Composition capable of efficiently proliferating Witzia ciliaris and application of composition

Abstract

The invention discloses a composition capable of efficiently proliferating Witzia ciliaris and application of the composition. The composition disclosed by the invention can be used for promoting the proliferation of the Witzia ciliaris so as to regulate the intestinal health, inhibit harmful bacteria in the intestinal tract and the like.

Description

Technical Field

[0001] The present disclosure relates to the fields of common food, health food, medicine and cosmetics, and in particular to a compound capable of efficiently proliferating and coagulating Weizmannella and its application. Background Art

[0002] Probiotics are microorganisms that are beneficial to humans and animals. Commonly known probiotics include Weizmannella coagulans, Lactobacillus and Bifidobacterium. They are living organisms that need to pass the test of strong acid and bile salts. Only some of them can survive and reach the intestines, replenish probiotics, colonize the human intestines, inhibit the growth of harmful bacteria, and improve the intestinal microenvironment. Therefore, how to promote their proliferation and industrialization has become the focus of current research and development workers. Summary of the invention

[0003] To solve the above problems, the present invention takes Weizmannella coagulans as the research object, adopts an in vitro simulated fermentation method, detects the growth curve of microorganisms in real time, and screens out litchi fermentation products and prebiotic compositions that can synergistically enhance the proliferation of Weizmannella coagulans.

[0004] A first aspect of the present invention provides a composition comprising litchi fermented product and prebiotics.

[0005] In some embodiments, the preparation method comprises the following steps: 1) primary fermentation, which comprises adding a first carbon source to litchi juice and litchi pomace, inoculating a first bacterial agent, and sealing and standing for fermentation at 20-45°C for 24-96 hours; 2) squeezing and filtering the product after the primary fermentation to obtain a primary fermentation filtrate; 3) secondary fermentation, which comprises adding a second carbon source to the primary fermentation filtrate, inoculating a second bacterial agent, and standing or aerating and culturing at 15-40°C for 3-15 days to obtain a secondary fermentation liquid; 4) filtering and sterilizing the secondary fermentation liquid to obtain a litchi fermentation product.

[0006] In certain embodiments, the first carbon source is selected from one or more of fructose, sucrose, brown sugar, maltose, and arabinose.

[0007] In certain embodiments, the first bacterial agent is a mixed bacterial agent of yeast and lactic acid bacteria.

[0008] In certain embodiments, the second carbon source is selected from one or more of lactose, brown sugar, glucose, molasses or honey.

[0009] In certain embodiments, the second bacterial agent is acetic acid bacteria.

[0010] In certain embodiments, the prebiotic is selected from one or more of isomaltooligosaccharides, galacto-oligosaccharides, and stachyose.

[0011] In certain embodiments, the composition consists of litchi fermentate and prebiotics.

[0012] In certain embodiments, the composition contains 28-95 parts by weight of the litchi fermented product and 5-72 parts by weight of the prebiotics.

[0013] In certain embodiments, the composition contains 28.08-94.28 parts by weight of the litchi fermented product, and 5.72-71.92 parts by weight of the prebiotics.

[0014] In certain embodiments, the composition contains 34-88 parts by weight of the litchi fermented product and 12-66 parts by weight of the prebiotics.

[0015] In certain embodiments, the composition contains 34.98-87.4 parts by weight of the litchi fermented product and 12.6-65.02 parts by weight of the prebiotics.

[0016] In certain embodiments, the composition contains 44-78 parts by weight of the litchi fermented product and 22-56 parts by weight of the prebiotics.

[0017] In certain embodiments, the composition contains 44.55-77.8 parts by weight of the litchi fermented product, and 22.2-55.45 parts by weight of the prebiotics.

[0018] In certain embodiments, the composition further comprises coagulated Weizmannella powder.

[0019] In certain embodiments, the composition contains 28-95 parts by weight of the litchi fermented product and 5-72 parts by weight of the prebiotics.

[0020] In certain embodiments, the composition contains 28.08-94.28 parts by weight of the litchi fermented product, and 5.72-71.92 parts by weight of the prebiotics.

[0021] In certain embodiments, the composition contains 34-88 parts by weight of the litchi fermented product and 12-66 parts by weight of the prebiotics.

