阅读说明：本技术 岩藻寡糖在制备肠道益生元中的应用 (Application of fucooligosaccharide in preparation of intestinal prebiotics ) 是由 邹祥 王振宇 马巍 李姗姗 徐兴然 于 2020-07-15 设计创作，主要内容包括：本发明公开了岩藻寡糖在制备肠道益生元中的应用,利用岩藻寡糖具有促进碳水化合物代谢、糖的生物合成与代谢、降低膜运输功能、增强消化系统功能或降低疾病感染的功能,促进肠道内短链脂肪酸产生,改善肠道微生物菌群水平,用于制备肠道益生元,用于人体或畜禽动物,具有广泛的应用前景。(The invention discloses application of fucoidan oligosaccharide in preparation of intestinal prebiotics, wherein the fucoidan oligosaccharide has the functions of promoting carbohydrate metabolism, biosynthesis and metabolism of sugar, reducing membrane transport function, enhancing digestive system function or reducing disease infection, is used for promoting the generation of short-chain fatty acid in intestinal tracts and improving the level of intestinal microbial flora, is used for preparing the intestinal prebiotics, is used for human bodies or livestock and poultry animals, and has wide application prospect.)

1. Application of fucooligosaccharide in preparing intestinal prebiotics is provided.

2. Use according to claim 1, characterized in that: the fucooligosaccharide is applied to the preparation of prebiotics for promoting carbohydrate metabolism, biosynthesis and metabolism of sugar, reducing membrane transport function, enhancing digestive system function or reducing disease infection.

3. Use according to claim 1 or 2, characterized in that: the fucooligosaccharide is prepared by hydrolyzing exopolysaccharide produced by fermentation of Kosakonia sp.CCTCCMM2018092.

4. Use according to claim 3, characterized in that: and the trifluoroacetic acid hydrolysis comprises the steps of adding extracellular polysaccharide with the mass fraction of 5% into trifluoroacetic acid to enable the final concentration of the trifluoroacetic acid to be 0.1M, hydrolyzing for at least 1 hour at 100 ℃, and then removing the trifluoroacetic acid by using a 200Da nanofiltration membrane to obtain the fucooligosaccharide.

5. Application of fucooligosaccharide in promoting production of intestinal tract short chain fatty acid is provided.

6. Use according to claim 3, characterized in that: the short-chain fatty acid is acetic acid or propionic acid.

7. Use of fucoidan for improving intestinal microbial flora level is provided.

8. Use according to claim 7, characterized in that: the improving intestinal microbial flora level comprises increasing bacteroidetes level and reducing firmicutes and proteobacteria level.

9. Use according to claim 7, characterized in that: the improving intestinal microflora level is to increase the levels of Parabacteroides, Bacteroides norcolmanii and Prevotella, and reduce the levels of Lactobacillus and Bacteroides.

Technical Field

The invention relates to the technical field of biology, in particular to application of fucooligosaccharide in preparation of intestinal prebiotics.

Background

Fucose is a rare 6-deoxyhexose in the L-configuration, commonly found in microbial exopolysaccharides, brown algae, and mammals. Fucose modifications have been identified as being associated with a number of biological functions, including immunomodulation and cancer. Sulfate-containing fucoidans rich in brown algae have anticoagulant, antithrombotic, immunomodulatory, anticancer, and antiproliferative activities. However, the yield of fucoidan in plants or algae is low, and its composition varies with climate and season. The fucose-rich exopolysaccharides produced by the microorganisms are considered to be a better alternative in view of the advantages of microorganisms having higher growth rates and easier control of production conditions. In recent decades, other fucose-containing exopolysaccharides with the highest yield have been reported to be produced by Enterobacter (Enterobacter A47) at a yield of 13.23 g/L. The various physicochemical properties of the fucose-rich exopolysaccharide, such as rheological properties, adhesive properties, emulsifying capacity, and the use of biodegradable films, show important commercial value.

In previous studies, we isolated a Kosakonia strain and identified it as Kosakonia sp.cctcc M2018092, elucidating the genome-wide sequence and genetic characteristics of this strain (Complete genome sequence of Kosakonia sp.strain CCTCC 2018092, a fusase-Rich exopolysporac producer. singfengfengfeng Niu, 2019). The function of the capsular exopolysaccharide hydrolyzed fucooligosaccharides produced by Kosakonia sp was not studied.

Disclosure of Invention

In view of the above, an object of the present invention is to provide an application of fucooligosaccharide in preparation of intestinal prebiotics; the invention also aims to provide the application of the fucooligosaccharide in promoting the generation of short-chain fatty acids in intestinal tracts; the invention also aims to provide the application of the fucooligosaccharide in improving the level of the intestinal microbial flora.

