阅读说明：本技术 一种食用菌复合多糖的制备工艺 (Preparation process of edible fungus composite polysaccharide ) 是由 曾文正 洪鹏飞 李宝蓉 陈亮 洪龙杰 郑碧容 于 2018-08-09 设计创作，主要内容包括：本发明涉及一种食用菌复合多糖的制备工艺,属于食用菌加工技术领域；称取经过研磨的食用菌干粉5g于烧杯中,加入100mL蒸馏水,用5mol/L盐酸水解,料液比为1：20,加热6h；经过离心取出上清液,加入活性炭,在60℃水浴振荡半小时左右脱色完全.加入混合试剂,三氯甲烷∶正丁醇,去除粗多糖中的蛋白质,并用葡聚糖凝胶对提取液中复合多糖进行纯化；采用类似实验的方法制得多糖锌的配合物,工艺简单,操作方便,且后续进行多种实验来确认配合物结合率与温度等参数,可得到最佳反应条件,从而可提高食用菌复合多糖制备的品质。(The invention relates to a preparation process of edible fungus composite polysaccharide, belonging to the technical field of edible fungus processing; weighing 5g of ground edible fungus dry powder in a beaker, adding 100mL of distilled water, and hydrolyzing with 5mol/L hydrochloric acid, wherein the material-liquid ratio is 1: 20, heating for 6 hours; centrifuging to obtain supernatant, adding active carbon, oscillating in water bath at 60 deg.C for half an hour to decolorize completely, adding mixed reagent, chloroform and n-butanol, removing protein in crude polysaccharide, and purifying the complex polysaccharide with dextran gel; the complex of polysaccharide zinc is prepared by adopting a method similar to an experiment, the process is simple, the operation is convenient, and a plurality of experiments are carried out subsequently to confirm parameters such as the complex binding rate, the temperature and the like, so that the optimal reaction condition can be obtained, and the quality of the edible fungus composite polysaccharide preparation can be improved.)

1. A preparation process of edible fungus composite polysaccharide is characterized by comprising the following steps: the preparation process of the edible fungus composite polysaccharide comprises the following steps:

(1) weighing 5g of ground edible fungus dry powder in a beaker, adding 100mL of distilled water, and hydrolyzing with 5mol/L hydrochloric acid, wherein the material-liquid ratio is 1: 20, heating for 6 hours, and standing for about three hours;

(2) performing microwave treatment for four minutes under the condition that the microwave power is 80 percent, and extracting mixed polysaccharide;

(3) centrifuging to obtain supernatant, adding active carbon, oscillating in water bath at 60 deg.C for half an hour to decolorize completely, adding mixed reagent, chloroform and n-butanol, removing protein in crude polysaccharide, and purifying the complex polysaccharide with dextran gel;

(4) centrifuging the purified solution at 3000r/min, freeze drying to obtain mixed polysaccharide,

(5) mixing and dissolving compound polysaccharide and zinc acetate in a certain mass ratio in 200mL of water, controlling the reaction temperature of the solution, adjusting the pH value, reacting for 10-30 minutes, centrifuging for about 10 minutes at 4000r/min, and taking supernatant for reduced pressure distillation;

(6) precipitating with ethanol, pre-freezing the residual liquid after vacuum distillation, and freeze-drying;

(7) weighing the mass of the mixture after drying;

(8) and hydrolyzing and separating to obtain the complex of polysaccharide zinc.

2. The preparation process of the edible fungus composite polysaccharide as claimed in claim 1, wherein the preparation process comprises the following steps: and (2) slowly cooling when standing in the step (1), and ensuring that the temperature is controlled to be about 25 ℃ for three hours.

3. The preparation process of the edible fungus composite polysaccharide as claimed in claim 1, wherein the preparation process comprises the following steps: the mixed reagent in the step (3) is chloroform to n-butyl alcohol 4: 1.

4. the preparation process of the edible fungus composite polysaccharide as claimed in claim 1, wherein the preparation process comprises the following steps:

the detection method of the complex polysaccharide complex binding rate comprises the following steps:

(a) sucking 10mL of water sample into a 25mL colorimetric tube, if the zinc content of the water sample exceeds 5mg, taking a proper amount of water sample, and diluting the water sample to 10mL by using pure water;

(b) taking 8 colorimetric tubes, adding 1mg/mL zinc, standard use solutions of 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0mL, adding pure water to 10mL respectively,

(c) adding 5mL of acetic acid-sodium acetate buffer solution into each colorimetric tube, mixing uniformly, adding 1mL of sodium thiosulfate solution respectively, mixing uniformly,

(d) then adding 2.5mL of dithizone solution, shaking up, and standing for about half an hour to obtain the product.

