Preparation method of encapsulated Lactobacillus plantarum J26 and its application in weight loss products

Abstract

The present invention discloses a preparation method of an embedded Lactobacillus plantarum J26 and its application in a weight loss product, comprising the following steps: (1) preparing colostrum: mixing a Lactobacillus plantarum J26 bacterial solution with an internal aqueous phase solution, then adding medium-chain fatty acid triglycerides and polyglycerol ricinoleate, and performing a primary emulsification to obtain W / O colostrum; (2) preparing a double emulsion: uniformly mixing a modified octenylsuccinic anhydride cassava starch solution and Tween 80 as an external aqueous phase solution, then adding W / O colostrum, and performing a secondary emulsification to obtain a W / O / W double emulsion; (3) freeze-drying the W / O / W double emulsion to obtain the embedded Lactobacillus plantarum J26. The present invention constructs a multi-layer composite microencapsulated Lactobacillus plantarum J26 system through a multi-layer composite encapsulation-freeze-drying combined technology to achieve stable co-encapsulation of Lactobacillus plantarum J26 with a weight loss effect, which has a good weight loss effect.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to a preparation method of an embedded Lactobacillus plantarum J26 and application of the same in weight-loss products. Background Art

[0002] Probiotics are a class of microorganisms that can maintain their numbers and benefit host health. Studies have shown that consuming foods containing probiotics can have multiple health-promoting effects, leading to increasing attention for the development of probiotic products. However, probiotic products are subject to a series of adverse effects during the process from production to transportation, distribution, and oral administration through the upper digestive tract to the intestine. This significantly reduces the number of viable bacteria that reach the intestine, limiting the physiological effects of probiotics.

[0003] Lactobacillus plantarum J26 has great potential in the development of functional foods due to its weight-loss effects. However, its low environmental stability and bioavailability limit its uptake and utilization in the human small intestine. To overcome this problem, the prior art has encapsulated probiotics in microcapsules to protect them from harmful external environmental factors, thereby increasing their survival rate. In recent years, extensive research has been conducted on probiotic microencapsulation technology, resulting in significant progress in the development of microencapsulation methods and the use of encapsulation materials.

[0004] Chinese patent application number 201910133308.6 discloses a double-layer microencapsulated prebiotic and probiotic composition and its preparation method. The invention uses microencapsulation protection technology, with natural polymer sodium alginate, glucose and prebiotics as the outer wall material, poly-L-arginine and prebiotics as the inner wall material, and probiotics and skim milk powder as the core material. The specific double-layer microcapsule structure of the invention provides double protection for unstable probiotics from the influence of the external environment and adverse factors, effectively prevents the reduction of probiotic activity during use and storage, and maximizes the stability, activity and effective colonization rate of the composition, and has high embedding efficiency and good embedding effect. Chinese patent application number 202011208280.7 discloses a method for preparing multi-layer coated probiotic microcapsules. The preparation method includes coating the probiotics before microencapsulation and coating them after fermentation, thereby preparing a method for preparing multi-layer composite coated probiotic microcapsules. The preparation method of the present invention plays a great role in maintaining the survival and stability of probiotic preparations. In addition, the preparation method of the present invention can more conveniently realize the large-scale production of probiotic microencapsulated products, and is suitable for the preparation of probiotic microcapsules in multiple fields such as feed, food, and medicine.

[0005] However, the microencapsulation method also has defects. For example, during the encapsulation process and freeze-drying process, bacterial powder or liquid can easily adhere to the outer wall of the encapsulation layer, resulting in waste of strain raw materials and microbial contamination; traditional single-layer encapsulation not only has a low yield but also has a poor encapsulation effect, and cannot provide good protection for the bacteria; although three-layer and above microcapsule encapsulation technology improves the stability of the bacteria, the process is cumbersome and time-consuming, and the probiotic strains cannot be quickly colonized and released in the intestines.

[0006] Water-in-oil-in-water (W1 / O / W2) double emulsion is a common emulsion system formed by dispersing a water-in-oil (W1 / O) primary emulsion in another continuous phase, water (W2). Since it can simultaneously encapsulate substances of different polarities (hydrophilic and lipophilic), it shows great application prospects in the co-encapsulation and delivery of active ingredients. Compared with traditional W / O emulsions, its "two membranes and three phases" structure allows the hydrophilic active ingredients in the inner aqueous phase (W1) to be more effectively protected and delayed in release, and it also has better dispersibility in water-soluble systems (such as human gastrointestinal digestion). The construction of a double emulsion requires a two-step emulsification process, which includes the use of lipophilic and hydrophilic emulsifiers or surfactants to stabilize the two oil-water interface films of the W1 / O / W2 emulsion and reduce interfacial tension.

[0007] In view of this, the present invention provides a preparation method of embedded Lactobacillus plantarum J26. Summary of the Invention

[0008] In view of the deficiencies in the prior art, the object of the present invention is to provide a preparation method of an embedded Lactobacillus plantarum J26 and its application in weight loss products. A multi-layer composite microencapsulated Lactobacillus plantarum J26 system is constructed by a multi-layer composite encapsulation-freeze-drying combined technology to achieve the stable co-encapsulation of Lactobacillus plantarum J26 with a weight loss effect, which has a good weight loss effect.

[0009] To achieve the above object, the present invention provides the following technical solutions:

[0010] A preparation method of an embedded Lactobacillus plantarum J26 comprises the following steps:

[0011] (1) Preparation of colostrum: Lactobacillus plantarum J26 bacterial solution was mixed with the internal aqueous phase solution, followed by addition of medium-chain fatty acid triglycerides and polyglycerol ricinoleate, and emulsification was performed once in a high-speed dispersing emulsifier to obtain W / O colostrum;

[0012] (2) Preparation of a double emulsion: a modified octenylsuccinic anhydride cassava starch solution and Tween 80 are uniformly mixed as an external aqueous phase solution, and then the W / O colostrum obtained in step (1) is added, and secondary emulsification is performed in a high-speed dispersing homogenizer to obtain a W / O / W double emulsion;

[0013] (3) The W / O / W emulsion obtained in step (2) was placed in a -20°C refrigerator for pre-freezing for 12 hours, and then freeze-dried to obtain the embedded Lactobacillus plantarum J26.

[0014] Preferably, the preparation method of the inner aqueous phase solution in step (1) is as follows: brown rice polysaccharide, carboxymethyl cellulose and maltodextrin are added to deionized water and stirred evenly to obtain the inner aqueous phase solution.

[0015] Preferably, the mass ratio of the brown rice polysaccharide, carboxymethyl cellulose, maltodextrin and deionized water is 5-10:1-3:1-2:150-200; the stirring temperature is 40-50° C. and the stirring time is 1-2 h.

