Fermentation composition for reducing trimethylamine, preparation method and application

Abstract

The invention discloses a fermentation composition for reducing trimethylamine as well as a preparation method and application thereof, and belongs to the technical field of probiotic compositions. The fermentation composition is prepared from the following raw materials in parts by weight: 800 to 1000 parts of raw milk, 250 to 300 parts of cereal auxiliary materials and 0.3 to 0.7 part of bifidobacterium bifidum i771 bacterial powder, the cereal auxiliary materials comprise buckwheat, radix puerariae and soybeans. The bifidobacterium bifidum i771 provided by the invention has the effect of directly cracking TMA, the selected cereal auxiliary materials can also indirectly achieve the effects of reducing TMA and preventing TMAO generation in vivo in manners of regulating intestinal flora and the like, and the bifidobacterium bifidum i771 and the cereal auxiliary materials have a synergistic effect in the aspect of reducing TMA. The invention is suitable for preparing trimethylamine-reduced fermented foods, such as fermented dairy products.

Description

Technical Field

[0001] The invention belongs to the technical field of probiotic compositions, and relates to a nortrimethylamine composition, in particular to a nortrimethylamine fermentation composition, a preparation method and an application thereof. Background Art

[0002] Trimethylamine N-oxide (TMAO) is a metabolite of intestinal flora generated by the oxidation of trimethylamine (TMA) by liver enzymes in the liver. In recent years, studies have found that plasma TMAO levels play an important role in the occurrence and development of cardiovascular disease and osteoporosis. Regulating plasma TMAO levels provides a new idea and target for the treatment and prevention of cardiovascular and osteoporosis.

[0003] There have been studies on the use of probiotics such as Bifidobacterium longum in reducing trimethylamine. For example, there is a Chinese invention patent with application number "202210097947.3" and the name "A strain of Bifidobacterium longum for reducing plasma oxidized trimethylamine and cecal trimethylamine levels and its application", but there is currently no research on compositions specifically for reducing trimethylamine levels. Summary of the invention

[0004] The purpose of the present invention is to provide a fermentation composition of nortrimethylamine, a preparation method and application thereof, so as to solve the problem that there is currently a blank market for nortrimethylamine compositions.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is: A trimethylamine-reducing fermentation composition, the raw materials for making the effective component of the composition, in parts by weight, include 800-1000 parts of raw milk, 250-300 parts of cereal auxiliary materials and 0.3-0.7 parts of Bifidobacterium bifidum i771 bacterial powder; The cereal auxiliary materials include buckwheat, kudzu root and soybean.

[0006] Among them, Bifidobacterium bifidum i771 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on May 20, 2020, with the deposit number: CGMCC NO.19855. The bacterium was first disclosed in the Chinese invention patent with application number "202011361327.3" and named "Bifidobacterium bifidum i771, its isolation and purification method and application".

[0007] As a limitation, the weight ratio of buckwheat, kudzu root and soybean is 2-3:1-2:1-2.

[0008] Among them, flavonoids such as rutin and quercetin contained in buckwheat can prevent vascular dysfunction and liver damage caused by TMAO; and the dietary fiber content of buckwheat is rich, which is 10 times that of ordinary refined rice, which helps to maintain intestinal health; the resistant starch in buckwheat is fermented in the colon to produce short-chain fatty acids, which helps to lower the pH value of the colon, regulate intestinal flora, and reduce the production of TMA; Puerarin contained in Pueraria root can inhibit Prevotella in intestinal flora, weaken its ability to metabolize TMA and generate TMAO, and has the effect of reducing trimethylamine; Puerarin can also regulate the dynamic balance between osteoclasts and osteoblasts, and play a positive role in the prevention and treatment of osteoporosis; the isoflavones contained in Pueraria root can promote the absorption and utilization of calcium, which helps to prevent osteoporosis; at the same time, through probiotic fermentation, Pueraria root and probiotics work synergistically, on the one hand, it can increase the total flavonoid content in the product, and on the other hand, it can increase the extracellular polysaccharide content of probiotics. Extracellular polysaccharides can maintain the activity of probiotics, so that probiotics can enter the body to play a role in regulating the intestines, thereby reducing the generation of TMA; Soy is a source of plant protein and can be a low-choline and low-carnitine food choice, which helps reduce the production of TMAO in the body. At the same time, based on fermentation, its fatty acid content is increased, accompanied by the potential benefits of soy milk in lowering cholesterol and preventing cardiovascular disease.

[0009] As another limitation, the amount of live bacteria of the Bifidobacterium bifidum i771 powder is 1×10 11 CFU / g.

[0010] The present invention also provides a method for preparing the above-mentioned trimethylamine-reducing fermentation composition, which comprises the steps of pasteurizing, homogenizing, sterilizing and cooling raw milk, adding cereal auxiliary materials and Bifidobacterium bifidum i771 bacterial powder for fermentation, breaking the emulsion, freeze-drying and pulverizing to obtain the fermentation composition.

