, lactobacillus plantarum LP550 is found in vitamin D 2 +MRS and vitamin D 3 The total acid level in the +mrs broth was not significantly different from the acid production level in the MRS broth, and further analysis by HPLC showed a significant increase in short chain fatty acids, in particular a significant increase in butyrate content (table 1), the above results indicated: vitamin D 2 Or vitamin D 3 Has little influence on the production of the total acid of lactobacillus plantarum LP550, but can improve the content of short-chain fatty acid and vitamin D 2 +MRS increased by 16.4% compared with MRS, vitamin D 3 +MRS is increased by 27% compared with MRS, especially the butyric acid content, vitamin D, playing an important role in the intestinal myoaxis and the intestinal bone axis is significantly increased 2 +MRS, vitamin D 3 +MRS was increased by 4.7 and 7.4 times, respectively, compared to MRS.
TABLE 1 short chain fatty acid detection results
Annotation: the quantitative unit of short-chain fatty acid is μg/ml.
(3) Bacteriostasis test of Lactobacillus plantarum LP550 on pathogenic bacteria
Bacteriostasis experiment for pathogenic bacteria: pouring 10mL of water agar medium in a sterile plate, cooling to solidify, placing on oxford cup, and collecting suspension of indicator bacteria (helicobacter pylori ATCC 26695, streptococcus mutans CGMCC1.2499, staphylococcus aureus CMCC26003, clostridium perfringens ATCC13124, candida albicans)Bacteria ATCC 10231) were added to agar media cooled to 50 ℃ and grown in response to the indicator bacteria to give an indicator bacteria concentration of 10 6 CFU/mL, mixing, pouring onto bottom water agar, solidifying, taking out oxford cup with forceps to form holes, adding 200 μl of sample to be tested (MRS culture medium, vitamin D) into each hole 2 +MRS Medium, vitamin D 3 +MRS medium), diffusion for 30min, and culturing at 37deg.C for 15-24 hr. And (3) observing whether a bacteriostasis ring appears around the culture hole, measuring the diameter of the culture hole by using a vernier caliper, recording the diameter of the bacteriostasis ring, and finally evaluating the bacteriostasis activity according to the existence and the size of the bacteriostasis ring.
TABLE 2 antibacterial ability analysis of Lactobacillus plantarum LP550 fermentation broths obtained by different media culture
Streptococcus mutans CGMCC1.2499 16.17+ -0.68.23.32+ -0.67.22.08+ -0.63
Staphylococcus aureus CMCC26003 14.81±0.75.21.18±0.63.20.75±0.42
Clostridium perfringens ATCC 13124.72±0.44.22.57±0.88.20.98±0.45
The results are shown in Table 2, wherein vitamin D was added to MRS medium relative to Lactobacillus plantarum LP550 obtained by fermentation of MRS medium 2 Or vitamin D 3 The bacteriostasis circle of the lactobacillus plantarum LP550 obtained by post fermentation on helicobacter pylori ATCC26695, streptococcus mutans CGMCC1.2499, staphylococcus aureus CMCC26003, clostridium perfringens ATCC 13124 and candida albicans ATCC 10231 is more than 20mm, which shows that: vitamin D 2 And vitamin D 3 The antibacterial capacity of the lactobacillus plantarum LP550 on pathogenic microorganisms is synergistically promoted, and the antibacterial capacity of the lactobacillus plantarum LP550 is improved by more than 11 percent on the basis of a conventional MRS culture medium, especially vitaminsD 2 Synergistic promotion of Lactobacillus plantarum LP550 with an increase in antibacterial capacity of Candida albicans ATCC 10231 up to 49% and vitamin D 3 The antibacterial capacity of the lactobacillus plantarum LP550 on helicobacter pylori ATCC26695 is improved by 42% in a synergistic way; binding lactobacillus plantarum in vitamin D 2 +MRS medium, vitamin D 3 The bacterial count in the +MRS medium is increased, but the total acid change is not obvious, which indicates that the lactobacillus plantarum LP550 is used for vitamin D 2 +MRS medium, vitamin D 3 The content of antibacterial substances such as antibacterial peptide in the +MRS culture medium is improved, so that the antibacterial performance of lactobacillus plantarum LP550 is obviously improved.
