阅读说明：本技术 大豆不溶性膳食纤维及其应用 (Soybean insoluble dietary fiber and application thereof ) 是由 于寒松 高俊鹏 王赛男 房佳琦 卢佳宏 代伟长 刘俊梅 朴春红 王玉华 其他发明人 于 2020-04-03 设计创作，主要内容包括：本发明公开了大豆不溶性膳食纤维,它是由下述方法制备的,它包括：取大豆经过脱皮、脱脂,提取分离蛋白,得湿豆渣；挤压脱水；淀粉水解；蛋白酶水解,水洗,过滤,冻干,得大豆不溶性膳食纤维。通过动物实验,得到的结果说明：1)长期摄入HCIDF能有效预防高脂饮食引起的小鼠体重增加；2)摄入大豆不溶性膳食纤维,可以调节血清TC、TG、LDL和HDL水平,对高脂饮食引起的血清脂质代谢紊乱具有积极预防作用；3)摄入大豆不溶性膳食纤维可减少腹部和肾周脂肪重量,减少小鼠肝损伤；4)大豆不溶性膳食纤维(HCIDF)可以通过调控与脂肪代谢相关的基因表达水平来预防小鼠由高脂饮食引起的肥胖。(The invention discloses soybean insoluble dietary fiber, which is prepared by the following method, wherein soybean is subjected to peeling and degreasing, protein extraction and separation to obtain wet soybean dregs, extrusion dehydration, starch hydrolysis, protease hydrolysis, water washing, filtration and freeze drying to obtain the soybean insoluble dietary fiber, and animal experiments prove that 1) the mouse weight increase caused by high-fat diet can be effectively prevented by taking the soybean insoluble dietary fiber for a long time, 2) the serum TC, TG, L D L and HD L levels can be regulated by taking the soybean insoluble dietary fiber, and the soybean insoluble dietary fiber has a positive prevention effect on the serum lipid metabolism disorder caused by the high-fat diet, 3) the mouse obesity caused by the high-fat diet can be prevented by taking the soybean insoluble dietary fiber to reduce the fat weight of the abdomen and the kidney periphery and reduce the liver injury of the mouse, and 4) the soybean insoluble dietary fiber (HCIDF) can be used for preventing the mouse obesity caused by the high-fat diet by regulating the gene expression level related to the fat metabolism.)

1. A soybean insoluble dietary fiber prepared by a process comprising:

1) peeling and degreasing soybeans, and extracting protein isolate to obtain wet bean dregs;

2) squeezing and dehydrating to obtain crude dietary fiber;

3) hydrolyzing starch;

4) hydrolyzing by protease;

5) washing with water, precipitating with ethanol, filtering, and lyophilizing.

2. The application of claim 1, wherein the starch hydrolysis in step 3) comprises adding water to the crude dietary fiber obtained in step 2), adding thermally stable α -amylase solution with a concentration of 1.25g/5ml, stirring, shaking at 95-100 deg.C for 30-40 min, and cooling.

3. The application of claim 2, wherein the protease hydrolysis in step 4) comprises adjusting pH to 6.8-7.2, adding 0.125g/25ml protease solution in water bath at 55-65 ℃, shaking for 25-35 min, adding 3mol/m L acetic acid solution while stirring, adjusting pH to 4-5, adding 0.25g/12.5ml amyloglucosidase enzymatic hydrolysate, and further shaking for 25-35 min.

4. Use according to claim 3, characterized in that: the step 5) of washing is to add distilled water of 65-75 ℃ for washing; the alcohol precipitation is ethanol precipitation.

5. The use of soybean insoluble dietary fiber for intervening in fatty acid synthesis pathway.

6. Application of soybean insoluble dietary fiber in preparing fatty acid reducing medicine is provided.

Technical Field

The invention belongs to the technical field of health-care food, and particularly relates to soybean insoluble dietary fiber and application thereof.