5. The preparation process of the edible fungus composite polysaccharide as claimed in claim 1, wherein the preparation process comprises the following steps: the method for selecting, measuring and measuring the complex polysaccharide coordination temperature comprises the following steps:

respectively weighing 6 parts of composite polysaccharide and zinc acetate according to a certain mass ratio, mixing and dissolving in 200mL of water, adjusting the pH value of the solution to 9.0, controlling the reaction time to 65min, and controlling the reaction temperatures to be 30, 40, 50, 60, 70 and 80 ℃ respectively;

(II) carrying out alcohol precipitation, pre-freezing a small amount of residual liquid after reduced pressure distillation, and carrying out freeze drying;

(III) weighing the mass of the mixture after drying;

(IV) hydrolyzing and separating to obtain a polysaccharide zinc complex;

and (V) detecting the optimal temperature according to the detection method of the complex polysaccharide complex binding rate.

6. The preparation process of the edible fungus composite polysaccharide as claimed in claim 1, wherein the preparation process comprises the following steps: the method for determining the molecular mass of the compound polysaccharide in the lentinus edodes comprises the following steps:

accurately weighing 0.05g of glucan standard products with different molecular masses, dissolving by using a mobile phase, fixing the volume to 5mL, drawing a standard coordinate curve by establishing a molecular mass logarithm value according to the separation retention time of the glucan standard products with different molecular masses and the mass of corresponding molecules, and obtaining the retention time values of different molecular mass peaks after the samples to be detected are separated, wherein the final mass concentration is 10mg/mL, and the sample to be detected is drawn according to the molecular mass logarithm value established by the separation retention time of the glucan standard products with different molecular masses and the mass of corresponding molecules, and the molecular mass of the glucan can be calculated by a molecular mass standard.

Technical Field

The invention relates to a preparation process of edible fungus composite polysaccharide, belonging to the technical field of edible fungus processing.

Background

The compound polysaccharide extracted from Lentinus Edodes, Tremella, and Hericium Erinaceus has effects of relieving stomach ache and flatulence; the compound polysaccharide extracted from Grifola frondosa and Coprinus comatus has the effects of assisting in lowering blood pressure and lowering blood sugar. The ability of the compound polysaccharide to remove free radicals is higher than that of the edible fungus polysaccharide extracted singly.

The fungal polysaccharide has the function of regulating the immunity of the organism, and the immunity of the organism is improved mainly through the functions of a plurality of layers of mononuclear macrophages, a cellular immune system and a humoral immune system. Some fungus polysaccharides (tremella polysaccharide, lentinan, corious versicolor polysaccharide, etc.) can enhance PFE reaction of normal mice sensitized by sheep red blood cells, thereby improving humoral immune reaction and promoting somatic cell immunity and humoral immune function.

The edible fungus polysaccharide has wide application value, for example, the lentinus edodes contains polysaccharide required by human body, the polysaccharide has extremely important biological function, and the polysaccharide has close relation with the regulation of immunologic function, the recognition of cells and cells, the transfer of substances among cells, the diagnosis and treatment of cancer and the like.

Therefore, the preparation process of the edible fungus compound polysaccharide is the subject of current research.

Disclosure of Invention

The invention aims to provide a preparation process of edible fungus composite polysaccharide, which is reasonable in design and convenient to operate, aiming at the defects and shortcomings of the prior art.

In order to achieve the purpose, the invention adopts the technical scheme that: the preparation process of the edible fungus composite polysaccharide comprises the following steps:

1. weighing 5g of ground edible mushroom (shiitake mushroom and oyster mushroom) dry powder in a beaker, adding 100mL of distilled water, and hydrolyzing with 5mol/L hydrochloric acid, wherein the material-liquid ratio is 1: 20, heating for 6 hours, and standing for about three hours;

2. performing microwave treatment for four minutes under the condition that the microwave power is 80 percent, and extracting mixed polysaccharide;

3. centrifuging to obtain supernatant, adding active carbon, decolorizing completely in 60 deg.C water bath for half an hour, adding mixed reagent (chloroform: n-butanol), removing protein in crude polysaccharide, and purifying the complex polysaccharide with dextran gel;

4. centrifuging the purified solution at 3000r/min, freeze drying to obtain mixed polysaccharide,

5. mixing and dissolving compound polysaccharide and zinc acetate in a certain mass ratio in 200mL of water, controlling the reaction temperature of the solution, adjusting the pH value, reacting for 10-30 minutes, centrifuging for about 10 minutes at 4000r/min, and taking supernatant for reduced pressure distillation;

6. precipitating with ethanol, pre-freezing the residual liquid, and freeze drying;

7. weighing the mass of the mixture after drying;

8. hydrolyzing and separating to obtain the complex of polysaccharide zinc.

Preferably, the temperature is slowly reduced when the mixture is kept still in the step 1, and the temperature is kept at about 25 ℃ for three hours.