4) Tolerance test of Lactobacillus plantarum LP550 to gastric acid and bile salts of human body
(1) Gastric acid tolerance assay of strains
Preparing simulated gastric juice: 2.0g/L NaCl, regulating pH to 2.0, 2.5, 3.0 and 4.0 with HCl, autoclaving, and pepsin 3.2g/L, wherein the pepsin is used at present in experiment; preparing simulated intestinal juice: 6.8g/L of potassium dihydrogen phosphate, regulating the pH value to 7.5 by NaOH, performing high-pressure sterilization, and adding 10.0g/L of trypsin which is currently used in experiments; inoculating LP550 strain preserved by glycerol pipe into MRS culture medium with 10% inoculum size, activating at 37deg.C for 24 hr; an equal amount of LP550 bacterial liquid is added into 50mL simulated gastric fluid of the system, initial viable bacteria are recorded, and the viable bacteria number is measured after the constant temperature culture for 3 hours at 37 ℃. The detected live lactic acid bacteria were counted and the survival rate was calculated, strain survival rate=test group/control group×100%.
Gastric acid begins to secrete when food enters the stomach. The pH value of gastric acid concentration secreted by normal stomach of human body is about 0.5-1.5. The pH value is about 7.0-7.2 when the stomach is emptied, and rapidly drops to 2-3 when food enters the stomach. After meals, gastric juice is diluted and the pH rises to about 3.5. Experimental results show that at pH2.0, the survival rate of LP550 is 49.52%; at pH2.50, LP550 survival rate was 74.37%; at pH3.0, the survival rate of LP550 was 95.43%, indicating that the LP550 strain had good gastric acid tolerance.
(2) Strain bile salt tolerance assay
The LP550 strain was inoculated into MRS liquid medium at an inoculum size of 5%, and subjected to activation culture at 37℃for 24 hours, followed by two consecutive activations. Inoculating the activated LP550 bacteria liquid into MRS liquid culture medium according to the inoculation amount of 5%, and standing and culturing for 15h at 37 ℃ in a constant temperature incubator. And (3) centrifuging the cultured bacterial liquid at 5000rpm for 10min to collect bacterial bodies, and vibrating the bacterial bodies uniformly with sterile physiological saline.
Adding the bacterial liquid with uniform vibration into MRS culture medium with bile salt concentration of 1.0g/L, 2.0g/L, 3.0g/L and 0.0g/L (initial bacterial liquid) according to the addition amount of 10%, and taking the bile salt concentration of 0.0g/L as a control group. Then incubated in a constant temperature incubator at 37℃for 3h. Taking out the incubated bacterial liquid, immediately diluting according to 10 times, adding sterile physiological saline, beating and uniformly mixing, and detecting the number of lactic acid bacteria; counting the detected live lactobacillus and calculating the survival rate, wherein the calculation formula is as follows:
strain survival (%) = test/control x 100%.
LP550 bile salt tolerance data indicate: the survival rates of the strains were 100% and 98.13% when the bile salt concentrations were 1.0g/L and 2.0g/L, respectively, but the survival rates of the strains still reached 95.76% when the bile salt concentrations reached 3.0 g/L. The concentration of bile salts in the intestinal tract is not more than 3.0g/L, which indicates that the LP550 strain has better bile salt tolerance.
In conclusion, lactobacillus plantarum LP550 is capable of metabolizing and utilizing vitamin D in the culture medium 2 Or vitamin D 3 At the same time, vitamin D 2 And vitamin D 3 The method can also promote the growth of lactobacillus plantarum LP550 and the formation of metabolites such as antibacterial peptide, and promote the formation of metabolites such as short chain fatty acid and the like in the growth process of lactobacillus plantarum LP550, and lactobacillus plantarum LP550 has good tolerance to gastric juice, intestinal juice and bile salts, and the main function based on vitamin D is to promote the absorption of calcium and phosphorus by small intestine mucosa cells, improve the concentration of calcium and phosphorus in blood, be favorable for new bone formation and calcification, and provide a new direction for the development and research of new application of lactobacillus plantarum LP 550.
