Preparation method and application of oyster peptide for improving human fertility quality

Abstract

The invention discloses a preparation method and application of oyster peptide for improving human fertility quality. Fresh oysters are used as raw materials, protease hydrolysis is utilized to release small molecule peptides, and then separation, purification and activity research are performed to explore the good utilization value of the oysters in the aspect of improving the reproductive function; the invention aims at innovating an oyster peptide extraction method to obtain the oyster peptide with high biological activity and high absorptivity, and discloses and introduces the application of the oyster peptide in the aspect of high human fertility quality. Meanwhile, the oyster peptide has the effects of improving the reproductive function, supplementing nutrition, mineral substances, protein, vitamins and the like, can be added into food, health care products, medicines and the like, and is a multifunctional biological peptide.

Description

Technical Field

[0001] The invention belongs to the technical field of oyster peptide preparation, and in particular relates to a method for preparing oyster peptide for improving human reproductive quality and application. Background Art

[0002] Oysters are one of the first health-care foods listed as both medicine and food in my country. In ancient times, oyster meat was called "oyster yellow". The Compendium of Materia Medica records: "Oyster meat is sweet, warm and non-toxic. Cooked and eaten, it can cure deficiency, regulate the middle, relieve erysipelas and women's blood and qi. Eaten raw with ginger and vinegar, it can cure erysipelas, restlessness after drinking, and quench thirst." Modern scientific research has confirmed that oyster meat is rich in high-quality protein, glycogen, taurine, multiple vitamins, and trace elements such as zinc, selenium, iron, and iodine.

[0003] Oysters, as a food that can be used as both medicine and food, are not only a delicious seafood that is popular among people, but also have the effect of strengthening the body. Oyster powder extract can significantly reduce liver damage and the content of transaminase, reduce the degree of damage to liver cells, and can also effectively prevent the occurrence of various cardiovascular diseases such as hypertension, arteriosclerosis, and stroke caused by vascular endothelial damage. At the same time, the oyster polysaccharides contained in oyster peptides can enhance the body's immunity. Oyster peptides obtained through the deep processing of oyster meat, as an emerging functional food ingredient, are developing rapidly in the fields of health food and medicine.

[0004] Scientists are constantly working to extract ingredients from natural foods that are beneficial to human health. Oyster peptides are a rare source of high-quality peptides. Oyster meat has high nutritional value and is rich in nutrients such as protein, glycogen, amino acids, and trace elements. A large number of studies have shown that oyster meat extracts have multiple biological activities such as anti-oxidation, enhancing immunity, anti-fatigue, anti-skin photoaging, lowering blood pressure, lowering blood sugar, sobering up and protecting the liver. In recent years, people's attention and pursuit of human health and population fertility have increased day by day. The incidence of male infertility has increased year by year, and the fertility rate in various regions of the world has also shown a downward trend year by year. How to effectively prevent and treat male reproductive dysfunction has become a serious problem that people face and need to be solved urgently.

[0005] At present, there are few research reports on the protective effect of oyster meat extract on male reproductive damage, and the relevant mechanism of action is not very clear. Therefore, the present invention specifically studies the extraction of components in oyster peptides that improve reproductive function, and provides a method for preparing oyster peptides that improve human reproductive quality. Summary of the invention