[0016] Preferably, the number of viable Lactobacillus plantarum J26 in the Lactobacillus plantarum J26 bacterial solution in step (1) is 1.5×10 10 CFU / mL~2.0×10 10 CFU / mL, the mass ratio of the Lactobacillus plantarum J26 bacterial solution, the inner aqueous phase solution, the medium-chain fatty acid triglyceride, and the polyglycerol ricinoleate is 20:100-150:120-200:5-10; the rotation speed of the primary emulsification is 8000-12000 r / min, and the time is 10-15 min.

[0017] Preferably, the preparation method of the modified octenylsuccinic anhydride cassava starch solution in step (2) is as follows:

[0018] Nano-silicon dioxide is added to deionized water, followed by sodium lauryl sulfate, and the mixture is stirred evenly. Chitosan acetic acid solution is then added and stirred for reaction. After the reaction is completed, the mixture is filtered, washed, and dried to obtain pretreated silicon dioxide. Cassava starch is added to deionized water to form starch milk, 5 wt% sodium hydroxide is added to adjust the pH to 8-9, and then an octenylsuccinic anhydride ethanol solution is added dropwise for heating reaction. The pH is maintained at 8-9 during the reaction. After the reaction is completed, 5 wt% hydrochloric acid is added to adjust the pH to 6.5-7, and α-amylase is subsequently added for enzymatic hydrolysis for 20-30 minutes to obtain an octenylsuccinic anhydride cassava starch solution. The pretreated silicon dioxide is added to the octenylsuccinic anhydride cassava starch solution, followed by β-cyclodextrin for constant temperature reaction. After the reaction is completed, the modified octenylsuccinic anhydride cassava starch solution is obtained.

[0019] Preferably, the mass ratio of the nano-silica, sodium lauryl sulfate, and chitosan acetic acid solution is 20:0.1-0.2:100-200, the concentration of chitosan in the chitosan acetic acid solution is 5%, and the concentration of acetic acid is 10%, the temperature of the stirring reaction is 60-70°C, and the reaction time is 2-4h; the mass ratio of the cassava starch, octenylsuccinic anhydride ethanol solution, and α-amylase is 50:200-300:1-3, the concentration of the octenylsuccinic anhydride ethanol solution is 6-9wt%, the temperature of the heating reaction is 40-50°C, and the reaction time is 4-8h; the mass ratio of the pretreated silica, octenylsuccinic anhydride cassava starch solution, and β-cyclodextrin is 3:150-200:10-15; the temperature of the isothermal reaction is 50-60°C, and the reaction time is 1-2h.

[0020] Preferably, the mass ratio of the modified octenylsuccinic anhydride cassava starch solution, Tween 80, and W / O colostrum in step (2) is 100:4-8:7-13.

[0021] Preferably, the rotation speed of the secondary emulsification in step (2) is 2000-3000 r / min, and the time is 15-20 min.

[0022] Preferably, the freeze-drying process in step (3) is as follows: the freeze-drying temperature is -80 to -60°C, the vacuum degree is 5.0 to 10.0 Pa, and the time is 12 to 16 hours.

[0023] The present invention also protects an embedded Lactobacillus plantarum J26 prepared by the method.

[0024] The present invention also protects the use of the embedded Lactobacillus plantarum J26 in weight loss products

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] (1) The embedded Lactobacillus plantarum J26 provided by the present invention uses brown rice polysaccharide, carboxymethyl cellulose and maltodextrin as the inner aqueous phase, utilizes the adhesiveness of brown rice polysaccharide to combine with Lactobacillus plantarum J26, and Lactobacillus plantarum J26 decomposes brown rice polysaccharide, decomposing the macromolecular brown rice polysaccharide into small molecular polysaccharide structures that are easy to digest and absorb. At the same time, the brown rice polysaccharide is utilized by Lactobacillus plantarum J26 as a carbon source to achieve the effect of proliferation. In addition, the addition of carboxymethyl cellulose and maltodextrin can effectively inhibit the migration and diffusion of the inner aqueous phase to the outer aqueous phase, thereby improving the stability of the double emulsion during storage. The modified octenyl succinic anhydride cassava starch solution obtained by reacting cassava starch with octenyl succinic anhydride as raw materials has the advantages of amphiphilicity, low cost and safety, and can improve the embedding rate and stability of the microcapsules. Finally, the liquid emulsion is converted into solid microcapsule particles by the freeze-drying method, thereby changing the product properties, facilitating transportation and storage, and meeting the needs of consumers. In addition, freeze-drying can reduce the loss of biological activity of the encapsulated substance during processing.

[0027] (2) The embedded Lactobacillus plantarum J26 provided by the present invention has a high hydroxyl content on the SiO2 surface and a large specific surface area. It can be chemically bonded with polymer compounds to give the capsule wall material special properties, such as improved mechanical properties and density, thereby improving the stability of the embedded Lactobacillus plantarum J26. By pretreating the nano-SiO2, its dispersibility is improved on the one hand, and chitosan is coated on the surface of the nano-SiO2, thereby improving the binding force between the nano-SiO2 and octenylsuccinic anhydride cassava starch. In addition, α-amylase is added to cause moderate enzymatic hydrolysis of the octenylsuccinic anhydride cassava starch, thereby reducing the viscosity of the octenylsuccinic anhydride cassava starch. The addition of nano-SiO2 causes hydrogen bonds and coordination bonds to form between the octenylsuccinic anhydride cassava starch and the nano-SiO2, making the intermolecular structure more compact and extending the migration path of the gas through the outer aqueous phase layer, thereby enhancing the comprehensive performance of the multilayer composite film. In addition, the addition of a small amount of β-cyclodextrin can improve the encapsulation and sustained-release properties of the microcapsule for Lactobacillus plantarum J26, thereby making the microcapsule have higher thermal stability.

[0028] (3) The encapsulated Lactobacillus plantarum J26 provided by the present invention uses different wall materials to encapsulate Lactobacillus plantarum J26. The raw materials of each layer act synergistically with each other. The prepared microcapsules have excellent encapsulation and sustained-release properties for Lactobacillus plantarum J26, thereby improving its stability and bioavailability. The microcapsules formed by the double emulsion have a multi-layer core-shell structure. The lipophilic components in the oil phase can play a good protective role on the delivered Lactobacillus plantarum J26 in the inner aqueous phase, avoiding oxidation by oxygen dissolved in the outer aqueous phase. The modified octenylsuccinic anhydride cassava starch in the outer aqueous phase coats the primary emulsion, further improving the acid resistance, bile salt resistance and storage stability of the encapsulated Lactobacillus plantarum J26. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a graph showing the test results of storage time and viable cell count of the embedded Lactobacillus plantarum J26 and free Lactobacillus plantarum J26 prepared in Example 1 in pure milk at 4°C;

[0030] Figure 2 This is a graph showing the test results of storage time and viable cell count of the encapsulated Lactobacillus plantarum J26 and free Lactobacillus plantarum J2 prepared in Example 1 in pure milk at room temperature;

