Lactobacillus paracasei LP007 probiotic liquid with effect of reducing blood fat and preparation method of lactobacillus paracasei LP007 probiotic liquid

Abstract

The invention discloses a lactobacillus paracasei LP007 probiotic liquid with a blood fat reducing effect and a preparation method of the lactobacillus paracasei LP007 probiotic liquid. On the basis of the casei paracasei LP007 and in combination with functional components such as the red yeast rice extract, the gamma-aminobutyric acid and the DHA, the probiotic liquid with the remarkable blood fat reducing effect is successfully prepared. Experiments show that the probiotic liquid can effectively reduce serum total cholesterol, low-density lipoprotein cholesterol and triglyceride, improve the level of high-density lipoprotein cholesterol and optimize lipid metabolism, and in addition, the formula can also improve intestinal flora balance, enhance the antioxidant capacity of the body, protect the liver to a certain extent, reduce fatty degeneration and improve the immunity of the human body. Compared with a contrast group without red yeast rice and GABA, the probiotic liquid has remarkable advantages in lipid-lowering effect, has good taste and biological stability, and is suitable for being drunk for a long time as a functional health product.

Description

Technical Field

[0001] The present invention relates to the fields of probiotic biomedicine and health products, and particularly to a Lactobacillus paracasei LP007 probiotic liquid with lipid-lowering effect and a preparation method thereof. Background Art

[0002] Probiotic liquid is a liquid preparation containing live probiotics, usually cultured from specific lactic acid bacteria, bifidobacteria or other beneficial microorganisms, aiming to regulate the intestinal flora, promote digestive health, and enhance immunity. In its preparation process, probiotics are fermented and cultured to form a liquid with a high viable bacteria concentration, and appropriate nutritional substrates or functional ingredients, such as oligosaccharides, vitamins or Chinese herbal medicine extracts, can be added to improve its efficacy and stability. Probiotic liquid can be used in the fields of food, beverage or health products, and is commonly found in yogurt, fermented beverages and functional nutritional supplements.

[0003] A strain of Lactobacillus paracasei with probiotic characteristics isolated from pickles, named Lactobacillus paracasei PC804, with the Chinese patent publication number: CN115261264A, has been deposited in the General Microbiological Culture Collection Center of the China Committee for Culture Collection of Microorganisms, with the deposit number CGMCC No. 23917. Lactobacillus paracasei PC804 in the present invention has a high survival rate in gastrointestinal fluid, good bile salt tolerance, and strong survival ability in the human body. Lactobacillus paracasei PC804 has probiotic characteristics of enhancing immune resistance, increasing the diversity of intestinal flora, and enhancing the stability of intestinal flora. In addition, PC804 can be well applied to fermented dairy products as a probiotic.

[0004] In the actual use process of the above patent and the prior art, the following problems exist: the lipid-lowering effect is weak, and during the fermentation process, it is at a fixed angle and in a stirring and mixing manner, resulting in reduced fermentation efficiency and effect. Summary of the Invention

[0005] Therefore, in order to solve the above deficiencies, the present invention provides a Lactobacillus paracasei LP007 probiotic liquid with lipid-lowering effect and a preparation method thereof.

[0006] In order to achieve the above object, the present invention adopts the following technical solution: a Lactobacillus paracasei LP007 probiotic liquid with lipid-lowering effect and a preparation method thereof, the ratio of the probiotic liquid is: Lactobacillus paracasei LP007 is 1×10 9 - 5×10¹ 0CFU / mL, 2 - 8 g / L of fructooligosaccharides, 50 - 100 mg / L of γ-aminobutyric acid, 150 - 300 mg / L of monascus rice extract, 20 - 50 mg / L of DHA, 1 - 5 g / L of casein hydrolyzate, 0.2 - 0.5 mg / L of vitamin B group (B6, B12), 10 - 20 g / L of lactose and 850 - 970 mL / L of purified water. The Lactobacillus paracasei LP007 is deposited in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, with the deposit number: CGMCC NO.32193.

