阅读说明：本技术 一种具有降血糖作用的黄大茶多糖 (A radix et rhizoma Rhei tea polysaccharide with blood sugar lowering effect ) 是由 陈彦 王志 姚程颖 潘文娟 于 2020-09-02 设计创作，主要内容包括：本发明公开了一种具有降血糖作用的黄大茶多糖制备方法,其特征在于：黄大茶多糖LYT-2是以霍山产的黄大茶为原料,经过EM菌固态发酵,利用深度共熔溶剂技术提取制备；该多糖化合物分子量为4.38×10<Sup>4</Sup> Da,含有4.98%结合蛋白,单糖组成按摩尔比为甘露糖:葡萄糖:阿拉伯糖=2.16:4.83:1。体内实验数据显示LYT-2能明显降低高糖高脂小鼠的体重和血糖水平。结果表明LYT-2在食品、医药保健等方面具有良好的应用前景。(The invention discloses a preparation method of a large yellow tea polysaccharide with a hypoglycemic effect, which is characterized by comprising the following steps: the yellow tea polysaccharide LYT-2 is prepared by taking yellow tea produced in Hovenia huoshanensis as a raw material, performing solid state fermentation by EM (effective microorganisms), and extracting by using a deep eutectic solvent technology; the polysaccharide compound has a molecular weight of 4.38 × 10 4 Da, containing 4.98% binding proteins, and monosaccharide composition in molar ratio mannose to glucose to arabinose =2.16:4.83: 1. The in vivo experimental data show that LYT-2 can significantly reduce the body weight and blood sugar level of high-sugar and high-fat mice. The results show that the LYT-2 has good application prospect in the aspects of food, medicine, health care and the like.)

1. The big yellow tea polysaccharide with the function of reducing blood sugar is characterized in that: the polysaccharide is derived from LYT-2.

2. The polysaccharide of yellow broadleaf tea having hypoglycemic effect according to claim 1, wherein: the molecular weight of the polysaccharide LYT-2 of the big yellow tea is 4.38 × 10⁴Da, containing 4.98% of binding protein, and monosaccharide composition according to a molar ratio of mannose to glucose to arabinose =2.16:4.83: 1.

3. The polysaccharide of yellow broadleaf tea having hypoglycemic effect according to claim 1 or 2, wherein: the preparation method of the rhubarb tea polysaccharide specifically comprises the following steps:

A. crushing the large Huoshan Huang tea purchased in the market to 60-80 meshes by solid fermentation, adding EM (effective microorganisms) powder according to 1% of the mass of the tea powder, spraying distilled water with the mass of 0.5-1 time, uniformly stirring, filling into an anaerobic fermentation bag, sealing, keeping the temperature at 20-27 ℃, and fermenting for 7-9 days;

B. the extraction and purification are carried out to prepare the large yellow tea polysaccharide by adopting a deep eutectic solvent extraction method, the extraction solvent is food-grade choline chloride and glycerol, the mass ratio of the food-grade choline chloride to the glycerol is 1:2, the material-liquid ratio is 1: 20-50, the extraction time is 120-1200 min, the extraction temperature is 50-60 ℃, the extract is subjected to centrifugation, concentration, alcohol precipitation, dialysis, separation and purification, freeze drying and other steps to obtain the large yellow tea polysaccharide LYT-2, namely the large yellow tea polysaccharide with the hypoglycemic effect.

Technical Field

The invention relates to a tea deep-processing product, in particular to an active polysaccharide component with the blood sugar reducing effect obtained by taking probiotics fermented yellow big tea as a raw material, belonging to the related fields of food, health care and the like.

Background

Diabetes, a "rich disease," is a metabolic disease characterized by hyperglycemia. At present, no method for radically treating diabetes exists, the treatment of diabetes at present is controlled by medicaments, and the use of the medicaments causes great damage to heart, liver, spleen, stomach and kidney. The natural active substances comprise polysaccharide, protein, sterol and the like, have various biological activities and low side effects, are widely applied to the fields of health care and disease treatment, and have wide application prospects.

The yellow big tea belongs to yellow tea, is also called Wanxi yellow big tea, is produced in places such as Huoshan, Jinzhai, Yuexi and the like in Anhui province, and has a plurality of health care functions besides unique tea fragrance, but the material basis of the activity efficacy is not clear. The polysaccharide is one of main functional components of the yellow big tea, and the structure and the function of the polysaccharide can be changed under the action of probiotics. The EM bacteria is a microbial viable bacteria preparation compounded by more than 10 microorganisms belonging to 80 species, mainly photosynthetic bacteria, lactic acid bacteria, saccharomycetes and actinomycetes, and can change the substance composition of a target product through biotransformation, thereby achieving stronger pharmacological effect.