Example 2
A composition comprising Lactobacillus plantarum LP550 metazoan containing 100 hundred million/g Lactobacillus plantarum cells of example 1, vitaminBiotin D 3 The content of the metazoan is 400IU/g, the free amino acid is more than 1mg/g, and the preparation method of the metazoan comprises the following steps: lactobacillus plantarum LP550 from example 1 was used in vitamin D 3 Fermenting in +MRS liquid culture medium to obtain fermentation broth, concentrating, spray drying to obtain fermented broth containing thallus and vitamin D 3 Solid powders of metabolites.
Example 3
Use of lactobacillus plantarum LP550 and its progeny for the preparation of a functional food or medicament for improving musculoskeletal health and improving exercise capacity.
Sample to be tested: (1) the metaplasia lactobacillus plantarum LP550 (called metaplasia LP550 for short): lactobacillus plantarum LP550 metazoan (100 hundred million cells/g, D in example 2 3 The content of the free amino acid is 400IU/g and is more than 1 mg/g), and the preparation method comprises the following steps: lactobacillus plantarum LP550 in vitamin D 3 Fermenting in +MRS liquid culture medium to obtain fermentation broth, concentrating, spray drying to obtain thallus and vitamin D 3 Solid powders of metabolites. (2) Compound vitamin D 3 Metagen LP550 (abbreviated as "compound VD") 3 ): in example 1, lactobacillus plantarum LP550 broth was obtained by fermentation in MRS medium, concentrated, spray-dried to form solid powder containing cells and metabolites, and then mixed with vitamin D 3 Mixing the finished products, and adding vitamin D into the compounded product 3 The content of the lactobacillus plantarum LP550 bacteria is 400, and the number of the lactobacillus plantarum LP550 bacteria is 100 hundred million/g.
Research and test of lactobacillus plantarum LP550 on improving musculoskeletal health and improving exercise capacity
Test grouping: 30 SPF-class SD rats purchased from Chengdu Corp were randomly divided into 5 groups by body mass, namely a normal group (CON group), a sham operation group (SHM group), an OS group, and a compound vitamin D 3 Metagen LP550 group (OS+compound VD) 3 Group) and the OS metaplasia lactobacillus plantarum LP550 (os+metaplasia LP550 group). Wherein OS group, OS+ compound VD 3 The rats in group and OS+ metazoan LP550 group were model-built by castration method, the SHM group was model-built by sham operation, and all the rats in group except CON group were intra-operatively injected with 8 ten thousand units/unit of penicillin sodium daily for 3 days. After 7d and 7d of postoperative recovery, starting to compound OS group and OS +VD 3 Group and os+ metazoan LP550 group rats were annotated 1mg (/ kg·d) of dexamethasone sodium phosphate injection (DXM) for 2 weeks. After the molding is finished, the medicine intervention is started: CON group, SHM group and OS group rats were perfused with 10mL/kg physiological saline daily, OS+ formulated VD 3 Group continuous 12-week gastric lavage compound vitamin D 3 Metagen LP550 (1×10) 8 Personal/per mouse/day), group OS+ metant LP550 continuous 12 weeks gavage of metant Lactobacillus plantarum LP550 (1×10) in example 2 8 Personal/per mouse/day), the data were analyzed after the experimental results were completed, compared to the CON group, a P<0.05; in contrast to the SHM group, b P<0.05; in contrast to the set of OS's, c P<0.05; compounding VD with OS + 3 In a group comparison the number of groups, d P<0.05。
(1) rat general condition observation and weight change analysis
Feeding activity, mental state, abnormal behavior, etc. of each group of rats were observed and recorded daily during the gavage. After the end of the intervention, the Body Weights (BW) of each group of rats were measured weekly by an electronic scale.
