阅读说明：本技术 一种核桃小分子多肽的发酵制备方法 (Fermentation preparation method of walnut micromolecule polypeptide ) 是由 盛军 赵存朝 王娅 刘佳 彭磊 于 2021-01-27 设计创作，主要内容包括：本发明公开了核桃小分子多肽发酵制备方法技术领域的一种核桃小分子多肽的发酵制备方法,将核桃仁利用低温物理压榨技术进行压榨,压榨完成后可得到核桃油和核桃粕两种产物,将核桃粕收集后将这些核桃粕导入到粉碎机中进行研磨粉碎处理,经粉碎后过40目筛,然后准备若干个250mL烧杯,每个烧杯盛装40.00g核桃粕,然后将这些烧杯放置在灭菌条件下进行无菌操作添加无菌水,利用乳酸菌结合生物发酵技术进行固态发酵,且针对多肽发酵的影响条件进行优化处理,对每一组试验对象进行多肽的产量检测并进行分别记录,以确定影响多肽发酵的因素,能够确定多肽产量最多的最佳条件,提高多肽发酵产量。(The invention discloses a fermentation preparation method of walnut micromolecule polypeptide in the technical field of fermentation preparation methods of walnut micromolecule polypeptide, which comprises the steps of squeezing walnut kernels by using a low-temperature physical squeezing technology to obtain two products of walnut oil and walnut dregs after squeezing is finished, collecting the walnut dregs, guiding the walnut dregs into a grinder to be ground and crushed, sieving the crushed walnut dregs by a 40-mesh sieve, preparing a plurality of 250mL beakers, containing 40.00g of walnut dregs, placing the beakers under a sterilization condition to perform aseptic operation and adding aseptic water, performing solid state fermentation by using lactic acid bacteria in combination with a biological fermentation technology, optimizing the influence conditions of polypeptide fermentation, detecting and respectively recording the output of polypeptide of each group of test objects to determine the factors influencing the polypeptide fermentation, and determining the optimal conditions of the maximum output of the polypeptide, the polypeptide fermentation yield is improved.)

1. A fermentation preparation method of walnut micromolecule polypeptide is characterized by comprising the following steps: the fermentation method of the walnut micromolecule polypeptide comprises the following steps: firstly, preprocessing walnut kernels, namely squeezing the walnut kernels by using a low-temperature physical squeezing technology to obtain two products of walnut oil and walnut meal after squeezing, separating the two products, collecting the walnut oil for eating, guiding the walnut meal into a grinder for grinding and smashing treatment, sieving the walnut meal with a 40-mesh sieve after smashing, then preparing a plurality of 250mL beakers, containing 40.00g of walnut meal in each beaker, then placing the beakers under a sterilization condition for aseptic operation and adding sterile water, inoculating fermentation seed liquid according to the inoculation amount, stirring and stirring for 5 minutes by using a glass rod to mix and mix materials in the beakers uniformly, then sealing the beakers by using gauze, and performing solid state fermentation by using lactic acid bacteria in combination with a biological fermentation technology to complete the process.

2. The method for preparing the walnut small molecule polypeptide by fermentation according to claim 1, which is characterized in that: the method for preparing the walnut micromolecule polypeptide by fermentation comprises the following specific steps:

the method comprises the following steps: taking out the sample after the fermentation in the claim 1, and placing the sample in an oven to dry for 48 hours at the temperature of 55 ℃;

step two: after drying, crushing the materials, and then sieving the materials by a 40-mesh sieve to be used as a basic raw material for a test for later use;

step three: the fermentation time (1, 2, 3, 4, 5, 6, 7, 8 and 9 days) is taken as a single research parameter, the optimal fermentation time parameter is researched, the test result is detected after the respective fermentation time, the detection result takes the yield of the polypeptide as an index, the yield of the polypeptide is respectively recorded after the fermentation time is finished each time, and the optimal fermentation time is 3 days after the test;