[0031] Figure 3 This is a graph showing the test results of storage time and pH of the embedded Lactobacillus plantarum J26 and free Lactobacillus plantarum J26 prepared in Example 1 in pure milk at 4°C;

[0032] Figure 4 This is a graph showing the test results of storage time and pH value of the embedded Lactobacillus plantarum J26 and free Lactobacillus plantarum J26 prepared in Example 1 in pure milk at room temperature;

[0033] Figure 5 This is a graph showing the test results of storage time and acidity of the embedded Lactobacillus plantarum J26 and free Lactobacillus plantarum J26 prepared in Example 1 in pure milk at 4°C;

[0034] Figure 6 This is a graph showing the test results of storage time and acidity of the embedded Lactobacillus plantarum J26 and free Lactobacillus plantarum J26 prepared in Example 1 in pure milk at room temperature;

[0035] Figure 7 This is a graph showing the test results of weight gain of mice;

[0036] Figure 8 This is the test result diagram of mouse body fat percentage;

[0037] Figure 9 This is the test result of TG content in mouse liver;

[0038] Figure 10 This is the test result of TC content in mouse liver;

[0039] Figure 11 Schematic diagram of H&E staining of mouse WAT;

[0040] Figure 12 This figure shows the measurement results of the average size of white adipose tissue in mice. DETAILED DESCRIPTION

[0041] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0042] In the present invention, Lactobacillus plantarum J26 is abbreviated as L. plantarum J26, LJ26 or J26; Lactobacillus plantarum TD109 is abbreviated as TD109; Lactobacillus plantarum J26 and Lactobacillus plantarum TD109 are both alternative names for Lactobacillus plantarum NDC75017. Sequence identification shows that Lactobacillus plantarum J26 and Lactobacillus plantarum TD109 are the same strain as Lactobacillus plantarum NDC75017 with a deposit number of CGMCC No. 5448 recorded in Chinese invention patent 201210552498.3. The applicant submitted documents certifying that Lactobacillus plantarum J26 and Lactobacillus plantarum TD109 are Lactobacillus plantarum NDC75017 in the relevant supporting documents. Therefore, Lactobacillus plantarum J26 is used in the present invention to represent Lactobacillus plantarum NDC75017.

[0043] The brown rice polysaccharide used in the present invention is prepared according to the method disclosed in Chinese Patent Application No. 201710232957.2. Specifically, the process involves removing the endosperm from food-grade germinated brown rice using a rice mill to obtain brown rice bran. The brown rice bran is then subjected to two extraction and defatting steps using food-grade n-hexane. The two defatted brown rice brans are combined to obtain defatted brown rice bran. The defatted brown rice bran is then ground to a 45-mesh size. The defatted brown rice bran is then ultrasonically extracted for 60 minutes in a 40-10°C water bath at an ultrasonic power of 140W at a solid-liquid ratio of 1 kg:L. The extract is then allowed to stand, filtered to remove the precipitate, and the filtrate is concentrated at 40°C to obtain the brown rice polysaccharide.

[0044] The nano-silica was purchased from Wuhan Kemik Biopharmaceutical Technology Co., Ltd., with a particle size of 50-100 nm.

[0045] Example 1

[0046] A preparation method of an embedded Lactobacillus plantarum J26 comprises the following steps:

[0047] (1) Preparation of colostrum: 20g of colostrum with a viable bacterial count of 1.7×10 10CFU / mL of Lactobacillus plantarum J26 bacterial liquid was mixed with 100 g of the internal aqueous phase solution, followed by the addition of 120 g of medium-chain fatty acid triglycerides and 5 g of polyglycerol ricinoleate, and emulsification was performed once in a high-speed dispersing emulsifier at a speed of 8000 r / min for 15 min to obtain W / O colostrum;

[0048] (2) Preparation of a double emulsion: 100 g of a modified octenylsuccinic anhydride cassava starch solution and 4 g of Tween 80 were mixed uniformly as the external aqueous phase solution, and then 7 g of the W / O colostrum obtained in step (1) was added, and secondary emulsification was performed in a high-speed dispersing homogenizer at a speed of 2000 r / min for 20 min to obtain a W / O / W double emulsion;

[0049] (3) The W / O / W emulsion obtained in step (2) was placed in a -20°C refrigerator for pre-freezing for 12 hours, and then freeze-dried at a temperature of -80°C, a vacuum degree of 5.0 Pa, and a time of 12 hours to obtain the encapsulated Lactobacillus plantarum J26.

[0050] The preparation method of the inner aqueous phase solution in step (1) is as follows: 5 g of brown rice polysaccharide, 1 g of carboxymethyl cellulose, and 1 g of maltodextrin are added to 150 g of deionized water, and stirred at 40° C. for 2 h to obtain the inner aqueous phase solution;

[0051] The preparation method of the modified octenylsuccinic anhydride cassava starch solution in step (2) is as follows:

[0052] 20g of nano-silica was added to 200mL of deionized water, followed by 0.1g of sodium lauryl sulfate, and after stirring evenly, 100g of chitosan acetic acid solution was added. The concentration of chitosan in the chitosan acetic acid solution was 5% and the concentration of acetic acid was 10%. The mixture was stirred at 60°C for 4h. After the reaction was completed, it was filtered, washed, and dried to obtain pretreated silica; 50g of cassava starch was added to 500mL of deionized water to form starch milk, 5wt% of sodium hydroxide was added to adjust the pH to 8, and then 200g of 6wt% octenylsuccinic anhydride ethanol solution was added dropwise. The mixture was heated at 40°C for 8h, and the pH was maintained at 8 during the reaction. After the reaction was completed, 5wt% of hydrochloric acid was added to adjust the pH to 6.5, and then 10g of α-amylase was added. The mixture was hydrolyzed for 30min and the enzyme was inactivated to obtain octenylsuccinic anhydride cassava starch solution; 3g of pretreated silica was added to 150g of octenylsuccinic anhydride cassava starch solution, and then 10g β-cyclodextrin, and react at a constant temperature of 50° C. for 2 hours. After the reaction is completed, the modified octenylsuccinic anhydride cassava starch solution is obtained.

[0053] Example 2

[0054] A preparation method of an embedded Lactobacillus plantarum J26 comprises the following steps:

[0055] (1) Preparation of colostrum: 20g of colostrum with a viable bacterial count of 1.5×10 10 CFU / mL of Lactobacillus plantarum J26 bacterial liquid was mixed with 130 g of the internal aqueous phase solution, followed by the addition of 150 g of medium-chain fatty acid triglycerides and 7 g of polyglycerol ricinoleate, and the mixture was emulsified in a high-speed dispersing emulsifier at a speed of 10,000 r / min for 15 min to obtain W / O colostrum;

[0056] (2) Preparation of a double emulsion: 100 g of a modified octenylsuccinic anhydride cassava starch solution and 6 g of Tween 80 were mixed uniformly as an external aqueous phase solution, followed by addition of 10 g of the W / O colostrum obtained in step (1), and secondary emulsification was performed in a high-speed dispersing homogenizer at a speed of 2500 r / min for 20 min to obtain a W / O / W double emulsion;

[0057] (3) The W / O / W emulsion obtained in step (2) was placed in a -20°C refrigerator for pre-freezing for 12 hours, and then freeze-dried at a freeze-drying temperature of -70°C, a vacuum degree of 10.0 Pa, and a time of 14 hours to obtain the encapsulated Lactobacillus plantarum J26.

