Black fungus polypeptide as well as preparation method and application thereof in lowering blood pressure

Abstract

The invention provides a black fungus polypeptide as well as a preparation method and application thereof in lowering blood pressure, and belongs to the technical field of biology. On one hand, the invention provides a black fungus sporocarp and pleurotus geesteranus mycelium composite edible fungus polypeptide product; in the second aspect, the invention further provides a preparation method of the black fungus sporocarp and pleurotus geesteranus mycelium composite edible fungus polypeptide. In the third aspect, the invention further provides application of the black fungus sporocarp and pleurotus geesteranus mycelium composite edible fungus polypeptide in preparation of food or medicine with the effects of preventing or treating hypertension and / or resisting fatigue.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a black fungus polypeptide, a preparation method and an application thereof in lowering blood pressure. Background Art

[0018] At present, the drugs used in clinical treatment of hypertension are mainly diuretics, beta-blockers, calcium antagonists, ACE inhibitors, and angiotensin receptor blockers, among which ACE inhibitors are the most commonly used. Traditional ACE inhibitors are synthetic drugs, such as enalapril and captopril. While these drugs are effective, they are often accompanied by side effects, such as dizziness, cough, nausea, and increased heart rate. Therefore, efficient, stable, and safe ACE inhibitors have become a new research trend in the field of food and medicine. From the perspective of edibility and safety, food-based bioactive peptides are becoming more and more important. At present, a large number of studies have shown that food-derived protein hydrolysates have a regulatory effect on animal blood pressure levels.

[0019] In view of this, the present invention provides a black fungus polypeptide and a preparation method and application for lowering blood pressure. Summary of the invention

[0020] The purpose of the present invention is to provide a black fungus polypeptide and a preparation method and application thereof in lowering blood pressure.

[0021] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are adopted:

[0022] In a first aspect, the present invention provides a composite edible fungus polypeptide of black fungus fruiting bodies and Pleurotus geesteranus mycelium. The composite edible fungus polypeptide is obtained from black fungus fruiting bodies and Pleurotus geesteranus mycelium through a fermentation method.

[0023] Furthermore, the black fungus fruiting body and Pleurotus geesteranus mycelium composite edible fungus polypeptide is fermented by conventionally inoculating the mother strain of Bifidobacterium breve and then performing liquid fermentation.

[0024] In a second aspect, the present invention provides a method for preparing a composite edible fungus polypeptide, the preparation method comprising inoculating Bifidobacterium breve into a composition of black fungus fruiting bodies and Pleurotus geesteranus mycelium extract residues and soybean meal, fermenting and then enzymolyzing to obtain the composite edible fungus polypeptide.

[0025] Furthermore, the preparation method comprises five steps: (1) reflux extraction of black fungus fruiting bodies and Pleurotus geesteranus mycelium; (2) adding water and soybean meal to the extraction residue, inoculating Bifidobacterium breve, and conducting closed culture; (3) enzymatic hydrolysis; and (4) ultrafiltration.

[0026] Furthermore, step (1) of the preparation method is: mixing black fungus fruiting bodies and Pleurotus geesteranus mycelium powder in a weight ratio of 3:1, adding 85-95% ethanol, with a liquid-to-solid ratio of 1:12-18, and reflux extraction twice, each time for 4 hours.

[0027] Furthermore, step (2) of the preparation method is: drying the black fungus fruiting body and Pleurotus geesteranus mycelium extract residue at 60° C., adding 12 to 18 times the weight of water and 0.4 to 0.6 times the weight of soybean meal, adjusting the pH value to 6.5 to 6.9, sterilizing at 121° C. for 20 minutes, inoculating Bifidobacterium breve at 37 to 41° C., and culturing in a closed manner for 2 to 4 days.

[0028] Furthermore, step (3) of the preparation method is: adding 1000-1500 U / L of cellulase and 800-1000 U / L of subtilisin, 37-39° C., pH 6.8-7.2, and enzymolysis time 7-9 hours.

[0029] Furthermore, step (4) of the preparation method is: the supernatant is ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons, and the filtrate is retained by a nanofiltration membrane with a pore size of 500 Daltons to obtain the polypeptide.