[0011] As a limitation, the pasteurization is performed at a temperature of 73-77°C and a time of 10-20s; the homogenization is performed at a temperature of 65-70°C and a pressure of 16-18MPa; the sterilization is performed at a temperature of 93-98°C and a time of 300-310s; and the cooling is performed at a temperature of 40-42°C.

[0012] As another limitation, the fermentation is carried out at a temperature of 37-42°C until the pH value reaches 4.8.

[0013] The present invention also provides a trimethylamine-reducing dairy product for the middle-aged and elderly. The raw materials for making the trimethylamine-reducing dairy product include, by weight, the above-mentioned fermentation composition for trimethylamine reduction, 250-300 parts of skim milk powder, 230-260 parts of desalted whey powder and 10-15 parts of calcium carbonate.

[0014] Due to the adoption of the above technical solution, the present invention has the following technical advances compared with the prior art: ① The present invention provides a trimethylamine-reducing fermentation composition, wherein Bifidobacterium bifidum i771 has the effect of directly lysing TMA, and the selected cereal auxiliary materials can also indirectly reduce TMA and prevent TMAO formation in vivo by regulating intestinal flora, and the two have a synergistic effect in reducing TMA; ② The present invention provides a method for preparing a trimethylamine-reducing fermentation composition, which makes the raw and auxiliary materials easier to digest and absorb through fermentation, and also improves the functional components. The preparation process is simple, does not require deep processing of the raw materials, effectively maintains the nutrition of the raw materials, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 It is the test result of trimethylamine content in Example 1. DETAILED DESCRIPTION

[0016] The present invention is further described in detail below by specific examples. It should be understood that the described examples are only used to explain the present invention, and are not intended to limit the present invention.

[0017] Unless otherwise specified, the experimental methods used in the following examples are conventional methods in the art.

[0018] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0019] In the following embodiments: Lactobacillus paracasei N1115, which was deposited in the China General Microbiological Culture Collection Center on March 17, 2011, with the deposit number: CGMCC No.4691. The strain was first disclosed in a Chinese invention patent with the application number of "201110357058.8" and the name of "Lactobacillus casei N1115, its immunomodulatory effect and application"; Bifidobacterium bifidum i771, this strain was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on May 20, 2020, with the deposit number: CGMCC NO.19855. This strain was first disclosed in a Chinese invention patent with application number "202011361327.3" and named "Bifidobacterium bifidum i771, its isolation and purification method and application".

[0020] Example 1 This example prepares a fermentation composition for reducing trimethylamine, and provides a control experiment to verify the effect of reducing trimethylamine, which specifically includes the following steps performed in sequence: S1. Preparation of fermentation composition After acceptance and standardization, the raw milk was separated from the impurities by a milk purifier. 1000 kg of raw milk was pasteurized at 75 °C for 15 seconds, then heated to 65 °C, homogenized at 17.5 MPa, and sterilized at 93 °C for 300 seconds. After cooling to 41 °C, 0.3 kg of Bifidobacterium bifidum i771 powder (live bacteria count 1×10 11 CFU / g) and 250 kg of cereal auxiliary materials (consisting of buckwheat, kudzu root and soybean in a weight ratio of 2:1:2) were stirred until dissolved, fermented at 42°C until the pH value reached 4.8, stirred to break the emulsion, freeze-dried, crushed and sieved to obtain the fermentation composition ɑ1.

[0021] S2. Trimethylamine test 1) Sample preparation Sample 1: The amount of viable bacteria is 1×10 11 CFU / g of Bifidobacterium bifidum i771 powder; Sample 2: The amount of viable bacteria is 1×10 11 CFU / g of Lactobacillus paracasei N1115 powder; Sample 3: Cereal adjunct (consisting of buckwheat, kudzu root and soybean in a weight ratio of 2:1:2); Sample 4: fermented composition ɑ1 prepared in step S1; Sample 5: The preparation method of Sample 5 is basically the same as the method for preparing the fermentation composition ɑ1 in step S1, the only difference being that the powder of Lactobacillus paracasei N1115 is substituted for the powder of Bifidobacterium bifidum i771, and the amounts of other raw materials and control parameters are the same.

[0022] 2) Experimental process Weigh 1g of fecal sample from healthy people into a 15mL centrifuge tube, add sterile saline to make a 10% (w / v) fecal dilution, centrifuge to remove impurities, use a sterile syringe to draw 0.5mL of fecal dilution and add it to YCFA basal medium, add equal amounts of samples 1-5, and culture at 37℃ for 24h. Set up a control group without adding samples; After the culture is completed, trimethylamine is tested according to the national standard "GB 5009.179-2016 National Food Safety Standard Determination of Trimethylamine in Food", and the results are as follows: Figure 1 As shown; Depend on Figure 1It can be seen that the Bifidobacterium bifidum i771 powder and the Lactobacillus paracasei N1115 powder still have the effect of directly reducing the content of trimethylamine in vitro, but the cereal auxiliary material cannot reduce the content of trimethylamine in vitro; however, the fermentation composition ɑ1 made of Bifidobacterium bifidum i771 and cereal auxiliary materials provided in this embodiment has a significant effect on reducing trimethylamine, which is better than the use of Bifidobacterium bifidum i771 powder alone. In contrast, the composition made of Lactobacillus paracasei N1115 and cereal auxiliary materials has a poor effect on reducing trimethylamine, which is significantly different from the fermentation composition ɑ1 or the use of Lactobacillus paracasei N1115 powder alone.