Background

Obesity is a chronic metabolic disease caused by various factors, such as High Fat Diet (HFD), and is associated with many metabolic disorders, including dyslipidemia, type 2 diabetes, nonalcoholic fatty liver and cardiovascular diseases, with the development of social science and economy, the acceleration of human rhythms of life and the modification of dietary structure cause excessive intake of high fat foods, leading to an increasing incidence and mortality of obesity in many countries around the world, and prevention and treatment of obesity has become the first public safety and health problem in the 21 st century, it is reported that intake of dietary fibers from whole grains, vegetables and fruits can reduce the risk of obesity and metabolic diseases, regulate parameters related to controlling metabolic syndrome, thereby improving glucose homeostasis, lipid metabolism, etc. obesity has been one of health problems of worldwide concern, long-term intake of HFD can lead to an increase in body weight, liver lipid accumulation (mainly triglyceride (mainly) is associated with fatty acid synthesis and fatty acid oxidation levels, liver lipid accumulation (mainly triglyceride (diacylglycerol acyltransferase) is associated with lipid synthesis and fatty acid oxidation levels, and lipid metabolism activation of diacylglycerol acyltransferase 1 (diacylglycerol acyltransferase) is a major gene that activates diacylglycerol acyltransferase 1-folate synthase-5, and diacylglycerol synthase-lyase (stearoyl-transferase) in tissues, and lipid metabolism-acyltransferase, thus, which can be a gene activating genes expressing diacylglycerol-5, and activating genes of diacylglycerol-5, such as stearoyl-5, and cholesterol-activating genes in tissues, and lipid metabolism.

Okara is a by-product produced during the processing of soybeans. The production of bean dregs has been increasing year by year, and most of them are from asian countries such as china, japan and korea. China is the major producing and largest consuming country of soybeans producing about 7000 million tons of bean dregs per year. However, the utilization of the okara has not been completely maximized, and a large amount of okara is discarded due to high water content, perishability, bad flavor and texture, thereby causing serious resource waste and a great environmental problem.

The bean dregs contain rich nutrient substances, especially Dietary Fiber (DF) which is known as the seventh nutrient substance, and can account for 50-60% of the dry matter of the bean dregs, wherein the Insoluble Dietary Fiber (IDF) accounts for more than 90% of the total amount of the dietary fiber. The IDF comprises cellulose, lignin, most of hemicellulose and the like, and has good health-care functions, such as promotion of intestinal peristalsis, reduction of blood cholesterol, regulation of blood sugar and the like, due to special structural properties of the IDF, such as looseness, porosity, large amount of-OH-COOH and other hydrophilic groups. DF plays a very important role as a key nutrient in the dietary framework of human health, in many physiological processes and in the prevention of different metabolic diseases. The reports of some current researches on dietary fibers show that the dietary fibers are fully supplemented, so that the balance of intestinal flora can be maintained, the blood fat and cholesterol levels can be reduced, the blood sugar in a human body can be maintained to be stable, and the constipation and the colon cancer can be prevented from happening. Since the dietary level of human beings is continuously becoming fine, resulting in a severe dietary intake deficiency of dietary fiber, people are gradually aware of the dietary importance of dietary fiber, and thus the development of dietary fiber has become a hot problem of current research.

The extraction method of dietary fiber mainly comprises a physical method, a chemical method, an enzymatic method, a composite method and the like. The research reports about the dietary fiber extraction method at home and abroad, the physical method: shaohuo and the like are subjected to ultrahigh pressure treatment and high-temperature cooking treatment on bean dregs, so that the extraction rate of the soluble dietary fibers is improved. The chemical method comprises the following steps: extracting insoluble dietary fiber from pericarpium Granati by alkaline method with extraction rate of 30.42%; zhang Jun extracts dietary fiber from pericarpium Citri Grandis with extraction rate of 48.93% by alkaline process; the extraction rate of insoluble dietary fiber from the dragon fruit peel by using an alkaline method, such as the extraction of the dried Liuqian and the like, is 30.29 percent; the extraction rate of insoluble dietary fiber in bean dregs extracted by alkali method of Ullah, etc. is 33.3%. An enzyme method comprises the following steps: the extraction rate of insoluble dietary fiber from Zhangzhou cherry tomato by an enzyme method is 59.4 percent; the extraction rate of insoluble dietary fiber from shaddock peel by enzyme method in Vandaparin spring is 57.5%; the extraction rate of dietary fiber extracted from the osmunda japonica by the enzyme method of the plum, the phoenix and the like is 74.28%; insoluble dietary fibers are extracted by Zhang Jintang, etc., Zhao, Daou, etc. through an enzyme method, and the quality of the insoluble dietary fibers is higher. The compounding method comprises the following steps: the extraction rate of insoluble dietary fiber extracted from ginger residue by enzyme method and chemical method is 66.21%; the extraction rate of insoluble dietary fiber from pepper seeds by using an ultrasonic-assisted enzyme method is 68.15%. In conclusion, the physical method is not suitable for extracting insoluble dietary fiber, and the chemical method has great influence on the quality of the dietary fiber and lower relative extraction rate.

Disclosure of Invention

The invention provides soybean insoluble dietary fiber and application of the soybean insoluble dietary fiber in intervening fatty acid synthesis pathway in order to solve the problem of soybean byproduct waste.

