High-extraction-rate hericium erinaceus protein extraction method

Abstract

The invention discloses a high-extraction-rate hericium erinaceus protein extraction method which comprises the following steps: step 1, taking hericium erinaceus powder according to a solid-liquid ratio of 1g: (20-30mL), and adding distilled water to obtain a hericium erinaceus powder suspension; and 2, adjusting the pH value of the hericium erinaceus powder suspension to 11-13 by using a NaOH solution, carrying out heat preservation for 2-3 hours under a water bath condition of 40-60 DEG C, cooling, carrying out suction filtration, collecting filtrate, adjusting the pH value of the filtrate to an isoelectric point 4.5 by using an HCl solution, carrying out centrifugal separation to obtain a precipitate, and carrying out freeze drying to obtain the hericium erinaceus protein. The preparation method is simple, the extraction cost of the hericium erinaceus protein is reduced, the hericium erinaceus protein is more suitable for large-scale industrial production, the extraction cost of the hericium erinaceus protein is reduced, and the application range of the hericium erinaceus protein is widened.

Description

Technical Field

[0001] The invention relates to protein extraction, in particular to a method for extracting Hericium erinaceus protein with a high extraction rate. Background Art

[0002] Hericium erinaceus, as a precious medicinal and edible fungus, is rich in a variety of biologically active ingredients. Among them, Hericium erinaceus protein has extremely high nutritional and medicinal value. For example, it shows significant effects in anti-oxidation, immunomodulation and anti-tumor. Therefore, Hericium erinaceus protein has broad application prospects in food, health care products and pharmaceutical industries.

[0003] However, the existing Hericium erinaceus protein extraction method has the problem of low extraction rate. The extraction rate of Hericium erinaceus protein is generally 27.3%, resulting in the failure to fully and effectively utilize Hericium erinaceus, serious waste, and relatively high production costs, which limits its large-scale industrial production and wider application.

[0004] Therefore, it is extremely important to develop a method that can significantly improve the protein extraction rate of Hericium erinaceus, which is of great significance to the development of Hericium erinaceus related industries. Summary of the invention

[0005] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a method for extracting Hericium erinaceus protein with a high extraction rate, which not only has a simple preparation method but also reduces the extraction cost of Hericium erinaceus protein.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for extracting Hericium erinaceus protein with a high extraction rate comprises the following steps:

[0008] Step 1, according to the material-liquid ratio of 1g: (20-30mL), take Hericium erinaceus powder and add distilled water to obtain Hericium erinaceus powder suspension;

[0009] Step 2, adjusting the pH value of the Hericium erinaceus powder suspension to 11-13 with a NaOH solution, and then keeping it warm for 2-3 hours in a water bath at 40-60° C., and after cooling, filtering and collecting the filtrate, adjusting the pH value of the filtrate to the isoelectric point of 4.5 with an HCl solution, and then centrifuging and separating the precipitate, and freeze-drying to obtain Hericium erinaceus protein.

[0010] Furthermore, the material-liquid ratio of step 1 is 1 g:25 mL.

[0011] Furthermore, in step 2, the concentrations of the NaOH solution and the HCl solution are both 0.1 mol / L.

[0012] Furthermore, the pH value in step 2 is 12.

[0013] Furthermore, the water bath temperature in step 2 is 60° C. and the insulation time is 2.5 h.

[0014] Furthermore, the centrifugal separation in step 2 is performed using a centrifuge with a rotation speed of 5000 r / min.

[0015] Compared with the prior art, the present invention has the following technical effects:

[0016] The invention improves the extraction rate of Hericium erinaceus protein and reduces the production cost by reasonably controlling the material-liquid ratio, pH value, water bath temperature and time, which is specifically embodied in the following aspects: firstly, by controlling the material-liquid ratio, the dissolution of Hericium erinaceus protein is promoted, thereby improving the extraction rate; secondly, in an alkaline environment, negatively charged protein molecules are more likely to attract protons, so they are more likely to combine with water molecules to form a hydration film, so that the solubility of the protein is increased, which is beneficial to the dissolution of the protein, thereby improving the extraction rate; thirdly, by regulating the temperature, the protein molecular structure is increased and activated, and the interaction between the protein and the water molecules is increased, thereby increasing the solubility of the protein, promoting the dissolution and separation of the protein, thereby improving the extraction rate; fourthly, by adjusting the extraction time, the protein molecular structure is gradually unfolded, the internal hydrophilic groups are exposed to the outside, which is beneficial to the dissolution of the protein, and improving the protein extraction rate.

