Auricularia auricula-judae polypeptide, preparation method and application for reducing blood pressure
The black fungus polypeptide prepared by fermentation solves the side effects of existing ACE inhibitors, achieving safe and effective blood pressure reduction and anti-fatigue effects, and providing a food-derived alternative to ACE inhibitors.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- INST OF MICROBIOLOGY HEILONGJIANG ACADEMY OF SCI
- Filing Date
- 2025-03-31
- Publication Date
- 2026-07-07
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Abstract
Description
Technical Field
[0001] This invention relates to the field of biotechnology, and more specifically, to a black fungus polypeptide, its preparation method, and its application in lowering blood pressure. Background Technology
[0002] Currently, the main drugs used clinically to treat hypertension include diuretics, beta-blockers, calcium channel blockers, ACE inhibitors, and angiotensin receptor blockers, with ACE inhibitors being the most widely used. Traditional ACE inhibitors are synthetic drugs, such as enalapril and captopril. While these drugs are highly effective, they are often accompanied by side effects such as dizziness, cough, nausea, and increased heart rate. Therefore, highly effective, stable, and safe ACE inhibitors have become a new research trend in the food and pharmaceutical fields. From the perspective of edibility and safety, food-based bioactive peptides are becoming increasingly important, and numerous studies have shown that food-derived protein hydrolysates have a regulatory effect on blood pressure levels in animals.
[0003] In view of this, the present invention provides a black fungus polypeptide, its preparation method, and its application in lowering blood pressure. Summary of the Invention
[0004] The purpose of this invention is to provide a black fungus polypeptide, its preparation method, and its application in lowering blood pressure.
[0005] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:
[0006] In a first aspect, the present invention provides a compound edible fungus polypeptide derived from the fruiting body of black fungus and the mycelium of oyster mushroom, wherein the compound edible fungus polypeptide is obtained by fermentation of the fruiting body of black fungus and the mycelium of oyster mushroom.
[0007] Furthermore, the black fungus fruiting body and oyster mushroom mycelium complex edible fungi polypeptides are fermented using a method that involves inoculating the mother culture of Bifidobacterium breve into a liquid culture and then fermenting it.
[0008] Secondly, the present invention provides a method for preparing compound edible fungi polypeptides. The method involves inoculating a mixture of black fungus fruiting body and oyster mushroom mycelium extract residue with soybean meal, followed by fermentation and enzymatic hydrolysis to obtain compound edible fungi polypeptides.
[0009] Furthermore, the five steps of the preparation method are: (1) reflux extraction of black fungus fruiting bodies and oyster mushroom mycelium; (2) adding water and soybean meal to the extraction residue, inoculating with Bifidobacterium breve, and culturing in a sealed environment; (3) enzymatic hydrolysis; and (4) ultrafiltration.
[0010] Further, step (1) of the preparation method is as follows: mix the fruiting body of black fungus and the mycelial powder of oyster mushroom in a weight ratio of 3:1, add 85-95% ethanol, the material-liquid ratio is 1:12-18, and reflux extract twice, each time for 4 hours.
[0011] Further, step (2) of the preparation method is as follows: after drying the extract residue of black fungus fruiting body and oyster mushroom mycelium at 60°C, add 12 to 18 times the weight of water and 0.4 to 0.6 times the weight of soybean meal, adjust the pH value to 6.5 to 6.9, sterilize at 121°C for 20 min, inoculate with Bifidobacterium breve at 37 to 41°C, and culture in a sealed container for 2 to 4 days.
[0012] Further, step (3) of the preparation method is as follows: add 1000-1500 U / L of cellulase and 800-1000 U / L of subtilisin, at 37-39°C, pH 6.8-7.2, and for 7-9 hours of enzymatic hydrolysis.
[0013] Further, step (4) of the preparation method is as follows: the supernatant is ultrafiltered with an ultrafiltration membrane with a pore size of 2000 Daltons, and the filtrate is retained with a nanofiltration membrane with a pore size of 500 Daltons to obtain the polypeptide.
[0014] Furthermore, the specific steps of this preparation method are as follows: Mix fine powders of black fungus fruiting bodies and oyster mushroom mycelium at a weight ratio of 3:1, add 85-95% ethanol (solid-liquid ratio 1:12-18), reflux extract twice, 4 hours each time, filter, and recover the ethanol to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium; dry the extract residue of black fungus fruiting bodies and oyster mushroom mycelium at 60℃, add 12-18 times the weight of water and 0.4-0.6 times the weight of soybean meal, and adjust the pH to 6.5-6. 9. Sterilize at 121℃ for 20 min, inoculate with Bifidobacterium breve at 37-41℃, and culture in a sealed container for 2-4 days. Add 1000-1500 U / L cellulase and 800-1000 U / L subtilisin, and culture at 37-39℃ with pH 6.8-7.2 for 7-9 hours. Centrifuge at 4000 r / min for 20 min, collect the supernatant, and ultrafilter using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate is then filtered through a nanofiltration membrane with a pore size of 500 Daltons to obtain the polypeptide.