The hot water extraction method is a traditional extraction method of polysaccharide, and has the defects of low extraction rate, high temperature, easy damage of active substances, more impurities and the like. Deep eutectic solvents (DESS) are formed by mechanically stirring or heating and mixing hydrogen bond acceptors and hydrogen bond donor compounds according to a certain proportion, can directionally promote the release and dissolution of polysaccharide components, and compared with the traditional extraction method, the DESs have the advantages of low extraction temperature, short time, less impurities and the like.

Disclosure of Invention

The invention aims to provide a large yellow tea polysaccharide with the function of reducing blood sugar, which has a remarkable function of reducing blood sugar in vivo and has huge application potential in the fields of medical care and the like; the invention firstly combines probiotics and yellow tea for fermentation to obtain a novel structural polysaccharide, and then extracts and prepares the polysaccharide by using a deep eutectic solvent extraction technology, thus being a simple and mild preparation method with high yield.

In order to realize the purpose of the invention, the following technical scheme is adopted:

a polysaccharide of radix et rhizoma Rhei tea with blood sugar lowering effect: the polysaccharide is derived from Camellia sinensisLYT-2 with molecular weight of 4.38 × 10⁴Da, containing 4.98% of binding protein, and monosaccharide composition according to a molar ratio of mannose to glucose to arabinose =2.16:4.83: 1.

The preparation method of the yellow big tea polysaccharide with the hypoglycemic effect comprises the following steps:

A. crushing the large Huoshan Huang tea purchased in the market to 60-80 meshes by solid fermentation, adding EM (effective microorganisms) powder according to 1% of the mass of the tea powder, spraying distilled water with the mass of 0.5-1 time, uniformly stirring, filling into an anaerobic fermentation bag, sealing, keeping the temperature at 20-27 ℃, and fermenting for 7-9 days;

B. the extraction and purification are carried out by adopting a deep eutectic solvent extraction method to extract the fermented rhubarb tea polysaccharide, wherein the extraction solvent is food-grade choline chloride and glycerol, the mass ratio of the food-grade choline chloride to the glycerol is 1:2, the material-liquid ratio is 1: 20-50, the extraction time is 120-1200 min, the extraction temperature is 50-60 ℃, and the extractive solution is subjected to centrifugation, concentration, alcohol precipitation, dialysis, chromatography, freeze drying and other steps to obtain the rhubarb tea polysaccharide LYT-2, namely the polysaccharide with the hypoglycemic effect.

The EM bacterial powder in the invention is in a conventional specification sold on the market.

The invention has the beneficial effects that:

the invention takes the yellow big tea as the basic raw material, and improves the polysaccharide yield and the biological activity by combining the probiotic fermentation with the deep eutectic solvent extraction method, and the product yield can reach more than 6 percent. The invention firstly combines the probiotics and the yellow tea for fermentation to generate a new polysaccharide, and then extracts and prepares the polysaccharide by using a deep eutectic solvent extraction technology, thereby being a simple and mild preparation method with high yield.

The in vivo blood sugar reduction experiment shows that the LYT-2 obtained by the invention can obviously reduce the blood sugar content in a diabetes model mouse induced by STZ and the weight of the mouse, and the blood sugar value of the experimental group mouse is reduced by 34.33 percent compared with the blood sugar value of the model group under the condition of 400mg/kg stomach irrigation. LYT-2 has obvious blood sugar reducing effect and wide market application prospect.

Drawings

FIG. 1 shows the polysaccharide component of the yellow camellia sinensis with hypoglycemic effect and the relative molecular mass detection result in example 5 of the present invention, wherein the chromatographic conditions are Agilent 1260 HPLC-ELSD, TSKgel G5000PWxL (7.8 mm. times.300 mm);

FIG. 2 shows the results of the measurement of the monosaccharide composition of the polysaccharide of Camellia sinensis with hypoglycemic activity in example 5 of the present invention, wherein the chromatographic conditions are Agilent 1260 HPLC, Eclipse XDB-C₁₈(2.1mm ×150mm)；

FIG. 3 is a measurement result of the weight of STZ-induced high-sugar and high-fat model mice reduced by the yellow camellia sinensis polysaccharide with hypoglycemic effect in example 5 of the invention, and experiments show that the weight reduction of the STZ-induced high-sugar and high-fat model mice can be obviously relieved under the condition of a gavage dose of 400 mg/kg;

fig. 4 is a measurement result of blood sugar of a model mouse with high sugar and high fat induced by STZ of the yellow camellia sinensis polysaccharide having the effect of reducing blood sugar in example 5 of the invention, and an experimental result shows that the yellow camellia sinensis polysaccharide LYT-2 can significantly reduce the blood sugar content in the model mouse with diabetes.

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