[0058] The preparation method of the inner aqueous phase solution in step (1) is as follows: 5 g of brown rice polysaccharide, 1 g of carboxymethyl cellulose, and 1 g of maltodextrin are added to 150 g of deionized water, and stirred at 40° C. for 2 h to obtain the inner aqueous phase solution;

[0059] The preparation method of the modified octenylsuccinic anhydride cassava starch solution in step (2) is as follows:

[0060] 20g of nano-silica was added to 200mL of deionized water, followed by 0.15g of sodium lauryl sulfate, and after stirring evenly, 150g of chitosan acetic acid solution was added, wherein the concentration of chitosan in the chitosan acetic acid solution was 5% and the concentration of acetic acid was 10%. The mixture was stirred at 65°C for 3h. After the reaction was completed, it was filtered, washed, and dried to obtain pretreated silica; 50g of cassava starch was added to 500mL of deionized water to form starch milk, 5wt% of sodium hydroxide was added to adjust the pH to 8.5, and then 250g of 7wt% octenylsuccinic anhydride ethanol solution was added dropwise, and the mixture was heated at 45°C for 6h. The pH was maintained at 8-9 during the reaction. After the reaction was completed, 5wt% of hydrochloric acid was added to adjust the pH to 6.5, and then 12g of α-amylase was added. The mixture was hydrolyzed for 25min and the enzyme was inactivated to obtain octenylsuccinic anhydride cassava starch solution; 3g of pretreated silica was added to 170g of octenylsuccinic anhydride cassava starch solution, followed by 12g of β-cyclodextrin, and react at a constant temperature of 55° C. for 1.5 hours. After the reaction is completed, the modified octenylsuccinic anhydride cassava starch solution is obtained.

[0061] Example 3

[0062] A preparation method of an embedded Lactobacillus plantarum J26 comprises the following steps:

[0063] (1) Preparation of colostrum: 20g of colostrum with a viable bacterial count of 2.0×10 10 CFU / mL of Lactobacillus plantarum J26 bacterial liquid was mixed with 130 g of the internal aqueous phase solution, followed by the addition of 170 g of medium-chain fatty acid triglycerides and 8 g of polyglycerol ricinoleate, and emulsification was performed in a high-speed dispersing emulsifier at a speed of 10,000 r / min for 10 min to obtain W / O colostrum;

[0064] (2) Preparation of a double emulsion: 100 g of a modified octenylsuccinic anhydride cassava starch solution and 6 g of Tween 80 were mixed uniformly as an external aqueous phase solution, followed by addition of 11 g of the W / O colostrum obtained in step (1), and secondary emulsification was performed in a high-speed dispersing homogenizer at a speed of 2500 r / min for 15 min to obtain a W / O / W double emulsion;

[0065] (3) The W / O / W emulsion obtained in step (2) was placed in a -20°C refrigerator for pre-freezing for 12 hours, and then freeze-dried at a freeze-drying temperature of -70°C, a vacuum degree of 5.0 Pa, and a time of 14 hours to obtain the embedded Lactobacillus plantarum J26.

[0066] The preparation method of the inner aqueous phase solution in step (1) is as follows: 10 g of brown rice polysaccharide, 3 g of carboxymethyl cellulose, and 2 g of maltodextrin are added to 200 g of deionized water, and stirred at 50° C. for 1 h to obtain the inner aqueous phase solution;

[0067] The preparation method of the modified octenylsuccinic anhydride cassava starch solution in step (2) is as follows:

[0068] 20g of nano-silica was added to 200mL of deionized water, followed by 0.15g of sodium lauryl sulfate, and after stirring evenly, 150g of chitosan acetic acid solution was added, wherein the concentration of chitosan in the chitosan acetic acid solution was 5% and the concentration of acetic acid was 10%. The mixture was stirred at 65°C for 3h. After the reaction was completed, it was filtered, washed, and dried to obtain pretreated silica; 50g of cassava starch was added to 500mL of deionized water to form starch milk, 5wt% of sodium hydroxide was added to adjust the pH to 8.5, and then 250g of 8wt% octenylsuccinic anhydride ethanol solution was added dropwise, and the mixture was heated at 45°C for 6h. The pH was maintained at 8.5 during the reaction. After the reaction was completed, 5wt% of hydrochloric acid was added to adjust the pH to 7, and then 13g of α-amylase was added. The mixture was hydrolyzed for 25min and the enzyme was inactivated to obtain octenylsuccinic anhydride cassava starch solution; 3g of pretreated silica was added to 180g of octenylsuccinic anhydride cassava starch solution, followed by 13g of β-cyclodextrin, and react at a constant temperature of 55° C. for 1 hour. After the reaction is completed, the modified octenylsuccinic anhydride cassava starch solution is obtained.

[0069] Example 4

[0070] A preparation method of an embedded Lactobacillus plantarum J26 comprises the following steps:

[0071] (1) Preparation of colostrum: 20g of colostrum with a viable bacterial count of 1.8×10 10 CFU / mL of Lactobacillus plantarum J26 bacterial liquid was mixed with 150 g of the internal aqueous phase solution, followed by the addition of 200 g of medium-chain fatty acid triglycerides and 10 g of polyglycerol ricinoleate, and emulsification was performed in a high-speed dispersing emulsifier at a speed of 12,000 r / min for 10 min to obtain W / O colostrum;

[0072] (2) Preparation of a double emulsion: 100 g of a modified octenylsuccinic anhydride cassava starch solution and 8 g of Tween 80 were mixed uniformly as an external aqueous phase solution, and then 13 g of the W / O colostrum obtained in step (1) was added, and secondary emulsification was performed in a high-speed dispersing homogenizer at a speed of 3000 r / min for 15 min to obtain a W / O / W double emulsion;

[0073] (3) The W / O / W emulsion obtained in step (2) was placed in a -20°C refrigerator for pre-freezing for 12 hours, and then freeze-dried at a freeze-drying temperature of -60°C, a vacuum degree of 5.0 Pa, and a time of 16 hours to obtain the encapsulated Lactobacillus plantarum J26.