[0022] In certain embodiments, the composition contains 34.98-87.4 parts by weight of the litchi fermented product and 12.6-65.02 parts by weight of the prebiotics.

[0023] In certain embodiments, the composition contains 44-78 parts by weight of the litchi fermented product and 22-56 parts by weight of the prebiotics.

[0024] In certain embodiments, the composition contains 44.55-77.8 parts by weight of the litchi fermented product, and 22.2-55.45 parts by weight of the prebiotics.

[0025] The second aspect of the present invention provides a method for preparing a composition, comprising the following steps: 1) preparing litchi fermented product; 2) mixing litchi fermented product and prebiotics; and optionally, further adding Weizmannella coagulans.

[0026] In some embodiments, the preparation method comprises the following steps: 1) primary fermentation, which comprises adding a first carbon source to litchi juice and litchi pomace, inoculating a first bacterial agent, and sealing and standing for fermentation at 20-45°C for 24-96 hours; 2) squeezing and filtering the product after the primary fermentation to obtain a primary fermentation filtrate; 3) secondary fermentation, which comprises adding a second carbon source to the primary fermentation filtrate, inoculating a second bacterial agent, and standing or aerating and culturing at 15-40°C for 3-15 days to obtain a secondary fermentation liquid; 4) filtering and sterilizing the secondary fermentation liquid to obtain a litchi fermentation product.

[0027] In some implementations of fermentation, a pre-treatment step is also included before step 1); preferably, the pre-treatment is to wash about 5 tons of litchi fruits and squeeze them to obtain litchi juice and litchi pomace.

[0028] In certain embodiments, the primary fermentation is sealed and allowed to stand for fermentation at about 25° C. for about 48 hours.

[0029] In certain embodiments, the first carbon source is a mixture of fructose:sucrose:brown sugar; preferably, the ratio of fructose:sucrose:brown sugar is 1:1:1.

[0030] In certain embodiments, the first carbon source is added in an amount of about 3% (w / w).

[0031] In certain embodiments, the first bacterial agent is a composite bacterial agent of yeast: lactic acid bacteria, preferably, the ratio of yeast: lactic acid bacteria is 500:1.

[0032] In certain embodiments, the yeast is a composite bacterial agent of baker's yeast and sake yeast; preferably, the ratio of baker's yeast to sake yeast is 1:1.

[0033] In certain embodiments, the lactic acid bacteria is a composite bacterial agent of Lactobacillus plantarum, Lactobacillus gasseri and Lactobacillus sakei; preferably, the ratio of Lactobacillus plantarum: Lactobacillus gasseri: Lactobacillus sakei is 1:1:1.

[0034] In some embodiments, the first bacterial agent is added in an amount of about 8‰ of the total mass of litchi.

[0035] In certain embodiments, in step 2), a filter press is used to squeeze to obtain a primary fermentation filtrate; preferably, the mass of the primary fermentation filtrate is greater than 60% of the total mass of the litchi fruit.

[0036] In certain embodiments, the step 3) secondary fermentation is cultured with aeration at about 40° C. for about 9 days.

[0037] In certain embodiments, the secondary fermentation culture method in step 3) is aeration culture with an aeration volume of 5 mL / min / mL of fermentation liquid.

[0038] In certain embodiments, the second carbon source is honey; preferably, the added amount of the second carbon source is 20% of the total mass of the primary fermentation filtrate.

[0039] In certain embodiments, the second bacterial agent is a composite bacterial agent of acetic acid bacteria of malodorous acetic acid bacteria: acetobacter acetic acid: and acetobacter pasteurianus. Preferably, the ratio of acetic acid bacteria of malodorous acetic acid bacteria: acetobacter acetic acid: and acetobacter pasteurianus is 1:1:1.

[0040] In certain embodiments, the amount of the second bacterial agent added is about 0.9‰ of the total mass of the primary fermentation filtrate.

[0041] The third aspect of the present invention provides use of the composition described in the first aspect of the present invention in regulating intestinal flora; preferably, the use is for non-therapeutic purposes; preferably, regulating intestinal flora includes promoting the proliferation of Weizmannella coagulans.

[0042] The fourth aspect of the present invention provides the use of the composition described in the first aspect of the present invention in the preparation of food, health care products or medicines with intestinal regulation function.

[0043] In certain embodiments, the content of the litchi fermented product and the prebiotics in the food, health product or medicine is 100 mg / 100 g to 1×10 4 mg / 100g.

