Preparation process and system of gastrodia elata polypeptide

Abstract

The invention relates to the technical field of gastrodia elata polypeptide preparation, in particular to a gastrodia elata polypeptide preparation process and system.The gastrodia elata polypeptide preparation process comprises the steps that gastrodia elata solids are used according to the solid-liquid ratio of 1: 10, the liquid is 0.5% NaCL solution, water bath is conducted for 8 h after homogenization, the water solution temperature is 70-80 DEG C, the filtered solution is concentrated under reduced pressure till thick paste with the solid content being 25-35% is obtained, proteolytic enzyme with the weight ratio being 1% is added into the thick paste, enzymolysis is conducted at the temperature of 40-50 DEG C, enzymolysis is conducted for 2-3 h, and the gastrodia elata polypeptide is obtained. The enzymolysis time is 2-3 h, after centrifugal separation of enzymatic hydrolysate, ultrafiltration is conducted for 30 min at the speed of 8000 rpm, the gastrodia elata polypeptide with the molecular weight of 5.8-7.8 KD is obtained, resource utilization of gastrodia elata extraction waste is achieved, the obtained gastrodia elata polypeptide is moderate in molecular weight and has good biological activity, no organic solvent is needed in the whole process, and the gastrodia elata polypeptide is environmentally friendly, simple in technology, convenient to operate and suitable for industrial production.

Description

TECHNICAL FIELD

[0001] The present application relates to the preparation technology field of Gastrodia elata polypeptide, and particularly relates to a preparation process and system of Gastrodia elata polypeptide. BACKGROUND

[0002] Gastrodia elata polypeptide is a plant macromolecular polypeptide extracted from Gastrodia elata, and has biological pharmacological activity. For example, the Gastrodia elata polypeptide has broad-spectrum bacteriostatic activity, and shows extremely high bacteriostatic and bactericidal effects on common bacteria, fungi causing gynecological diseases and green pyocyanic bacillus causing wound infection. The Gastrodia elata polypeptide also shows killing power on influenza virus, hepatitis B virus and H1N1 influenza virus, and can be used for immune enhancement treatment against human papilloma virus (HPV).

[0003] At present, a large amount of solid waste is generated in the extraction process of Gastrodia elata extract and crude polysaccharide, and the solid waste contains rich protein resources. At present, the treatment method for the solid waste is mainly direct disposal, which not only causes resource waste, but also may pollute the environment.

[0004] Therefore, the present application provides a preparation process and system of Gastrodia elata polypeptide, which uses the solid waste after Gastrodia elata extraction to prepare high-value-added Gastrodia elata polypeptide. SUMMARY

[0005] The present application aims to provide a preparation process and system of Gastrodia elata polypeptide, which solves the problem that a large amount of solid waste is generated in the extraction process of Gastrodia elata extract and crude polysaccharide, and the solid waste is directly disposed, which not only causes resource waste, but also may pollute the environment.

[0006] To achieve the above-mentioned purpose, the present application provides a preparation process of Gastrodia elata polypeptide, which comprises the following steps:

[0007] Step S1: mixing the solid material after water boiling of Gastrodia elata alcohol extraction with a NaCl solution, homogenizing, and then water bathing at T1 temperature;

[0008] Step S2: pressure filtering the solution obtained in step S1, and reducing pressure to concentrate the solution into thick paste;

[0009] Step S3: adding hydrolytic enzyme to the thick paste, and enzymolysis at T2 temperature to obtain enzyme solution;

[0010] Step S4: centrifuging the enzyme solution, and then performing ultrafiltration treatment to obtain Gastrodia elata polypeptide.

[0011] In step S1, the mass percentage concentration of the NaCl solution is 0.5%, and the weight ratio of the Gastrodia elata solid material to the 0.5% NaCl solution is 1:10; the temperature of T1 is 70-80 DEG C; and the water bathing time is 8h.

[0012] The solid content mass ratio of the thick paste is 25% to 35% in step S2.

[0013] The hydrolytic enzyme is a proteolytic enzyme, the weight ratio is 1%, the temperature of T2 is 40 to 50 DEG C, and the enzymolysis time is 2 to 3 hours in step S3.

[0014] The centrifugal speed of the ultrafiltration is 8000 rpm, and the time is 30 minutes in step S4.

[0015] The molecular weight of the obtained Gastrodia elata polypeptide is 5.8 to 7.8 KD in step S4.

[0016] The application further provides a system for preparing the Gastrodia elata polypeptide, which comprises sequentially connected water bath extraction module, concentration module, enzymolysis module and separation and purification module.

[0017] The application provides a preparation process and system for Gastrodia elata polypeptide. BRIEF DESCRIPTION OF DRAWINGS

[0018] The accompanying drawings for describing the technical schemes in the embodiments of the application or the prior art are briefly introduced as follows.

[0019] Figure 1is a step flow chart of the preparation process of the Gastrodia elata polypeptide provided by the present application.

[0020] Figure 2 is a step flow chart of the preparation process of the Gastrodia elata polypeptide provided by the present application.