[0017] In particular, when the solid-liquid ratio is 1g:25mL, the pH value is 12, the water bath temperature is 60 and the time is 2.5h, the extraction rate of Hericium erinaceus protein can reach 60.98±1.2%, which is much higher than the existing extraction rate of 27.3%. Therefore, the extraction method of the present invention is more suitable for large-scale industrial production, which helps to reduce the extraction cost of Hericium erinaceus protein and broaden the application scope of Hericium erinaceus protein. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 : Effect of different material-liquid ratios on the extraction rate of Hericium erinaceus protein;

[0019] Figure 2 :The effect of different pH values ​​on the extraction rate of Hericium erinaceus protein;

[0020] Figure 3 :The effect of different water bath temperatures on the extraction rate of Hericium erinaceus protein;

[0021] Figure 4 :Effect of different extraction time on the extraction rate of Hericium erinaceus protein. DETAILED DESCRIPTION

[0022] The specific contents of the present invention are further explained in detail below in conjunction with embodiments.

[0023] Example 1

[0024] Step 1, according to the material-liquid ratio of 1g:25mL, take Hericium erinaceus powder and add distilled water to obtain Hericium erinaceus powder suspension;

[0025] Step 2, adjusting the pH value of the Hericium erinaceus powder suspension to 12 with a 0.1 mol / L NaOH solution, and then keeping it warm in a water bath at 60°C for 2.5 hours. After cooling, filtering and collecting the filtrate, adjusting the pH value of the filtrate to an isoelectric point of 4.5 with a 0.1 mol / L HCl solution, and then centrifuging and separating the precipitate using a centrifuge with a rotation speed of 5000 r / min, and freeze-drying to obtain Hericium erinaceus protein.

[0026] Example 2

[0027] The only difference from Example 1 is that the material-liquid ratio in step 1 is 1 g:20 mL.

[0028] Example 3

[0029] The only difference from Example 1 is that the material-liquid ratio in step 1 is 1 g:30 mL.

[0030] Example 4

[0031] The only difference from Example 1 is that the pH value in step 2 is 11.

[0032] Example 5

[0033] The only difference from Example 1 is that the pH value in step 2 is 13.

[0034] Example 6

[0035] The only difference from Example 1 is that the water bath temperature in step 2 is 40°C.

[0036] Example 7

[0037] The only difference from Example 1 is that the water bath temperature in step 2 is 50°C.

[0038] Example 8

[0039] The only difference from Example 1 is that the insulation time in step 2 is 2 h.

[0040] Example 9

[0041] The only difference from Example 1 is that the insulation time in step 2 is 3 h.

[0042] Comparative Example 1

[0043] The only difference from Example 1 is that the material-liquid ratio in step 1 is 1 g:10 mL.

[0044] Comparative Example 2

[0045] The only difference from Example 1 is that the material-liquid ratio in step 1 is 1 g:15 mL.

[0046] Comparative Example 3

[0047] The only difference from Example 1 is that the pH value in step 2 is 9.

[0048] Comparative Example 4

[0049] The only difference from Example 1 is that the pH value in step 2 is 10.

[0050] Comparative Example 5

[0051] The only difference from Example 1 is that the water bath temperature in step 2 is 30°C.

[0052] Comparative Example 6

[0053] The only difference from Example 1 is that the water bath temperature in step 2 is 70°C.

[0054] Comparative Example 7

[0055] The only difference from Example 1 is that the insulation time in step 2 is 1 h.

[0056] Comparative Example 8

[0057] The only difference from Example 1 is that the insulation time in step 2 is 1.5 h.

[0058] Figure 1 The following are the curves of the extraction rate of Hericium erinaceus protein when the solid-liquid ratio is 1g:25mL, 1g:20mL, 1g:30mL, 1g:10mL and 1g:15mL for Examples 1 to 3 and Comparative Examples 1 to 2, respectively. It can be seen that, at first, as the solid-liquid ratio increases, the extraction rate of Hericium erinaceus protein also increases accordingly. This is because the contact area between the protein and the water molecules increases, which promotes the interaction between the molecules and is beneficial to the dissolution of the protein. However, when the solid-liquid ratio reaches 1g:25mL, the extraction rate decreases instead. This is because the combination of the protein and the water molecules in the solution has reached saturation. In short, when the solid-liquid ratio of Hericium erinaceus powder and distilled water is 1g:(20-30mL), the extraction rate of Hericium erinaceus protein can reach more than 30%, and when the solid-liquid ratio is 1g:25mL, the extraction rate of Hericium erinaceus protein reaches 36.65%, indicating that 1g:25mL is the best solid-liquid ratio.