[0044] In certain embodiments, the food, health product or medicine contains 1×10 9 CFU / 100g to 1×10 12 CFU / 100 g of Weizmannella coagulans.

[0045] The fifth aspect of the present invention provides use of the composition according to the first aspect of the present invention in promoting the proliferation of Weizmannella coagulans in vivo or in vitro.

[0046] The sixth aspect of the present invention provides a method for promoting the proliferation of Weizmannella coagulans, the method comprising adding the composition of the first aspect of the present invention to the living environment of Weizmannella coagulans.

[0047] The seventh aspect of the present invention provides a functional product, which includes the composition described in the first aspect of the present invention; preferably, the functional product is a food, a health product or a medicine.

[0048] In certain embodiments, the content of the litchi fermentation product and the prebiotics in the functional product is 100 mg / 100 g to 1×10 4 mg / 100g.

[0049] In some embodiments, the functional product contains 1×10 9 CFU / 100g to 1×10 12 CFU / 100 g of Weizmannella coagulans.

[0050] In various embodiments herein, Weizmannella coagulans is deposited in the General Microbiological Center of China National Microbiological Culture Collection Administration, and the registration number of the collection center is: CGMCC No.21801.

[0051] The advantages of the present invention over the prior art are:

[0052] 1) It was found that litchi fermentation products can be used as a carbon source to proliferate Weizmannella flocculata, and its proliferation effect is twice that of glucose.

[0053] 2) It was found that litchi fermentation products combined with isomaltooligosaccharide have a synergistic effect on the proliferation of Weizmannella coagulans, which is better than any single material.

[0054] 3) The combination of litchi fermented products and prebiotics can play a role in regulating intestinal health and inhibiting harmful intestinal bacteria. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 Effects of litchi fermentation products and common prebiotics on the growth curve of Weizmannella coagulans.

[0056] Figure 2 Effects of litchi fermentation products, isomaltooligosaccharides and their combination on the growth curve of Weizmannella coagulans. DETAILED DESCRIPTION

[0057] The following definitions are provided to facilitate understanding of the present invention by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art to which the present invention belongs. Preferred materials and methods are described herein, but any methods and materials similar to or equivalent to those described herein may be used in the practice of the present invention's tests. It should also be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to be limiting.

[0058] Definition of terms

[0059] Unless otherwise indicated or defined, all terms used have the usual meaning in the art, which will be understood by those skilled in the art. In addition, unless otherwise specified, all methods, steps, techniques and operations not specifically described in detail can and have been performed in a manner known per se, which will be understood by those skilled in the art.

[0060] As used herein, the term "prebiotic" refers to some organic substances that are not digested and absorbed by the host but can selectively promote the metabolism and proliferation of beneficial bacteria in the body, thereby improving the health of the host. The most basic prebiotic is carbohydrate, but by definition, it does not exclude non-carbohydrate substances used as prebiotics. Commonly used prebiotics include oligosaccharides and bifidus factors. Oligosaccharides include oligofructose, oligogalactose, oligoxylose, oligomaltosaccharides, soy oligosaccharides, inulin, etc. Prebiotics can provide an effective carbon source for endogenous probiotics, stimulate their growth and control, and indirectly protect intestinal health and promote the absorption of nutrients, and generally do not produce immune responses such as allergies. Prebiotics are non-organic living bodies that arrive at the intestine in an undigested form, and there is no problem of viability.

[0061] As used herein, "probiotics" are a class of active microorganisms that are beneficial to the host by colonizing in the human body and changing the composition of the flora in a certain part of the host. They promote nutrient absorption and maintain intestinal health by regulating the host mucosal and systemic immune function or by regulating the balance of intestinal flora, thereby producing single microorganisms or mixed microorganisms with clear composition that are beneficial to health. Commonly known probiotics include Weizmannella coagulans, Lactobacillus, and Bifidobacterium.