[0015] Furthermore, the specific steps of this preparation method are as follows: Black fungus fruiting bodies and Pleurotus ostreatus mycelium powder are mixed at a weight ratio of 3:1, 90% ethanol is added, the material-to-liquid ratio is 1:15, and the mixture is refluxed twice, 4 hours each time. After filtration, the ethanol is recovered to obtain an ethanol extract of black fungus fruiting bodies and Pleurotus ostreatus mycelium. The extract residue of black fungus fruiting bodies and Pleurotus ostreatus mycelium is dried at 60℃, then 15 times its weight of water and 0.5 times its weight of soybean meal are added, the pH is adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. Cellulase 1200 U / L and Bacillus subtilis protease 900 U / L are added, and the mixture is hydrolyzed at 38℃, pH 7.0, for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant is collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate is then retained using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0016] Thirdly, the present invention also provides the application of a complex edible fungus polypeptide of black fungus fruiting body and oyster mushroom mycelium in the preparation of a drug for the prevention or treatment of hypertension and / or anti-fatigue.
[0017] Compared with the prior art, the beneficial effects of the present invention include:
[0018] Compared with existing technologies, the complex edible fungi polypeptides of black fungus fruiting bodies and oyster mushroom mycelium and their preparation methods have been significantly improved. Detailed Implementation
[0019] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.
[0020] The features and performance of the present invention will be further described in detail below with reference to embodiments:
[0021] Example 1
[0022] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 85% ethanol was added. The material-to-liquid ratio was 1:18. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residue of black fungus fruiting bodies and oyster mushroom mycelium was dried at 60℃ and then mixed with 12 times the weight of water and 0.4 times the weight of soybean meal. The pH was adjusted to 6.9, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 37℃, and cultured in a sealed container for 4 days. Cellulase 1000 U / L and Bacillus subtilis protease 1000 U / L were added. The mixture was enzymatically hydrolyzed at 37℃, pH 7.2, for 7 hours, and centrifuged at 4000 r / min for 20 minutes. The supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0023] Example 2
[0024] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 95% ethanol was added. The material-to-liquid ratio was 1:12. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residue of black fungus fruiting bodies and oyster mushroom mycelium was dried at 60℃ and then 18 times its weight of water and 0.6 times its weight of soybean meal were added. The pH was adjusted to 6.5, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 41℃, and cultured in a sealed container for 2 days. Cellulase 1500 U / L and Bacillus subtilis protease 800 U / L were added. The mixture was enzymatically hydrolyzed at 39℃, pH 6.8, for 9 hours, and centrifuged at 4000 r / min for 20 minutes. The supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0025] Example 3
[0026] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residue of black fungus fruiting bodies and oyster mushroom mycelium was dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. Cellulase 1200 U / L and Bacillus subtilis protease 900 U / L were added. The mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0027] Comparative Example 1
[0028] Fine powders of *Auricularia auricula-judae* mycelium and *Pleurotus ostreatus* fruiting bodies were mixed at a weight ratio of 3:1, and 90% ethanol was added. The solid-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of *Auricularia auricula-judae* mycelium and *Pleurotus ostreatus* fruiting bodies. The extract residues were dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, and the mixture was sterilized at 121℃ for 20 minutes. *Bifidobacterium breve* was inoculated at 39℃ and cultured in a sealed container for 3 days. Cellulase 1200 U / L and *Bacillus subtilis* protease 900 U / L were added. The mixture was hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0029] Comparative Example 2
[0030] Fine powders of *Auricularia auricula-judae* mycelium and *Pleurotus ostreatus* mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of *Auricularia auricula-judae* mycelium and *Pleurotus ostreatus* mycelium. The extract residue of *Auricularia auricula-judae* mycelium and *Pleurotus ostreatus* mycelium was dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with *Bifidobacterium breve* at 39℃, and cultured in a sealed container for 3 days. Cellulase 1200 U / L and Bacillus subtilis protease 900 U / L were added, and the mixture was enzymatically hydrolyzed at 38℃, pH 7.0, for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0031] Comparative Example 3
[0032] Fine powders of Auricularia auricula-judae fruiting bodies and Pleurotus ostreatus fruiting bodies were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of Auricularia auricula-judae and Pleurotus ostreatus fruiting bodies. The extract residues of Auricularia auricula-judae and Pleurotus ostreatus fruiting bodies were dried at 60℃, and 15 times the weight of water and 0.5 times the weight of soybean meal were added. The pH was adjusted to 6.7, and the mixture was sterilized at 121℃ for 20 minutes. Bifidobacterium breve was inoculated at 39℃ and cultured in a sealed container for 3 days. Cellulase 1200 U / L and Bacillus subtilis protease 900 U / L were added. The mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0033] Comparative Example 4