A soybean insoluble dietary fiber prepared by a process comprising:

1) peeling and degreasing soybeans, and extracting protein isolate to obtain wet bean dregs;

2) squeezing and dehydrating to obtain crude dietary fiber;

3) hydrolyzing starch;

4) hydrolyzing by protease;

5) washing with water, precipitating with ethanol, filtering, and lyophilizing;

the starch hydrolysis in the step 3) comprises the steps of adding water into the crude dietary fiber obtained in the step 2), adding thermally stable α -amylase liquid with the concentration of 1.25g/5ml, stirring, shaking for 30-40 min at the temperature of 95-100 ℃, and cooling;

adjusting the pH value to 6.8-7.2, performing water bath at 55-65 ℃, adding a protease dissolving solution with the concentration of 0.125g/25ml, oscillating for 25-35 min, adding a 3mol/m L acetic acid solution while stirring, adjusting the pH value to 4-5, adding an amyloglucosidase enzymatic hydrolysate with the concentration of 0.25g/12.5ml, and continuing oscillating for 25-35 min;

the step 5) of washing is to add distilled water of 65-75 ℃ for washing; the alcohol precipitation is ethanol precipitation.

Use of soybean insoluble dietary fiber for interfering in fatty acid synthesis pathway.

The invention provides soybean insoluble dietary fiber, which is prepared by a method comprising the steps of taking soybeans, peeling, degreasing, extracting and separating protein to obtain wet soybean dregs, squeezing and dehydrating, hydrolyzing starch, hydrolyzing protease, washing with water, filtering, freeze-drying to obtain the soybean insoluble dietary fiber, intervening High Fat Diet (HFD) induced C57B L/6J mice by the soybean Insoluble Dietary Fiber (IDFS), continuously measuring 20w, and researching the influence of the IDFS on the metabolism of serum and liver lipid of the miceP<0.05). In addition, IDFS can down-regulate Fatty Acid Synthase (FAS) and diacylglycerol acyltransferase 1 (DGAT 1), diacylglycerol acyltransferase 2 (d) in liver after dryingExpression of iaCylglycerol acyl transferase-2, DGAT 2), stearoyl-CoA desaturase 1 (stearoyl-coenzyme A dehydrogenase-1, SCD 1) ((see examples for FIGS.)P<0.05), and simultaneously up-regulating the expression of peroxisome proliferator-activated receptor α (peroxisome proliferators-activator- α α), carnitine palmitoyl transferase 1A (CPT 1A) and uncoupling protein 2 (uncoupling protein, UCP 2) of thermogenesis-related genes (fatty acid β)P<The results show that 1) the long-term intake of HCIDF can effectively prevent the weight increase of mice caused by high-fat diet without having significant influence on the appetite of the body, 2) the intake of soybean insoluble dietary fiber can adjust the TC, TG, L D L and HD L levels of serum and has positive prevention effect on the serum lipid metabolism disorder caused by high-fat diet, 3) the intake of soybean insoluble dietary fiber can reduce the fat weight of abdomen and perirenal, 4) the intake of soybean insoluble dietary fiber can reduce the liver injury of the mice, and 5) the soybean insoluble dietary fiber (HCIDF) can prevent the obesity of the mice caused by the high-fat diet by regulating the gene expression level related to the fat metabolism.

Drawings

FIG. 1 effect of different enzyme addition amounts on the degree of proteolysis;

FIG. 2 the effect of different enzymatic temperatures on the degree of proteolysis;

FIG. 3 the effect of different enzymatic pH values on the degree of proteolysis;

FIG. 4 effect of different enzymatic hydrolysis times on the degree of proteolysis;

FIG. 5 Effect of HCIDF on HFD-induced mice, body weight, food intake, calorie intake; (a) body weight, (b) food intake, (c) calorie intake;

FIG. 6 results of HCIDF effect on serum levels of HFD-induced mice; (a) total cholesterol, (b) triglycerides; (c) high density lipoprotein; (d) low density lipoprotein;

FIG. 7 shows the effect of HCIDF on the liver index of HFD-induced mice; (a) total cholesterol, (b) triglycerides;

FIG. 8 effect of HCIDF on HFD-induced liver coefficients in mice;

figure 9 effect of HCIDF on mouse liver pathology morphology (× 200);

FIG. 10 HCIDF effect on HFD-induced mouse liver; (a) a triglyceride synthesis-related gene; (b) lipid metabolism-related gene expression.

Detailed Description