[0062] As used herein, "isomaltooligosaccharide", abbreviated as "IMO", is also known as isomaltooligosaccharide, isomaltooligosaccharide, and branched oligosaccharide. "Isomaltooligosaccharide" is a type of starch sugar, and its main components are isomaltose, panose, isomaltotriose, and oligosaccharides above tetrasaccharide, which are bound by α-1,6 glycosidic bonds between glucose molecules. Isomaltooligosaccharide can effectively promote the growth and reproduction of beneficial bacteria-bifidobacteria in the human body, so it is also called "bifidobacterium growth promoting factor", referred to as "bifidobacterium factor". The method for industrial production of isomaltooligosaccharide includes the following steps: using starch as a raw material, after emulsification and liquefaction, fungal amylase and glucosyltransferase are added to the liquefied liquid for saccharification and transglycoside reaction. After a certain period of time, a sugar solution with isomaltose, isomaltotriose and panose as the main components is produced. After decolorization, filtration, separation and concentration, an isomaltooligosaccharide finished syrup with a solid content of more than 75% is made. According to needs, the syrup concentrated to a certain concentration can be spray-dried to prepare isomaltooligosaccharide powder with the same composition as the syrup.

[0063] As used herein, "galacto-oligosaccharide", abbreviated as "GOS", is a functional oligosaccharide with natural properties. Its molecular structure is generally 1 to 7 galactose groups connected to galactose or glucose molecules, that is, Gal-(Gal)n-Glc / Gal (n is 0-6). Galacto-oligosaccharides (GOS) are mainly composed of trans-oligosaccharides and monosaccharides. The main components of trans-oligosaccharides include lactose, galactose, disaccharides, trisaccharides, tetrasaccharides, pentasaccharides, etc., while the main components of monosaccharides are glucose and galactose. In industry, lactose is usually used as a raw material and is produced by the action of β-galactosidase.

[0064] Example

[0065] The present invention will be more readily understood with reference to the following examples, which are included only to illustrate certain aspects and embodiments of the present invention and are not intended to limit the present invention.

[0066] Unless otherwise stated, the reagents used in this example are all commercially available materials or conventional materials.

[0067] Example 1 Experimental Materials

[0068] 1.1 Probiotics: bacterial powder composed of Weizmannella coagulans. Weizmannella coagulans is commercially available and deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with the registration number of the collection center being: CGMCC No.21801.

[0069] 1.2 Prebiotics: isomaltooligosaccharides (IMO), galacto-oligosaccharides, stachyose, inulin, polydextrose, fructooligosaccharides, resistant dextrin, and xylo-oligosaccharides.

[0070] 1.3 Preparation of litchi fermentation products

[0071] The preparation of litchi fermented product is carried out according to the method described in the authorized patent ZL202210565739.1. For example, the preparation of litchi fermented product can be carried out according to the method of Example 5 in the patent, and the specific steps are as follows: (1) Pretreatment: 5 tons of litchi fruit are washed and squeezed;

[0072] (2) Primary fermentation: add 1% sugar to the mixture of fruit juice and pomace, inoculate 1% bacterial agent, and ferment at 25°C in a sealed container for 48 hours;

[0073] (3) Filtration: using a filter press to obtain a primary fermentation filtrate, wherein the mass of the primary fermentation filtrate is greater than 60% of the total mass of the litchi fruit;

[0074] (4) Secondary fermentation: add sugar 2 to the primary fermentation filtrate, inoculate bacterial agent 2, and culture at 40°C for 9 days with aeration;

[0075] (5) Filtration and sterilization.

[0076] Sugar 1 is fructose: sucrose: brown sugar in a ratio of 1:1:1; the added amount of Sugar 1 is 3% (w / w); the bacterial agent 1 is yeast: lactic acid bacteria in a ratio of 500:1; the yeast is baker's yeast: sake yeast in a ratio of 1:1; the lactic acid bacteria are Lactobacillus plantarum: Lactobacillus gasseri: Lactobacillus sake in a ratio of 1:1:1; the added amount of the bacterial agent 1 is 8‰ of the total mass of litchi.

[0077] Sugar 2 is honey; the addition amount of Sugar 2 is 20% of the total mass of the primary fermentation filtrate; the bacterial agent 2 is 1:1:1 of acetic acid bacteria: Acetobacter aceticus: Acetobacter pasteurianus; the addition amount of bacterial agent 2 is 0.9‰ of the total mass of the primary fermentation filtrate; the culture method is aeration culture with an aeration volume of 5 mL / min / mL of fermentation liquid.

