阅读说明：本技术 一种具有免疫活性的白及多糖及其制备方法与应用 (Bletilla striata polysaccharide with immunocompetence and preparation method and application thereof ) 是由 周美 王道平 潘卫东 万科 王瑜 张妮 李金玉 陈长芬 于 2019-11-15 设计创作，主要内容包括：本发明公开了一种具有免疫活性的白及多糖及其制备方法与应用。采用果胶酶酶解法提取制备具有免疫活性的白及白糖,提取工艺条件为：酶添加量0.5-2％,酶解温度：35-60℃,酶解时间：50-100min,pH值为：4-7。在该酶解条件下白及多糖得率为58.32％；通过本发明制备的白及多糖对DPPH自由基、OH自由基和ABTS+自由基的清除率分别可达到43.6％、74.8％、51.7％,对Raw264.7细胞生长有一定的促进作用,具有免疫调节活性,其浓度为400μg/mL时免疫诱导活性效果显著。本发明制备的白及多糖可应用到免疫增强药物或增强免疫力的保健食品的开发中,为白及多糖在食品和医药领域的开发利用及工业化生产提供理论基础。(The invention discloses a bletilla striata polysaccharide with immune activity and a preparation method and application thereof. Extracting and preparing bletilla white sugar with immunocompetence by adopting a pectinase enzymolysis method, wherein the extraction process conditions are as follows: the addition amount of enzyme is 0.5-2%, and the enzymolysis temperature is as follows: 35-60 ℃, enzymolysis time: 50-100min, the pH value is: 4-7. Under the enzymolysis condition, the yield of the bletilla striata polysaccharide is 58.32 percent; the bletilla polysaccharide prepared by the method has the clearance rates of DPPH free radicals, OH free radicals and ABTS + free radicals reaching 43.6%, 74.8% and 51.7% respectively, has a certain promotion effect on the growth of Raw264.7 cells, has immunoregulation activity, and has an obvious immune induction activity effect when the concentration of the bletilla polysaccharide is 400 mu g/mL. The bletilla polysaccharide prepared by the invention can be applied to the development of immunopotentiating drugs or health-care foods for enhancing immunity, and provides a theoretical basis for the development and utilization and industrial production of the bletilla polysaccharide in the fields of food and medicine.)

1. The bletilla striata polysaccharide with immune activity is characterized in that the clearance rates of the bletilla striata polysaccharide to DPPH free radicals, OH free radicals and ABTS + free radicals can respectively reach 43.6%, 74.8% and 51.7%; has promoting effect and immunoregulation activity on cell Raw264.7 growth.

2. The immunologically active bletilla polysaccharide of claim 1, wherein the immunologically active bletilla polysaccharide is prepared by an enzymatic hydrolysis process comprising the steps of: the common bletilla pseudobulb is crushed, subjected to constant-temperature enzymolysis and inactivated, wherein the conditions of the constant-temperature enzymolysis are as follows: the addition amount of enzyme is 0.5-2%, and the enzymolysis temperature is as follows: 35-60 ℃, enzymolysis time: 50-100min, the pH value is: 4-7.

3. The immunologically active bletilla polysaccharide of claim 1, wherein the immunologically active bletilla polysaccharide is prepared by an enzymatic hydrolysis process comprising the steps of: the common bletilla pseudobulb is crushed, enzymolyzed at constant temperature, and subjected to enzyme deactivation and centrifugation, wherein the conditions of the enzymolysis at constant temperature are as follows: enzyme addition amount of 1.4%, enzymolysis temperature: enzymolysis time at 55 ℃: 87min, and the pH value is: 5.43.

4. the immunocompetent rhizoma bletillae polysaccharide as claimed in claim 2 or 3, wherein the enzyme preparation adopted by the enzymolysis method is pectinase, and the enzyme activity is 2 ten thousand u/g.

5. The immunologically active bletilla striata polysaccharide as claimed in claim 2 or 3, wherein the step of enzyme deactivation centrifugation is performed by incubating bletilla striata enzymatic hydrolysate in a boiling water bath for 3-5 minutes; centrifuging at the rotation speed of 5000-.

6. The bletilla striata polysaccharide with immune activity according to any one of claims 1 to 5, wherein the bletilla striata polysaccharide is applied to the development of an immune enhancing medicament or an immune enhancing health food.

Technical Field

The invention relates to the field of polysaccharide extraction, and particularly relates to a bletilla polysaccharide with immune activity, and a preparation method and application thereof.