[0074] The preparation method of the inner aqueous phase solution in step (1) is as follows: 10 g of brown rice polysaccharide, 2 g of carboxymethyl cellulose, and 2 g of maltodextrin are added to 200 g of deionized water, and stirred at 50° C. for 1 h to obtain the inner aqueous phase solution;

[0075] The preparation method of the modified octenylsuccinic anhydride cassava starch solution in step (2) is as follows:

[0076] 20g of nano-silica was added to 200mL of deionized water, followed by 0.2g of sodium lauryl sulfate, and after stirring evenly, 200g of chitosan acetic acid solution was added, wherein the concentration of chitosan in the chitosan acetic acid solution was 5% and the concentration of acetic acid was 10%. The mixture was stirred at 70°C for 2h. After the reaction was completed, it was filtered, washed, and dried to obtain pretreated silica; 50g of cassava starch was added to 500mL of deionized water to form starch milk, 5wt% of sodium hydroxide was added to adjust the pH to 9, and then 300g of 9wt% octenylsuccinic anhydride ethanol solution was added dropwise. The mixture was heated at 50°C for 4h, and the pH was maintained at 9 during the reaction. After the reaction was completed, 5wt% of hydrochloric acid was added to adjust the pH to 7, and then 15g of α-amylase was added. The mixture was enzymatically hydrolyzed for 20min and the enzyme was inactivated to obtain octenylsuccinic anhydride cassava starch solution; 3g of pretreated silica was added to 200g of octenylsuccinic anhydride cassava starch solution, and then 15g β-cyclodextrin, and react at a constant temperature of 60° C. for 1 hour. After the reaction is completed, the modified octenylsuccinic anhydride cassava starch solution is obtained.

[0077] Comparative Example 1

[0078] A preparation method of an embedded Lactobacillus plantarum J26 comprises the following steps:

[0079] (1) Preparation of colostrum: 20g of colostrum with a viable bacterial count of 1.7×10 10 CFU / mL of Lactobacillus plantarum J26 bacterial liquid was mixed with 100 g of the internal aqueous phase solution, followed by the addition of 120 g of medium-chain fatty acid triglycerides and 5 g of polyglycerol ricinoleate, and emulsification was performed once in a high-speed dispersing emulsifier at a speed of 8000 r / min for 15 min to obtain W / O colostrum;

[0080] (2) Preparation of a double emulsion: 100 g of a modified octenylsuccinic anhydride cassava starch solution and 4 g of Tween 80 were mixed uniformly as the external aqueous phase solution, and then 7 g of the W / O colostrum obtained in step (1) was added, and secondary emulsification was performed in a high-speed dispersing homogenizer at a speed of 2000 r / min for 20 min to obtain a W / O / W double emulsion;

[0081] (3) The W / O / W emulsion obtained in step (2) was placed in a -20°C refrigerator for pre-freezing for 12 hours, and then freeze-dried at a freeze-drying temperature of -80°C, a vacuum degree of 5.0 Pa, and a time of 12 hours to obtain the embedded Lactobacillus plantarum J26.

[0082] The preparation method of the inner aqueous phase solution in step (1) is as follows: 5 g of brown rice polysaccharide is added to 150 g of deionized water, and stirred at 40° C. for 2 h to obtain the inner aqueous phase solution;

[0083] The preparation method of the modified octenylsuccinic anhydride cassava starch solution in step (2) is as follows:

[0084] 20g of nano-silica was added to 200mL of deionized water, followed by 0.1g of sodium lauryl sulfate, and after stirring evenly, 100g of chitosan acetic acid solution was added. The concentration of chitosan in the chitosan acetic acid solution was 5% and the concentration of acetic acid was 10%. The mixture was stirred at 60°C for 4h. After the reaction was completed, it was filtered, washed, and dried to obtain pretreated silica; 50g of cassava starch was added to 500mL of deionized water to form starch milk, 5wt% of sodium hydroxide was added to adjust the pH to 8, and then 200g of 6wt% octenylsuccinic anhydride ethanol solution was added dropwise. The mixture was heated at 40°C for 8h, and the pH was maintained at 8 during the reaction. After the reaction was completed, 5wt% of hydrochloric acid was added to adjust the pH to 6.5, and then 10g of α-amylase was added. The mixture was hydrolyzed for 30min and the enzyme was inactivated to obtain octenylsuccinic anhydride cassava starch solution; 3g of pretreated silica was added to 150g of octenylsuccinic anhydride cassava starch solution, and then 10g β-cyclodextrin, and react at a constant temperature of 50° C. for 2 hours. After the reaction is completed, the modified octenylsuccinic anhydride cassava starch solution is obtained.

[0085] Comparative Example 2

[0086] A preparation method of an embedded Lactobacillus plantarum J26 comprises the following steps:

[0087] (1) Preparation of colostrum: 20g of colostrum with a viable bacterial count of 1.7×10 10 CFU / mL of Lactobacillus plantarum J26 bacterial liquid was mixed with 100 g of the internal aqueous phase solution, followed by the addition of 120 g of medium-chain fatty acid triglycerides and 5 g of polyglycerol ricinoleate, and emulsification was performed once in a high-speed dispersing emulsifier at a speed of 8000 r / min for 15 min to obtain W / O colostrum;

[0088] (2) Preparation of a double emulsion: 100 g of a modified octenylsuccinic anhydride cassava starch solution and 4 g of Tween 80 were mixed uniformly as the external aqueous phase solution, and then 7 g of the W / O colostrum obtained in step (1) was added, and secondary emulsification was performed in a high-speed dispersing homogenizer at a speed of 2000 r / min for 20 min to obtain a W / O / W double emulsion;

[0089] (3) The W / O / W emulsion obtained in step (2) was placed in a -20°C refrigerator for pre-freezing for 12 hours, and then freeze-dried at a freeze-drying temperature of -80°C, a vacuum degree of 5.0 Pa, and a time of 12 hours to obtain the embedded Lactobacillus plantarum J26.

[0090] The preparation method of the inner aqueous phase solution in step (1) is as follows: 5 g of brown rice polysaccharide, 1 g of carboxymethyl cellulose, and 1 g of maltodextrin are added to 150 g of deionized water, and stirred at 40° C. for 2 h to obtain the inner aqueous phase solution;

[0091] The preparation method of the modified octenylsuccinic anhydride cassava starch solution in step (2) is as follows:

[0092] 50 g of cassava starch was added to 500 mL of deionized water to form a starch milk, 5 wt % of sodium hydroxide was added to adjust the pH to 8, and then 200 g of a 6 wt % octenylsuccinic anhydride ethanol solution was added dropwise. The mixture was heated at 40° C. for 8 h, and the pH was maintained at 8 during the reaction. After the reaction was completed, 5 wt % of hydrochloric acid was added to adjust the pH to 6.5, and then 10 g of α-amylase was added. The mixture was enzymatically hydrolyzed for 30 min and the enzyme was inactivated to obtain the modified octenylsuccinic anhydride cassava starch solution.