[0078] 1.4 Culture medium

[0079] 0.1% peptone water: accurately weigh 1.0 g of peptone and dissolve it in 1000 mL of deionized water. Adjust the pH to 7.0 with hydrochloric acid / NaOH. Aliquot 10 mL into each test tube and sterilize it with high pressure steam at 121°C for 15 min for later use.

[0080] Trace mineral solution: Accurately weigh 0.5 g sodium chloride, 0.9 g ferrous sulfate heptahydrate, 0.8 g manganese sulfate monohydrate, 0.08 g zinc sulfate heptahydrate, 0.08 g copper sulfate pentahydrate, and 0.08 g cobalt sulfate heptahydrate and add to 50 mL of deionized water.

[0081] Proliferation liquid culture medium: Accurately weigh 5 g yeast extract, 5 g peptone, 20 g D-glucose, 0.5 g dipotassium hydrogen phosphate, 0.5 g potassium dihydrogen phosphate, 0.3 g magnesium sulfate and 1 mL trace mineral solution and dissolve them in 1000 mL deionized water. Adjust the pH to 7.2 with hydrochloric acid / NaOH and sterilize by high-pressure steam at 121°C for 15 min.

[0082] Carbon-free fermentation liquid medium: Accurately weigh 5 g of peptone, 0.5 g of dipotassium hydrogen phosphate, 0.5 g of potassium dihydrogen phosphate, 0.3 g of magnesium sulfate and 1 mL of trace mineral solution and dissolve them in 1000 mL of deionized water, and adjust the pH to 7.2 with hydrochloric acid / NaOH.

[0083] Example 2 Effects of litchi fermentation products and common prebiotics on the proliferation of Weizmannella coagulans 2.1 Method of adding litchi fermentation products and prebiotics: 10 mL of carbon source-free fermentation liquid culture medium was filled into each test tube, and 200 mg of corresponding different carbon sources (litchi fermentation products and prebiotics, see Table 1) were added to each test tube. After they were fully dissolved, the pH was adjusted to 7.2, and then sterilized with high-pressure steam at 121°C for 20 min for use.

[0084] The coagulated Weizmannella strains stored in a freeze-drying tube at -80°C were inoculated into a proliferation liquid culture medium at an inoculum rate of 3.33% (v / v), and cultured in a shaking incubator at 40°C for 24 hours to obtain a strain mother solution.

[0085] The mother liquid of the strain was centrifuged at 1000r / min for 10min at 4°C, the supernatant was discarded, and the mixture was redissolved in sterile water, and the mixture was fully shaken and mixed, and the mixture was repeated twice, and the proliferation medium was washed away. The obtained bacterial liquid was inoculated into the fermentation liquid medium (different carbon sources, see Table 1) at an inoculum rate of 1% (v / v), and cultured at 40°C for 72h. Each group of samples was measured in triplicate.

[0086] 2.2 Proliferation-promoting effect detection: The OD600nm value of the fermentation liquid was used as the evaluation index. 200 μL of fermentation liquid was added to a 96-well plate, and then its absorbance at 600nm was measured using an ELISA reader. The temperature was set to 40°C, the time was 72 hours, and the OD value was measured every 30 minutes. The change curve of OD600nm under different fermentation times was drawn.

[0087] The test results are shown in Table 1 below:

[0088] Table 1 Effect of different carbon sources on the proliferation of Weizmannella coagulans

[0089] Experimental materials 0h 12h 24h 36h 48h 60h 72h 2% Lychee ferment 0.0021 0.1531 0.3216 0.4159 0.4664 0.4739 0.4626 2% Isomaltooligosaccharide 0.0025 0.1098 0.2235 0.2799 0.3044 0.3158 0.3258 2% Stachyose 0.0008 0.1027 0.2333 0.3111 0.3375 0.3362 0.3327 2% Galacto-oligosaccharide 0.0016 0.0653 0.2428 0.3039 0.3178 0.3121 0.3110 2% Glucose 0.0006 0.1043 0.2489 0.3185 0.3396 0.3438 0.3467 2% Polydextrose 0.0012 0.0834 0.2207 0.2386 0.2430 0.2402 0.2377 2% Inulin 0.0015 0.1197 0.2283 0.2663 0.2762 0.2759 0.2741 2% Fructooligosaccharides 0.0008 0.1003 0.2290 0.2682 0.2787 0.2780 0.2787 2% L-arabinose 0.0001 0.1043 0.2011 0.2312 0.2398 0.2327 0.2309 2% resistant dextrin 0.0063 0.1097 0.2275 0.2643 0.2738 0.2738 0.2695 2% Xylo-oligosaccharide 0.0001 0.0929 0.1956 0.2240 0.2245 0.2170 0.2143

