Lactobacillus plantarum with high proliferation activity and application thereof

Abstract

The invention discloses lactobacillus plantarum with high proliferation activity and application thereof, and belongs to the technical field of lactobacillus, the lactobacillus plantarum is preserved in China General Microbiological Culture Collection Center (CGMCC) on November 29, 2024, and the preservation number is CGMCC No.32857. The lactobacillus plantarum has the advantages that the lactobacillus plantarum is high in proliferation activity; the lactobacillus plantarum single cream is obtained by preparing a TPY liquid culture medium, adding nutritional ingredients, performing anaerobic culture, performing single cream fermentation and inoculating lactobacillus plantarum subjected to anaerobic culture into single cream for fermentation. By optimizing the proliferation conditions of the lactobacillus, the influence of different nutritional ingredients and different oxygen conditions on the proliferation of the lactobacillus is explored. The exploration is helpful for determining optimal lactobacillus proliferation conditions, so that the growth rate and metabolic activity of lactobacillus are improved. The culture efficiency of the lactobacillus is improved, and the proliferation efficiency of the lactobacillus in the TPY liquid culture medium can be remarkably improved by accurately controlling the components of the culture medium and oxygen conditions.

Description

Technical Field

[0001] The invention relates to the technical field of lactobacillus, and in particular to a plant lactobacillus with high proliferation activity and application thereof. Background Art

[0002] Cream is rich in nutritional value, and its fat content is much higher than the milk we usually consume. Not only that, cream also contains a lot of vitamins A and D, which is a good vitamin supplement for young children in their growth period. Secondly, cream contains a lot of protein, which can promote muscle growth and development.

[0003] Chinese patent CN117085079B discloses that the fermented product of Lactobacillus plantarum SF-L38 can be directly applied to the skin surface to effectively remove free radicals, inhibit the formation of reactive oxygen species, and reduce the degradation of extracellular matrix. It has the significant functions of alleviating ultraviolet damage and repairing skin photoaging, which is convenient and fast, and the effect is fast and lasting. Chinese patent CN118480475A discloses that Lactobacillus plantarum LP-1-679 can increase the lactic acid accumulation and malic acid consumption rate when fermenting fruits and vegetables with high L-malic acid content, and has good genetic stability.

[0004] In traditional microbial culture technology, the culture medium cannot fully meet the specific strains' delicate needs for key nutrients such as amino acids in some cases. This lack of nutrient supply directly limits the growth rate and metabolic activity of the strains, which in turn affects their performance during the fermentation process, such as acid production and flavor generation, which poses a challenge to improving the quality and stability of the final product.

[0005] On the other hand, as a popular dairy product, the diversity of cream's ingredients and taste not only comes from the differences in raw milk, but is also deeply influenced by the processing technology. Consumers' increasing requirements for food quality, especially their preference for delicate taste and flavor, means that there is still room for improvement in the taste of existing cream products, such as seeking a more balanced creaminess, reducing unnecessary impurities, or adding specific flavor elements to better adapt to different dietary cultures and personal taste preferences.

[0006] Based on this, the present invention designs a strain of Lactobacillus plantarum with high proliferation activity and application thereof to solve the above problems. Summary of the invention

[0007] In view of the above-mentioned shortcomings of the prior art, the present invention provides a strain of Lactobacillus plantarum with high proliferation activity and application thereof.

[0008] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0009] A strain of plant lactobacillus with high proliferation activity, which was deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration on November 29, 2024, with the deposit number: CGMCC No.32857.

[0010] The application of a plant lactobacillus with high proliferation activity comprises the following steps:

[0011] Step 1: Prepare TPY liquid medium;

[0012] Step 2, adding nutrients: adding whey protein hydrolysate and tributyrin to the culture medium, performing high-pressure steam sterilization, and inoculating the plant lactobacillus described in claim 1 into the culture medium;

[0013] Step 3, anaerobic culture: culturing and proliferating the inoculated Lactobacillus plantarum under anaerobic conditions;

[0014] Step 4, storing the Lactobacillus plantarum proliferated in step 3 in low-temperature glycerol for later use;

[0015] Step 5: Resuscitate the strains and then culture them in a constant temperature incubator. Inoculate the single colonies on the plate into MRS liquid culture medium and culture them anaerobically in a constant temperature incubator.

