CN116286462A

CN116286462A - Lactobacillus plantarum with comprehensive performance in motion improvement and application thereof

Info

Publication number: CN116286462A
Application number: CN202211589419.6A
Authority: CN
Inventors: 杨锁华; 黄启彰; 李孟谦; 徐艺洳; 程怡蓁; 李珈嘉
Original assignee: Synbiotech Biotechnology Yangzhou Co ltd
Current assignee: Synbiotech Biotechnology Yangzhou Co ltd
Priority date: 2022-12-11
Filing date: 2022-12-11
Publication date: 2023-06-23

Abstract

The invention discloses a lactobacillus plantarum (Lactobacillus plantarum) TWK10 with comprehensive performance in the aspect of improving exercise, which is characterized in that the strain is preserved in China general microbiological culture Collection center (China center for type culture Collection) at the year of 2022, 6 and 27, and the preservation address is the North Chen West Lu No. 1, 3 in the Chaoyang area of Beijing city, and the preservation number is CGMCC No.25190. And mixing the lactobacillus plantarum with pea protein to prepare the composition. The composition has the effects of improving amino acid absorption rate, improving anaerobic power and muscle strength, and improving body constitution and muscle mass.

Description

Lactobacillus plantarum with comprehensive performance in motion improvement and application thereof

Technical Field

The invention relates to lactobacillus plantarum, in particular to lactobacillus plantarum with comprehensive performance in the aspect of improving exercise and application thereof. Belongs to the technical field of biological medicine processing.

Background

In addition to scientific and bitter training, the physical performance requirements of athletes are higher and higher, and the athletes must have reasonable nutritional support, and long-term high-strength or excessive exercise training, unreasonable nutrition, rest and the like can lead the athletes to generate high stress and high metabolism, and the athletes must have higher tissue and organ adaptability and energy metabolism level, otherwise, various adverse physical and psychological health problems such as infection, asthma, gastrointestinal diseases, inflammatory disorder, immunosuppression, oxidative stress, chronic fatigue, depression anxiety and the like can be induced, and especially in the athletes with three long-distance items such as marathons, super-endurance and iron men, gastrointestinal dysfunction, endotoxemia and the like become important negative factors for affecting the health of the athletes and finally affecting athletic performance.

For people engaged in high exercise, it is often necessary to supplement a large amount of protein in order to maintain and enhance muscle strength, repair damaged muscle proteins. The protein-supplementing sports nutritional food takes protein or protein hydrolysate as a main component, aims to enhance the immunity of human bodies, improve the antioxidant capacity, promote the synthesis of skeletal muscle protein, inhibit muscle decomposition and accelerate the recovery of physical energy after training, and is an ideal sports nutritional food. There are different protein supplementation strategies depending on the type of protein and different sports.

The microorganisms in human bodies are various and have a number of trillions, and part of the microorganisms are beneficial to the health of the human bodies, and are called probiotics, and mainly play a role in maintaining the balance of the human bodies. The probiotics can be used as nutritional supplements for preventing or alleviating respiratory tract infections and gastrointestinal symptoms in athletes, thereby promoting healthy physical development in athletes.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide lactobacillus plantarum with comprehensive performance in the aspect of improving exercise and application thereof.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

lactobacillus plantarum (Lactobacillus plantarum) TWK10 (also called lactobacillus plantarum (Lactiplantibacillus plantarum) TWK 10) with comprehensive performance in motion improvement, which is preserved in China general microbiological culture Collection center (China center for type culture Collection) for 2022, 6 and 27 days, and has a preservation address of Beijing, chaoyang area, north Chen West Lu No. 1, 3 and a preservation number of CGMCC No.25190.

( And (3) injection: the "Commission Wei Jian" requirements for update of the "food-available strain List" and "infant food-available strain List" (2022, 4) ", were adjusted )

The lactobacillus plantarum is applied to the preparation of foods or medicines for improving comprehensive performance in sports.

A food or medicine for improving sports comprehensive performance contains Lactobacillus plantarum as effective component.

A composition with comprehensive performance in improving exercise is prepared from the freeze-dried powder of lactobacillus plantarum and pea protein according to a mass ratio of 0.5-1: 99 to 99.5, wherein the viable count in the freeze-dried powder is more than or equal to 1.0x10 ¹¹ cfu/g。

Preferably, the ratio of the pea protein mass to the viable count of the lactobacillus plantarum contained in the freeze-dried powder is 20g: 1.0X10 ¹⁰ cfu。

The invention has the beneficial effects that:

the invention provides a lactobacillus plantarum with comprehensive performance in improving exercise, and the lactobacillus plantarum is mixed with pea protein to prepare a composition. The composition has the effects of improving amino acid absorption rate, improving anaerobic power and muscle strength, and improving body constitution and muscle mass.