[0034] Fine powders of white fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of white fungus fruiting bodies and oyster mushroom mycelium. The extract residue of white fungus fruiting bodies and oyster mushroom mycelium was dried at 60℃ and then 15 times its weight of water and 0.5 times its weight of soybean meal were added. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. Cellulase 1200 U / L and Bacillus subtilis protease 900 U / L were added, and the mixture was enzymatically hydrolyzed at 38℃, pH 7.0, for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0035] Comparative Example 5
[0036] Black fungus fruiting bodies and oyster mushroom mycelium powder were mixed at a weight ratio of 3:1. 15 times the weight of water and 0.5 times the weight of soybean meal were added, and the pH was adjusted to 6.7. The mixture was sterilized at 121℃ for 20 min, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. Cellulase 1200 U / L and Bacillus subtilis protease 900 U / L were added, and the mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 min, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered through a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0037] Comparative Example 6
[0038] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice for 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residues of black fungus fruiting bodies and oyster mushroom mycelium were dried at 60℃ and then mixed with 15 times their weight of water, 1200 U / L of cellulase, and 900 U / L of Bacillus subtilis protease. The mixture was hydrolyzed at 38℃, pH 7.0, for 8 hours, and centrifuged at 4000 r / min for 20 min. The supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0039] Comparative Example 7
[0040] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residue of black fungus fruiting bodies and oyster mushroom mycelium was dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain peptides.
[0041] Comparative Example 8
[0042] The fruiting bodies of black fungus and the mycelial powder of oyster mushroom were mixed at a weight ratio of 3:1. 15 times the weight of water and 0.5 times the weight of soybean meal were added, the pH was adjusted to 6.7, sterilized at 121℃ for 20 min, centrifuged at 4000 r / min for 20 min, the supernatant was collected, and ultrafiltration was performed using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered through a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0043] Comparative Example 9
[0044] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residues of black fungus fruiting bodies and oyster mushroom mycelium were dried at 60℃, and 15 times the weight of water and 0.5 times the weight of glucose were added to adjust the pH to 6.7. The mixture was sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. Cellulase 1200 U / L and Bacillus subtilis protease 900 U / L were added. The mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0045] Comparative Example 10
[0046] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residue of black fungus fruiting bodies and oyster mushroom mycelium was dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium longum at 39℃, and cultured in a sealed environment for 3 days. Cellulase 1200 U / L and Bacillus subtilis protease 900 U / L were added. The mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0047] Comparative Example 11
[0048] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residues of black fungus fruiting bodies and oyster mushroom mycelium were dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. 900 U / L of Bacillus subtilis protease was added, and the mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0049] Comparative Example 12
[0050] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residues of black fungus fruiting bodies and oyster mushroom mycelium were dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. Cellulase 1200 U / L was added, and the mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0051] Comparative Example 13
[0052] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice, 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residue of black fungus fruiting bodies and oyster mushroom mycelium was dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. Hemicellulase 1200 U / L and Bacillus subtilis protease 900 U / L were added. The mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0053] Comparative Example 14
[0054] Fine powders of black fungus fruiting bodies and oyster mushroom mycelium were mixed at a weight ratio of 3:1, and 90% ethanol was added. The material-to-liquid ratio was 1:15. The mixture was refluxed twice for 4 hours each time. After filtration, the ethanol was recovered to obtain ethanol extracts of black fungus fruiting bodies and oyster mushroom mycelium. The extract residues of black fungus fruiting bodies and oyster mushroom mycelium were dried at 60℃ and then mixed with 15 times the weight of water and 0.5 times the weight of soybean meal. The pH was adjusted to 6.7, sterilized at 121℃ for 20 minutes, inoculated with Bifidobacterium breve at 39℃, and cultured in a sealed container for 3 days. Cellulase 1200 U / L and bromelain 900 U / L were added, and the mixture was enzymatically hydrolyzed at 38℃ and pH 7.0 for 8 hours. After centrifugation at 4000 r / min for 20 minutes, the supernatant was collected and ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate was then filtered using a nanofiltration membrane with a pore size of 500 Daltons to obtain polypeptides.