[0016] Step 6: Cream fermentation: melt and stir the packaged cream, perform homogenization and sterilization, and add emulsifier;

[0017] Step 7: inoculate the Lactobacillus plantarum cultured anaerobicy in step 5 into the cream treated in step 6 for fermentation to obtain Lactobacillus plantarum cream.

[0018] Furthermore, the Lactobacillus plantarum proliferated in step 3 is stored in glycerol at minus 80°C for later use.

[0019] Furthermore, step seven specifically includes: inoculating the cream processed in step six with the Lactobacillus plantarum cultured anaerobically in step five at an inoculation amount of 1-3%, fermenting at 32-42° C. for 18-30 hours, and obtaining the Lactobacillus plantarum cream.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The present invention optimizes the lactobacillus proliferation conditions and explores the effects of different nutrients (such as Hilma 8350 whey protein hydrolyzate and tributyrin) and different oxygen conditions (aerobic and anaerobic) on the lactobacillus proliferation. This exploration helps to determine the optimal lactobacillus proliferation conditions, thereby improving the growth rate and metabolic activity of lactobacillus.

[0022] The present invention improves the lactobacillus culture efficiency, and by precisely controlling the culture medium components and oxygen conditions, the present invention can significantly improve the proliferation efficiency of lactic acid bacteria in TPY liquid culture medium. This not only shortens the lactobacillus culture cycle, but also improves the stability and repeatability of the culture process, providing strong support for industrial production.

[0023] The present invention enhances the application performance of lactobacillus. The optimized proliferation conditions not only promote the growth of lactic acid bacteria, but also improve their performance in the fermentation process, such as increasing acid production capacity and flavor generation. These performance improvements help enhance the competitiveness of lactic acid bacteria in the fields of dairy fermentation, food preservation and biotechnology applications.

[0024] The cream product prepared by the high-proliferation-activity Lactobacillus plantarum of the present invention can better meet the consumers' increasing requirements for food quality, has an effectively improved taste, and can better adapt to different dietary cultures and personal taste preferences. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the prior art descriptions are briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0026] Figure 1 This is a colony morphology diagram of a strain of Lactobacillus plantarum with high proliferation activity according to the present invention;

[0027] Figure 2 This is a microscopic photograph of a strain of Lactobacillus plantarum with high proliferation activity according to the present invention;

[0028] Figure 3 A phylogenetic tree of a strain of Lactobacillus plantarum with high proliferation activity according to the present invention;

[0029] Figure 4 This is a graph showing the change in turbidity value of a strain of Lactobacillus plantarum with high proliferation activity according to the present invention;

[0030] Figure 5 This is a sample of pasteurized cream after adding Lactobacillus plantarum;

[0031] Figure 6 is the blank control chromatogram;

[0032] Figure 7 This is the chromatogram after 18h of fermentation. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0034] Embodiment 1: This embodiment provides a kind of plant lactobacillus, comprising the following steps:

[0035] Take 2g of naturally fermented cream from Inner Mongolia Autonomous Region and dilute it to 10% with sterile water. -6 (i.e. sterile water: cream is 10 6 :1), continue to add sterile water and dilute to 10 -7 , continue to add sterile water and dilute to 10 -8 ;

[0036] The samples after gradient dilution were spread on MRS solid medium plates and cultured in a 37°C incubator for 24 h;

[0037] After 24 hours, single colonies with good growth status were picked up with an inoculation loop and placed in TPY liquid culture medium for shaking culture to obtain Lactobacillus plantarum.

[0038] The plant lactobacillus was deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration on November 29, 2024, with the deposit number: CGMCC No.32857.