[0023] Example 2 This embodiment prepares a fermentation composition of trimethylamine, which specifically includes the following steps performed in sequence: After acceptance and standardization, the raw milk was separated from the impurities by a milk purifier. 950 kg of raw milk was pasteurized at 73 °C for 20 s, then heated to 68 °C, homogenized at 16 MPa, and sterilized at 95 °C for 310 s. After cooling to 40 °C, 0.7 kg of Bifidobacterium bifidum i771 powder (with a live bacterial count of 1 × 10 11 CFU / g) and 280kg of cereal auxiliary materials (consisting of buckwheat, kudzu root and soybean in a weight ratio of 2:1:1) were stirred until dissolved, fermented at 37°C until the pH value reached 4.8, stirred to break the emulsion, freeze-dried, crushed and sieved to obtain the fermentation composition ɑ2.

[0024] Example 3 This embodiment prepares a fermentation composition of trimethylamine, which specifically includes the following steps performed in sequence: After acceptance and standardization, the raw milk was separated from the impurities by a milk purifier. 800 kg of raw milk was pasteurized at 77 °C for 10 seconds, then heated to 70 °C, homogenized at 18 MPa, and sterilized at 98 °C for 305 seconds. After cooling to 42 °C, 0.5 kg of Bifidobacterium bifidum i771 powder (with a live bacterial count of 1 × 10 11 CFU / g) and 300 kg of cereal auxiliary materials (consisting of buckwheat, kudzu root and soybean in a weight ratio of 3:2:1) were stirred until dissolved, fermented at 40°C until the pH value reached 4.8, stirred to break the emulsion, freeze-dried, crushed and sieved to obtain the fermentation composition ɑ3.

[0025] Example 4 The present embodiment provides a trimethylamine-lowering dairy product for middle-aged and elderly people, and the raw materials for making the dairy product include the fermentation composition ɑ1 prepared in Example 1, 250 kg of skim milk powder, 230 kg of desalted whey powder and 10 kg of calcium carbonate.

[0026] Example 5 The present embodiment provides a trimethylamine-lowering dairy product for middle-aged and elderly people, and the raw materials for making the dairy product include the fermentation composition ɑ2 prepared in Example 2, 280 kg of skim milk powder, 240 kg of desalted whey powder and 13 kg of calcium carbonate.

[0027] Example 6 The present embodiment provides a trimethylamine-lowering dairy product for middle-aged and elderly people, and the raw materials for making the dairy product include the fermentation composition ɑ3 prepared in Example 3, 300 kg of skim milk powder, 260 kg of desalted whey powder and 15 kg of calcium carbonate.

Claims

1. A fermentation composition of trimethylamine, characterized in that: The raw materials for making the effective ingredients thereof include, by weight, 800-1000 parts of raw milk, 250-300 parts of cereal auxiliary materials and 0.3-0.7 parts of Bifidobacterium bifidum i771 bacterial powder; The cereal auxiliary materials include buckwheat, kudzu root and soybean.

2. A fermentation composition for reducing trimethylamine according to claim 1, characterized in that: The weight ratio of buckwheat, kudzu root and soybean is 2-3:1-2:1-2.

3. A fermentation composition for reducing trimethylamine according to claim 1 or 2, characterized in that: The live bacteria amount of the Bifidobacterium bifidum i771 powder is 1×10 11 CFU / g.

4. The method for preparing a fermentation composition of trimethylamine according to any one of claims 1 to 3, characterized in that: The method comprises the following steps: pasteurizing, homogenizing, sterilizing and cooling raw milk, adding cereal auxiliary materials and Bifidobacterium bifidum i771 bacterial powder to ferment, breaking the milk, freeze drying and crushing to obtain the fermentation composition.

5. The preparation method according to claim 4, characterized in that: The pasteurization is carried out at a temperature of 73-77°C and a time of 10-20s; the homogenization is carried out at a temperature of 65-70°C and a pressure of 16-18MPa; the sterilization is carried out at a temperature of 93-98°C and a time of 300-310s; and the cooling is carried out at a temperature of 40-42°C.

6. The preparation method according to claim 4 or 5, characterized in that: The fermentation temperature is 37-42°C until the pH value reaches 4.

8.

7. A dairy product for middle-aged and elderly people that reduces trimethylamine, characterized in that: The raw materials for making the product include, by weight, the fermented composition of the trimethylamine-reducing agent described in any one of claims 1 to 3, 250 to 300 parts of skim milk powder, 230 to 260 parts of desalted whey powder and 10 to 15 parts of calcium carbonate.

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