[0030] Furthermore, the preparation method comprises the following specific steps: mixing black fungus fruiting bodies and Pleurotus geesteranus mycelium powder in a weight ratio of 3:1, adding 85-95% ethanol, with a liquid-to-solid ratio of 1:12-18, performing reflux extraction twice, each time for 4 hours, filtering, and recovering ethanol to obtain black fungus fruiting bodies and Pleurotus geesteranus mycelium ethanol extracts; drying the black fungus fruiting bodies and Pleurotus geesteranus mycelium extract residues at 60°C, adding 12-18 times the weight of water and 0.4-0.6 times the weight of soybean meal, and adjusting the pH value to 6.5-6. 9. Sterilize at 121°C for 20 min, inoculate Bifidobacterium breve at 37-41°C, culture in a closed manner for 2-4 days, add 1000-1500 U / L of cellulase and 800-1000 U / L of subtilisin, culture at 37-39°C, pH 6.8-7.2, enzymatic hydrolysis time 7-9 hours, centrifuge at 4000 r / min for 20 min, collect the supernatant, ultrafilter with an ultrafiltration membrane with a pore size of 2000 Daltons, and use a nanofiltration membrane with a pore size of 500 Daltons to retain the filtrate to obtain the polypeptide.

[0031] Furthermore, the preparation method comprises the following specific steps: mixing black fungus fruiting bodies and Pleurotus geesteranus mycelium powder in a weight ratio of 3:1, adding 90% ethanol, with a liquid-to-solid ratio of 1:15, refluxing extraction for 2 times, each time for 4 hours, filtering, and recovering ethanol to obtain black fungus fruiting bodies and Pleurotus geesteranus mycelium alcohol extracts; drying the black fungus fruiting bodies and Pleurotus geesteranus mycelium extract residues at 60°C, adding 15 times the weight of water and 0.5 times the weight of soybean meal, adjusting the pH value to 6.7, sterilizing at 121°C for 20 minutes, inoculating Bifidobacterium breve at 39°C, culturing in a closed manner for 3 days, adding 1200U / L of cellulase and 900U / L of subtilisin, 38°C, pH 7.0, enzymolysis time for 8 hours, centrifuging at 4000r / min for 20 minutes, collecting the supernatant, ultrafiltering with an ultrafiltration membrane with a pore size of 2000 Daltons, and retaining the filtrate with a nanofiltration membrane with a pore size of 500 Daltons to obtain a polypeptide.

[0032] In a third aspect, the present invention also provides an application of a composite edible fungus polypeptide of Auricularia auricula fruiting body and Pleurotus geesteranus mycelium in the preparation of food or medicine for preventing or treating hypertension and / or resisting fatigue.

[0033] Compared with the prior art, the beneficial effects of the present invention include:

[0034] Compared with the prior art, the composite edible fungus polypeptide of black fungus fruiting body and Pleurotus geesteranus mycelium and the preparation method thereof are significantly improved. DETAILED DESCRIPTION

[0035] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. If specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0036] The features and performance of the present invention are further described in detail below in conjunction with the embodiments:

[0037] Example 1

[0038] The black fungus fruit body and the pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 85% ethanol is added, the liquid-to-solid ratio is 1:18, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 12 times the weight of water and 0.4 times the weight of soybean meal are added, the pH value is adjusted to 6.9, sterilized at 121°C for 20 minutes, inoculated with breve bifidobacterium at 37°C, and cultured in a closed manner for 4 days, 1000U / L of cellulase and 1000U / L of subtilisin are added, 37°C, pH is 7.2, the enzymatic hydrolysis time is 7 hours, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0039] Example 2

[0040] The black fungus fruit body and the pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 95% ethanol is added, the liquid-to-solid ratio is 1:12, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 18 times the weight of water and 0.6 times the weight of soybean meal are added, the pH value is adjusted to 6.5, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 41°C, cultured in a closed manner for 2 days, 1500U / L of cellulase and 800U / L of subtilisin are added, the mixture is subjected to enzymatic hydrolysis at 39°C and pH 6.8 for 9 hours, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0041] Example 3

[0042] The black fungus fruit body and the pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with breve bifidobacterium at 39°C, and cultured in a closed manner for 3 days, 1200U / L of cellulase and 900U / L of subtilisin are added, the mixture is heated at 38°C, the pH is 7.0, the enzymolysis time is 8 hours, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0043] Comparative Example 1

[0044] The black fungus mycelium and the Pleurotus geesteranus fruiting body fine powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus mycelium and Pleurotus geesteranus ethanol extract; the black fungus mycelium and Pleurotus geesteranus fruiting body extraction residues are dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, and cultured in a closed manner for 3 days, 1200 U / L of cellulase and 900 U / L of subtilisin are added, the mixture is subjected to enzymolysis at 38°C and pH 7.0 for 8 hours, centrifuged at 4000 r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0045] Comparative Example 2