[0093] The embedding efficiency of test examples 1-4 and comparative examples 1-2 is as follows:

[0094] Calculation of encapsulation yield: After washing the prepared encapsulated Lactobacillus plantarum J26 three times with sterile saline, 1 g of wet capsules was placed in 10 mL of encapsulation solution, and the capsules were decapsulated at 37±1°C with constant temperature shaking (210 rpm, 40 min), and the viable bacteria were counted.

[0095] The formula for calculating the number of viable bacteria in 1g wet capsule after decapsulation is: N1=G1×V1

[0096] Embedding yield (%) = N1×M / (N0×V0)×100%

[0097] Where: G1—number of viable bacteria in 1 mL of cystic fluid (CFU / mL); V1—volume of cystic fluid (mL); N1—number of viable bacteria in wet capsules after cystic decapsulation (CFU / g); N0—number of viable bacteria in the original bacterial solution before embedding (CFU / mL); V0—volume of the original bacterial solution used to prepare microcapsules (mL); M—total weight of the obtained wet capsules (g).

[0098] The test results are shown in Table 1 below:

[0099] Table 1 Encapsulation rate of encapsulated Lactobacillus plantarum J26

[0100]

[0101] The embedded Lactobacillus plantarum J26 prepared in Example 1 and Comparative Examples 1-2 was subjected to stability tests, as follows:

[0102] (1) Acid resistance test of embedded Lactobacillus plantarum J26

[0103] Only after probiotics reach the intestinal tract in sufficient quantity, will they produce certain treatment and health-care effects on the host. Probiotic food is taken by the human body and often needs to be digested by the stomach in the process leading to the intestinal tract, and probiotics have poor acid resistance and are easily affected by gastric acid and cause death. Take the embedded plant lactobacillus J261g prepared by Example 1 and Comparative Examples 1-2 and put it into the simulated artificial gastric juice prepared to carry out acid resistance test, wherein the volume of artificial gastric juice is 9mL, the required temperature condition of the test is 37 ℃, and the processing time is set to 4 time periods, namely 0, 1h, 2h and 3h. After processing according to the above time periods, the microcapsules are filtered and collected, and then washed and put into the prepared decapsulation liquid for decapsulation, wherein the volume of the decapsulation liquid is 9mL, and after decapsulation is completed, the number of viable bacteria is measured and calculated respectively. In addition, 1mL of free plant lactobacillus J26 bacterial suspension is used for control test respectively, and the original bacterial liquid volume used when preparing the bacterial suspension is the same as the original bacterial liquid volume used for the microcapsules. The test results are as shown in Table 2 below:

[0104] Table 2 Acid resistance test results of embedded Lactobacillus plantarum J26

[0105]

[0106] As can be seen from Table 2, after gastric juice is processed, due to the highly acidic environment of gastric juice, the viable count of plant lactobacillus J26 has the trend of minimizing, and the viable count of free probiotic bacteria reduces.After free probiotic bacteria was processed 2 hours in gastric juice, viable count appears to significantly reduce.Comparative Example 1 is owing to inner layer not adding carboxymethyl cellulose, maltodextrin, thereby causes plant lactobacillus J26 to migrate and diffuse to outer aqueous phase easily, and Comparative Example 2 is owing to outer layer not adding nano-SiO2 and beta-cyclodextrin, thereby causes the stability of capsule to descend.But under equal conditions, the viable count of plant lactobacillus J26 embedded through multi-layer composite microcapsule is obviously higher than control group and Comparative Example 1-2, illustrates that the plant lactobacillus J26 microcapsule of preparation has certain repellency to the poisoning of the gastric juice of low pH, and resists the infringement of gastric juice through the probiotic activity of multi-layer composite embedding.

[0107] (2) Bile salt tolerance test of embedded Lactobacillus plantarum J26

[0108] Take 1g of the embedded plant lactobacillus J26 obtained in Example 1 and Comparative Examples 1-2 respectively and put it into the bile salt solution prepared for bile salt tolerance test, wherein the volume of the artificial bile salt solution is 9mL, the temperature condition required for the test is 37°C, and the processing time is set to 3 time periods, namely 0, 1h and 2h. After the treatment is completed according to the above time period, the microcapsules are filtered and collected, and then washed and put into the prepared decapsulation liquid for decapsulation, wherein the volume of the decapsulation liquid is 9mL, and the number of viable bacteria is determined and calculated after decapsulation is completed. In addition, 1mL of free plant lactobacillus J26 bacterial suspension is used for control test, and the volume of the original bacterial liquid used when preparing the bacterial suspension is the same as the volume of the original bacterial liquid used for the microcapsule. The test results are shown in Table 3 below:

[0109] Table 3 Results of bile salt tolerance test of embedded Lactobacillus plantarum J26

[0110]

[0111] As can be seen from Table 3, after bile salt solution treatment, plant lactobacillus J26 has a decreasing trend, and the viable count of free probiotics decreases. After free probiotics are treated for 1 hour in bile salts, the viable count is greatly reduced. Under the same conditions, the viable count of the embedded type plant lactobacillus J26 prepared by the present invention is significantly greater than that of the control group and comparative example 1-2. Show: the embedded type plant lactobacillus J26 prepared by the present invention can effectively improve its bile salt tolerance.

[0112] (3) Enteric solubility test of embedded Lactobacillus plantarum J26

[0113] Take 1g of embedded plant lactobacillus J26 and place it in the artificial intestinal fluid that has been prepared for enteric solubility test. The volume of artificial intestinal fluid is 9mL. Then place it in a constant temperature and speed shaker for processing. The temperature of the constant temperature shaker is adjusted to 37℃, the shaking speed is set to 210r / min, and the processing time is set to the following 4 time periods, including 20min, 40min, 60min and 80min respectively. After that, fixed-point sampling is carried out to measure and calculate the number of viable bacteria dissolved. In addition, 1mL of free plant lactobacillus J26 bacterial suspension is used for control test. The original bacterial liquid volume used when preparing the bacterial suspension is the same as the original bacterial liquid volume used for the microcapsule. The test results are shown in Table 4 below:

[0114] Table 4 Enteric solubility test results of embedded Lactobacillus plantarum J26

[0115]

[0116] As can be seen from Table 4, after 40 minutes of treatment with artificial simulated intestinal fluid, the viable count of the microcapsules detected in the embedded Lactobacillus plantarum J26 gradually decreased. Further treatment with artificial simulated intestinal fluid showed a continued decrease in the viable count of Lactobacillus plantarum J26. The encapsulated Lactobacillus plantarum J26 prepared by the present invention is well protected and has good enteric solubility.

[0117] (4) Storage stability test of encapsulated Lactobacillus plantarum J26 in food

[0118] A control experiment was conducted by mixing the encapsulated Lactobacillus plantarum J26 prepared in Example 1 with pure milk at a ratio of 1 / 9 (mass / volume). A free Lactobacillus plantarum J26 suspension was used as the control group. Both the experimental and control groups were stored at 4°C and room temperature. The viable cell count, acidity, and pH of the products were measured over a specified time period to determine changes in these indicators. The test results are shown in Figure 2. Figures 1-6 .