[0090] As shown in Table 1, with the increase of fermentation time, the effect of litchi fermentation on the proliferation of Weizmannella coagulans was significantly better than that of the positive control group (glucose group). Among them, isomaltooligosaccharide, galacto-oligosaccharide and stachyose had similar effects on the proliferation of Weizmannella coagulans as the positive control group (glucose).

[0091] Example 3 Effects of different carbon sources or carbon source combinations on the proliferation of Weizmannella coagulans

[0092] The experimental method of Example 3 is the same as that of Example 2, and the specific experimental groups are set as follows (Table 2): Experimental group 1 is 1% litchi fermentation + 1% isomaltooligosaccharide, experimental group 2 is 1% litchi fermentation + 1% stachyose, experimental group 3 is 1% litchi fermentation + 1% galacto-oligosaccharide, experimental group 4 is 1% litchi fermentation + 1% resistant dextrin, experimental group 5 is 2% litchi fermentation, experimental group 6 is 2% isomaltooligosaccharide, experimental group 7 is 2% stachyose, experimental group 8 is 2% galacto-oligosaccharide, and experimental group 9 is 2% resistant dextrin. The results are shown in Table 2.

[0093] Table 2 Proliferation promoting effect of different carbon sources or carbon source combinations on Weizmannella coagulans

[0094]

[0095] From the above results, it can be seen that the effect of 1% litchi fermentation combined with 1% isomaltooligosaccharide on the proliferation of Weizmannella coagulans is better than that of any of the two materials alone, which shows that the two have a synergistic effect on the proliferation of Weizmannella coagulans. However, the effect of 1% litchi fermentation combined with 1% resistant dextrin on the proliferation of Weizmannella coagulans is lower than that of any of the two materials alone, indicating that the two materials have a synergistic effect.

[0096] Example 4 Optimal Point Fitting

[0097] 4.1 Obtaining the Optimal Ratio

[0098] Experimental group settings:

[0099] According to Minitab software, a two-material mixing experiment design was performed to obtain the five groups of litchi fermentation products and oligosaccharide ratios shown in Table 3 below. The proliferation ability of the five groups of ratios was tested for Weizmannella procoagulans, with three replicates in each group, and the corresponding OD values ​​of the five groups of experiments were obtained. 600 The experimental results are brought into the software to perform mixture analysis, including mixture design analysis, response tracking diagram and response optimizer, so as to predict the optimal ratio.

[0100] Table 3 Ratios of different compositions

[0101] Experimental Group Addition ratio of litchi fermentation Isomaltooligosaccharide addition ratio 1 100％ 0 2 0 100％ 3 50％ 50％ 4 75％ 25％ 5 25％ 75％

[0102] Among them, the detection experiment of the proliferation ability of pro-coagulant Weizmannella includes the following steps:

[0103] (1) Anaerobic culture in small bottles, fermentation liquid medium without carbon source + 2% mixture of litchi fermentation product and oligosaccharide (5 ratios, see Table 3)

[0104] (2) Cultivation with automatic growth curve instrument, liquid medium without carbon source + 2% mixture of litchi fermentation product and isomaltooligosaccharide. (5 ratios, see Table 3)

[0105] Detection indicators: real-time growth curve (automatic growth curve instrument, requiring an uninoculated system as a control).

[0106] According to the OD of different ratios in Table 3 600 The results show that the optimal ratio at 2% carbon source concentration is 61.62%:38.38%.

[0107] 4.2 Verification of the optimal ratio

[0108] The optimal ratio obtained in 4.1 was experimentally verified. The inventor designed the ratios of each group in Table 4 based on the optimal ratio of 61.62%:38.38% obtained in 4.1. The proliferation ability of the pro-coagulant Weizmannella in each group in Table 4 was tested experimentally. The test experimental steps were the same as those described in 4.1.

[0109] Finally, the combination of litchi fermentation products and prebiotics was screened out based on the bacterial flora simulated in vitro fermentation.