[0046] The black fungus mycelium and the pleurotus geesteranus mycelium fine powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain black fungus mycelium and pleurotus geesteranus mycelium alcohol extracts; the black fungus mycelium and pleurotus geesteranus mycelium extract residues are dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with breve bifidobacterium at 39°C, cultured in a closed manner for 3 days, 1200 U / L of cellulase and 900 U / L of subtilisin are added, the mixture is subjected to enzymolysis at 38°C and pH 7.0 for 8 hours, centrifuged at 4000 r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0047] Comparative Example 3

[0048] The black fungus fruit body and the Pleurotus geesteranus fruit body fine powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and the Pleurotus geesteranus fruit body alcohol extract; the black fungus fruit body and the Pleurotus geesteranus fruit body extraction residue are dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, and cultured in a closed manner for 3 days, 1200U / L of cellulase and 900U / L of subtilisin are added, 38°C, pH is 7.0, the enzymatic hydrolysis time is 8 hours, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0049] Comparative Example 4

[0050] The white fungus fruiting body and the pleurotus geesteranus mycelium fine powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the white fungus fruiting body and pleurotus geesteranus mycelium alcohol extract; the white fungus fruiting body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with breve bifidobacterium at 39°C, and cultured in a closed manner for 3 days, 1200U / L of cellulase and 900U / L of subtilisin are added, 38°C, pH is 7.0, the enzymatic hydrolysis time is 8 hours, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, ultrafiltered with an ultrafiltration membrane with a pore size of 2000 Daltons, and the filtrate is intercepted with a nanofiltration membrane with a pore size of 500 Daltons to obtain a polypeptide.

[0051] Comparative Example 5

[0052] The black fungus fruiting body and the Pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, and cultured in a closed manner for 3 days, 1200 U / L of cellulase and 900 U / L of subtilisin are added, the temperature is 38°C, the pH is 7.0, the enzymatic hydrolysis time is 8 hours, and the mixture is centrifuged at 4000 r / min for 20 minutes, the supernatant is collected, and ultrafiltration is performed using an ultrafiltration membrane with a pore size of 2000 Daltons. The filtrate is retained by a nanofiltration membrane with a pore size of 500 Daltons to obtain a polypeptide.

[0053] Comparative Example 6

[0054] The black fungus fruit body and the Pleurotus geesteranus mycelium fine powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and Pleurotus geesteranus mycelium ethanol extract; the black fungus fruit body and Pleurotus geesteranus mycelium extraction residue are dried at 60°C, 15 times the weight of water, 1200 U / L of cellulase, 900 U / L of subtilisin are added, 38°C, pH 7.0, enzymolysis time is 8 hours, centrifugation is performed at 4000 r / min for 20 minutes, the supernatant is collected, ultrafiltration is performed 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 a polypeptide.

[0055] Comparative Example 7

[0056] The black fungus fruit body and the Pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and Pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and Pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, cultured in a closed manner for 3 days, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, ultrafiltered with an ultrafiltration membrane with a pore size of 2000 Daltons, and the filtrate is intercepted with a nanofiltration membrane with a pore size of 500 Daltons to obtain a polypeptide.

[0057] Comparative Example 8

[0058] The black fungus fruiting body and the Pleurotus geesteranus mycelium powder were mixed in 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 value was adjusted to 6.7, sterilized at 121°C for 20 minutes, centrifuged at 4000 r / min for 20 minutes, the supernatant was collected, ultrafiltered with an ultrafiltration membrane with a pore size of 2000 Daltons, and the filtrate was retained by a nanofiltration membrane with a pore size of 500 Daltons to obtain the polypeptide.

[0059] Comparative Example 9

[0060] The black fungus fruit body and the pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of glucose are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, and cultured in a closed manner for 3 days, 1200U / L of cellulase and 900U / L of subtilisin are added, the mixture is subjected to enzymolysis at 38°C and pH 7.0 for 8 hours, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0061] Comparative Example 10

[0062] The black fungus fruit body and the pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with long bifidobacterium at 39°C, and cultured in a closed manner for 3 days, 1200U / L of cellulase and 900U / L of subtilisin are added, 38°C, pH is 7.0, the enzymatic hydrolysis time is 8 hours, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0063] Comparative Example 11

[0064] The black fungus fruit body and the pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, cultured in a closed manner for 3 days, 900 U / L of subtilisin is added, 38°C, pH is 7.0, the enzymatic hydrolysis time is 8 hours, centrifuged at 4000 r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0065] Comparative Example 12

