CN114403450B - Application of black fungus polysaccharide additive in inhibiting body metabolism odor production - Google Patents

Abstract

The invention belongs to the field of food, medicine or feed, and relates to a new application of a black fungus polysaccharide additive, in particular to an application of the black fungus polysaccharide additive in inhibiting the generation of metabolic odor of an organism. The auricularia auricula polysaccharide additive comprises an auricularia auricula extract with the auricularia auricula polysaccharide content of 5-99% (dry weight ratio), and can also comprise a drying agent. The auricularia auricula polysaccharide additive can be applied to food or health food, and the daily intake of the auricularia auricula polysaccharide additive is 0.05 g to 0.2 g per kilogram of body weight. The auricularia auricula polysaccharide additive can be used in animal feed, and the dosage of the auricularia auricula polysaccharide additive accounts for 5-200 mg of dry weight of each gram of feed. The Auricularia auricula polysaccharide additive can effectively reduce intestinal microorganism metabolism odor such as NH ₃ And/or H ₂ The S can be developed into health products, functional foods, medical foods, medicines or feeds for inhibiting the generation of body metabolic odor.

Description

Application of black fungus polysaccharide additive in inhibiting body metabolism odor production

technical field

The invention belongs to the field of health products, functional food, food, medicine, medical food or feed, and relates to a new application of black fungus polysaccharide additives, which are applied to inhibit the generation of metabolic odor of the body.

Background technique

With the improvement of people's living standards, the intake ratio of protein diet is gradually increasing. According to the Food and Agriculture Organization of the United Nations, the dietary protein levels of Western European populations far exceed the recommended intake. Dietary protein fermentation is considered harmful to the gut, and excessive protein intake can lead to high levels of residual nitrogen in the colon and become a metabolic substrate for colonic microbes, producing gaseous compounds, amines, and indoles, among others. Among them, the accumulation of H ₂ S and NH ₃ in the colon is toxic to intestinal epithelial cells, and the production of amines and phenolic compounds may also affect the health of the host by changing the composition of intestinal microorganisms.

Undigested carbohydrates in the small intestine can serve as energy substrates for bacterial fermentation in the large intestine, reduce protein catabolism, and affect the microbial community, thereby reducing damage to intestinal health. Glycans extracted from chicory, a non-digestible carbohydrate, can exert a prebiotic effect by replacing fermented proteins or improving the structure of intestinal flora to reduce the level of malodors produced by protein fermentation.

However, there is currently a lack of research on the improvement of odor emissions by improperly adjusted diets with prebiotics, and the application of polysaccharides such as inulin as prebiotics is mostly concentrated in the feed addition of animal husbandry. Black fungus polysaccharide is the main carbohydrate component in black fungus, and it is also an important bioactive substance. There is no report on the effect of black fungus polysaccharide on reducing odor emission. The functional properties of prebiotics can be verified by in vitro fermentation experiments. Based on this, the present invention digests proteins and prebiotics in vitro to obtain a simulated large intestine fermentation substrate, and cultures under anaerobic conditions by adding fecal bacteria liquid to confirm that black fungus polysaccharides Prebiotic action that suppresses odor production.

Contents of the invention

The technical problem solved by the invention is to provide a new application of the black fungus polysaccharide additive. It specifically relates to the use of black fungus polysaccharide additives in inhibiting the production of body metabolic odor.

Further, the black fungus polysaccharide additive of the present invention comprises black fungus extract with a black fungus polysaccharide content of 5%-99% (dry weight ratio), and may also include a desiccant; the odor includes NH _and /or or _H2S .

Further, the black fungus polysaccharide additive is made into food or health product, and the daily intake is 0.05g-0.2g per kilogram of body weight.

Further, the black fungus polysaccharide additive is made into medicines, and the preparations of the medicines include tablets, powders, granules, teas, capsules, soft capsules, oral liquids, pills, ointments, and liquors.

Further, the black fungus polysaccharide additive is made into animal feed, and its dosage accounts for 5-200 mg per gram of feed dry weight.

Further, the black fungus polysaccharide additive can effectively reduce the production of intestinal microbial metabolic odor such as NH ₃ and/or H ₂ S.

In order to realize the above-mentioned purposes, the present invention specifically uses soybean protein as a model fermentation substrate and inulin as a positive control fermentation substrate through in vitro digestion-in vitro fermentation experiments to study the odor produced by black fungus polysaccharides on fecal bacteria fermentation model substrates inhibitory effect. It was found that the fecal fungus fully fermented the black fungus polysaccharide as a substrate, effectively reducing the content of odor such as NH ₃ and H ₂ S, thus obtaining the use of black fungus polysaccharide additives that can inhibit the body's metabolic odor.