[0039] Comparative Example 1: This comparative example provides a lactobacillus proliferation condition, adding 0.25 g of Hilma 8350 whey protein hydrolyzate (addition amount is 0.25%) to 100 mL of TPY liquid culture medium, sterilizing the TPY liquid culture medium with high pressure steam (121° C. for 20 min), inoculating the plant lactobacillus described in Example 1 into the culture medium at an inoculum amount of 3%, and detecting the turbidity value (A) of the lactobacillus liquid culture medium at different time periods (0 h, 2 h, 4 h, 6 h, 8 h, 12 h, 14 h) by a spectrophotometer under anaerobic conditions. 600 ) to determine its proliferation (such as Figure 4 shown).

[0040] Comparative Example 2: This comparative example provides a lactobacillus proliferation condition, adding 0.25 g of Hilma 8350 whey protein hydrolyzate (addition amount is 0.25%) to 100 mL of TPY liquid culture medium, sterilizing the TPY liquid culture medium with high pressure steam (121° C. for 20 min), inoculating the plant lactobacillus described in Example 1 into the culture medium at an inoculum amount of 3%, and detecting the turbidity value (A) of the lactobacillus liquid culture medium at different time periods (0 h, 2 h, 4 h, 6 h, 8 h, 12 h, 14 h) by a spectrophotometer under aerobic conditions. 600 ) to determine its proliferation (such as Figure 4 shown).

[0041] Comparative Example 3: This comparative example provides a lactobacillus proliferation condition, adding 0.25g of Hilma 8350 whey protein hydrolyzate (added in an amount of 0.25%) and 0.15g of tributyrin (added in an amount of 0.15%) to 100mL of TPY liquid culture medium, sterilizing the TPY liquid culture medium with high pressure steam (121°C for 20min), inoculating the plant lactobacillus described in Example 1 into the culture medium at an inoculum amount of 3%, and detecting the turbidity value (A) of the lactobacillus liquid culture medium at different time periods (0h, 2h, 4h, 6h, 8h, 12h, 14h) by a spectrophotometer under aerobic conditions. 600 ) to determine its proliferation (such as Figure 4 shown).

[0042] Comparative Example 4: This comparative example provides a lactobacillus proliferation condition, adding 0.25 g of Hilma 8350 whey protein hydrolyzate (addition amount is 0.25%) to 100 mL of TPY liquid culture medium, sterilizing the TPY liquid culture medium with high pressure steam (121° C. for 20 min), inoculating the plant lactobacillus described in Example 1 into the culture medium at an inoculum amount of 3%, and detecting the turbidity value (A) of the lactobacillus liquid culture medium at different time periods (0 h, 2 h, 4 h, 6 h, 8 h, 12 h, 14 h) by a spectrophotometer under anaerobic conditions. 600 ) to determine its proliferation (such as Figure 4 shown).

[0043] Comparative Example 5: This comparative example provides a lactobacillus proliferation condition, adding 0.25g of Hilma 8350 whey protein hydrolyzate (added in an amount of 0.25%) and 0.15g of tributyrin (added in an amount of 0.15%) to 100mL of TPY liquid culture medium, sterilizing the TPY liquid culture medium with high pressure steam (121°C for 20min), inoculating the plant lactobacillus described in Example 1 into the culture medium at an inoculum amount of 3%, and detecting the turbidity value (A) of the lactobacillus liquid culture medium at different time periods (0h, 2h, 4h, 6h, 8h, 12h, 14h) by a spectrophotometer under anaerobic conditions. 600 ) to determine its proliferation (such as Figure 4 shown).