[0007] Preferably, the specific steps are as follows:

[0008] S1 Medium Preparation: Dissolve 12 - 18 g / L of lactose in 60% volume of purified water, stir evenly, add 4 - 6 g / L of fructooligosaccharides, continue stirring, add DHA, 50 - 80 mg / L of γ-aminobutyric acid, 0.3 - 0.4 mg / L of vitamin B group, adjust the pH to 6.5 - 6.8, sterilize at 121℃ under high pressure for 15 min, and cool to 37℃ for standby;

[0009] S2 Bacterial Liquid Culture: Activate Lactobacillus paracasei LP007: Use sterile MRS medium, culture at 37℃ for 12 - 18 h until it reaches 1×10 9 CFU / mL. Under sterile conditions, inoculate 5% (v / v) into the fermentation broth, culture in a shaker at 37℃ for 12 - 18 h, monitor the growth of the bacteria, and ensure the concentration is 1×10 9 -3×10¹ 0 CFU / mL;

[0010] S3 Fermentation Process: With an inoculation amount of 5%, transfer the bacterial liquid to a fermenter. The fermenter is installed on an auxiliary fermentation device for adjusting the fermentation angle position. Then maintain at 37℃, stir at 150 - 200 rpm, introduce 5% CO2, ferment for 24 - 48 h, monitor the pH, and finally it is about 4.5 - 5.0 to ensure the metabolic balance of lactic acid bacteria;

[0011] S4 Addition and Mixing of Lipid-lowering Components: Measure 150 - 300 mg / L of monascus rice extract and 1 - 5 g / L of casein hydrolyzate, slowly add them to the fermentation broth at 4℃, stir at 1000 rpm for 10 - 15 min, and perform homogenization at 120 MPa for 2 times;

[0012] S5 Sterilization and Packaging: Perform pasteurization at 65 - 75℃ for 15 min to ensure the survival rate of bacteria >90%, filter to remove particulate impurities, and perform aseptic filling and encapsulation into bottles.

[0013] Preferably, the auxiliary fermentation device includes an adjusting mechanism for supporting and adjusting the angular position, a shaking mechanism provided at the top of the adjusting mechanism and having a shaking function, and a clamping member provided at the top of the shaking mechanism for clamping the fermentation tank.

[0014] Preferably, the adjusting mechanism includes a chassis, a support rod provided at the top of the chassis, a support plate installed at the top of the support rod, a bottom frame provided at the right end of the top of the support plate, adjusting knobs screwed into the front and rear ends of the bottom frame, a top frame connected to the top of the adjusting knobs. The top frame has the same structure as the bottom frame and openings are respectively formed at the front and rear positions of the right ends of both. A connecting ball for connecting the right ends of the top frame and the bottom frame, a fixing rod provided inside the opening, a spring for connecting two groups of fixing rods, and a support plate provided on the left side of the top frame. The top of the support plate is connected to the shaking mechanism.

[0015] Preferably, the bottom frame is in an L shape.

[0016] Preferably, the right end of the bottom of the bottom frame is in movable cooperation with the connecting ball.

[0017] Preferably, the shaking mechanism includes a bottom box provided at the bottom of the support plate, a guiding frame provided at the top of the bottom box, a sliding mechanism installed inside the guiding frame, and a support plate provided at the top of the sliding mechanism. The sliding mechanism drives the support plate to move, and the top of the support plate is connected to the clamping member.

[0018] Preferably, the sliding mechanism includes a sliding plate movably embedded in the inner side of the guiding frame, connecting members provided at both ends of the top of the sliding plate for connecting the support plate, a swinging member provided inside the sliding plate, a connecting member connected to the right end of the swinging member, a support provided at the right end of the bottom of the guiding frame, and a connecting shaft passing through the support and connecting the connecting member. The top of the connecting shaft is connected to the left end of the connecting member.

[0019] Preferably, a driving motor is provided at the right end inside the bottom box. The output shaft at the bottom of the driving motor is connected to a driving belt pulley. The driving belt pulley rotates synchronously with a driven belt pulley through a synchronous belt on the outside. A rotating shaft is provided in the middle of the driven belt pulley. The top of the rotating shaft is connected to the connecting shaft, and the connecting shaft rotates with the rotating shaft.