During the experiment, no mice in each group died. The OS group of rats had a dry skin and hair, a messy hair and a reduced gloss compared with the other groups. OS+Compositon VD 3 Group, OS+ metagen LP550 group rats had a softer skin and hair than OS group, and normal spirit and activity.
The body weight changes of rats in the different test groups are shown in table 3. No significant differences between each dose group and control group, SHM group (P>0.05 A) is provided; the weight of the OS group is slightly smaller than that of the CON group and the SHM group, and the OS group is consistent with weight loss which is one of clinical manifestations of patients with musculoskeletal co-loss; OS+Compositon VD 3 Compared with the OS group, the weight of the group and the OS+metaplasia LP550 group has an ascending trend, but the ascending amplitude is not obvious, which indicates that the lactobacillus plantarum LP550 and the LP550 metaplasia have a trend of relieving the weight reduction of OS rats, and the vitamin D is orally taken for a long time 3 The live strain of the lactobacillus plantarum LP550 or the metaplasia thereof can effectively prevent the sarcopenia of animals, and has good growth safety.
Table 3 weight change of rats in each experimental group
Annotation: in contrast to the CON-group, a P<0.05; in contrast to the SHM group, b P<0.05; in contrast to the set of OS's, c P<0.05; compounding VD with OS + 3 In a group comparison the number of groups, d P<0.05。
(2) analysis of bone Density changes in rats of each group
The test method comprises the following steps: after the intervention, before the material is obtained, the rats in each group are anesthetized, and the total bone density (Bone mineral density, BMD) and lumbar BMD of the rats in each group are measured by a dual-energy X-ray bone densitometer.
Referring to fig. 6, the test results indicate that: compared with CON group and SHM group, OS group rats showed significantly decreased total body BMD and lumbar vertebrae BMD (P<0.05 The successful modeling is shown, and the OS has obvious influence on the muscle and bone health of rats; compared with the OS group, the OS+compound VD 3 Group os+ metaplasia LP550 group rats showed significant increase in whole body BMD and lumbar BMD (P<0.05 Indicated that: using MRS medium and vitamin D 3 The metaplasia of the lactobacillus plantarum LP550 obtained by the +MRS culture medium can obviously improve the bone density of the rat with the muscular-bone co-reduction disease (OS group), thereby effectively improving the muscular-bone health of the OS rat.
Compounding VD with OS + 3 Group comparison, os+ metagen LP550 group significantly increased whole body BMD and lumbar BMD (P<0.05 Description of the following): metazoan and vitamin D formed by lactobacillus plantarum LP550 fermentation broth obtained by fermenting and culturing relative to conventional MRS culture medium 3 Compound formed composition for metabolizing vitamin D 3 The metazoan of Lactobacillus plantarum LP550 was more effective in improving bone mineral density in OS rats, and could be used with Lactobacillus plantarum LP550 in the utilization of vitamin D 3 In the growth metabolism process of (2), metabolites which are not only the growth metabolites of the conventional MRS culture medium but also promote the increase of the bone density of the OS rat are synthesized, so that the increase promotion effect of the metabolites on the bone density of the OS rat is more obvious And is known.
(3) Comparative analysis of forelimb grip, SMI and RSMI in rats of each group
The test method comprises the following steps: after the intervention was completed, rat forelimb grip was measured using a grip tester for a fixed time per week (Forelimb grip strength). Rats were allowed to acclimate on the grab instrument for 5 minutes prior to testing, then the rat tail level was grabbed and pulled slowly back until the rat forelimbs were unable to grab the rail, and the maximum grab force reading on the instrument was recorded. Three measurements were made and recorded, with the average of the three grips being used as an indicator reflecting the muscle strength of the forelimb. The dual energy X-ray densitometer measures the total skeletal muscle mass (Lean mass, LM) and the skeletal muscle mass of the extremities (appendicular Lean mass, ALM) of each group of rats. And calculates the rat skeletal muscle mass index (smi=lm/BW) and the limb skeletal muscle mass index (rsmi=alm/BW).