[0044] Experimental Example 1: Morphological observation experiment;

[0045] Gram stain the cultured colonies for preliminary morphological observation: take a glass slide, add a drop of PBS solution to the center of the slide, use a burned inoculation loop to pick up a small amount of bacterial moss from the probiotic plate and add it to the PBS solution, mix well and apply it into a thin film. Then heat the coated surface slightly on an alcohol lamp to dry it, then add 1 drop of crystal violet to the coated surface of the slide, stain for 1 minute, use filter paper to absorb the staining solution, rinse with fine water until the washing solution is colorless, and shake off the water on the slide. Use Lugol's iodine solution to mordant for about 1 minute and wash with water. Use filter paper to absorb the residual water on the slide, tilt the slide, and use a dropper to add 95% anhydrous ethanol to decolorize against a white background. When the ethanol flowing out is no longer purple, wash with water immediately to stop decolorization, shake off the water on the slide, and finally use safranin dye to re-stain. After absorbing the water with absorbent paper, you can observe rod-shaped Gram-positive bacteria (such as Figure 2 shown).

[0046] Experimental Example 2: DNA sequencing;

[0047] The above-mentioned bacterial colony was inoculated into TPY liquid culture medium, and the 16S rDNA of the strain was sequenced using 16S rDNA universal primers. After BLAST (biological macromolecular sequence alignment search tool) retrieval and comparison were performed by NCBI (National Center for Biotechnology Information, USA), it was found that the strain provided by the present invention was Lactobacillus plantarum, and an evolutionary tree was constructed using MEGA software, as shown in FIG. Figure 3 As shown;

[0048] 16S rDNA sequencing results:

[0049]

[0050] Experimental Example 3: The plant lactobacillus after proliferation in Comparative Example 5 was placed in a glycerol storage at -80°C to resuscitate the strain (the plant lactobacillus described in Example 1) (the thawed glycerol tube was streaked in three zones on an MRS solid culture medium to obtain a single colony). The streaked plate was sealed with a sealing film and then placed in a constant temperature incubator at 37°C for 24 hours.

[0051] A single colony on the plate was inoculated into MRS liquid culture medium and cultured anaerobically in a constant temperature incubator at 37°C for 24 hours to achieve optimal activity.

[0052] The cream was melted and placed in a blender, and then the cream was homogenized at a high speed and Tween 80 emulsifier was added. After homogenization, the cream was dispensed into 50 mL small fermentation bottles, and then the dispensed fermentation bottles were pasteurized to obtain blank samples before inoculation and fermentation. The blank samples were inoculated with anaerobic cultured lactobacilli for fermentation to obtain plant lactobacillus cream.

[0053] Three single factor conditions, namely fermentation time, fermentation temperature and inoculation amount, were set for investigation.

[0054] Fermentation time: Inoculate the pasteurized fermentation bottle when it is cooled to about 37°C, with an inoculation volume of 2%. Select the fermentation time of 18h, 24h, and 30h as the time gradient, make 3 parallels for each gradient, and keep a blank for detection. After the inoculation is completed, seal it and place it in a 37°C constant temperature incubator for fermentation. After the fermentation is completed, immediately place it in a -80°C refrigerator to stop the fermentation, so as to facilitate the subsequent determination of physical and chemical indicators.

[0055] Fermentation temperature: The pasteurized fermentation bottle was cooled to 37℃ for inoculation, with an inoculation volume of 2%. The fermentation temperature was selected as 32℃, 37℃, and 42℃ as the temperature gradient. Three parallels were made for each gradient, and one blank was reserved for detection. After the inoculation, the bottle was sealed and placed in a constant temperature incubator at 32℃, 37℃, and 42℃ for fermentation for 24 hours. After the fermentation was completed, it was immediately placed in a refrigerator at -80℃ to stop the fermentation, so as to facilitate the subsequent determination of physical and chemical indicators.

[0056] Inoculation amount: The pasteurized fermentation bottle was cooled to 37℃ for inoculation. The inoculation amount was selected as 1%, 2%, and 3% as the inoculation amount gradient. Three parallels were made for each gradient, and one blank was reserved for testing. After the inoculation was completed, it was sealed and placed in a 37℃ constant temperature incubator for fermentation for 24 hours. After the fermentation was completed, it was immediately placed in a -80℃ refrigerator to stop the fermentation, so as to facilitate the subsequent determination of physical and chemical indicators.