step four: after the fermentation time is determined (3 days), the optimal raw material particle size fermentation parameter is researched by taking the raw material crushed particle size (not crushed, 20, 40 and 60 meshes) as a single research parameter under the condition of the optimal fermentation time, the test result is detected after the fermentation time of 3 days, the detection result takes the yield of the polypeptide as an index, the yield of the polypeptide is detected and respectively recorded for each group of test objects, and the test shows that the optimal fermentation raw material particle size is 40 meshes under the condition of the optimal fermentation time of 3 days;

step five: after the fermentation time (3 days) and the particle size of the raw material (40-mesh sieve) are determined, grouping tests are carried out under the conditions by taking the fermentation temperatures (28, 31, 34 and 37 ℃) as single research parameters to research the optimal fermentation temperature parameters, test results are detected after the tests are finished, the test results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and recorded for each group of test objects, and the test shows that the optimal fermentation temperature is 28 ℃ under the conditions of the fermentation time (3 days) and the particle size of the raw material (40-mesh sieve);

step six: after the fermentation time (3 days), the raw material grain size (40-mesh sieve) and the fermentation temperature (28 ℃) are determined, grouping tests are carried out under the conditions by taking the inoculation amount (5, 10, 15, 20 and 25%) as a single research parameter to research the optimal fermentation inoculation amount, test results are detected after the tests are finished, the test results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and recorded for each group of test objects, and the optimal fermentation inoculation amount is 5% under the conditions of the fermentation time (3 days), the raw material grain size (40-mesh sieve) and the fermentation temperature (28 ℃);

step seven: after determining the fermentation time (3 days), the raw material particle size (40 mesh screen), the fermentation temperature (28 degrees) and the fermentation inoculum size (5%), the composite strain ratio was determined under these conditions-lactobacillus plantarum: streptococcus thermophilus: bacillus subtilis (2:2:1, 2:1:1, 1:1:2 and 1:2:2) is a single research parameter grouping test to research the optimal fermentation inoculation amount, test results are detected after the tests are finished, the detection results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and recorded for each group of test objects, and the optimal ratio of lactobacillus is 1:1:1 under the conditions of fermentation time (3 days), raw material particle size (40 meshes), fermentation temperature (28 ℃) and fermentation inoculation amount (5%);

step eight: after the fermentation time (3 days), the raw material grain size (40 meshes), the fermentation temperature (28 degrees), the fermentation inoculation amount (5 percent) and the lactic acid bacteria ratio (1:1:1), under the conditions, the grain-liquid ratio (1:0.5, 1:0.75, 1:1, 1:1.25 and 1:1.5) is taken as a single research parameter to carry out grouping tests to research the optimal fermentation inoculation amount, after the tests are finished, test results are detected, the test results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and respectively recorded for each group of test objects, and the test shows that the optimal grain-liquid ratio is 1:1.25 under the conditions of the fermentation time (3 days), the raw material grain size (40 meshes), the fermentation temperature (28 degrees), the fermentation inoculation amount (5 percent) and the lactic acid bacteria ratio (1:1: 1: 1);

step nine: when the walnut micromolecule polypeptide is prepared by fermentation, the optimal fermentation conditions are as follows: the fermentation time is 3 days, the grain size of the raw material is 40 meshes, the fermentation temperature is 28 ℃, the inoculation amount is 5%, the ratio of lactic acid bacteria is 1:1:1 and the ratio of material to liquid is 1:1.25, and under the condition, the polypeptide fermentation amount is the most.

3. The method for preparing the walnut small molecule polypeptide by fermentation according to claim 1, which is characterized in that: the walnut oil and the walnut meal are separated by the filter screen in a shaking mode, and the filter screen is kept still for a period of time, so that the walnut oil cannot be adhered to the filter screen, and the extraction rate of the walnut oil is improved.

4. The fermentation preparation method of walnut small molecule polypeptide according to claim 3, characterized in that: when the beaker is filled with walnut meal, a precise small balance is required for weighing; the sterilization conditions are controlled as follows: the sterilization temperature is 121 ℃, and the sterilization time is 20 minutes; the mouth of the beaker is sealed by a rubber band when the gauze is sealed.

5. The method for preparing the walnut small molecule polypeptide by fermentation according to claim 2, which is characterized in that: in the test process in the third step, fermentation time of a single factor is used as a variable, a table is registered, nine groups of polypeptide content test data under different fermentation times are obtained, a line drawing is carried out on the data, and the optimal fermentation time is visually judged.