Referring to fig. 7 and 8, the OS group rats had significantly reduced forelimb grip compared to the CON and SHM groups (P<0.05 SMI and RSMI also significantly decreased (P)<0.05 A) is provided; compared with the OS group, the OS+compound VD 3 Group and os+ metazoan LP550 group rats had significantly increased forelimb grip, SMI and RSMI (P<0.05 And the OS+ metagen LP550 group is better than the OS+ compound VD 3 Indicating OS+built VD 3 The group and the OS+ metazoan LP550 group can significantly improve the forelimb grasping power and skeletal muscle mass of OS rats, thereby improving musculoskeletal health.
Compared to CON group, os+ metagen LP550 group rats had significantly increased forelimb grip, SMI, and RSMI (P < 0.05), indicating: the metaplasia LP550 not only can promote the recovery of the forelimb holding power, SMI and RSMI of the OS rat, but also can promote the further promotion of the forelimb holding power and skeletal muscle quantity of the rat, namely, the metaplasia LP550 has the effects of improving the muscle bone health and promoting the movement ability of the OS rat or healthy rat, and can effectively prevent and treat sarcopenia or osteoporosis or sarcoskeletal co-reduction syndrome.
(4) Running experiments on rats of each group
The test method comprises the following steps: rats were tested for endurance performance 12 weeks after continuous feeding and were allowed to adapt to treadmill training 30 minutes after each feeding 1 week prior to testing. Placing the rat in a small chamber of the running machine, setting the rotating speed to be 20r/min, setting the current intensity of an electric stimulation area to be 1.2A, and recording the running time and the running distance from the rat to the exhaustion, wherein the exhaustion judgment standard is that the rat is at rest for more than 10S in an electric shock area.
As shown in FIG. 9, the running time and running distance of the OS group rats were significantly shortened compared to the control group (P<0.05 Indicating that OS has a significant impact on the running ability of rats; compared with the running time and running distance of the OS group rats, the OS+compound VD 3 The group and the OS+ metazoan LP550 group can obviously prolong the running time and the running distance of rats (P)<0.05 And the running time and running distance of the rat with the OS+metagen LP550 group are also obviously better than those of the rat with the OS+compound VD 3 Group, elucidation, metaplasia LP550 helps to improve the physical exercise capacity of rats.
Compared with the CON group, the os+ metagen LP550 group rats had significantly prolonged running time and running distance (P < 0.05), indicating that: the metaplasia LP550 not only can promote the recovery of the exercise capacity of the OS rat, but also can promote the further improvement of the exercise capacity of the rat, namely, the metaplasia LP550 has the effect of improving the exercise capacity of the organism for the OS rat or the healthy rat, thereby effectively preventing and treating sarcopenia, osteoporosis or musculoskeletal co-reduction syndrome.
(5) Detection and analysis of butyric acid in feces of rats of each group
Fresh feces from each group of rats were collected 12 weeks after continuous feeding of the rats, sampled and tested as required by HPLC on a machine. The test results are shown in FIG. 10, in which the butyric acid content of rats in the model group was significantly reduced compared to the CON group (P<0.01). After 12 weeks of intervention, OS+ compound VD 3 The butyric acid content in the rat feces of the group and the OS+ metaplasia LP550 group is obviously increased, and simultaneously, the OS+ metaplasia LP550 group is obviously better than the OS+ compound VD 3 Group (P)<0.01). The postnatal of the lactobacillus plantarum LP550 is shown to be capable of improving the butyric acid content in the intestinal tract of a rat, which is used for meeting the energy supply requirement of colon mucosa epithelial cells, so as to improve the intestinal flora of the rat, further promote the absorption of vitamin D and lactobacillus plantarum LP550 metabolites by the intestinal tract, and achieve the purpose of effectively preventing and treating the musculoskeletal total loss syndrome (OS).