[0119] Depend on Figure 1 and Figure 2 As can be seen, the viable counts of both free and encapsulated Lactobacillus plantarum J26 in pure milk showed a downward trend with prolonged storage time. The viable count of free Lactobacillus plantarum J26 decreased more rapidly than that of encapsulated Lactobacillus plantarum J26. At room temperature, the viable counts of both free and encapsulated Lactobacillus plantarum J26 decreased faster than those stored at 4°C.

[0120] Depend on Figure 3-Figure 6 As can be seen, with extended storage time, the pH of pure milk supplemented with both free and encapsulated Lactobacillus plantarum J26 decreased, while the acidity increased. At 4°C, the acidity and pH of pure milk changed more slowly than at room temperature, and the changes in acidity and pH of pure milk supplemented with encapsulated Lactobacillus plantarum J26 were slower than those with free Lactobacillus plantarum J26. This may be due to the more rapid metabolic rate of Lactobacillus plantarum J26 at room temperature, leading to a faster pH decrease and acidity increase at room temperature than at 4°C. Pure milk, with its near-neutral pH, does not significantly damage Lactobacillus plantarum J26. While the viable count of encapsulated Lactobacillus plantarum J26 may be affected with extended storage, the impact is minimal.

[0121] The encapsulated Lactobacillus plantarum J26 prepared in Example 1 was used to conduct a weight loss test to alleviate obesity in mice induced by a high-fat diet.

[0122] (1) Grouping of experimental animals and establishment of models

[0123] Forty male C57BL / 6J mice, approximately 5 weeks old and weighing 20 ± 2 g, were used as experimental animals. The mice were housed at a temperature of 22 ± 2°C, a humidity of 55 ± 5%, and a 12-hour day / night cycle. The mice had free access to food, water, and chow. After purchase, the mice were acclimated to the environment for one week using a basal diet with free access to water. The mice were randomly divided into four groups (n = 10 / group): a normal control group (ND), a high-fat diet group (HFD), a J26 group (HFD plus encapsulated Lactobacillus plantarum J26), and a positive drug control group (PC) (HFD plus the positive drug orlistat). The ND group was fed a basal diet, while the remaining groups were fed a high-fat diet. The basal diet was purchased from Shenyang Maohua Biotechnology Co., Ltd., and the high-fat diet was purchased from Beijing Keao Xieli Feed Co., Ltd. The formula consisted of 8% normal diet, 20% lard, 10% soybean oil, 10% sucrose, 10% maltodextrin, 10% egg yolk powder, 1.8% cholesterol, and 0.2% bile salts. The mice in the normal group and high-fat group were gavaged with 0.2 mL of sterile PBS solution, and the mice in the other groups were gavaged with 0.2 mL of 10 9 The mice were gavaged with CFU / mL of encapsulated Lactobacillus plantarum J26 and positive drug solutions for 8 consecutive weeks. Body weights were measured weekly and changes in the mice were recorded.

[0124] (2) Experimental sample preparation

[0125] At the end of the eighth week of oral gavage, whole blood was collected from mice that had fasted for 12 hours by enucleation. The blood was then allowed to stand at room temperature for 2-4 hours, and serum was collected and stored at -20°C until further use. Mice were sacrificed by cervical dislocation, and the liver, scapular fat, epididymal fat, subcutaneous fat, and visceral fat were quickly removed and weighed. Portions of the liver and adipose tissue were fixed in 4% paraformaldehyde for sectioning and staining. The remaining tissue was quickly frozen in liquid nitrogen and transferred to a -80°C freezer for long-term storage.

[0126] (3) Determination of obesity weight and body fat percentage in mice

[0127] Weigh the mice weekly, calculate the weight gain, and weigh the fat weight of the mice.

[0128] (4) Determination of TC and TG content in the liver

[0129] The total cholesterol and triglyceride assay kits produced by Nanjing Jiancheng were used to detect the TC and TG contents in mouse liver.

[0130] (5) Adipose tissue morphology

[0131] At the end of the 12th week, the mice were killed and their livers were removed for observation of morphology and color. After weighing, subcutaneous fat, visceral fat, epididymal fat, and scapular fat were removed for H&E staining.

[0132] (6) Data processing

[0133] The experimental data were analyzed using origin 9.0, SPSS22.0 and Jade6 software, and the experimental results were expressed as "mean + standard deviation".

[0134] The weight loss results are as follows:

[0135] Lactobacillus has been shown to be closely related to the regulation of glucose and lipid metabolism. In this study, an obese mouse model was induced by a high-fat diet. The mice were gavaged with encapsulated Lactobacillus plantarum J26 for 8 consecutive weeks during the high-fat diet period. The mental state of the mice was observed daily and their weight was recorded. The mice in the ND group had smooth hair, agile movements, and good mental state. The mice in the HFD group had greasy hair, slow movements, and lethargy. After the intervention of encapsulated Lactobacillus plantarum J26, the mice's hair became smooth again, they could move freely, and their mental state was significantly improved. Figure 7 As shown in the data, compared with the HFD group, the embedded Lactobacillus plantarum J26 significantly inhibited the weight gain of mice (p<0.01), and the inhibitory effect was close to that of the positive drug group. Oral administration of the embedded Lactobacillus plantarum J26 did not cause adverse effects on the mice.

[0136] The degree of obesity of mice is measured based on body fat percentage, which reflects the fat accumulation in the mouse body through the fat content per unit body weight. Figure 8 It can be seen that the high-fat diet significantly increased the fat content in the mice. After gavage with encapsulated Lactobacillus plantarum J26, the body fat rate of the mice showed a downward trend, which could significantly reduce the body fat rate of the mice (p<0.001), indicating that the intervention of encapsulated Lactobacillus plantarum J26 can effectively alleviate the degree of fat accumulation in obese mice.

[0137] The liver is the site of triglyceride synthesis, and long-term intake of a high-fat diet leads to an increase in fat content in the body. Figure 9 As can be seen, compared with the ND group, the high-fat diet significantly increased the TG content in the mouse liver (p<0.01). After gavage with the embedded Lactobacillus plantarum J26, the TG content in the mouse liver showed a downward trend, with the TG content in the liver decreasing by 33.15%, close to the TG content in the liver of normal mice, indicating that the embedded Lactobacillus plantarum J26 can significantly improve the liver fat accumulation caused by the high-fat diet, and the effect is close to that of the positive drug group and the ND group.