6. The method for preparing the walnut small molecule polypeptide by fermentation according to claim 2, which is characterized in that: in the test process of the fourth step, the raw material crushed particle size of a single factor is used as a variable, a table is registered, four groups of polypeptide content test data under different raw material crushed particle sizes are obtained, a line drawing is carried out on the data, and the optimal raw material crushed particle size is visually judged.

7. The method for preparing the walnut small molecule polypeptide by fermentation according to claim 2, which is characterized in that: and step five, taking the fermentation temperature of a single factor as a variable in the test process, registering a table to obtain four groups of polypeptide content test data under different fermentation temperatures, drawing a line drawing on the data, and visually judging the optimal fermentation temperature.

8. The method for preparing the walnut small molecule polypeptide by fermentation according to claim 2, which is characterized in that: and step six, taking the inoculation amount of a single factor as a variable in the test process, registering a table, obtaining five groups of polypeptide content test data under different inoculation amounts, drawing a line drawing on the data, and visually judging the optimal inoculation amount.

9. The method for preparing the walnut small molecule polypeptide by fermentation according to claim 2, which is characterized in that: in the test process in the seventh step, the ratio of the composite strains of a single factor, namely lactobacillus plantarum: streptococcus thermophilus: the bacillus subtilis is used as a variable, a table is registered, five groups of polypeptide content experimental data under different composite strain proportions are obtained, a line drawing is drawn on the data, and the optimal composite strain proportion is visually judged.

10. The method for preparing the walnut small molecule polypeptide by fermentation according to claim 2, which is characterized in that: and step eight, in the test process, the material-liquid ratio of a single factor is used as a variable, a table is registered, five groups of polypeptide content test data under different fermentation times are obtained, a line drawing is carried out on the data, and the optimal material-liquid ratio is visually judged.

Technical Field

The invention relates to the technical field of walnut micromolecule polypeptide fermentation preparation methods, in particular to a fermentation preparation method of walnut micromolecule polypeptide.

Background

The insomnia is difficult to ensure sleep quality during night sleep, difficult to fall asleep or short in sleep time, light dreaminess and easy to wake up, and the main reason of insomnia is considered to be that nerves in brain are in an overstrain state or a mood fluctuation state for a long time, such as long-time work, high-intensity psychological pressure, frequent shift, overfatigue and the like, long-term insomnia causes listlessness, nerve disorder, immunity reduction, memory hypofunction and the like of a patient, and cerebral hemorrhage, hypertension, hemiplegia, diabetes and other diseases are more likely to occur, so that the death probability of the patient is increased, in order to relieve cerebral cortex and inhibit imbalance, a relaxation therapy and a drug therapy are usually adopted, wherein the best component for treating insomnia, namely small molecular polypeptide, can achieve good effects of enhancing immunity and soothing nerves and helping sleep, and plays an important role in treating insomnia, however, in the current fermentation method of walnut small molecular polypeptides in the market, the fermentation conditions are immature, the fermentation conditions are inaccurate, the fermentation yield of the small molecular polypeptides is small, and therefore the effect of the small molecular polypeptides cannot be fully exerted.

Disclosure of Invention

The invention aims to provide a fermentation preparation method of walnut small-molecule polypeptide, which aims to solve the problems in the background art.

In order to achieve the purpose, the invention provides the following technical scheme: a fermentation preparation method of walnut micromolecule polypeptide comprises the following steps: firstly, preprocessing walnut kernels, namely squeezing the walnut kernels by using a low-temperature physical squeezing technology to obtain two products of walnut oil and walnut meal after squeezing, separating the two products, collecting the walnut oil for eating, guiding the walnut meal into a grinder for grinding and smashing treatment, sieving the walnut meal with a 40-mesh sieve after smashing, then preparing a plurality of 250mL beakers, containing 40.00g of walnut meal in each beaker, then placing the beakers under a sterilization condition for aseptic operation and adding sterile water, inoculating fermentation seed liquid according to the inoculation amount, stirring and stirring for 5 minutes by using a glass rod to mix and mix materials in the beakers uniformly, then sealing the beakers by using gauze, and performing solid state fermentation by using lactic acid bacteria in combination with a biological fermentation technology to complete the process.