In order to achieve the purpose, the invention provides the following technical scheme:

1. application of fucooligosaccharide in preparing intestinal prebiotics is provided.

The fucoidin has the functions of promoting carbohydrate metabolism, sugar biosynthesis and metabolism, reducing membrane transport function, enhancing the function of a digestive system or reducing disease infection; the application of the prebiotics in preparing the prebiotics for promoting carbohydrate metabolism, sugar biosynthesis and metabolism, reducing membrane transport function, enhancing digestive system function or reducing disease infection.

In the invention, the fucooligosaccharide is prepared by hydrolyzing extracellular polysaccharide generated by fermentation of Kosakonia sp.CCTCC M2018092 by trifluoroacetic acid or enzymolysis and the like.

Preferably, the trifluoroacetic acid is hydrolyzed by adding 5% of exopolysaccharide by mass fraction into trifluoroacetic acid to make the final concentration of the trifluoroacetic acid 0.1M, hydrolyzing at 100 ℃ for at least 1 hour, and then removing the trifluoroacetic acid by using a 200Da nanofiltration membrane to obtain the fucooligosaccharide.

2. Application of fucooligosaccharide in promoting production of intestinal tract short chain fatty acid is provided.

Preferably, the short chain fatty acid is acetic acid or propionic acid.

3. Use of fucoidan for improving intestinal microbial flora level is provided.

Preferably, the improving gut microflora levels are increasing bacteroidetes levels, decreasing firmicutes and proteobacteria levels.

Preferably, the improving the level of the intestinal microflora is increasing the level of parabacteroides, norcolobacterium and prevotella, and decreasing the level of lactobacillus and bacteroides.

The invention has the beneficial effects that: the invention provides a fucoidin oligosaccharide with a probiotic function, and discloses the fucoidin oligosaccharide with the functions of promoting carbohydrate metabolism, biosynthesis and metabolism of sugar, reducing a membrane transportation function, enhancing a digestive system function or reducing disease infection for the first time, and the fucoidin oligosaccharide can promote the generation of short-chain fatty acid in an intestinal tract and improve the level of intestinal microbial flora, so that the fucoidin oligosaccharide can be prepared and can be applied to the fields of human health, livestock and poultry animal culture and the like.

Drawings

In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:

FIG. 1 is a GC-MS total ion chromatogram of a monosaccharide standard.

FIG. 2 is a GC-MS total ion flow chromatogram of the oligosaccharide complete acid hydrolysis product.

FIG. 3 is a graph of the effect of fucooligosaccharides on pH of the fermentation broth.

Figure 4 is the effect of fucooligosaccharides on short chain fatty acids.

FIG. 5 shows the levels of the phyla of test tube fermentation intestinal flora (SGCON is a control group, SGFOP is a group added with fucooligosaccharides).

FIG. 6 shows the levels of test tube fermentation gut flora (SGCON for control group, SGFOP for fucoidan-added group).

FIG. 7 is the preparation of mucosal pellets.

FIG. 8 shows the results of mucosal globule simulation (A: lumen; B: mucosa; S, H, J resolution represents ascending, transverse, descending colon, the latter numbers represent days of fermentation, w represents stop of sample addition).

FIG. 9 shows the results of the simulated intestinal tract (A: level of phylum of intestine; B: level of genus of intestine; C: level of phylum of mucosa; D: level of genus of mucosa; S: ascending colon; H: transverse colon; J: descending colon; 0,3, w respectively represent the control group, and fucoidan addition was stopped on day 3 of fucoidan addition).

Fig. 10 is the effect of fucoidan on the metabolic activity of the intestinal flora (left histogram is the proportion of the metabolic pathway in all secondary metabolic pathways, right is the significance value of the two groups comparison, blue represents the strong metabolic pathway of the control group, orange represents the strong metabolic pathway of the 0.1% fucoidan experimental group, and the metabolic pathways are arranged from small to large according to the significance value).

FIG. 11 is the results of evaluation of safety of fucoidan oligosaccharide (A: the effect of fucoidan oligosaccharide on weight; B: the effect of fucoidan oligosaccharide on brain and heart; C: the effect of fucoidan oligosaccharide on colon length; D: the effect of fucoidan oligosaccharide on liver weight; E: the effect of fucoidan oligosaccharide on liver function and kidney function).

FIG. 12 shows the results of measurement of short-chain fatty acids in mouse feces (A: HPLC; B: GC; C: GC-MS; D: statistical results of short-chain fatty acid content).

Detailed Description

The present invention is further described with reference to the following drawings and specific examples so that those skilled in the art can better understand the present invention and can practice the present invention, but the examples are not intended to limit the present invention.