Drawings

FIG. 1 is a method of calculating muscle thickness;

fig. 2 is a method of calculating the thickness of muscle fascia.

Preservation information

Classification naming: lactobacillus plantarum TWK10

Latin Wen Xueming: lactobacillus plantarum

Preservation unit name: china general microbiological culture Collection center (China general microbiological culture collection center)

Deposit unit address: beijing city, chaoyang area, north Chenxi Lu No. 1 and 3

Preservation date: 2022, 6, 27

Preservation number: CGMCC No.25190

Detailed Description

The present invention will be further illustrated by the following examples, which are given by way of illustration only and are not intended to be limiting.

Test sample for experiment

1. Sample name and composition:

lactobacillus plantarum TWK10: lactiplantibacillus plantarum (Lactobacillus plantarum) TWK10 (the strain has been preserved in China general microbiological culture Collection center (China center for type culture Collection) for 6.27 days, with a preservation address of CGMCC No.25190, no. 3, north Chen West Lu No. 1, korea, beijing, city).

Pea protein:

c9 (suppliers: hanxin science and technology Co., ltd.)

2. Test sample:

according to the purpose of the experiment, the experimental group was designed as follows:

(1) Control group (Control): pea protein without TWK10 (20 g/day).

(2) TWK10 group (TWK 10): containing TWK10 (1X 10) ¹⁰ CFU/day) pea protein (20 g/day).

3. The time of replenishment of the test sample:

(1) Untrained 4 days: and (5) drinking 2 hours before sleeping.

(2) Training for 3 days: after training, the beverage was drunk within 30 minutes (uniformly infused by laboratory).

(3) And analyzing the concentration of amino acid in blood, and immediately finishing drinking before on-site blood sampling.

(II) Experimental design and flow

1. Subject recruitment:

(1) Inclusion conditions: 32 are 20 to 45 years old, male and female halves, healthy, with exercise habits (exercise at least 3 times per week and at least more than 30 minutes each time).

(2) Exclusion conditions: smoking, drinking, cardiovascular disease, neuromuscular disease, metabolic disease, asthma, pregnant woman, lactation, BMI >30.

2. Experimental design and subject grouping:

at the time of recruitment of subjects, the subjects were investigated in what form of exercise they did at ordinary times, several times per week, several minutes each time, and after evaluation, those who reached inclusion condition were included in the protocol. The experiments were conducted using a double-blind design (double-blind) and subjects were randomized into 2 groups: (1) Pea proteins (Control) without TWK10, (2) TWK10 group (TWK 10). Two groups of subjects were supplemented with test samples for 28 consecutive days and were paired 3 exercises per week. The samples were subjected to exercise testing, blood and stool sample collection, body composition analysis and muscle mass analysis before and after intervention.

Each subject had to eat the test food (twk10+pea protein) or the control food (Pea protein) separately and a diet was recorded 3 days before the test and 3 days before the end of the recording after 4 weeks of sample intervention, and the calories ingested were calculated.

The subjects were kept on a normal diet and had to take nutritional supplements during the experiment and had to drink alcohol, eugerd, yandol, etc. probiotic-related products, e.g. and other medications, 1 week prior to exercise testing and blood drawing, to avoid affecting the experimental data.

Before and after testing and blood sampling, the subjects must be allowed to empty at least 8 hours before the test. And physical activities other than training are avoided 3 days before each exercise test.

(III) exercise training

Three exercises per week are all provided with professional exercise coaches and detailed training textbooks as shown in table 1.

Table 1 training class table

(IV) subject profile

Measuring the basic height, weight, resting heart rate and blood pressure of the subject.