[0020] Advantages of the present invention:

[0021] The present invention is based on Lactobacillus paracasei LP007 and combines functional ingredients such as red yeast rice extract, γ-aminobutyric acid, and DHA to successfully prepare a probiotic liquid with a significant lipid-lowering effect. Experiments show that this probiotic liquid can effectively reduce serum total cholesterol, low-density lipoprotein cholesterol, and triglycerides, while increasing the level of high-density lipoprotein cholesterol, optimizing lipid metabolism. In addition, this formula can also improve the balance of the intestinal flora, enhance the body's antioxidant capacity, and provide protection to the liver to a certain extent, reducing fatty degeneration. Compared with the control group without red yeast rice and GABA added, this probiotic liquid has a significant advantage in lipid-lowering effect, and has a good taste and biological stability, making it suitable for long-term drinking as a functional health product.

[0022] In the present invention, an auxiliary fermentation device is used in cooperation during the fermentation process. A regulating mechanism and a shaking mechanism are arranged inside the device. A clamping member is arranged at the top of the shaking mechanism, and the clamping member can clamp and place the fermentation tank. Cooperating with the shaking mechanism at the bottom, the fermentation tank is stably and efficiently shaken. And the regulating mechanism at the bottom can be tilted at an angle position manually, which is convenient for stably shaking the liquid inside the fermentation tank evenly, effectively improving the fermentation effect and efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 is a schematic structural diagram of the auxiliary fermentation device of the present invention;

[0024] Figure 2 is a schematic structural diagram of the regulating mechanism of the present invention;

[0025] Figure 3 is a partial structural schematic diagram of the regulating mechanism of the present invention;

[0026] Figure 4 is a schematic structural diagram of the shaking mechanism of the present invention;

[0027] Figure 5 is a schematic structural diagram of the sliding mechanism of the present invention;

[0028] Figure 6 is a schematic structural diagram of another perspective of the sliding mechanism of the present invention;

[0029] Figure 7 is a schematic structural diagram of the interior of the bottom box of the present invention.

[0030] Wherein: adjusting mechanism - 1, shaking mechanism - 2, clamping member - 3, chassis - 11, support rod - 12, support plate - 13, chassis - 14, adjusting knob - 15, top frame - 16, connecting ball - 17, fixed rod - 18, spring - 19, support plate - 110, bottom box - 21, guiding frame - 22, sliding mechanism - 23, support plate - 24, sliding plate - 231, connecting member - 232, swinging member - 233, connecting member - 234, support - 235, connecting shaft - 236, driving motor - 211, driving pulley - 212, synchronous belt - 213, driven pulley - 214, rotating shaft - 215. Detailed implementation mode

[0031] In order to further explain the technical solution of the present invention, the following will be elaborated in detail through specific embodiments.

[0032] Embodiment 1

[0033] The present invention provides a probiotic liquid of Lacticaseibacillus paracasei LP007 with lipid - lowering effect. The proportion of the probiotic liquid is as follows: Lacticaseibacillus paracasei LP007 is 1×10 9 - 5×10¹ 0 CFU / mL, fructooligosaccharide 2 - 8 g / L, γ - aminobutyric acid 50 - 100 mg / L, monascus extract 150 - 300 mg / L, DHA 20 - 50 mg / L, casein hydrolyzate 1 - 5 g / L, vitamin B group (B6, B12) 0.2 - 0.5 mg / L, lactose 10 - 20 g / L, and purified water 850 - 970 mL / L. Lacticaseibacillus paracasei LP007 is preserved in the China General Microbiological Culture Collection Center, with the preservation number: CGMCC N.32193, preservation address: China General Microbiological Culture Collection Center, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, preservation result: survived;