[0066] The black fungus fruit body and the pleurotus geesteranus mycelium fine powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, cultured in a closed manner for 3 days, 1200 U / L of cellulase is added, 38°C, pH is 7.0, the enzymatic hydrolysis time is 8 hours, centrifuged at 4000 r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0067] Comparative Example 13

[0068] The black fungus fruit body and the pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, and cultured in a closed manner for 3 days, 1200 U / L of hemicellulase and 900 U / L of subtilisin are added, the mixture is subjected to enzymolysis at 38°C and pH 7.0 for 8 hours, centrifuged at 4000 r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0069] Comparative Example 14

[0070] The black fungus fruit body and the pleurotus geesteranus mycelium powder are mixed in a weight ratio of 3:1, 90% ethanol is added, the liquid-to-solid ratio is 1:15, reflux extraction is performed twice, each time for 4 hours, filtering, and recovering ethanol to obtain the black fungus fruit body and pleurotus geesteranus mycelium alcohol extract; the black fungus fruit body and pleurotus geesteranus mycelium extraction residue is dried at 60°C, 15 times the weight of water and 0.5 times the weight of soybean meal are added, the pH value is adjusted to 6.7, sterilized at 121°C for 20 minutes, inoculated with Bifidobacterium breve at 39°C, and cultured in a closed manner for 3 days, 1200U / L of cellulase and 900U / L of bromelain are added, the mixture is subjected to enzymolysis at 38°C and pH 7.0 for 8 hours, centrifuged at 4000r / min for 20 minutes, the supernatant is collected, 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 a polypeptide.

[0071] The beneficial effects of the present invention are as follows:

[0072] 1. Product pressure reduction

[0073] Spontaneously hypertensive rats (SHR), male, 6 weeks old, weighing 168.48±3.95g.

[0074] SHR rats were randomly divided into a blank group, a positive control group (captopril group), each group of the embodiment, and each group of the comparative example, with 10 rats in each group. All rats were adapted to feeding for 1 week before the experiment. Administration began 1 week later, and 1000 mg of captopril was dissolved in 1000 ml of normal saline. Each group of the embodiment and each group of the comparative example was gavaged with the corresponding polypeptide sample 20 ml / kg / day every day (corresponding to the converted black fungus fruiting body and Pleurotus geesteranus mycelium fine powder of 500 mg / kg / day). The positive control group was gavaged with an equal amount of captopril solution every day. The blank control group was gavaged with an equal amount of normal saline every day. The drug was administered once a day at regular intervals for 30 consecutive days, and the systolic blood pressure of the rats was measured 1 hour after administration on the 30th day.

[0075] The experiment used a non-invasive blood pressure measurement system and tail pulse method to measure the systolic blood pressure of rats. The Medlab rat non-invasive blood pressure measurement system was installed, the computer was turned on, and the biological signal acquisition software was started. The rat was heated in a constant temperature chamber. When it was heated to 34°C, the rat was placed in a fixator, and the pressure cuff and pulse transducer were placed on the rat's tail artery to keep it quiet. The measurement began when the pulse wave appeared.

[0076] The blood pressure reduction is shown in Table 1.

[0077] Table 1 Product voltage reduction (X±SD)

[0078]

[0079]

[0080] 2. Anti-fatigue effect of the product

[0081] Mice, weighing 18-22g, half male and half female, were randomly divided into a normal control group, each group of the embodiment and each group of the comparative example after adaptive feeding for 3 days, with 10 mice in each group. Each group of the embodiment and the comparative example was gavaged with the corresponding polypeptide sample 20ml / kg / day (corresponding to the converted black fungus fruiting body and Pleurotus geesteranus mycelium fine powder of 500mg / kg / day), the normal control group was gavaged with the same amount of physiological saline every day, and each group was free to eat every day. The gavage was continued for 30 days. One hour after the administration on the 30th day, the tail root of each mouse was rolled with a lead sheet weighing 5% of its own body weight, and placed in a bucket with a water temperature of (23±1)℃ and a water depth of 30cm for swimming experiment. The time from the beginning of the mouse's swimming to exhaustion (the mouse sank into the water for 10s and still could not rush to the surface of the water) was recorded as the weighted swimming time of the mouse.