(V) blood amino acid concentration

Blood samples were collected from subjects at 5 time points for 0, 30, 60, 120 and 180 minutes after pea protein supplementation, both before and at the end of the experiment. Centrifuging at 4deg.C and 12,000Xg for 15 min, collecting serum sample, adding ultrapure water with the same amount, mixing with amino acid isotope internal standard substance solution, adding isopropanol containing 1% formic acid, mixing, standing at-20deg.C for 20min, centrifuging to obtain supernatant, adding borate buffer (pH 8.6), mixing, adding AQC acetonitrile solution, and heating at 55deg.C for 10 min for derivatization. Finally diluted 100-fold with ultrapure water and analyzed by UPLC/MS/MS (Gray et al, 2017). The experiment was performed using a liquid chromatography tandem mass spectrometer (liquid chromatography tandem mass spectrometry, LC-MS/MS), wherein the liquid chromatography system was an Acquity UPLC, the column used was a Waters C18 column (2.1 x 150mm,1.8 μm), the temperature of the column was set at 45 ℃, the mobile phase used was water and 100% acetonitrile, both containing 0.1% formic acid by mass, the flow rate was 0.6mL/min, and the volume of the test solution injected was 2 μl. In addition, the tandem Mass spectrometer uses Xex TQ-S triple quadrupole Mass spectrometer as a Mass spectrometry mode, ESI+ parameters of 30 free amino acids are shown in Table 2, an ion pair with the highest signal intensity is selected as a quantitative ion, and data collection and quantitative calculation are performed by using Mass Lynx 4.1 software.

TABLE 2 ESI+ mode parameters for 30 free amino acids

The area under the amino acid concentration curved surface is calculated by a trapezoid formula to obtain [ (upper bottom: front time point + lower bottom: rear time point)/(time)/2 ], and the area under the amino acid concentration curved surface is calculated and summed up after individual calculation for 0-30min, 30-60min, 60-120min and 120-180 min. Cmax is the maximum concentration that was found at 5 time points in total of 0, 30, 60, 120 and 180 minutes after pea protein supplementation. Tmax is the time required to reach the maximum concentration, and thus the time at which the maximum value of the transmission Cmax is located is referred to as the Tmax value. The AUC and Cmax of BCAA, EAA, total amino acids are calculated by summing up the above terms. And Tmax for BCAA, EAA, total amino acids is averaged after summation.

Sixth athletic performance test

Wingate anaerobic test (Wingate Anaerobic test)

Full force sprinting was performed in 30 seconds using an anaerobic power cycle (894E,Monark Exercise AB,Sweden). At the beginning of the test, the test subjects were gradually increased to 120rpm, at which time a weight of 7.5% of the weight set (e.g., a 60 kg person would weigh 4.5 kg of resistance) would automatically fall onto the flywheel of the bicycle to create additional resistance, and the full force sprint was continuously required until 30 seconds. And recording the subject's fatigue index after testing using a motion conscious scale. The measured parameters are Mean power output, which is the power output average at the time of the test, relative Mean power (relative average power, i.e., average power/body weight), peak power, which is the Peak value of the output power during the test, maximum value, relative Peak power (relative Peak power, i.e., peak power/body weight), time to Peak power (which is the time taken from the start of the Peak power to reach during the test, the smaller the value the better the explosive force), fatigue index (Peak power minus valley power divided by Peak power, which indicates the greater the degree of power loss, the more fatigued), respectively.

2. Isotonicity test (Isometric Mid-Thigh, IMPT)

IMTP is a reliable and effective test that can be used to measure maximum force using custom IMTP test equipment (IMTP Rack, kairos Strength, USA) and a force plate (Type 9260aa, kistler, switzerland). All subjects stand equally wide apart with the bars placed between the thighs, the trunk upright, the spine neutral, the knee and hip angles 140 °, and the barbell pulled with maximum effort for 3-5 seconds. Before testing, the testers must be familiar with the IMPT test method, and the correct force and stable weight frequency are ensured through practice. Each test was repeated at 2 minute intervals to reduce detection errors associated with posture changes. The measured parameters are Peak Force (Peak Force, maximum Force), relative Peak Force (Peak Force relative, peak Force/body weight), pRFD (Force rate, i.e. [ Time spent starting motion To Peak Force increment/motion To Peak Force), time To Peak Force (i.e., time spent starting motion To Peak Force), time To Peak RFD (i.e., time spent starting motion To maximum Force rate), respectively.

3. Horizontal speculation test (One repetition maximum (1-RM) test Press test)

The device used was a smith machine (MG-PL 62, matrix Fitness, USA), the test mode was specified to the subject before the test, and sufficient warming should be performed. For example, at least 5-10 light to medium weight actions should be performed. After calm, the subject should rest for two to four minutes before any horizontal conjecture trial is attempted. The subject first lies flat on the bench with his feet flat on the floor and his buttocks and shoulders in contact with the bench. The distance to hold the barbell is slightly wider than the shoulder width (so that the elbow is at right angles to the lowest point). The test typically begins with the arms fully extended, placing weight directly over the chest. The weight descends at a controlled rate and with a smooth motion, just touching the chest, and then returns to the starting position. The maximum bench press test, the upper body muscle strength endurance test, was used with resistance set to a percentage of body weight and increased to the maximum muscle strength that the subject can produce with a single contraction.