[0110] The optimal ratio of litchi fermented product to isomaltooligosaccharide is 61.62%:38.38%.

[0111] Table 4

[0112] Specific experimental examples Addition ratio of litchi fermentation Isomaltooligosaccharide Addition Ratio OD value mm Comparative Example 1 0.00％ 100.00％ 0.3321 Comparative Example 2 0.52％ 99.48％ 0.3356 Comparative Example 3 5.00％ 95.00％ 0.3648 Experimental Example 1 29.63％ 70.37％ 0.4852 Experimental Example 2 45.71％ 54.29％ 0.5274 Experimental Example 3 55.81％ 44.19％ 0.5391 Experimental Example 4 61.22％ 38.78％ 0.5407 Experimental Example 5 64.15％ 35.85％ 0.5402 Experimental Example 6 67.80％ 32.20％ 0.5383 Experimental Example 7 70.31％ 29.69％ 0.5361 Comparative Example 4 100.00％ 0.00％ 0.4567 Comparative Example 6 99.68％ 0.32％ 0.4581 Comparative Example 7 96.88％ 3.13％ 0.4696 Experimental Example 8 83.78％ 16.22％ 0.5122 Experimental Example 9 72.09％ 27.91％ 0.5341 Experimental Example 10 65.96％ 34.04％ 0.5394 Experimental Example 11 55.36％ 44.64％ 0.5388 Experimental Example 12 61.62％ 38.38％ 0.5408 Comparative Example 5 22.03％ 77.97％ 0.4551 Experimental Example 13 28.08％ 71.92％ 0.4796 Experimental Example 14 94.28％ 5.72％ 0.4797 Experimental Example 15 34.98％ 65.02％ 0.5025 Experimental Example 16 87.4％ 12.6％ 0.5024 Experimental Example 17 44.55％ 55.45％ 0.5253 Experimental Example 18 77.8％ 22.2％ 0.5254

[0113] The total amount of litchi fermentation product and isomaltooligosaccharide is 200 mg (in a 10 ml system). As can be seen from Table 4, the OD value corresponding to Experimental Example 12 is the highest, which is the best addition ratio, which is consistent with the results of software analysis. Therefore, preferably, the mass ratio of litchi fermentation product and isomaltooligosaccharide in the composition is (28.08-94.28): (5.72-71.92), at this time, the corresponding OD value of the composition is increased by 5% compared with the single material of litchi fermentation product, more preferably, the mass ratio of litchi fermentation product and isomaltooligosaccharide in the composition is (34.98-87.4): (12.6-65.02), at this time, the corresponding OD value of the composition is increased by 10% compared with the single material of litchi fermentation product, and further optimized, the mass ratio of litchi fermentation product and isomaltooligosaccharide in the composition is (44.55-77.8): (22.2-55.45), at this time, the corresponding OD value of the composition is increased by 15% compared with the single material of litchi fermentation product.

Claims

1. A composition, characterized in that Includes lychee ferment and prebiotics.

2. The composition according to claim 1, wherein the preparation method of the litchi fermented product comprises the following steps: 1) primary fermentation, which comprises adding a first carbon source to litchi juice and litchi pomace, inoculating a first bacterial agent, and sealing and standing for fermentation at 20-45°C for 24-96 hours; 2) squeezing and filtering the product after the primary fermentation to obtain a primary fermentation filtrate; 3) secondary fermentation, which comprises adding a second carbon source to the primary fermentation filtrate, inoculating a second bacterial agent, and standing or aerating and culturing at 15-40°C for 3-15 days to obtain a secondary fermentation liquid; 4) filtering and sterilizing the secondary fermentation liquid to obtain a litchi fermented product; preferably, the first carbon source is selected from one or more of fructose, sucrose, brown sugar, maltose, and arabinose; Preferably, the first bacterial agent is a mixed bacterial agent of yeast and lactic acid bacteria; Preferably, the second carbon source is selected from one or more of lactose, brown sugar, glucose, molasses or honey; Preferably, the second bacterial agent is acetic acid bacteria.

3. The composition according to claim 1 or 2, wherein the prebiotic is selected from one or more of isomaltooligosaccharides, galacto-oligosaccharides and stachyose.