[0021] Figure 3 is a step flow chart of the preparation process of the Gastrodia elata polypeptide provided by the present application.

[0022] Figure 4 is a step flow chart of the preparation process of the Gastrodia elata polypeptide provided by the present application. DETAILED DESCRIPTION

[0023] The embodiments of the present application are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar notations represent the same or similar elements or elements having the same or similar functions throughout. The embodiments described below by referring to the drawings are exemplary and are intended to explain the present application, and cannot be understood as a limitation of the present application.

[0024] Please refer to Figure 1 , Figure 1 is a step flow chart of the preparation process of the Gastrodia elata polypeptide provided by the present application.

[0025] The present application provides a preparation process of Gastrodia elata polypeptide, comprising the following steps:

[0026] Step S1: mixing the solid material after water boiling of Gastrodia elata alcohol extraction with NaCl solution, homogenizing, and then water bathing at T1 temperature;

[0027] Step S2: after pressure filtration of the solution obtained in step S1, the solution is concentrated into thick paste under reduced pressure;

[0028] Step S3: adding hydrolytic enzyme to the thick paste, and enzymolysis at T2 temperature to obtain an enzymolysis solution;

[0029] Step S4: centrifugal separation of the enzymolysis solution, and then ultrafiltration treatment to obtain Gastrodia elata polypeptide.

[0030] In the embodiment, the application prepares Gastrodia elata polypeptide through sequentially connected water bath extraction module, concentration module, enzymatic hydrolysis module and separation and purification module. Firstly, the water bath extraction module precisely realizes that Gastrodia elata solid is mixed with 0.5% NaCL solution at a material-liquid ratio of 1:10 in a water bath at 70-80°C for 8 hours. Then, the concentrated solution after the water bath is reduced pressure concentrated into thick paste with solid content of 25%-35% through the concentration module. Then, the enzymatic hydrolysis module controls the addition amount of 1% proteolytic enzyme to be hydrolyzed at 40-50°C for 2-3 hours. Finally, the separation and purification module is centrifuged, and then ultrafiltration is performed at 8000 rpm for 30 min to obtain Gastrodia elata polypeptide with a molecular weight of 5.8-7.8 KD. Therefore, the process of generating Gastrodia elata polypeptide by enzymatic hydrolysis of the solidified material after Gastrodia elata alcohol extraction and boiling is feasible, and the economic value is considerable.

[0031] In step S1, the material-liquid ratio of 1:10 ensures that there is enough NaCL solution to contact the Gastrodia elata solid, so that the solid can be fully dispersed and soaked, which is beneficial to the subsequent extraction process. The temperature environment of 70-80°C can increase the solubility of polypeptide and other components in the Gastrodia elata solid, so that they are more easily dissolved from the solid into the NaCL solution. At the same time, the ionic environment provided by the 0.5% NaCl solution helps to maintain the osmotic pressure balance inside and outside the cell, promotes the release of cell contents, and further improves the extraction efficiency of polypeptide and other components.

[0032] In step S2, the solution is concentrated to thick paste with solid content of 25%-35%, which can enrich Gastrodia elata polypeptide, remove part of impurities, facilitate subsequent enzymatic hydrolysis and separation and purification operations, avoid damage to the activity of polypeptide, and ensure stable product quality.

[0033] In step S3, the proteolytic enzyme can maintain high enzyme activity at 40-50°C and prevent inactivation. Enzymatic hydrolysis for 2-3 hours can fully hydrolyze proteins and reduce side reactions.

[0034] The preparation process of Gastrodia elata polypeptide of the application uses Gastrodia elata solid at a material-liquid ratio of 1:10, 0.5% NaCL solution, 8h water bath after uniform mixing, water bath temperature 70-80°C, the solution is reduced pressure concentrated into thick paste with solid content of 25%-35% after filtration, 1% proteolytic enzyme is added to the thick paste, the enzymatic hydrolysis temperature is 40-50°C, the enzymatic hydrolysis time is 2-3h, the enzymatic hydrolysis liquid is centrifuged, and then ultrafiltration is performed at 8000 rpm for 30 min to obtain Gastrodia elata polypeptide with a molecular weight of 5.8-7.8 KD. The resource utilization of Gastrodia elata extraction waste is realized. The obtained Gastrodia elata polypeptide has moderate molecular weight and good biological activity. The whole process does not need to use organic solvent, is green and environmentally friendly, and has simple process, convenient operation and is suitable for industrialized production.

[0035] Please refer to Figures 2 to 4 , Figure 2is a step flow chart of example one in the preparation process of Gastrodia elata polypeptide provided by the present application, Figure 3 is a step flow chart of example two in the preparation process of Gastrodia elata polypeptide provided by the present application, Figure 4 is a step flow chart of example three in the preparation process of Gastrodia elata polypeptide provided by the present application.