Seven musculoskeletal ultrasound scanning measurements

The muscle thickness, fascia change length of Biceps Brachii muscle and Quadriceps femoris muscle were scanned using an ultrasonic scanner (H1300, BENQ, taipei, taiwan) and the detailed examination was as follows: measurement position reference (Mah & van Alfen, 2018) study, please first show that the experimental participant is lying down, the palms of both hands are upwards to present anatomical positions, biceps Brachii muscle measurement position is the straight line distance from the Acromion (Acromion) to the elbow transverse line (Antecubital crease), and the ultrasonic probe placement position is 1/3 of the distance from the elbow transverse line; quadriceps femoris muscle the probe is placed at a distance of 1/2 of the straight line from the anterior superior iliac spine (Anterior superior iliac spine, ASIS) to the superior border of the Patella (Patella). Data analysis was performed using ImageJ version 2.3.0/1.53f (National Institutes of Health, bethesda, maryland, USA). The muscle thickness analysis method is to calculate the sum of the thicknesses of the two ends of the muscle and divide the sum by two, as shown in fig. 1. Reference for muscular fascia calculation method (Nakamura et al 2011), calculation method is: muscle thickness Sin (θ), θ is the angle between the fascia and the horizontal line, as shown in fig. 2. The calculation method of the thickness of the muscle and the thickness variation of the muscle fascia is (thickness measured later-thickness measured earlier) =thickness variation.

Eight double-energy X-ray absorption bone density examination room

The X-ray transmitted through the detected part is received by the scintillation detector, and the obtained data is analyzed by the computer to calculate the muscle mass, body fat and bone density, and the muscle mass, body fat and bone density values of the detected person are compared with the computer report to make a diagnosis. This examination measures the muscle mass, body fat and bone density of the whole body, and the time required for each examination is reduced from 25 minutes to 10 minutes per site. In addition, examination of the patient with DEXA (Lunar iDXA, GE Healthcare, chicago, IL, USA) was performed to obtain about 1/100 of the exposure to a chest X-ray, which was quite safe. This examination is currently the most popular and popular instrument for measuring muscle mass, body fat and bone density due to its high speed, accuracy, sensitivity, ease of handling and non-invasive nature.

Nine personal intensity checklists (Checklist individual strength, CIS), fatigue vision scale (Visual analog fatigue scale, VAFS) and athletic subjective intensity scale (Rating of perceived exertion, RPE)

Four orientations for fatigue assessment were performed through a personal intensity checklist questionnaire: fatigue, attention, power and mobility. The scoring modes of each item of topic are as follows: a score of 1 is very good for-7 is very bad for, and so on. Wherein, 1, 3, 4, 9, 10, 13, 14, 16, 17, 18, 19 are entitled reverse questions, and are scored in reverse: a score of 1 is very non-conforming to a score of 7 is very conforming. The score totals of questions 1, 4, 6, 9, 12, 14, 16, and 20 are expressed as a score representing the degree of fatigue experienced by the subject; taking the total score of the questions 3, 8, 11, 13 and 19 as a sub-scale II, and representing the concentration degree; taking the total score of the questions 2, 5, 15 and 18 as a sub-scale three, and representing the score of the degree of the exercise; the score of questions 7, 10 and 17 is taken as a sub-scale IV and represents the score of the physical activity degree; further, the total of the scores of the questions 1 to 20 was calculated as the individual fatigue strength score, and a higher score indicates a fatiguer.

Before and after the exercise, a fatigue vision analog meter is carried out, a straight line with the actual length of 100mm is marked on the leftmost side, 0mm is marked on the rightmost side, 100mm is marked on the rightmost side, 0mm is interpreted to a subject to represent no fatigue, 100mm represents very high fatigue, the left end moves to the right to represent more and more fatigue, the subject draws a short line on the straight line vertically by taking a pen, then the measured cm value is recorded, and the value is converted into 0-100 minutes to be displayed. The graphic rating scale is one of the rating scales, and has the greatest characteristic that each characteristic or quality is represented by a horizontal line. The assessment method is to mark on-line. Typically, a few vertical lines are added to the line segment, and each intersection represents a category of different degrees, expressed as a total of 5 points without fatigue, mild, moderate, severe, and very tired. The numerical evaluation scale indicates that the fatigue degree is relaxed by the number of 1-10, and the higher the score is, the more fatigue is indicated.