Taken together, the OS group rats showed a significant decrease in whole body BMD, lumbar BMD, SMI and RSMI compared to CON and SHM groups, indicating that castration was combined with DXM can cause the occurrence of musculoskeletal co-subtraction syndrome (OS for short) in rats. Compared with the OS group, the OS+compound VD 3 And the whole body BMD, lumbar BMD, forelimb holding power, SMI, RSMI, running time and running distance of the rat in the OS+metaplasia LP550 group are obviously increased, meanwhile, the butyric acid content in the feces is obviously increased, and the corresponding performances of the metaplasia LP550 group are better than those of the compound VD 3 Group, indicating: the lactobacillus plantarum LP550 and the metazoan thereof can obviously improve the bone health of the OS rat, improve the exercise capacity and further effectively prevent and treat the musculoskeletal co-reduction syndrome.
Example 4
A composition of this example comprises a Lactobacillus plantarum LP550 metazoan containing 100 hundred million/g Lactobacillus plantarum cells of example 1, vitamin D 2 The content of the metazoan is 400IU/g, the free amino acid is more than 1mg/g, and the preparation method of the metazoan comprises the following steps: lactobacillus plantarum LP550 from example 1 was used in vitamin D 2 Fermenting in +MRS liquid culture medium to obtain fermentation broth, concentrating, spray drying to obtain thallus and vitamin D 2 Solid powders of metabolites.
The composition of this example was used for treatment of OS rats, and its weight loss inhibitory effect on OS rats, and its lifting effect on whole body BMD, lumbar BMD, forelimb grip, SMI, RSMI, running time and running distance of OS rats were slightly superior to those of the composition of example 2.
Example 5
A composition of this embodiment comprises a live strain of Lactobacillus plantarum LP550 and an inactivated strain of Lactobacillus plantarum LP550, wherein the live strain of Lactobacillus plantarum LP550 is vitamin D in embodiment 1 2 Lactobacillus plantarum LP550 obtained by culturing in +mrs liquid medium; the inactivated strain of lactobacillus plantarum LP550 is adopted in vitamin D 2 Culturing in +MRS liquid culture medium at 37deg.C for 48 hr to obtain lactobacillus plantarum LP550 bacterial liquid, sterilizing at 125deg.C for 5 min; centrifuging 8000g of the inactivated bacterial liquid for 5min to obtain an inactivated strain of lactobacillus plantarum LP550; the weight ratio of live strain of lactobacillus plantarum LP550 to inactivated strain of lactobacillus plantarum LP550 was 1:3.
A group of the present embodimentApplication of compound in preparing medicine for improving muscle and bone health and improving exercise capacity, and when in use, the dosage of lactobacillus plantarum LP550 in the compound is 1.0 x 10 10 CFU/day, inactivated Lactobacillus plantarum LP550 at 6.0x10 10 CFU/day。
Example 6
Application of lactobacillus plantarum LP550 in preparation of functional food or medicine for preventing and treating OS (operating system), wherein dosage of lactobacillus plantarum LP550 is 1.5X10 10 CFU/day。
Wherein lactobacillus plantarum LP550 was obtained by culture using MRS medium as described in example 1.
Example 7
Use of lactobacillus plantarum LP550 as a metazoan-improving agent in a functional food or medicament for improving musculoskeletal health and improving exercise capacity, wherein the metazoan amount of lactobacillus plantarum LP550 is 500mg/day.
The preparation method of the metaplasia lactobacillus plantarum LP550 comprises the following steps: lactobacillus plantarum LP550 of example 1 was fermented in MRS liquid medium to obtain a fermentation broth, which was concentrated and spray dried to form a solid powder containing bacterial cells and metabolites.
Example 8
A composition of this example comprises a live strain of Lactobacillus plantarum LP550 and a metazoan of Lactobacillus plantarum LP550 in a mass ratio of 1:1.
As can be seen from the antibacterial ability tests of the lactobacillus plantarum LP550 in the embodiments 1, 3) to 3), the live lactobacillus plantarum LP550 strain has good inhibition effect on common pathogenic bacteria such as helicobacter pylori ATCC 26695, streptococcus mutans CGMCC1.2499, staphylococcus aureus CMCC26003, clostridium perfringens ATCC13124, candida albicans ATCC10231 and the like.