[0059] Figure 2The following are the curves of the extraction rate of Hericium erinaceus protein when the pH values ​​of the Hericium erinaceus powder suspension are 12, 11, 13, 9 and 10 for Example 1, Example 4 to Example 5 and Comparative Examples 3 to Comparative Examples 4, respectively. It can be seen that at first the extraction rate of Hericium erinaceus protein increases with the increase of pH value. This is because in an alkaline environment, negatively charged protein molecules are more likely to attract protons, and proteins are more likely to combine with water molecules to form a hydration film, increasing protein solubility and facilitating protein dissolution. However, when the pH value is higher than 12, protein denaturation will occur, causing the protein to lose its activity, thereby resulting in a decrease in the extraction rate. In short, when the pH value is 11 to 13, the extraction rate of Hericium erinaceus protein can reach more than 33%, and when the pH value is 12, the extraction rate of Hericium erinaceus protein reaches 41.44%, indicating that an alkaline environment with a pH value of 12 is more conducive to the improvement of the extraction rate of Hericium erinaceus protein.

[0060] Figure 3 The extraction rate variation curves of Hericium erinaceus protein are shown in Example 1, Example 6 to Example 7 and Comparative Examples 5 to Comparative Examples 6, respectively, when the water bath temperature of the Hericium erinaceus powder suspension is 60°C, 40°C, 50°C, 30°C and 70°C. It can be seen that, at first, the protein extraction rate increases with increasing temperature. This is because the increase in temperature will activate the protein molecular structure and enhance the interaction between protein and water molecules, thereby increasing the solubility of the protein and promoting protein dissolution. However, as the temperature continues to rise to above 60°C, the solubility of the protein will decrease. This is because too high a temperature will cause the protein to denature, thereby reducing the extraction rate. In short, when the water bath temperature is 40°C to 60°C, the extraction rate of Hericium erinaceus protein can reach more than 18.5%, and when the water bath temperature is 60°C, the extraction rate of Hericium erinaceus protein reaches 20.08%, indicating that the optimal water bath temperature is 60°C.

[0061] Figure 4 The extraction rate variation curves of Hericium erinaceus protein are shown in Example 1, Example 8 to Example 9 and Comparative Examples 7 to Comparative Examples 8 when the holding time of the Hericium erinaceus powder suspension is 2.5h, 2h, 3h, 1h and 1.5h, respectively. It can be seen that with the increase of the holding time, the protein extraction rate first increases and then decreases. This may be due to the insufficient combination of protein and water at the beginning of the experiment, and only a small amount of protein interacts with water molecules. With the increase of time, the protein molecular structure gradually unfolds, and the internal hydrophilic groups are exposed to the outside, thereby increasing the solubility of the protein and improving the protein extraction rate. In short, when the holding time is 2-3h, the extraction rate of Hericium erinaceus protein can reach more than 25%, and when the holding time is 2.5h, the extraction rate of Hericium erinaceus protein reaches 28.10%, indicating that the optimal holding time is 2.5h.

[0062] Therefore, Example 1 selected the optimal experimental parameters, and carried out three parallel experiments under the conditions of a solid-liquid ratio of 1 g: 25 mL, a pH value of 12, a water bath temperature of 60 and a time of 2.5 h. The average extraction rate of Hericium erinaceus protein was calculated to be 60.98 ± 1.2%. At the same time, the solubility of the extracted Hericium erinaceus protein reached 61.06%.

Claims

1. A method for extracting Hericium erinaceus protein with a high extraction rate, characterized in that: The steps include: Step 1, according to the material-liquid ratio of 1g: (20-30mL), take Hericium erinaceus powder and add distilled water to obtain Hericium erinaceus powder suspension; Step 2, adjusting the pH value of the Hericium erinaceus powder suspension to 11-13 with a NaOH solution, and then keeping it warm for 2-3 hours in a water bath at 40-60° C., and after cooling, filtering and collecting the filtrate, adjusting the pH value of the filtrate to the isoelectric point of 4.5 with an HCl solution, and then centrifuging and separating the precipitate, and freeze-drying to obtain Hericium erinaceus protein.

2. The method for extracting Hericium erinaceus protein with a high extraction rate according to claim 1, characterized in that: The material-liquid ratio of step 1 is 1 g:25 mL.

3. The method for extracting Hericium erinaceus protein with a high extraction rate according to claim 1, characterized in that: The concentrations of the NaOH solution and the HCl solution in step 2 are both 0.1 mol / L.

4. The method for extracting Hericium erinaceus protein with a high extraction rate according to claim 1, characterized in that: The pH value in step 2 is 12.

5. The method for extracting Hericium erinaceus protein with a high extraction rate according to claim 1, characterized in that: The water bath temperature in step 2 is 60° C. and the insulation time is 2.5 h.

6. The method for extracting Hericium erinaceus protein with a high extraction rate according to claim 1, characterized in that: The centrifugal separation in step 2 is performed using a centrifuge with a rotation speed of 5000 r / min.

Images