[0057] like Figure 5 Shown is cream after pasteurization.

[0058] Table 1 Factor level table:

[0059]

[0060] Table 2 Orthogonal experiment table:

[0061]

[0062]

[0063] From the mean and range, we can see that the influence of each factor is from large to small, fermentation temperature > fermentation time > inoculation amount, and the optimal fermentation conditions are fermentation temperature 37°C, fermentation time 30h, and inoculation amount 2%. However, this combination did not appear in the orthogonal experiment. Therefore, the optimal fermentation conditions were verified, and the sensory score was 9.70, which was greater than the various combinations in the orthogonal experiment. In summary, the optimal fermentation conditions obtained by orthogonal experiment optimization are fermentation temperature 37°C, fermentation time 30h, and inoculation amount 2%.

[0064] This test adopts fuzzy mathematical sensory evaluation method. The evaluation team consists of 10 people, all of whom are food professionals; they evaluate the color, smell, texture and taste of fermented cream.

[0065] Table 3 Sensory evaluation criteria:

[0066]

[0067]

[0068] The sensory evaluation results are processed secondary using the “f function method” to avoid double peaks or even multiple peaks caused by uniform weight distribution or large differences in evaluation results.

[0069] Experimental Example 4: A sample with an inoculation amount of 3% and an uninoculated sample were placed in a constant temperature incubator at 37°C for fermentation for 24 hours. After the fermentation, the samples were measured according to the national standard GB 5009.239-2016 "Determination of Acidity of Foods".

[0070] According to the national standard, the reagents are first prepared, including sodium hydroxide standard solution (0.1000 mol / L), reference solution, phenolphthalein indicator solution, neutral ethanol-ether mixture, and distilled water without carbon dioxide.

[0071] Based on the national standard classification of different milk and other dairy products, select appropriate methods for analysis operations.

[0072] First, prepare the reference solution: add 2.0 mL of the reference solution to a conical flask containing an equal volume of the corresponding solution, gently rotate it to mix it, and obtain the standard reference color. If multiple similar products are to be measured, this reference solution can be used for the entire measurement process, but the time shall not exceed 2 hours.

[0073] Weigh 10g (accurate to 0.001g) of the mixed sample, place it in a 250mL conical flask, add 30mL of neutral ethanol-ether mixture, mix well, add 2.0mL of phenolphthalein indicator solution, mix well, and titrate with sodium hydroxide standard solution, rotating the flask while adding until the color is similar to that of the reference solution and does not fade within 5 seconds. The entire titration process should be completed within 45 seconds. During the titration process, blow nitrogen into the conical flask to prevent the solution from absorbing carbon dioxide in the air. Record the milliliters of sodium hydroxide standard titration solution consumed (V 2) , substitute it into the formula for calculation.

[0074] The acidity value of the cream sample is expressed in (°T) and is calculated according to the formula:

[0075] X 2 =C 2 ×(V 2 -V 0 )×100 / m 2 ×0.1

[0076] Where:

[0077] X 2 --- acidity of the sample, in degrees (°T) [measured as milliliters of 0.1 mol / L sodium hydroxide consumed by 100 g of sample, in milliliters per 100 g (mL / 100 g)];

[0078] C 2 ---The molar concentration of the sodium hydroxide standard solution, in moles per liter (mol / L);

[0079] V 2 ---The volume of sodium hydroxide standard solution consumed during titration, in milliliters (mL);

[0080] V 0 ---The volume of sodium hydroxide standard solution consumed in the blank experiment, in milliliters (mL);

[0081] 100——100g sample;

[0082] m 2 --- mass of the sample, in grams (g);

[0083] 0.1——Acidity Theoretical definition of the molar concentration of sodium hydroxide, the unit is mole per liter (mol / L).