The composition of the embodiment has antibacterial effect and is applied to preparation of functional foods or medicines for inhibiting pathogenic bacteria, wherein pathogenic helicobacter includes helicobacter pylori ATCC 26695, streptococcus mutans CGMCC1.2499, staphylococcus aureus CMCC26003, clostridium perfringens ATCC13124 and candida albicans ATCC10231. When in use, the active strain of the lactobacillus plantarum LP550 is 1 multiplied by 10 10 CFU/day, lactobacillus plantarum LP550Inactivated strain 2×10 10 CFU/day, the metagen dose is 100mg/day.
Example 9
A composition of this example, comprising a live strain of Lactobacillus plantarum LP550 and a Lactobacillus plantarum LP550 metazoan, was prepared as in example 4.
As can be seen from example 1, in vitamin D 2 The live strain of the lactobacillus plantarum LP550 cultured on the +MRS culture medium has better acid production and bacteriostasis capacity, and the composition of the embodiment is applied to the preparation of fermented foods and health-care foods as a starter.
The fermented food is pickle, and the application method of the composition as a starter in preparing pickle is as follows:
cleaning fresh vegetables, adding into 4-5 times of drinking water, adding edible glucose with total volume of 1% and edible sodium chloride with total volume of 0.5-2.0%, inoculating lactobacillus plantarum LP550 prepared in example 1 of the invention, and making its concentration reach 10 7 Fermenting at room temperature for 5-24 hr to obtain fermented sauerkraut containing Lactobacillus plantarum LP550 and Lactobacillus plantarum LP550 metazoan composition. The fermented pickle has crisp and unique flavor, contains lactobacillus plantarum LP550 thalli and metabolites, and has good safety and probiotic functions.
Example 10
Application of lactobacillus plantarum LP550 and metaplasia in functional food or medicine for improving bone health, wherein the dosage of lactobacillus plantarum LP550 is 5 x 10 10 CFU/day, the dosage of the metaplasia lactobacillus plantarum LP550 is 5 x 10 10 Personal/day.
Wherein Lactobacillus plantarum LP550 and its progeny are each as in example 1, vitamin D 3 +MRS medium, obtained as described in example 2.
Example 11
A composition of this example comprises, in parts by weight, lactobacillus plantarum LP550 powder (4.0X10 10 CFU/g) 10 parts by weight, lactobacillus rhamnosus S24 (4.0X10) 10 CFU/g) 8 parts by weight of Lactobacillus plantarum LP550 metazoan (cell number 4.0X10) 10 Each gram of the beverage comprises 2 parts by weight of auxiliary materials including 10 parts by weight of skimmed milk powder, 10 parts by weight of resistant starch, 10 parts by weight of maltodextrin, 10 parts by weight of sodium hyaluronate, 10 parts by weight of cherry extract, 10 parts by weight of kiwi fruit extract, 10 parts by weight of apple extract, 5 parts by weight of vitamin C and 3 parts by weight of folic acid.
Firstly, 10 parts by weight of skimmed milk powder, 10 parts by weight of resistant starch, 10 parts by weight of maltodextrin, 10 parts by weight of sodium hyaluronate, 10 parts by weight of cherry extract, 10 parts by weight of kiwi fruit extract, 10 parts by weight of apple extract, 5 parts by weight of vitamin C and 3 parts by weight of folic acid are weighed and uniformly mixed, and are granulated into wet granules by adopting a 20-mesh screen with the concentration of 30% alcohol wet method, the wet granules are dried for 3.5 hours at 55 ℃, after the granules are finished by adopting the 20-mesh screen, lactobacillus plantarum LP550 bacterial powder (4.0x10) is added 10 CFU/g) 10 parts, lactobacillus rhamnosus S24 (4.0X10) 10 CFU/g) 8 parts by weight of the post-metazoan (cell number 4.0X10) of Lactobacillus plantarum LP550 10 2 parts per gram) and 2 parts of magnesium stearate, and tabletting by a rotary tablet press after uniform mixing, thus obtaining the tablet with the lactobacillus plantarum LP550 dietary supplement.