Preferably, the mixed reagent in the step 3 is chloroform to n-butanol of 4: 1.

the method for selectively measuring the complex polysaccharide coordination temperature comprises the following steps: respectively weighing 6 parts of composite polysaccharide and zinc acetate according to a certain mass ratio, mixing and dissolving in 200mL of water, adjusting the pH value of the solution to 9.0, controlling the reaction time to 65min, controlling the reaction temperature to be 30, 40, 50, 60, 70 and 80 ℃, carrying out alcohol precipitation, pre-freezing a small amount of liquid left after reduced pressure distillation, carrying out freeze drying, weighing the mass of a matching substance after drying, carrying out hydrolysis separation to obtain a complex of polysaccharide and zinc, and detecting the optimal temperature according to a detection method of the matching binding rate of the composite polysaccharide.

After the method is adopted, the invention has the beneficial effects that: the preparation process of the edible fungus composite polysaccharide adopts a similar experiment method to prepare the complex of the polysaccharide zinc, has simple process and convenient operation, and can obtain the optimal reaction condition by subsequently carrying out various experiments to confirm parameters such as the combination rate, the temperature and the like of the complex, thereby improving the preparation quality of the edible fungus composite polysaccharide.

Detailed Description

The preparation process of the edible fungus composite polysaccharide comprises the following steps:

1. weighing 5g of ground dried mushroom powder into a beaker, adding 100mL of distilled water, and hydrolyzing with 5mol/L hydrochloric acid, wherein the material-liquid ratio is 1: 20, heating for 6 hours, and standing for about three hours;

2. performing microwave treatment for four minutes under the condition that the microwave power is 80 percent, and extracting mixed polysaccharide;

3. centrifuging to obtain supernatant, adding active carbon, shaking in water bath at 60 deg.C for half an hour to decolorize completely, adding mixed reagent (chloroform: n-butanol 4: 1), removing protein from crude polysaccharide, and purifying the complex polysaccharide with dextran gel;

4. centrifuging the purified solution at 3000r/min, and freeze drying to obtain mixed polysaccharide.

5. Mixing and dissolving compound polysaccharide and zinc acetate in a certain mass ratio in 200mL of water, controlling the reaction temperature of the solution, adjusting the pH value, reacting for 10-30 minutes, centrifuging for about 10 minutes at 4000r/min, and taking supernatant for reduced pressure distillation;

6. precipitating with ethanol, pre-freezing the residual liquid, and freeze drying;

7. weighing the mass of the mixture after drying;

8. hydrolyzing and separating to obtain the complex of polysaccharide zinc.

The detection method of the complex polysaccharide complex binding rate in the specific embodiment comprises the following steps:

a. sucking 10mL of water sample into a 25mL colorimetric tube, if the zinc content of the water sample exceeds 5mg, taking a proper amount of water sample, and diluting the water sample to 10mL by using pure water;

b. adding 1mg/mL zinc, standard solutions 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0mL, adding pure water to 10mL respectively,

c. adding 5mL of acetic acid-sodium acetate buffer solution into each colorimetric tube, mixing uniformly, adding 1mL of sodium thiosulfate solution respectively, mixing uniformly,

d. then adding 2.5mL of dithizone solution, shaking up, and standing for about half an hour to obtain the product.

The method for selectively measuring the complex polysaccharide coordination temperature comprises the following steps:

respectively weighing 6 parts of composite polysaccharide and zinc acetate in a certain mass ratio, mixing and dissolving in 200mL of water, adjusting the pH value of the solution to 9.0, controlling the reaction time to 65min, and controlling the reaction temperatures to be 30, 40, 50, 60, 70 and 80 ℃ respectively;

II, precipitating with ethanol, pre-freezing a small amount of residual liquid after reduced pressure distillation, and freeze-drying;

III, weighing the mass of the mixture after drying;

IV, hydrolyzing and separating to obtain a polysaccharide zinc complex;

and V, detecting the optimal temperature according to a detection method of the complex polysaccharide complex binding rate.

The method for determining the molecular mass of the compound polysaccharide in the lentinus edodes comprises the following steps:

accurately weighing 0.05g of glucan standard products with different molecular masses, dissolving by using a mobile phase, fixing the volume to 5mL, drawing a standard coordinate curve by establishing a molecular mass logarithm value according to the separation retention time of the glucan standard products with different molecular masses and the mass of corresponding molecules, and obtaining the retention time values of different molecular mass peaks after the samples to be detected are separated, wherein the final mass concentration is 10mg/mL, and the sample to be detected is drawn according to the molecular mass logarithm value established by the separation retention time of the glucan standard products with different molecular masses and the mass of corresponding molecules, and the molecular mass of the glucan can be calculated by a molecular mass standard.

The preparation process of the edible fungus composite polysaccharide adopts a method similar to an experiment to prepare the complex of the polysaccharide zinc, has simple process and convenient operation, and can obtain optimal reaction conditions by subsequently carrying out various experiments to confirm parameters such as the combination rate, the temperature and the like of the complex, thereby improving the preparation quality of the edible fungus composite polysaccharide.

The above description is only for the purpose of illustrating the technical solutions of the present invention and not for the purpose of limiting the same, and other modifications or equivalent substitutions made by those skilled in the art to the technical solutions of the present invention should be covered within the scope of the claims of the present invention without departing from the spirit and scope of the technical solutions of the present invention.