Preferably, the walnut small molecule polypeptide fermentation preparation method comprises the following specific steps:

the method comprises the following steps: taking out the sample after the fermentation in the claim 1, and placing the sample in an oven to dry for 48 hours at the temperature of 55 ℃;

step two: after drying, crushing the materials, and then sieving the materials by a 40-mesh sieve to be used as a basic raw material for a test for later use;

step three: the fermentation time (1, 2, 3, 4, 5, 6, 7, 8 and 9 days) is taken as a single research parameter, the optimal fermentation time parameter is researched, the test result is detected after the respective fermentation time, the detection result takes the yield of the polypeptide as an index, the yield of the polypeptide is respectively recorded after the fermentation time is finished each time, and the optimal fermentation time is 3 days after the test;

step four: after the fermentation time is determined (3 days), the optimal raw material particle size fermentation parameter is researched by taking the raw material crushed particle size (not crushed, 20, 40 and 60 meshes) as a single research parameter under the condition of the optimal fermentation time, the test result is detected after the fermentation time of 3 days, the detection result takes the yield of the polypeptide as an index, the yield of the polypeptide is detected and respectively recorded for each group of test objects, and the test shows that the optimal fermentation raw material particle size is 40 meshes under the condition of the optimal fermentation time of 3 days;

step five: after the fermentation time (3 days) and the particle size of the raw material (40-mesh sieve) are determined, grouping tests are carried out under the conditions by taking the fermentation temperatures (28, 31, 34 and 37 ℃) as single research parameters to research the optimal fermentation temperature parameters, test results are detected after the tests are finished, the test results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and recorded for each group of test objects, and the test shows that the optimal fermentation temperature is 28 ℃ under the conditions of the fermentation time (3 days) and the particle size of the raw material (40-mesh sieve);

step six: after the fermentation time (3 days), the raw material grain size (40-mesh sieve) and the fermentation temperature (28 ℃) are determined, grouping tests are carried out under the conditions by taking the inoculation amount (5, 10, 15, 20 and 25%) as a single research parameter to research the optimal fermentation inoculation amount, test results are detected after the tests are finished, the test results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and recorded for each group of test objects, and the optimal fermentation inoculation amount is 5% under the conditions of the fermentation time (3 days), the raw material grain size (40-mesh sieve) and the fermentation temperature (28 ℃);

step seven: after determining the fermentation time (3 days), the raw material particle size (40 mesh screen), the fermentation temperature (28 degrees) and the fermentation inoculum size (5%), the composite strain ratio was determined under these conditions-lactobacillus plantarum: streptococcus thermophilus: bacillus subtilis (2:2:1, 2:1:1, 1:1:2 and 1:2:2) is a single research parameter grouping test to research the optimal fermentation inoculation amount, test results are detected after the tests are finished, the detection results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and recorded for each group of test objects, and the optimal ratio of lactobacillus is 1:1:1 under the conditions of fermentation time (3 days), raw material particle size (40 meshes), fermentation temperature (28 ℃) and fermentation inoculation amount (5%);

step eight: after the fermentation time (3 days), the raw material grain size (40 meshes), the fermentation temperature (28 degrees), the fermentation inoculation amount (5 percent) and the lactic acid bacteria ratio (1:1:1), under the conditions, the grain-liquid ratio (1:0.5, 1:0.75, 1:1, 1:1.25 and 1:1.5) is taken as a single research parameter to carry out grouping tests to research the optimal fermentation inoculation amount, after the tests are finished, test results are detected, the test results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and respectively recorded for each group of test objects, and the test shows that the optimal grain-liquid ratio is 1:1.25 under the conditions of the fermentation time (3 days), the raw material grain size (40 meshes), the fermentation temperature (28 degrees), the fermentation inoculation amount (5 percent) and the lactic acid bacteria ratio (1:1: 1: 1);

step nine: when the walnut micromolecule polypeptide is prepared by fermentation, the optimal fermentation conditions are as follows: the fermentation time is 3 days, the grain size of the raw material is 40 meshes, the fermentation temperature is 28 ℃, the inoculation amount is 5%, the ratio of lactic acid bacteria is 1:1:1 and the ratio of material to liquid is 1:1.25, and under the condition, the polypeptide fermentation amount is the most.