The composition of this example is used to prepare a functional food that improves bone health and enhances exercise capacity.
Example 12
A composition of this example comprises, in parts by weight, lactobacillus plantarum LP550 (4.0X10) 10 CFU/g) 10 parts by weight, lactobacillus plantarum LP550 metazoan (4.0X10) 10 2 parts by weight of Lactobacillus fermentum GF1800 (4.0X10) 10 CFU/g) 8 parts by weight, maltodextrin 10 parts by weight, sorbitol 10 parts by weight, corn peptide 10 parts by weight, sodium hyaluronate 10 parts by weight, anserine 10 parts by weight, soybean peptide 10 parts by weight, xylooligosaccharide 4 parts by weight, broccoli seed aqueous extract 4 parts by weight, selenium-enriched yeast 2 parts by weight, folic acid 2 parts by weight, malic acid 2 parts by weight, glutathione 2 parts by weight, vitamin E2 parts by weight, and vitamin C2 parts by weight.
The raw materials of the composition are firstly sieved by a 40-mesh screen, then uniformly mixed according to a proportion, and packaged by a screw back-packaging machine to prepare 2-10 g/bag of solid beverage with the functions of improving muscle and bone health and improving exercise capacity.
Example 13
Application of lactobacillus plantarum LP550 and its metaplasia in medicines for preventing and treating OS, wherein the dosage of lactobacillus plantarum LP550 is 5 x 10 10 CFU/day, lactobacillus plantarum LP550 metazoan dose 1.2x10 11 Personal/day. The Lactobacillus plantarum LP550 was described in example 1 in vitamin D 3 +MRS medium, and the Lactobacillus plantarum metazoan are obtained by culturing as described in example 2.
Claims (8)
1. A lactobacillus plantarum is characterized in that the lactobacillus plantarum is named as lactobacillus plantarumLactobacillus plantarum) LP550, 4-month 23-year 2021, was preserved in China center for type culture collection of Wuhan, china, with a preservation number of CCTCC NO: m2021437.
2. A composition comprising a Lactobacillus plantarum LP550 metant in MRS medium or vitamin D 2 +MRS Medium or vitamin D 3 The fermentation broth obtained by fermenting Lactobacillus plantarum according to claim 1 in +MRS medium is concentrated and spray dried to obtain solid powder containing inactivated strain and its metabolites.
3. The composition of claim 2, wherein said vitamin D 2 +MRS medium, vitamin D 3 The +MRS culture medium is obtained by adding 1000-4000 IU vitamin D to MRS culture medium 2 Vitamin D of 1000-4000 IU 3 。
4. The composition of claim 2 or 3, further comprising the live strain of lactobacillus plantarum LP550, the inactivated strain of lactobacillus plantarum LP550, the metabolite of lactobacillus plantarum LP550, vitamin D of claim 1 2 Vitamin D 3 Lactobacillus rhamnosus and fermented milk stalksA mixture of one or more of the bacteria.
5. Use of lactobacillus plantarum LP550 as claimed in claim 1 or a composition as claimed in any of claims 2-4 for the manufacture of a functional food product for improving musculoskeletal health and/or improving exercise capacity.
6. Use of lactobacillus plantarum LP550 as claimed in claim 1 or a composition as claimed in any of claims 2 to 4 in the manufacture of a medicament for the prevention and treatment of sarcopenia or osteoporosis or sarcoossium co-subtraction syndrome.
7. Use of lactobacillus plantarum LP550 as claimed in claim 1 or a composition as claimed in claim 2, for the manufacture of a medicament for inhibiting pathogenic bacteria.
8. Use of lactobacillus plantarum LP550 of claim 1 or a composition of any of claims 2-4 as a starter in the preparation of a fermented food, a health food or a dietary supplement.
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