Aiming at currently no dietary supplements that are easy to obtain and suitable for suppressing odor generation, the present invention provides a black fungus polysaccharide additive for food, medicine, health food or New use in feed, compared with the traditional dietary supplement inulin, the black fungus polysaccharide used in the present invention has a better effect in reducing intestinal odor, and has better application in regulating intestinal fermentation as a dietary supplement potential.

Detailed ways

In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following examples further illustrate the content of the present invention, but should not be construed as limiting the present invention.

(1) Materials and Design

Buy commercially available soybean protein, inulin and black fungus polysaccharide. The polysaccharide content was determined by the phenol sulfuric acid method, and the purity of inulin and black fungus polysaccharide were 90% and 50%, respectively.

The fermentation system of this embodiment is 25mL and consists of feces liquid and substrate. According to the different fermentation substrates, it was divided into control group, model group, positive control group and black fungus low, middle and high dose group, with 12 repetitions in each group.

The specific amount of substrate added is shown in Table 1. The sugar content of the SI and SAAPM groups was 1%, and the sugar content of SAAPL and SAAPH were 0.5% and 1.5%, respectively.

Table 1 Substrate addition amount of each group

(2) The establishment of the fermentation system of the present invention

In this example, the in vitro digestion of the fermentation substrate is first carried out, that is, gastric juice-small intestinal juice digestion.

Prepare simulated artificial gastric juice and simulated artificial small intestine juice according to the Chinese Pharmacopoeia, weigh the substrates of each group as shown in Table 1, dissolve them in artificial gastric juice (1:8, w/v), and digest in a constant temperature shaker at 110rpm 37°C 2h, after cooling to room temperature, the pH was adjusted to neutral with 1M NaOH. Then add an equal amount of artificial small intestine juice, digest at 110 rpm at 37°C for 2 hours, freeze-dry and store at 4°C for later use.

Next, prepare the fermentation medium.

Each 1000mL basal salt medium for in vitro fermentation includes: 5.0g NaHCO ₃ , 0.9g NaCl, 0.9g (NH ₄ ) ₂ SO ₄ , 0.45g KH ₂ PO ₄ , 0.45g K ₂ HPO ₄ ·3H ₂ O, 0.03g CaCl ₂ 2H ₂ O, 0.02g MgCl ₂ , 0.01g MnSO ₄ 4H ₂ O, 0.01g CoCl ₂ 6H ₂ O, 0.01g FeSO ₄ 7H ₂ O and 1.0g cysteine, prepared at 121°C Autoclave for 20min.

Finally, prepare the feces liquid and complete the fermentation.

This example recruited three healthy subjects who had not received antibiotic or probiotic treatment for at least three months prior to sample collection. After collecting their fresh feces, they were immediately suspended in sterile anaerobic (5% H ₂ , 5% CO ₂ , 90% N ₂ ) basal salt medium at a concentration of 0.5% (w/v). Shake and mix the feces suspension and filter it with four layers of gauze to remove coarse particulate matter to obtain the fecal bacteria liquid. The fecal bacteria liquid and the digested substrate were anaerobically distributed into 50mL serum bottles and fermented at 37°C for 24h.

(3) Evaluation method and results of odor gas

Odor gas evaluation methods include the concentration and production of NH ₃ and H ₂ S in the gas, where the production is calculated by the gas production and concentration. The evaluation method of gas production is the excess gas volume generated in the fermentation bottle.

After the fermentation of each group of fermentation systems is completed, the sterile syringe is quickly inserted into the culture bottle, and the volume of the piston pushed by the gas is the gas production of the system. The gas collected in the sterile syringe is introduced into the NH ₃ or H ₂ S absorption solution through the three-way valve to measure the gas content. Nessler's reagent colorimetric method was used for NH ₃ determination, that is, 100 μL of NH ₃ absorption solution and 50 μL of Nessler's reagent were added to a 96-well plate, and the absorbance was measured at a wavelength of 420 nm. H ₂ S is determined by methylene blue spectrophotometry according to the national standard GB/T 11742-89. The specific results are shown in Table 2:

Table 2 Odor gas levels in fermentation systems of each group

Note: ^# means p<0.05 compared with the control group, ^* means p<0.05 compared with the model group.