[0055] The beneficial effects of this invention are as follows:
[0056] 1. Product pressure drop
[0057] Spontaneously hypertensive rats (SHR), male, 6 weeks old, weighing 168.48±3.95 g.
[0058] SHR rats were randomly divided into a blank control group, a positive control group (captopril group), each of the example groups, and each of the comparative groups, with 10 rats in each group. All rats were acclimatized for one week before the experiment. One week later, drug administration began, with 1000 mg of captopril dissolved in 1000 ml of physiological saline. Each example group and each comparative group was administered 20 ml / kg / day of the corresponding polypeptide sample by gavage (equivalent to 500 mg / kg / day of converted black fungus fruiting body and oyster mushroom mycelium powder). The positive control group was administered an equal volume of captopril solution by gavage daily. The blank control group was administered an equal volume of physiological saline by gavage daily. Drug administration was performed once daily for 30 consecutive days. Systolic blood pressure was measured one hour after administration on the 30th day.
[0059] A non-invasive blood pressure measurement system was used in the experiment, and the systolic blood pressure of rats was measured using the tail pulse method. The Medlab non-invasive rat blood pressure measurement system was installed, the computer was turned on, and the biosignal acquisition software was started. The rats were heated in a constant temperature chamber to 34°C. The rats were then placed in a restraint device, and the pulse cap and pulse transducer were placed at the tail artery of the rat. The rats were kept still, and the measurement began once a pulse wave appeared.
[0060] The blood pressure reduction is shown in Table 1.
[0061] Table 1. Product voltage drop (X±SD)
[0062]
[0063] 2. Product's anti-fatigue effect
[0064] Mice, weighing 18–22 g, half male and half female, were acclimatized for 3 days and then randomly divided into a normal control group, each of the example groups, and each of the comparative groups, with 10 mice in each group. Each example group and each comparative group was administered 20 ml / kg / day of the corresponding polypeptide sample by gavage (corresponding to 500 mg / kg / day of converted black fungus fruiting body and oyster mushroom mycelial powder), while the normal control group was administered an equal volume of physiological saline by gavage daily. All groups had free access to food daily. This gavage treatment continued for 30 days. One hour after administration on day 30, a lead sheet weighing 5% of the mouse's body weight was wrapped around the base of each mouse's tail, and the mice were placed in a bucket of water at a temperature of (23±1)℃ and a depth of 30 cm for a swimming experiment. The time from the start of swimming to exhaustion (the mouse sinking underwater for 10 seconds and still unable to surface) was recorded as the weighted swimming time.
[0065] Table 2 Results of fatigue resistance test
[0066]
[0067] All aspects not described in detail in this invention are common knowledge that can be readily understood by those skilled in the art. Although the invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which are obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of this invention fall within the scope of protection claimed by this invention.
Claims
1. A method for preparing a complex edible fungus polypeptide from black fungus fruiting bodies and oyster mushroom mycelium, characterized in that, The specific steps of the preparation method are as follows: Mix fine powders of *Auricularia auricula-judae* fruiting bodies and *Pleurotus ostreatus* mycelium at a weight ratio of 3:1, add 85-95% ethanol (solid-liquid ratio 1:12-18), reflux extract twice, 4 hours each time, filter, and recover the ethanol to obtain an ethanol extract of *Auricularia auricula-judae* fruiting bodies and *Pleurotus ostreatus* mycelium; dry the extract residue of *Auricularia auricula-judae* fruiting bodies and *Pleurotus ostreatus* mycelium at 60℃, add 12-18 times the weight of water and 0.4-0.6 times the weight of soybean meal, adjust the pH to 6.5-6.9, and... Sterilize at 21℃ for 20 min, inoculate with Bifidobacterium breve at 37-41℃, and culture in a sealed container for 2-4 days. Add 1000-1500 U / L cellulase and 800-1000 U / L subtilisin, and culture at 37-39℃, pH 6.8-7.2 for 7-9 hours. Centrifuge at 4000 r / min for 20 min, collect the supernatant, and ultrafilter using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate is then filtered through a nanofiltration membrane with a pore size of 500 Daltons to obtain the polypeptide.
2. The application of the black fungus fruiting body and oyster mushroom mycelium complex edible fungus polypeptide prepared by the method described in claim 1 in the preparation of drugs for the prevention or treatment of hypertension and / or anti-fatigue.