[0006] In view of the above problems, the present invention provides a method for preparing oyster peptides and applications for improving human reproductive quality.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is: A method for preparing oyster peptides for improving human fertility quality comprises the following steps: A. Take fresh oysters, use an oyster knife to open their shells and take out the oyster meat; B. Rinse the oyster meat under clean water; C. Then soak the oyster meat in light salt water for a while and drain; D. Prepare oyster meat homogenate using a low-temperature high-speed homogenizer; E. Determine the total protein content in the oyster slurry by Kjeldahl method and calculate the protein concentration (wt%); F. Add 4 times the mass of water to the oyster homogenate to make the protein concentration in the homogenate about 15%; adjust the pH value to 7.5; G. Add Aspergillus niger seed solution (cell concentration 107 CFU·mL or more), inoculation volume 5%, fermentation tank liquid volume 20%. Fermentation culture at 30°C, rotation speed 140rpm for 50h; H. After the fermentation is completed, the fermentation liquid is centrifuged at 8000 rpm for 20 min and the supernatant is taken; I. The supernatant was heated to 85°C for 20 min to inactivate the enzyme activity; J. Cooling to below 40° C., filtering with a 0.25 μm microporous filter membrane, and taking the filtrate to obtain the Aspergillus niger fermented oyster protein hydrolyzate; K. Decolorization and deodorization: Add activated carbon with a liquid content of 15% to the filtrate, stir evenly, and let it stand for 4 hours to decolorize and deodorize; centrifuge to remove the activated carbon; L. The decolorized and deodorized oyster protein hydrolyzate is filtered through an ultrafiltration membrane with a molecular weight cutoff of 3000, and the filtrate is then filtered through an ultrafiltration membrane with a molecular weight cutoff of 1000. The filtrate obtained by the second filtration is the oyster small molecule peptide solution, which is freeze-dried; M. The lyophilized powder was dissolved in PBS with pH 7.2 and 0.1 mol·L-1 and separated by Sephadex LH 20 gel chromatography column; the received eluate was detected by RP-HPLC in combination with EMS to detect small molecule peptides and determine their molecular weight; the measured molecular weight was 500-1500.

[0008] The oyster peptide prepared by the above method can be added to food, health products and medicines.

[0009] Compared with the prior art, the advantages and positive effects of the present invention are: The present invention uses peptide molecular biotechnology enzymatic hydrolysis and biofermentation engineering technology to develop an oyster peptide product that has the effects of improving reproductive function, supplementing nutrition, supplementing minerals, proteins, vitamins, etc., to address the current reproductive quality problems and at the same time strengthen the body. This bioactive peptide can be added to food, health products, medicines, etc. through different processes; The present invention uses Aspergillus niger as the strain to ferment oyster meat homogenate, and extracts oyster small molecule peptides through processes such as enzyme inactivation, microporous membrane filtration, decolorization and deodorization, ultrafiltration membrane filtration, and gel chromatography column separation, which can increase serum testosterone levels, improve sperm quality, and improve sperm motility, and are used to regulate physiological functions, improve malnutrition or sexual dysfunction symptoms, and help enhance male reproductive function. DETAILED DESCRIPTION

[0010] In order to more clearly understand the above-mentioned purpose, features and advantages of the present invention, the present invention is further described below in conjunction with embodiments. It should be noted that the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

[0011] In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention may also be implemented in other ways than those described herein. Therefore, the present invention is not limited to the specific embodiments of the following disclosure.

[0012] Example 1, the present invention discloses a method for preparing oyster peptides for improving human reproductive quality and its application. The present invention uses fresh oysters as raw materials, utilizes protease hydrolysis to release small molecule peptides, and then separates and purifies them and studies their activity to explore their good utilization value in improving reproductive function; the present invention intends to innovate the oyster peptide extraction method, obtain oyster peptides with high biological activity and high absorption rate, and publicly introduce the application of oyster peptides in improving human reproductive quality. At the same time, oyster peptides have the effects of improving reproductive function, supplementing nutrition, supplementing minerals, proteins, vitamins, etc., and can be added to food, health products, medicines, etc., and are a multifunctional biological peptide.

[0013] The present invention provides a method for preparing oyster peptide and its application. To achieve the above-mentioned purpose, the present invention adopts the following technical scheme: (1) Take fresh oysters, use an oyster knife to open their shells, take out the oyster meat, and rinse it under clean water.

[0014] (2) Soak the cleaned oyster meat in light salt water for a while and then drain.

[0015] (3) Prepare oyster meat homogenate using a low-temperature high-speed homogenizer.

[0016] (4) Prepare fermentation broth.

[0017] a. The total protein content in the oyster slurry was determined by Kjeldahl method, and the protein concentration (wt%) was calculated.

[0018] b. Add 4 times the mass of water to the oyster homogenate to make the protein concentration in the homogenate about 15%, and adjust the pH to 7.5.

[0019] c. Add Aspergillus niger seed liquid.

[0020] Among them, the bacterial concentration is above 107 CFU·mL and the inoculation amount is 5%.