It can be seen from the table that the addition of black fungus polysaccharide significantly reduces the production and concentration of NH ₃ and H ₂ S in the gas produced by the fermentation system. Compared with inulin, which has been reported to inhibit the generation of odor, black fungus polysaccharide reduces the The ability to reduce odor is better; under the three dosage levels, black fungus polysaccharide has excellent ability to reduce odor, and the ability to reduce odor of black fungus polysaccharide is better at low dosage.

(4) Evaluation methods and results of total sugar, reducing sugar and polysaccharide content before and after fermentation

The content of total sugar, reducing sugar and polysaccharide was measured in the supernatant of the fermentation broth after 0h and 24h of fermentation, and the standard curve was drawn with glucose as the standard.

The detection of total sugar and reducing sugar content was carried out by DNS colorimetric method. The determination of the total sugar content is as follows: add 1:0.75 (v/v) 6M HCl solution to a certain amount of fermentation broth supernatant, cool to room temperature after boiling water bath for 20 minutes, and then add 1:1 (v/v) 6M HCl solution. NaOH solution and 1:1.5 (v/v) DNS reagent were heated in a boiling water bath for 5 minutes, the absorbance value was measured at a wavelength of 540nm, and the total sugar content was obtained according to the standard curve. The measuring procedure of the reducing sugar content is: mixing the supernatant of the fermentation broth with the DNS reagent at a ratio of 1:1.5 (v/v), heating in a boiling water bath for 5 minutes, measuring the absorbance value at a wavelength of 540nm, and obtaining the reducing sugar content according to the standard curve.

The polysaccharide content was extracted and determined by water extraction and alcohol precipitation-phenol sulfuric acid method. That is, add 3 times the volume of absolute ethanol to the supernatant of the fermentation broth, let it stand overnight at 4°C, centrifuge at 4,200 rpm for 15 minutes, and wash the precipitate with absolute ethanol 3 times to obtain crude polysaccharide. Suspend the crude polysaccharide with deionized water, add 1:0.5 (v/v) 5% phenol solution and 1:2.5 (v/v) concentrated sulfuric acid, measure the absorbance value at 490nm after standing at room temperature for 20 minutes, according to the standard curve Much sugar content. The test results are shown in Table 3:

Table 3 Degradation levels of total sugar, reducing sugar and polysaccharide in each fermentation system before and after fermentation

Note: ^* indicates p<0.05 compared with the positive control group.

It can be seen from the table that compared with the positive control group, the consumption of total sugar, reducing sugar and polysaccharide in the black fungus middle dose group increased significantly; the consumption of total sugar and reducing sugar in the low dose black fungus group increased significantly, while the The significant reduction of the content was due to the consumption of polysaccharides; the consumption of total sugars and polysaccharides in the black fungus high-dose group increased significantly. It is known that the fermentation of protein/amino acid by the flora will produce odor, and the addition of inulin will be used by the flora to replace the fermented protein/amino acid to reduce the generation of odor. In the present invention, the polysaccharide or reducing sugar form of black fungus polysaccharide is consumed in a large amount during the 24h of fermentation and is significantly higher than that of inulin, indicating that the polysaccharide or reducing sugar form of black fungus polysaccharide is more easily utilized by the flora so as to play a better role in reducing odor produced effect.

The results show that the black fungus polysaccharide adopted in the present invention has a better effect of reducing odor and odor substances as a fermentation substrate, and has better application potential in regulating intestinal fermentation as a polysaccharide dietary supplement, and can be developed as Health care products, functional foods, foods, medicines, medical foods or feeds that inhibit the body's metabolic odor are effective and easy to obtain.

Claims (2)

1. The application of the auricularia auricula polysaccharide additive in inhibiting the generation of metabolic odor of an organism is characterized in that: the black fungus polysaccharide additive comprises a black fungus extract with the black fungus polysaccharide content of 5-99% (dry weight ratio), and can also comprise a drying agent; the auricularia auricula polysaccharide additive is prepared into food or health care products, and the daily intake of 0.05 g-0.2 g is applied to each kilogram of body weight; the auricularia auricula polysaccharide additive is prepared into animal feed, and the dosage of the auricularia auricula polysaccharide additive accounts for 5-200 mg of the dry weight of each gram of the feed; the odor is NH ₃ And/or H ₂ S。

2. Use according to claim 1, characterized in that: the auricularia auricula polysaccharide additive is prepared into a medicine, and the preparation of the medicine comprises tablets, powder, granules, tea, capsules, oral liquid, pills, paste and wine.