4. The composition according to any one of claims 1 to 3, which is composed of litchi fermented product and prebiotics; preferably, the weight proportion of the litchi fermented product in the composition is 28-95, and the weight proportion of the prebiotics is 5-72; Preferably, the weight percentage of the litchi fermented product in the composition is 28.08-94.28, and the weight percentage of the prebiotics is 5.72-71.92; Preferably, the composition contains 34-88 parts by weight of the litchi fermented product and 12-66 parts by weight of the prebiotics; Preferably, the weight percentage of the litchi fermented product in the composition is 34.98-87.4, and the weight percentage of the prebiotics is 12.6-65.02; Preferably, the composition contains 44-78 parts by weight of the litchi fermented product and 22-56 parts by weight of the prebiotics; Preferably, the weight percentage of the litchi fermented product in the composition is 44.55-77.8, and the weight percentage of the prebiotics is 22.2-55.

45.

5. The composition according to any one of claims 1 to 3, further comprising coagulated Weizmannella powder.

6. The composition according to claim 5, wherein the weight proportion of the litchi fermented product in the composition is 28-95, and the weight proportion of the prebiotics is 5-72; preferably, the weight proportion of the litchi fermented product in the composition is 28.08-94.28, and the weight proportion of the prebiotics is 5.72-71.92; preferably, the weight proportion of the litchi fermented product in the composition is 34-88, and the weight proportion of the prebiotics is 12-66; preferably, the weight proportion of the litchi fermented product in the composition is 34.98-87.4, and the weight proportion of the prebiotics is 12.6-65.02; preferably, the weight proportion of the litchi fermented product in the composition is 44-78, and the weight proportion of the prebiotics is 22-56; preferably, the weight proportion of the litchi fermented product in the composition is 44.55-77.8, and the weight proportion of the prebiotics is 22.2-55.

45.

7. A method for preparing a composition, characterized in that The method comprises the following steps: 1) preparing litchi fermented product; 2) mixing litchi fermented product and prebiotics; optionally, further adding Weizmannella coagulans; preferably, the preparation method of the litchi fermented product comprises the following steps: 1) primary fermentation, which comprises adding a first carbon source to litchi juice and litchi pomace, inoculating a first bacterial agent, and sealing and standing at 20-45° C. for fermentation for 24-96 hours; 2) squeezing and filtering the product after the primary fermentation to obtain a primary fermentation filtrate; 3) secondary fermentation, which comprises adding a second carbon source to the primary fermentation filtrate, inoculating a second bacterial agent, and standing at 15-40° C. or aeration culture for 3-15 days to obtain a secondary fermentation broth; 4) filtering and sterilizing the secondary fermentation broth to obtain litchi fermented product; Preferably, the first carbon source is selected from one or more of fructose, sucrose, brown sugar, maltose and arabinose; Preferably, the first bacterial agent is a mixed bacterial agent of yeast and lactic acid bacteria; Preferably, the second carbon source is selected from one or more of lactose, brown sugar, glucose, molasses or honey; Preferably, the second bacterial agent is acetic acid bacteria.

8. Use of the composition according to any one of claims 1 to 6 in regulating intestinal flora; preferably, the use is for non-therapeutic purposes; preferably, regulating intestinal flora includes promoting the proliferation of Weizmannella coagulans.

9. Use of the composition according to any one of claims 1 to 6 in the preparation of food, health products or medicines with intestinal regulation function; preferably, the content of litchi fermentation product and prebiotics in the food, health products or medicines is 100 mg / 100 g to 1×10 4 mg / 100g; preferably, the food, health product or medicine contains 1×10 9 CFU / 100g to 1×10 12 CFU / 100 g of Weizmannella coagulans.

10. Use of the composition according to any one of claims 1 to 6 for promoting the proliferation of Weizmannella coagulans in vivo or in vitro.

11. A method for promoting the proliferation of Weizmannella coagulans, characterized in that: Add the composition according to any one of claims 1 to 6 into the living environment of coagulant Weizmannella.

12. Functional products, characterized by The composition comprises any one of claims 1 to 6; preferably, the functional product is a food, a health product or a medicine; preferably, the content of the litchi fermentation product and the prebiotics in the functional product is 100 mg / 100 g to 1×10 4 mg / 100g; preferably, the functional product contains 1×10 9 CFU / 100g to 1×10 12 CFU / 100 g of Weizmannella coagulans.

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