[0084] The results were expressed as the arithmetic mean of two independent determination results obtained under repeatability conditions, with three significant figures retained.

[0085] The acidities were 18.0, 72.0, and 77.5, respectively. The data showed that after 24 hours of culture in a 37°C constant temperature incubator, the acidity of the sample with an inoculation amount of 3% increased significantly, and Lactobacillus plantarum produced acid during the fermentation process.

[0086] Experimental Example 6: Determination of metabolites in fermented cream;

[0087] The following samples of fermented cream were selected for fatty acid determination:

[0088] 1. Blank control group;

[0089] 2. 1% inoculum, ferment at 37℃ for 18 hours;

[0090] 3. 1% inoculum, ferment at 37℃ for 30 hours;

[0091] 4. 1% inoculum, ferment at 32℃ for 24 hours;

[0092] 5. 1% inoculum, ferment at 37℃ for 24 hours;

[0093] 6. Ferment with 1% inoculum at 42°C for 24 hours.

[0094] Gas chromatography-quadrupole mass spectrometry was used to detect metabolites in samples. Metabolites in biological samples were qualitatively and quantitatively analyzed by comparing with the retention time and molecular mass (molecular mass error <10ppm) of the standard.

[0095] Project Process:

[0096] 1. Preparation of standard products;

[0097] The following standard substances were purchased from Sigma: acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, hexanoic acid, and isohexanoic acid.

[0098] Weigh the standard samples required for this project accurately, add n-hexane solution to prepare 1 mg / mL or 3 mg / mL single standard stock solution, take appropriate amount of single standard stock solution and mix to prepare mixed standard solution for future use. Perform the test on the machine according to the concentration gradient shown in Table 4.

[0099] Table 4 Mixed standard gradient concentration information:

[0100]

[0101] Experimental method: First, take an appropriate amount of sample into a centrifuge tube, add 50 μL of 30% phosphoric acid solution, then add 300 μL of acetone solution, homogenize for 3 minutes, centrifuge at 12000 rpm for 10 minutes, take the supernatant solution, dilute it several times according to the actual situation, and then test it on the machine.

[0102] The operating parameters are as follows: The data acquisition instrument system mainly includes a gas chromatograph (Agilent 7820, USA) and a quadrupole mass spectrometer detection system (Agilent 5977, USA). The Agilent gas chromatograph system (Agilent 7820, Agilent Technologies, USA) was used, the chromatographic column was DB-FFAP (30m×0.25mm×0.25μm), the injection volume was 1μL, the split ratio was 10:1, the carrier gas was high-purity helium, the flow rate was 1.0mL / min, the initial temperature of the column oven was 70℃ for 5.0min, and the temperature was raised to 100℃ at a rate of 6℃ / min. The quadrupole mass spectrometer detection system (Agilent 5977, Agilent Technologies, USA) of Aiglent was used, equipped with an electron impact ion source (EI) and a MassHunter workstation. The optimized mass spectrometry analysis conditions were as follows: the injection port temperature was 260°C, the quadrupole temperature was 150°C, the scanning mode was single channel scanning (SIM), and the mass scanning range (m / z range) was 30-550.

[0103] Qualitative and quantitative analysis of metabolites: Quantitative analysis refers to the analysis of the content of metabolites, which is mainly achieved by drawing a standard curve of a standard substance. The concentration of the standard substance is used as the horizontal axis and the peak area is used as the vertical axis to obtain the mathematical relationship between the target compound and its peak area (linear, quadratic equation, logarithmic, etc.), and then the concentration of the corresponding compound in the unknown sample is calculated according to its peak area to achieve absolute quantitative analysis of the target compound.

[0104] The raw data were pre-processed using Quant-My-Way software, including baseline filtering, peak identification, peak matching, retention time correction, and peak alignment, to obtain a data matrix containing retention time, mass-to-charge ratio, and peak intensity. The total ion flow chromatogram (TIC) is a graph obtained by summing up the intensities of all ions in the mass spectrum at each time point and plotting them continuously. The horizontal axis is the retention time (Time, min), and the vertical axis is the ion flow intensity (Counts) of the ion detection.