[0021] (5) Fermentation.

[0022] a. Fermentation conditions: Fermentation tank liquid volume 20% The fermentation was carried out at 30°C and 140 rpm for 50 h.

[0023] (6) After fermentation is completed, centrifuge the fermentation liquid and take the supernatant.

[0024] a. Centrifugation conditions: 8000rpm, 20min (7) The obtained supernatant is heated to inactivate the enzyme activity, cooled to below 40°C, filtered, and the filtrate is collected to obtain the Aspergillus niger fermented oyster protein hydrolyzate.

[0025] a. Heating conditions: 85°C, 20 min b. Use 0.25μm microporous filter membrane to filter the cooled liquid (8) Decolorization and deodorization.

[0026] a. Add activated carbon with a liquid content of 15% to the filtrate and stir b. After stirring evenly, let it stand for 4 hours to decolorize and remove the fishy smell. c. Centrifugation to remove activated carbon (9) The decolorized and deodorized oyster protein hydrolyzate is filtered twice using an ultrafiltration membrane, and the resulting filtrate is an oyster small molecule peptide solution.

[0027] a. Filter the first time with an ultrafiltration membrane with a molecular weight cutoff of 3000 and take the filtrate b. Filter the final (10) The obtained solution is freeze-dried, and the oyster small molecule peptide is detected and its molecular weight is determined.

[0028] a. The lyophilized powder was dissolved in PBS (pH 7.2, 0.1 mol·L-1) and separated using a Sephadex LH 20 gel column.

[0029] b. The received eluate was used to detect small molecule peptides by RP-HPLC combined with EMS and to determine their molecular weight. The molecular weight was measured to be 500-1500.

[0030] The above description is only a preferred embodiment of the present invention and does not limit the present invention in other forms. Any technician familiar with the profession may use the technical content disclosed above to change or modify it into an equivalent embodiment with equivalent changes and apply it to other fields. However, any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present invention without departing from the content of the technical solution of the present invention still falls within the protection scope of the technical solution of the present invention.

Claims

1. A method for preparing oyster peptides for improving human fertility quality, characterized in that: The specific steps include: A. Take fresh oysters, use an oyster knife to open their shells and take out the oyster meat; B. Rinse the oyster meat under clean water; C. Then soak the oyster meat in light salt water for a while and drain; D. Prepare oyster meat homogenate using a low-temperature high-speed homogenizer; E. Determine the total protein content in the oyster slurry by Kjeldahl method and calculate the protein concentration (wt%); F. Add 4 times the mass of water to the oyster homogenate to make the protein concentration in the homogenate about 15%; adjust the pH value to 7.5; G. Add Aspergillus niger seed solution (cell concentration 107 CFU·mL or more), inoculation volume 5%, fermentation tank liquid volume 20%. Fermentation culture at 30°C, rotation speed 140rpm for 50h; H. After the fermentation is completed, the fermentation liquid is centrifuged at 8000 rpm for 20 min and the supernatant is taken; I. The supernatant was heated to 85°C for 20 min to inactivate the enzyme activity; J. Cooling to below 40° C., filtering with a 0.25 μm microporous filter membrane, and taking the filtrate to obtain the Aspergillus niger fermented oyster protein hydrolyzate; K. Decolorization and deodorization: Add activated carbon with a liquid content of 15% to the filtrate, stir evenly, and let it stand for 4 hours to decolorize and deodorize; Centrifuge to remove activated carbon; L. The decolorized and deodorized oyster protein hydrolyzate is filtered through an ultrafiltration membrane with a molecular weight cutoff of 3000, and the filtrate is then filtered through an ultrafiltration membrane with a molecular weight cutoff of 1000. The filtrate obtained by the second filtration is the oyster small molecule peptide solution, which is freeze-dried; M. The lyophilized powder was dissolved in PBS with pH 7.2 and 0.1 mol·L-1 and separated by Sephadex LH 20 gel chromatography column; the received eluate was detected by RP-HPLC in combination with EMS to detect small molecule peptides and determine their molecular weight; the measured molecular weight was 500-1500.

2. The oyster peptide prepared by the method of claim 1 can be added to food, health products and medicines.