[0034] The specific steps are as follows: S1 Medium preparation: Take 12 g / L of lactose and dissolve it in 60% volume of purified water, stir evenly, add 4 g / L of fructooligosaccharide, continue stirring, add 25 mg / L of DHA, 50 mg / L of γ - aminobutyric acid, 0.3 mg / L of vitamin B group, adjust the pH to 6.5, sterilize at 121℃ for 15 min, and cool to 37℃ for standby; S2 Bacterial liquid culture: Activate Lacticaseibacillus paracasei LP007: Use sterile MRS medium, culture at 37℃ for 12 h until 1×10 9 CFU / mL, under sterile conditions, inoculate 5% (v / v) into the fermentation broth, culture on a shaker at 37℃ for 12 h, monitor the growth of the bacteria, and ensure the concentration is 1×10 9CFU / mL; S3 fermentation process: With an inoculation amount of 5%, transfer the bacterial liquid to the fermenter. The fermenter is installed on the auxiliary fermentation device for adjusting the fermentation angle position. Then maintain at 37°C, stir at 150 rpm, introduce 5% CO2, and ferment for 24 h. Monitor the pH, which is finally about 4.5 to ensure the metabolic balance of lactic acid bacteria; S4 addition and mixing of lipid-lowering components: Measure 150 mg / L of red yeast rice extract and 2 g / L of casein hydrolyzate, slowly add them to the fermentation broth at a low temperature of 4°C, stir at 1000 rpm for 10 min, and perform homogenization at 120 MPa for 2 times; S5 sterilization and packaging: Pasteurize at 65°C for 15 min to ensure that the survival rate of bacteria is >90%, filter to remove particulate impurities, and perform aseptic filling and encapsulation into bottles.

[0035] Please refer to Figure 1 , the auxiliary fermentation device includes an adjusting mechanism 1 for adjusting the shaking angle. The top of the adjusting mechanism 1 is connected to the shaking mechanism 2. Four groups of clamping members 3 are installed on the top of the shaking mechanism 2. The four groups of clamping members 3 are convenient for clamping and lifting the fermenter, and are shaken by the shaking mechanism 2 at the bottom.

[0036] Please refer to Figure 2 and Figure 3 , the adjusting mechanism 1 includes a chassis 11 for support. Three groups of support rods 12 are vertically welded to the left end of the top of the chassis 11. The top of the support rods 12 is fixed with a support plate 13. The right end of the top of the support plate 13 is welded with a bottom frame 14. The bottom frame 14 is in an L shape. Adjusting knobs 15 are screwed into the front and rear ends inside the bottom frame 14. The top of the adjusting knob 15 is connected to a top frame 16. The top frame 16 has the same structure as the bottom frame 14. And the right ends inside the bottom frame 14 and the top frame 16 are connected by a connecting ball 17. The right end of the bottom of the bottom frame 14 can move on the connecting ball 17. Corresponding two groups of openings are provided at the upper end of the opening inside the top frame 16 and the lower end of the opening inside the bottom frame 14. And fixing rods 18 are fixed at the upper end of the opening inside the top frame 16 and the lower end of the opening inside the bottom frame 14. The two groups of fixing rods 18 are connected by a spring 19. A support plate 110 is fixed on the left side inside the top frame 16. The top of the support plate 110 is connected to the shaking mechanism 2.

[0037] Please refer to Figures 4 - 7 , the shaking mechanism 2 includes a bottom box 21 provided on the top of the support plate 110. The top of the bottom box 21 is connected to a guiding frame 22. A sliding mechanism 23 is arranged inside the guiding frame 22. The top of the sliding mechanism 23 is connected to a support plate 24. The support plate 24 can move left and right along with the sliding mechanism 23;

[0038] The sliding mechanism 23 includes a sliding plate 231 which is movably embedded inside the guiding frame 22. An opening is formed in the middle of the sliding plate 231. At the left and right ends of the top of the opening, connecting members 232 for connecting the support plate 24 are fixed. In the middle of the inner side of the opening, a swinging member 233 is attached. At the right end inside the swinging member 233, a connecting member 234 is connected. The connecting member 234 is placed on the top of a support 235. The front and rear positions on the top of the support 235 are fixed to the guiding frame 22. A connecting shaft 236 is arranged in the middle of the guiding frame 22. The top of the connecting shaft 236 is connected to the left end of the bottom of the connecting member 234. By rotating the connecting shaft 236, the connecting member 234 can perform eccentric rotation;

[0039] A driving motor 211 is arranged at the right end inside the bottom box 21. The output shaft at the bottom of the driving motor 211 is connected to a driving pulley 212. The driving pulley 212 rotates synchronously with a driven pulley 214 through a synchronous belt 213 on the outside. A rotating shaft 215 is arranged in the middle of the driven pulley 214. The top of the rotating shaft 215 is connected to the connecting shaft 236. The connecting shaft 236 rotates along with the rotating shaft 215.