[0105] The retention time of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, caproic acid, and isocaproic acid was obtained by using the standard, and then the peak areas of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, caproic acid, and isocaproic acid were obtained by comparing the chromatograms of the blank control and each sample, so that the final concentration can be calculated. As shown in Table 5, the changes in metabolites of the blank control chromatogram (unfermented) and each sample (after fermentation) were compared using the final concentration;

[0106] in, Figure 6 is the blank control chromatogram. Figure 7 This is the chromatogram after 18h of fermentation.

[0107] Table 5 Final concentration of metabolites in each sample:

[0108]

[0109]

[0110] After comparison with the blank control, it was found that the content of short-chain fatty acids including acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, caproic acid, and isocaproic acid after fermentation increased, among which acetic acid increased most significantly, followed by caproic acid, butyric acid, and propionic acid, and acetic acid increased most significantly under the condition of 1% inoculation at 37℃ for 18h. Compared with 1% inoculation at 37℃ for 24h, the content of propionic acid and isobutyric acid under the condition of 1% inoculation at 37℃ for 24h was higher than that under the condition of 1% inoculation at 37℃ for 18h. It can be estimated that plant lactobacillus mainly secretes acetic acid before 18h, and may tend to secrete propionic acid and butyric acid from 18 to 24h. These short-chain fatty acids have a positive regulatory effect on the body, such as acetic acid can regulate the acid-base balance of the intestine, propionic acid can reduce cholesterol and reduce fat storage, butyric acid can protect the intestinal mucosal barrier and regulate insulin sensitivity, etc.

[0111] By comparing the metabolite contents under different temperature gradients, it can be concluded that acetic acid, propionic acid, butyric acid and caproic acid vary greatly in the range of 32°C to 37°C. When the temperature rises from 37°C to 42°C, the metabolite content does not change as much as in the range of 32°C to 37°C. Despite this, acetic acid continues to increase, but isobutyric acid and isovaleric acid decrease.

[0112] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. Such modifications or replacements will not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A strain of Lactobacillus plantarum with high proliferation activity, characterized in that: The plant lactobacillus was deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration on November 29, 2024, with the deposit number: CGMCC No.32857.

2. Use of a strain of Lactobacillus plantarum with high proliferation activity as claimed in claim 1, characterized in that: The following steps are involved: Step 1: Prepare TPY liquid medium; Step 2, adding nutrients: adding whey protein hydrolysate and tributyrin to the culture medium, performing high-pressure steam sterilization, and inoculating the plant lactobacillus described in claim 1 into the culture medium; Step 3, anaerobic culture: culturing and proliferating the inoculated Lactobacillus plantarum under anaerobic conditions; Step 4, storing the Lactobacillus plantarum proliferated in step 3 in low-temperature glycerol for later use; Step 5: Resuscitate the strains and then culture them in a constant temperature incubator. Inoculate the single colonies on the plate into MRS liquid culture medium and culture them anaerobically in a constant temperature incubator. Step 6: Cream fermentation: melt and stir the packaged cream, perform homogenization and sterilization, and add emulsifier; Step 7: inoculate the Lactobacillus plantarum cultured anaerobicy in step 5 into the cream treated in step 6 for fermentation to obtain Lactobacillus plantarum cream.

3. The use of plant lactobacillus with high proliferation activity according to claim 2, characterized in that: The plant lactobacillus proliferated in step 3 is stored in glycerol at -80°C for later use.

4. The use of the plant lactobacillus with high proliferation activity according to claim 3, characterized in that: Step seven specifically comprises: inoculating the cream processed in step six with the Lactobacillus plantarum cultured anaerobically in step five at an inoculation amount of 1-3%, and fermenting at 32-42° C. for 18-30 hours to obtain Lactobacillus plantarum cream.

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