[0036] In the present application, three examples are also provided as follows:

[0037] Example one:

[0038] Step S1: take 100g of solid material after water boiling of Gastrodia elata alcohol extraction, add 1000ml of 0.5% NaCL solution, mix and homogenize, and then water bath at 70℃ for 8h;

[0039] Step S2: after pressure filtration of the solution obtained in step S1, the solution is concentrated under reduced pressure to a thick paste with solid content of 35%;

[0040] Step S3: add 1g of proteolytic enzyme to the thick paste, and enzymolysis at 50℃ for 2h to obtain an enzymolysis solution;

[0041] Step S4: after centrifugal separation of the enzymolysis solution, ultrafiltration treatment is carried out at 8000rpm for 30min to obtain Gastrodia elata polypeptide with molecular weight of about 5.8KD.

[0042] Example two:

[0043] Step S1: take 100g of solid material after water boiling of Gastrodia elata alcohol extraction, add 1000ml of 0.5% NaCL solution, mix and homogenize, and then water bath at 75℃ for 8h;

[0044] Step S2: after pressure filtration of the solution obtained in step S1, the solution is concentrated under reduced pressure to a thick paste with solid content of 30%;

[0045] Step S3: add 1g of proteolytic enzyme to the thick paste, and enzymolysis at 45℃ for 2.5h to obtain an enzymolysis solution;

[0046] Step S4: after centrifugal separation of the enzymolysis solution, ultrafiltration treatment is carried out at 8000rpm for 30min to obtain Gastrodia elata polypeptide with molecular weight of about 6.5KD.

[0047] Example three:

[0048] Step S1: take 100g of solid material after water boiling of Gastrodia elata alcohol extraction, add 1000ml of 0.5% NaCL solution, mix and homogenize, and then water bath at 80℃ for 8h;

[0049] Step S2: after pressure filtration of the solution obtained in step S1, the solution is concentrated under reduced pressure to a thick paste with solid content of 25%;

[0050] Step S3: 1g protease is added into the thick paste, and enzymolysis is carried out at 40℃ for 3h to obtain an enzymolysis solution;

[0051] Step S4: After centrifugal separation, the enzymolysis solution is subjected to ultrafiltration treatment at 8000rpm for 30min to obtain a Gastrodia elata polypeptide with a molecular weight of about 7.8KD.

[0052] The above only discloses one preferred embodiment of the present application, and of course cannot limit the scope of the present application, and those skilled in the art can understand that all or part of the above-mentioned processes can be implemented, and equivalent changes made according to the claims of the present application still belong to the scope of the present application.

Claims

1. A preparation process of Gastrodia elata polypeptide, characterized in that, It comprises the following steps: Step S1: the solid material after boiling the Gastrodia elata ethanol extract solution is mixed with NaCL solution, and then is uniformly mixed and water bathed at T1 temperature; Step S2: the solution obtained in step S1 is filtered under pressure, and the solution is concentrated under reduced pressure into a thick paste; Step S3: hydrolytic enzyme is added to the thick paste, and enzymolysis is carried out at T2 temperature to obtain an enzymolysis solution; Step S4: the enzymolysis solution is centrifuged and then subjected to ultrafiltration treatment to obtain Gastrodia elata polypeptide.

2. The preparation process of Gastrodia elata polypeptide according to claim 1, wherein in step S1, the mass percentage concentration of the NaCL solution is 0.5%, the ratio of the weight of the Gastrodia elata solid material to the weight of the 0.5% NaCL solution is 1:10, the temperature of T1 is 70-80°C, and the water bath time is 8h.

3. The preparation process of Gastrodia elata polypeptide according to claim 1, wherein in step S2, the solid content mass percentage of the thick paste is 25%-35%.

4. The preparation process of Gastrodia elata polypeptide according to claim 1, wherein in step S3, the hydrolytic enzyme is protein hydrolytic enzyme, the weight ratio is 1%, the temperature of T2 is 40-50°C, and the enzymolysis time is 2-3h.

5. The preparation process of Gastrodia elata polypeptide according to claim 1, wherein in step S4, the centrifugal speed of ultrafiltration is 8000rpm, and the time is 30min.

6. The preparation process of Gastrodia elata polypeptide according to claim 1, wherein in step S4, the molecular weight of the obtained Gastrodia elata polypeptide is 5.8-7.8KD.

7. A system for preparing Gastrodia elata polypeptide according to any one of claims 1-6, comprising water bath extraction module, concentration module, enzymolysis module and separation and purification module connected in sequence, wherein the water bath extraction module realizes mixing of Gastrodia elata solid material with 0.5% NaCL solution at a 1:10 solid-liquid ratio, water bathing at 70-80°C for 8h, the concentration module concentrates the solution after water bathing under reduced pressure into a thick paste with a solid content of 25%-35%, the enzymolysis module controls the addition amount of protein hydrolytic enzyme to carry out enzymolysis at 40-50°C for 2-3h, and the separation and purification module obtains Gastrodia elata polypeptide with a molecular weight of 5.8-7.8KD through centrifugation and ultrafiltration. ​ ​ ​ ​ ​ ​

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