[0040] The specific implementation process is as follows:

[0041] When the auxiliary fermentation device is needed to assist the fermentation process, first, the fermentation tank is clamped into the clamping member 3, and then the angle is adjusted. The user can rotate a set of adjusting knobs 15 according to the needs. During the rotation of the adjusting knobs 15, one end of the top frame 16 can be lifted or pulled down. The right end of the top frame 16 can be guided on the connecting ball 17 to realize the adjustment of the angular position of the top sliding mechanism 23;

[0042] After the adjustment, shaking is started to mix and stir the solution inside the fermentation tank. The driving motor 211 can be started to work. The driving motor 211 drives the driving pulley 212 to rotate. During the rotation of the driving pulley 212, the driven pulley 214 can be driven to rotate through the synchronous belt 213. The rotating shaft 215 in the middle of the driven pulley 214 can drive the connecting shaft 236 to rotate. The connecting shaft 236 can drive the connecting member 234 connected to the top to perform eccentric motion, so as to realize the reciprocating motion of the swinging member 233 inside the sliding plate 231. The sliding plate 231 can move reciprocally inside the guiding frame 22 under force. The support plate 24 connected to the top can drive the fermentation tank inside the clamping member 3 to shake.

[0043] Embodiment 2

[0044] A probiotic liquid of Lactobacillus paracasei LP007 with a blood lipid-lowering effect. The proportion of the probiotic liquid is: Lactobacillus paracasei LP007 is 5×10 93×10¹ CFU / mL, fructooligosaccharide 6 g / L, γ-aminobutyric acid 80 mg / L, monascus extract 250 mg / L, DHA 40 mg / L, casein hydrolyzate 3.5 g / L, vitamin B complex (B6, B12) 0.4 mg / L, and lactose 18 g / L. The specific preparation steps are the same as those in Example 1.

[0045] Example 3

[0046] A probiotic liquid of Lactobacillus paracasei LP007 with lipid-lowering effect. The ratio of the probiotic liquid is as follows: Lactobacillus paracasei LP007 is 3×10¹ 0 CFU / mL, fructooligosaccharide 5 g / L, γ-aminobutyric acid 60 mg / L, monascus extract 200 mg / L, DHA 200 mg / L, casein hydrolyzate 2.5 g / L, vitamin B complex (B6, B12) 0.35 mg / L, and lactose 15 g / L. The specific preparation steps are the same as those in Example 1.

[0047] A comparative experiment was conducted, and two comparative examples were added for comparison with three examples.

[0048] The components of the two comparative examples are as follows:

[0049] Ingredient Control Example 1 (g / L or CFU / mL) Control Example 2 (g / L or CFU / mL) Lactobacillus paracasei LP007 <![CDATA[1×10 9 CFU / mL]]> <![CDATA[5×10 9 CFU / mL]]> Fructooligosaccharide (FOS) 4 g / L 6 g / L γ-Aminobutyric acid (GABA) 0 mg / L (not added) 0 mg / L (not added) Red yeast rice extract 0 mg / L (not added) 0 mg / L (not added) DHA (from algal oil) 20 mg / L 30 mg / L Casein hydrolyzate 2 g / L 3 g / L Vitamin B complex (B6, B12) 0.2 mg / L 0.3 mg / L Lactose (or glucose) 10 g / L 15 g / L Purified water Appropriate amount Appropriate amount

[0050] SD rats aged 6 - 8 weeks with a body weight of 180 - 220 g were randomly divided into 5 groups of 12 rats each. All rats were adaptively fed for 7 days before the experiment and were given a high-fat diet containing 1.5% cholesterol, 10% lard, 0.5% bile salt, and 87% basal diet. The experimental groups are as follows:

[0051] Group Experimental content Treatment method Normal control group Normal diet, no additional intervention Normal diet + purified water High-fat control group High-fat diet, no probiotic intervention High-fat diet + purified water Example 1 High-fat diet + probiotic solution (low dose) <![CDATA[2 mL per day, bacterial concentration 1×10 9 CFU / mL]]> Example 2 High-fat diet + probiotic solution (medium dose) <![CDATA[2 mL per day, bacterial concentration 5×10 9 CFU / mL]]> Example 3 High-fat diet + probiotic solution (high dose) <![CDATA[2 mL per day, bacterial concentration 3×10¹ 0 CFU / mL]]> Control Example 1 High-fat diet + without red yeast rice / GABA 2 mL per day, without key ingredients Control Example 2 High-fat diet + only containing lactic acid bacteria 2 mL per day, only containing lactic acid bacteria, without lipid-lowering functional ingredients

[0052] Experimental period: 8 weeks. Blood was collected every 2 weeks to detect blood lipid levels, and the body weight changes of the rats were recorded.