[0082] Table 2 Anti-fatigue test results

[0083]

[0084]

[0085] The links not described in detail in the present invention are all common knowledge that can be selected by ordinary technicians in the field. Although the present invention has been described in detail above with general descriptions and specific implementation schemes, it is obvious to ordinary technicians in the field that some modifications or improvements can be made on the basis of the present invention. Therefore, these modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. A composite edible fungus polypeptide of black fungus fruiting body and Pleurotus geesteranus mycelium, characterized in that: The composite edible fungus polypeptide is obtained from black fungus fruiting bodies and Pleurotus geesteranus mycelium through a fermentation method.

2. The composite edible fungus polypeptide of black fungus fruiting body and Pleurotus geesteranus mycelium according to claim 1, characterized in that: The fermentation method is to carry out liquid fermentation after conventional inoculation of Bifidobacterium breve mother strain.

3. A method for preparing the composite edible fungus polypeptide according to claim 1, characterized in that: The preparation method comprises the following steps: inoculating Bifidobacterium breve into a composition of extract residues of Auricularia auricularia auricula and Pleurotus geesteranus mycelium and soybean meal, and performing enzymolysis after fermentation to obtain the composite edible fungus polypeptide.

4. The method for preparing a composite edible fungus polypeptide according to claim 3, characterized in that: The preparation method comprises five steps: (1) reflux extraction of black fungus fruiting bodies and Pleurotus geesteranus mycelium; (2) adding water and soybean meal to the extraction residue, inoculating Bifidobacterium breve, and conducting closed culture; (3) enzymatic hydrolysis; and (4) ultrafiltration.

5. The method for preparing a composite edible fungus polypeptide according to claim 4, characterized in that: The step (1) of the preparation method is: mixing black fungus fruiting bodies and Pleurotus geesteranus mycelium fine powder in a weight ratio of 3:1, adding 85-95% ethanol, with a liquid-to-solid ratio of 1:12-18, and reflux extraction twice, each time for 4 hours.

6. The method for preparing a composite edible fungus polypeptide according to claim 4, characterized in that: Step (2) of the preparation method comprises: drying the extract residue of black fungus fruiting bodies and Pleurotus geesteranus mycelium at 60° C., adding 12 to 18 times the weight of water and 0.4 to 0.6 times the weight of soybean meal, adjusting the pH value to 6.5 to 6.9, sterilizing at 121° C. for 20 minutes, inoculating Bifidobacterium breve at 37 to 41° C., and culturing in a closed manner for 2 to 4 days.

7. The method for preparing a composite edible fungus polypeptide according to claim 4, characterized in that: The step (3) of the preparation method is: adding 1000-1500 U / L of cellulase and 800-1000 U / L of subtilisin, 37-39°C, pH 6.8-7.2, and enzymolysis time 7-9 hours.

8. The method for preparing a composite edible fungus polypeptide according to claim 4, characterized in that: Step (4) of the preparation method is: the supernatant is ultrafiltered using an ultrafiltration membrane with a pore size of 2000 Daltons, and the filtrate is retained by a nanofiltration membrane with a pore size of 500 Daltons to obtain the polypeptide.

9. The method for preparing a composite edible fungus polypeptide according to claim 3, characterized in that: The preparation method comprises the following specific steps: mixing black fungus fruiting bodies and Pleurotus geesteranus mycelium powder in a weight ratio of 3:1, adding 85-95% ethanol, with a liquid-to-solid ratio of 1:12-18, performing reflux extraction twice, each time for 4 hours, filtering, and recovering ethanol to obtain black fungus fruiting bodies and Pleurotus geesteranus mycelium ethanol extracts; drying the black fungus fruiting bodies and Pleurotus geesteranus mycelium extract residues at 60° C., adding 12-18 times the weight of water and 0.4-0.6 times the weight of soybean meal, adjusting the pH value to 6.5-6.9, 1 Sterilize at 21°C for 20 min, inoculate Bifidobacterium breve at 37-41°C, culture in a closed manner for 2-4 days, add 1000-1500 U / L of cellulase and 800-1000 U / L of subtilisin, culture at 37-39°C, pH 6.8-7.2, enzymatic hydrolysis time 7-9 hours, centrifuge at 4000 r / min for 20 min, collect the supernatant, ultrafilter with an ultrafiltration membrane with a pore size of 2000 Daltons, and use a nanofiltration membrane with a pore size of 500 Daltons to retain the filtrate to obtain the polypeptide.

10. Use of the composite edible fungus polypeptide of Auricularia auricula fruiting body and Pleurotus geesteranus mycelium according to claim 1 in the preparation of food or medicine for preventing or treating hypertension and / or resisting fatigue.

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