[0138] Long-term intake of high-fat diet accelerates the decomposition of fat and inhibits lipid synthesis, which in turn produces excessive fatty acids and leads to an increase in the total cholesterol (TC) content in the liver. Figure 10As shown in the results, long-term high-fat diet significantly increased TC levels in the serum of mice (p<0.01). After oral administration of the embedded Lactobacillus plantarum J26 to mice, TC levels in the liver showed a downward trend, with TC levels decreasing by 42%, approaching the levels of the ND group and the positive drug group, indicating that oral administration of the embedded Lactobacillus plantarum J26 can improve the increase in cholesterol levels in mice caused by a high-fat diet and further reduce fat accumulation to a certain extent.

[0139] Obesity is caused by a regular high-fat diet, and excessive white fat accumulation is the main cause of obesity. White adipose tissue (WAT) contains large fat vacuoles, which mainly store excess energy in the body and supply it to the body. When WAT is given external stimuli such as intestinal flora and their metabolites, WAT exhibits a beige adipocyte phenotype. By performing H&E staining on mouse WAT, such as Figure 11 and Figure 12 As shown, the white fat cells in the ND group were smooth, round, tightly arranged, and normal in size. The average size of white fat cells was 2572.39 μm. 2 In the HFD group, larger fat vacuoles were visible to the naked eye in the white fat, and the volume of fat cells in the field of view increased significantly. The cell sizes were uneven and the differences were significant. A small amount of inflammatory cell infiltration was seen in the local interstitium, and the average size of white fat cells increased to 7782.43μm. 2 Compared with the ND group, the mice that received Lactobacillus plantarum J26 showed a highly significant increase (p<0.001). Compared with the HFD group, mice that received Lactobacillus plantarum J26 had smaller white adipocytes and fewer fat vacuoles. No inflammatory cell infiltration was observed. The average size of adipocytes decreased, and a small number of small, lipid-like, multi-locular adipocytes, also known as beige adipocytes, appeared. This phenomenon is called browning of white fat. After intervention with Lactobacillus plantarum J26, the average size of adipocytes reached 3638.16μm. 2 , significantly reduced adipocyte size (p<0.001), approaching the level of the positive drug group. These results indicate that encapsulated Lactobacillus plantarum J26 inhibits white adipocyte enlargement, reduces inflammatory responses in adipose tissue, and induces WAT to exhibit a browning fat phenotype.

[0140] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A preparation method of an embedded Lactobacillus plantarum J26, characterized in that, The following steps are involved: (1) Preparation of colostrum: Lactobacillus plantarum J26 bacterial solution was mixed with the internal aqueous phase solution, followed by addition of medium-chain fatty acid triglycerides and polyglycerol ricinoleate, and emulsification was performed once in a high-speed dispersing emulsifier to obtain W / O colostrum; (2) Preparation of a double emulsion: a modified octenylsuccinic anhydride cassava starch solution and Tween 80 are uniformly mixed as an external aqueous phase solution, and then the W / O colostrum obtained in step (1) is added, and secondary emulsification is performed in a high-speed dispersing homogenizer to obtain a W / O / W double emulsion; (3) The W / O / W emulsion obtained in step (2) was placed in a -20°C refrigerator for pre-freezing for 12 hours, and then freeze-dried to obtain the embedded Lactobacillus plantarum J26; The preparation method of the inner aqueous phase solution in step (1) is as follows: adding brown rice polysaccharide, carboxymethyl cellulose and maltodextrin into deionized water and stirring uniformly to obtain the inner aqueous phase solution; The preparation method of the modified octenylsuccinic anhydride cassava starch solution in step (2) is as follows: Add nano-silica to deionized water, then add sodium lauryl sulfate, stir evenly, then add chitosan acetic acid solution, carry out stirring reaction, filter, wash, and dry after the reaction is completed to obtain pretreated silica; add cassava starch to deionized water to form starch milk, add 5wt% sodium hydroxide to adjust the pH to 8-9, then add octenyl succinic anhydride ethanol solution dropwise, carry out heating reaction, maintain the pH at 8-9 during the reaction process, add 5wt% hydrochloric acid to adjust the pH to 6.5-7 after the reaction is completed, then add α-amylase and carry out enzymatic hydrolysis for 20-30 minutes to obtain octenyl succinic anhydride cassava starch solution; add the pretreated silica to the octenyl succinic anhydride cassava starch solution, then add β-cyclodextrin, carry out constant temperature reaction, and obtain the modified octenyl succinic anhydride cassava starch solution after the reaction is completed; The temperature of the isothermal reaction is 50-60° C., and the reaction time is 1-2 hours.

2. The preparation method of an embedded type Lactobacillus plantarum J26 according to claim 1, wherein The mass ratio of the brown rice polysaccharide, carboxymethyl cellulose, maltodextrin and deionized water is 5-10:1-3:1-2:150-200; the stirring temperature is 40-50° C. and the stirring time is 1-2 hours.

3. The preparation method of a kind of embedded type Lactobacillus plantarum J26 according to claim 1, wherein The number of viable Lactobacillus plantarum J26 in the Lactobacillus plantarum J26 bacterial solution in step (1) is 1.5×10 10 CFU / mL~2.0×10 10 CFU / mL, the mass ratio of the Lactobacillus plantarum J26 bacterial solution, the inner aqueous phase solution, the medium-chain fatty acid triglyceride, and the polyglycerol ricinoleate is 20:100-150:120-200:5-10; the rotation speed of the primary emulsification is 8000-12000 r / min, and the time is 10-15 min.

4. The preparation method of an embedded type Lactobacillus plantarum J26 according to claim 1, wherein The mass ratio of the nano-silica, sodium lauryl sulfate, and chitosan acetic acid solution is 20:0.1-0.2:100-200, the concentration of chitosan in the chitosan acetic acid solution is 5%, and the concentration of acetic acid is 10%. The temperature of the stirring reaction is 60-70°C, and the reaction time is 2-4 hours. The mass ratio of the cassava starch, octenylsuccinic anhydride ethanol solution, and α-amylase is 50:200-300:1-3, the concentration of the octenylsuccinic anhydride ethanol solution is 6-9wt%, the temperature of the heating reaction is 40-50°C, and the reaction time is 4-8 hours. The mass ratio of the pretreated silica, octenylsuccinic anhydride cassava starch solution, and β-cyclodextrin is 3:150-200:10-15.

5. The preparation method of an embedded type Lactobacillus plantarum J26 according to claim 1, wherein The mass ratio of the modified octenylsuccinic anhydride cassava starch solution, Tween 80, and W / O colostrum in step (2) is 100:4-8:7-13; the rotation speed of the secondary emulsification in step (2) is 2000-3000 r / min, and the time is 15-20 min.

6. The method for preparing an embedded Lactobacillus plantarum J26 according to claim 1, wherein The freeze-drying process in step (3) is as follows: the freeze-drying temperature is -80~-60°C, the vacuum degree is 5.0-10.0Pa, and the time is 12-16h.

7. An embedded Lactobacillus plantarum J26 prepared by the preparation method according to any one of claims 1 to 6.

8. Use of the encapsulated Lactobacillus plantarum J26 according to claim 7 in preparing a weight loss product.

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