[0053] Detection indicators: Blood lipid-related indicators, serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglyceride (TG), liver function indicators, alanine aminotransferase ALT, aspartate aminotransferase AST, and liver pathological section detection to detect the degree of fatty degeneration.

[0054] The changes in blood lipid levels (after 8 weeks) are as follows in the table:

[0055] Index (mmol / L) Normal control group High-fat control group Example 1 Example 2 Example 3 Control Example 1 Control Example 2 Total cholesterol (TC) 2.9±0.3 6.8±0.5 5.2±0.4 4.8±0.3 4.5±0.2 6.1±0.4 6.3±0.5 Low-density lipoprotein (LDL-C) 0.8±0.2 3.5±0.4 2.6±0.3 2.2±0.3 2.0±0.2 3.2±0.4 3.4±0.5 High-density lipoprotein (HDL-C) 1.5±0.2 0.9±0.2 1.2±0.1 1.3±0.2 1.4±0.1 1.0±0.2 1.0±0.2 Triglyceride (TG) 0.7±0.1 2.1±0.3 1.8±0.2 1.5±0.2 1.2±0.1 2.0±0.3 2.1±0.4

[0056] Key results: Example 3 had the best effect, with TC, LDL-C and TG significantly decreased, and HDL-C increased to 1.4mmol / L; Example 2 also performed well, with TC and LDL-C decreased by more than 25%; Comparative Examples 1 and 2 had poor effects, with only slight improvements (indicating that ingredients such as GABA and red yeast rice extract are crucial for lipid-lowering effects); the LDL-C level in the high-fat control group increased by nearly 4 times, verifying the effect of a high-fat diet on blood lipid levels.

[0057] The liver function indexes are analyzed as follows:

[0058] Index (U / L) Normal control group High-fat control group Example 1 Example 2 Example 3 Control Example 1 Control Example 2 ALT 22±4 55±6 43±5 38±4 33±3 50±6 52±5 AST 28±5 63±7 50±5 44±4 40±3 58±7 60±6

[0059] Key Results:

[0060] Example 3 significantly reduced ALT and AST, indicating that it had no obvious toxicity to the liver and might even protect the liver. Comparative Examples 1 and 2 had no significant difference from the high-fat control group, indicating that the use of lactic acid bacteria alone failed to effectively improve liver function;

[0061] Liver pathological analysis, tissue section results:

[0062] High-fat control group: diffuse fatty degeneration of hepatocytes and a large number of lipid droplets deposited;

[0063] Example 3: Lipid droplets decreased by 60%, and liver cells returned to normal;

[0064] Example 2: The lipid droplets were reduced by 45%, but some fatty degeneration still occurred;

[0065] Comparative Examples 1 and 2: There was no significant difference compared with the high-fat group, and there was still severe fat deposition.