In addition, subjective exercise fatigue feeling was also recorded by the exercise subjective intensity scale.

(ten) statistical analysis

All values are expressed as mean±sd, 16 subjects per group; analysis was performed using the statistical package software SPSS Statistics 25. Comparing before and after the intervention in the group, and if the data is parametric, adopting a Paired Student's t test; if the data is non-parametric, wilcoxon signed-rank test is used. For comparison between groups, unpaired Student's t test is used for data for parametric; the data were verified using Mann-Whitney U test for non-parameter. P <0.05 represents a significant difference.

The general characteristics and exercise habits of the subjects are shown in tables 3-1 and 3-2, and the results are shown in tables 4-9.

TABLE 3-1 general characteristics of the subject

TABLE 3-2 exercise habit of subject

TABLE 4 area under the curve (AUC) for individual amino acids, BCAA, EAA and total amino acids

Data are shown as mean ± standard deviation. Analysis of treatment effect by unpaired student t-test P<0.05，***P<0.001. Differences between groups before and after dosing were analyzed by paired student t-test. #P (#P)<0.05，###P<0.01，###P<0.001. Changes in AUC were analyzed using the Mann-Whitney U test.

TABLE 5 maximum concentrations of individual amino acids, BCAA, EAA and total amino acids (C _max )

TABLE 6 time to maximum concentration (Tmax) of individual amino acids, BCAA, EAA and total amino acids

TABLE 7 muscle thickness and fascia thickness of subject

Data are shown as mean ± SD. Treatment effect was analyzed by unpaired student t-test. P <0.05 was analyzed by paired student t-test for differences before and after dosing for each group,

**P<0.01，***P<0.001. the change in muscle thickness was calculated as the difference between before and after administration and analyzed for statistical significance using the Mann-Whitney U test.

P<0.001。

TABLE 8 physical Condition changes in a subject

TABLE 9 personal force (CIS) questionnaire checklist

Data are shown as mean ± standard deviation. The therapeutic effect was analyzed by Mann-Whitney U test. Differences between groups before and after dosing were analyzed by Wilcoxon signed rank test.

The numerical changes were calculated from the differences before and after dosing and analyzed for statistical significance using the Mann-Whitney U test. #P (#P)<0.05。.

The sport lactobacillus TWK10 can obviously increase the peak value and the maximum stress rate of the isotonic muscle strength compared with the prior intervention after being supplemented for four weeks continuously in the isotonic muscle strength expression part.

The athletic lactobacillus TWK10 can significantly increase the performance of the lying-pushing muscle strength after being supplemented for four weeks in the muscle strength performance part compared with the prior intervention.

The lactic acid bacteria TWK10 can significantly increase muscle thickness after continuous four-week supplementation of body components compared to prior to intervention.

Thus, the applicant believes that 20g of pea protein is supplemented per day with Lactobacillus kinesis TWK10 (1X 10) ¹⁰ CFU/day) in combination with three times per week resistance exercise training, with:

(1) Is helpful for improving the absorption rate of total BCAA, total EAA and total amino acid in blood;

(2) The maximum muscle strength, the isotonicity, the anaerobic power and other athletic performances are improved;

(3) Helping to increase muscle thickness.

While the foregoing describes the embodiments of the present invention, it is not intended to limit the scope of the present invention, and various modifications or variations may be made by those skilled in the art without the need for inventive effort on the basis of the technical solutions of the present invention.

Claims

1. Lactobacillus plantarum (Lactobacillus plantarum) TWK10 with comprehensive performance in the aspect of improving exercise is characterized in that the strain is preserved in China general microbiological culture Collection center (China center for type culture Collection) for 6 months and 27 days in 2022, wherein the preservation address is North Chen Xili No. 1 and 3 in the Korean region of Beijing city, and the preservation number is CGMCC No.25190.

2. Use of lactobacillus plantarum according to claim 1 for the preparation of a food or a medicament for improving the overall performance of sports.

3. A food or medicament for improving the comprehensive performance of sports, characterized in that the effective component is the lactobacillus plantarum of claim 1.

4. A composition for improving the overall performance of sports, characterized in thatThus, the mass ratio of the freeze-dried powder of the lactobacillus plantarum to the pea protein is 0.5-1: 99 to 99.5, wherein the viable count in the freeze-dried powder is more than or equal to 1.0x10 ¹¹ cfu/g。