Background

Modern pharmacological research shows that free radicals are the cause of various diseases and aging, such as cancer, cardiovascular diseases and the like, and antioxidants can effectively remove active oxygen free radicals, slow down human aging and prevent diseases; similarly, low immunity can increase the invasion of bacteria and viruses to human bodies, threaten human health, and the immunomodulator can improve the immune function of the organisms. Therefore, the search for the development of safe and non-toxic natural antioxidants and immunomodulators becomes the current research focus.

Bletilla striata is a dried tuber of Bletilla striata (Thunb.) Reiehb.f. of Orchidaceae, and is a perennial herb mainly produced in Guizhou, Sichuan, Hunan, Anhui, etc. Rhizoma Bletillae mainly contains polysaccharides, phenanthrenes, bibenzenes, terpenes, etc. It is reported that: the polysaccharide has wide biological activity, can eliminate free radicals which bring great harm to human bodies as an antioxidant, maintains health and prolongs life; the immune regulator can activate various immune cells, promote the generation of cytokines, increase the generation amount of antibodies, and improve the phagocytic capacity of macrophages and leukocytes; improve the lymphocyte transformation rate and the NK cell activity, and achieve the effect of enhancing the immunologic function. The bletilla polysaccharide is used as the main active ingredient of bletilla and becomes a natural active substance with great development potential due to the advantages of safety, high efficiency and small toxic and side effects. As an efficient extraction method, the enzymatic hydrolysis method can dissolve out more effective substances when degrading the product, and is widely used in the food and drug industries. Rhizoma Bletillae is rich in pectin, and its cell wall is degraded by pectinase to quickly dissolve intracellular polysaccharide component, thereby increasing polysaccharide extraction rate.

Disclosure of Invention

The invention aims to provide a bletilla striata polysaccharide with immunocompetence and a preparation method and application thereof, wherein the cell wall of the bletilla striata polysaccharide is degraded by pectinase, so that intracellular polysaccharide components can be effectively and rapidly dissolved out, and the extraction rate of the bletilla striata polysaccharide is increased. The prepared bletilla striata polysaccharide can be applied to drugs or health-care food for enhancing immunity, and the technical scheme adopted for achieving the purpose is as follows: a bletilla striata polysaccharide with immunocompetence, wherein the clearance rate of the bletilla striata polysaccharide to DPPH free radicals, OH free radicals and ABTS + free radicals can reach 43.6%, 74.8% and 51.7% respectively; has promoting effect and immunoregulation activity on cell Raw264.7 growth.

The method for preparing the bletilla striata polysaccharide comprises the following steps: the preparation method adopts an enzymolysis method, and the enzymolysis step comprises the following steps: the common bletilla pseudobulb is crushed, subjected to constant-temperature enzymolysis and inactivated, wherein the conditions of the constant-temperature enzymolysis are as follows: the addition amount of enzyme is 0.5-2%, and the enzymolysis temperature is as follows: 35-60 ℃, enzymolysis time: 50-100min, the pH value is: 4-7.

The preferable conditions for constant-temperature enzymolysis are as follows: enzyme addition amount of 1.4%, enzymolysis temperature: enzymolysis time at 55 ℃: 87min, and the pH value is: 5.43.

the enzyme preparation adopted by the enzymolysis method is pectinase, and the enzyme activity is 2 ten thousand u/g.

The step of enzyme deactivation and centrifugation refers to that bletilla striata enzymatic hydrolysate is subjected to heat preservation in a boiling water bath for 3-5 minutes; centrifuging at the rotation speed of 5000-.

The bletilla polysaccharide is applied to the development of immune enhancing medicines or health-care food for enhancing immunity.

Has the advantages that: the bletilla polysaccharide prepared by the method has the clearance rates of DPPH free radicals, OH free radicals and ABTS + free radicals reaching 43.6%, 74.8% and 51.7% respectively, has a certain promotion effect on the growth of Raw264.7 cells, has immunoregulation activity, and has an obvious immune induction activity effect when the concentration of the bletilla polysaccharide is 400 mu g/mL.

Drawings

FIG. 1 Effect of different concentrations of bletilla striata polysaccharide on the viability of RAW264.7 cells

FIG. 2 levels of TNF- α secretion from RAW264.7 cells induced by different concentrations of bletilla striata polysaccharide

Detailed Description