[0066] The above description is only a preferred example of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a probiotic liquid of Lactobacillus paracasei LP007 having a lipid-lowering effect, characterized in that: The specific steps are as follows: Preparation of S1 culture medium: Dissolve 12-18 g / L lactose in 60% volume of pure water, stir evenly, add 4-6 g / L fructooligosaccharide, continue stirring, add DHA, 50-80 mg / L γ-aminobutyric acid, 0.3-0.4 mg / L vitamin B group, adjust pH to 6.5-6.8, sterilize at 121℃ for 15 min, cool to 37℃ for use; S2 culture: Activated Lactobacillus paracasei LP007: Use sterile MRS medium and culture at 37℃ for 12-18h until 1×10 9 CFU / mL, inoculate 5% (v / v) into the fermentation broth under sterile conditions, culture at 37°C in a shaking incubator for 12-18h, monitor bacterial growth, and ensure that the concentration is 1×10 9 -3×10¹ 0 CFU / mL; S3 fermentation process: 5% inoculation amount is used to transfer the bacterial liquid to the fermentation tank, which is installed in the auxiliary fermentation device to adjust the fermentation angle and position, then maintained at 37°C, stirred at 150-200rpm, and 5% CO2 is introduced. Fermentation is carried out for 24-48h, and the pH is monitored, which is finally about 4.5-5.0 to ensure the metabolic balance of lactic acid bacteria; S4 lipid-lowering ingredient addition and mixing: 150-300 mg / L of red yeast rice extract and 1-5 g / L of casein hydrolyzed peptide were measured, and the fermentation liquid was slowly added at a low temperature of 4°C, stirred at 1000 rpm for 10-15 min, and homogenized at 120 MPa for 2 times; S5 Sterilization and packaging: pasteurization at 65-75℃ for 15min, ensuring bacterial survival rate>90%, filtering to remove particulate impurities, aseptic filling, and packaging into bottles; The auxiliary fermentation device includes an adjustment mechanism for supporting and adjusting the angle position, a shaking mechanism with a shaking function provided on the top of the adjustment mechanism, and a clamping member provided on the top of the shaking mechanism for clamping the fermentation tank; The adjustment mechanism includes a chassis, a support rod arranged at the top of the chassis, a support plate installed at the top of the support rod, a bottom frame arranged at the right end of the top of the support plate, an adjustment knob screwed into the front and rear ends of the bottom frame, a top frame connected to the top of the adjustment knob, the top frame and the bottom frame have the same structure and both have openings at the front and rear positions of the right ends, a connecting ball for connecting the top frame and the right end of the bottom frame, a fixing rod arranged inside the opening, a spring for connecting the two sets of fixing rods, and a support plate arranged on the left side of the top frame, and the top of the support plate is connected to a shaking mechanism; The shaking mechanism includes a bottom box arranged at the bottom of the pallet, a guide frame arranged at the top of the bottom box, a sliding mechanism installed inside the guide frame and a support plate arranged at the top of the sliding mechanism. The sliding mechanism drives the support plate to move, and the top of the support plate is connected to a clamp.

2. according to the preparation method of a kind of probiotic liquid of Lactobacillus paracasei LP007 with hypolipidemic effect of claim 1, it is characterized in that: The base frame is L-shaped.

3. according to the preparation method of a kind of probiotic liquid of Lactobacillus paracasei LP007 with hypolipidemic effect of claim 2, it is characterized in that: The right end of the bottom of the chassis is movably matched with the connecting ball.

4. A method for preparing a probiotic liquid of Lactobacillus paracasei LP007 with a lipid-lowering effect according to claim 1, characterized in that: The sliding mechanism includes a slide plate movably embedded in the inner side of the guide frame, connecting parts arranged at both ends of the top of the slide plate for connecting to the support plate, a swinging part arranged inside the slide plate, a connecting part connected to the right end of the swinging part, a support installed at the right end of the bottom of the guide frame, and a connecting shaft passing through the support and connecting the connecting part, wherein the top of the connecting shaft is connected to the left end of the connecting part.

5. A method for preparing a probiotic liquid of Lactobacillus paracasei LP007 having a lipid-lowering effect according to claim 4, characterized in that: A driving motor is arranged at the right end of the bottom box, and the bottom output shaft of the driving motor is connected to the driving pulley. The outer side of the driving pulley rotates synchronously with the driven pulley through a synchronous belt. A rotating shaft is arranged in the middle of the driven pulley, and the top of the rotating shaft is connected to a connecting shaft, and the connecting shaft rotates with the rotating shaft.

6. The probiotic liquid prepared by the preparation method of a probiotic liquid of Lactobacillus paracasei LP007 with hypolipidemic effect according to claim 1, characterized in that: The probiotic liquid ratio is: Lactobacillus paracasei LP007 is 1×10 9 - 5×10¹ 0 CFU / mL, oligofructose 2-8 g / L, γ-aminobutyric acid 50-100 mg / L, red yeast rice extract 150-300 mg / L, DHA 20-50 mg / L, casein hydrolyzed peptide 1-5 g / L, vitamin B group (B6, B12) 0.2-0.5 mg / L, lactose 10-20 g / L and purified water 850-970 mL / L. The Lactobacillus paracasei LP007 is deposited in the General Microbiology Center of China Microorganism Culture Collection Committee with the deposit number: CGMCC NO.32193.

Images