Preferably, the walnut oil and the walnut meal are separated by a filter screen in a shaking mode, and the filter screen is kept still for a period of time, so that the walnut oil cannot be adhered to the filter screen, and the extraction rate of the walnut oil is improved.

Preferably, when the beaker is filled with walnut meal, a precise small balance is required for weighing; the sterilization conditions are controlled as follows: the sterilization temperature is 121 ℃, and the sterilization time is 20 minutes; the mouth of the beaker is sealed by a rubber band when the gauze is sealed.

Preferably, in the third step of the test process, the fermentation time of a single factor is used as a variable, a table is registered, nine groups of experimental data of polypeptide content under different fermentation times are obtained, a line drawing is carried out on the data, and the optimal fermentation time is visually judged.

Preferably, in the test process in the fourth step, the raw material crushed particle size of a single factor is used as a variable, a table is registered, four groups of polypeptide content test data under different raw material crushed particle sizes are obtained, a line drawing is performed on the data, and the optimal raw material crushed particle size is visually judged.

Preferably, in the test process in the fifth step, the fermentation temperature of a single factor is used as a variable, a table is registered, four groups of polypeptide content test data under different fermentation temperatures are obtained, a line drawing is performed on the data, and the optimal fermentation temperature is visually judged.

Preferably, in the test process in the sixth step, the inoculation amount of a single factor is used as a variable, a table is registered, five groups of polypeptide content test data under different inoculation amounts are obtained, a line drawing is performed on the data, and the optimal inoculation amount is visually judged.

Preferably, in the seventh test, the ratio of the single-element composite strains, lactobacillus plantarum: streptococcus thermophilus: the bacillus subtilis is used as a variable, a table is registered, five groups of polypeptide content experimental data under different composite strain proportions are obtained, a line drawing is drawn on the data, and the optimal composite strain proportion is visually judged.

Preferably, in the test process in the step eight, the material-liquid ratio of a single factor is used as a variable, a table is registered, five groups of polypeptide content test data under different fermentation times are obtained, a line drawing is performed on the data, and the optimal material-liquid ratio is visually judged.

Compared with the prior art, the invention has the beneficial effects that:

1. the walnut kernels are squeezed by a low-temperature physical squeezing technology, two products of walnut oil and walnut meal can be obtained after squeezing is completed, the walnut oil and the walnut meal are separated, the walnut oil is collected and eaten, the walnut meal is collected and then is led into a grinder to be ground and smashed, the walnut meal is sieved by a 40-mesh sieve after being smashed, then a plurality of 250mL beakers are prepared, each beaker contains 40.00g of walnut meal, then the beakers are placed under a sterilization condition to be subjected to aseptic operation and added with sterile water, fermentation seed liquid is inoculated according to the inoculation amount and is stirred for 5 minutes by a glass rod, the materials in the beakers are mixed and uniformly mixed, then the beakers are sealed by gauze, and solid state fermentation is carried out by utilizing lactic acid bacteria in combination with a biological fermentation technology;

2. the invention is different from the traditional method for preparing polypeptide by an enzyme method, provides a new way for preparing walnut polypeptide, optimizes the influence conditions of polypeptide fermentation, and can optimize the fermentation time (1, 2, 3, 4, 5, 6, 7, 8 and 9 days), the crushed particle size of raw materials (20, 40 and 60 meshes), the fermentation temperature (28, 31, 34 and 37 degrees), the inoculation amount (5, 10, 15, 20 and 25 percent), the proportion of compound strains-lactobacillus plantarum: streptococcus thermophilus: the method comprises the following steps of (1) performing single research parameter group tests on bacillus subtilis (2:2:1, 2:1:1, 1:1:2, 1:2:2) and material-liquid ratios (1:0.5, 1:0.75, 1:1, 1:1.25 and 1:1.5) to research optimal fermentation parameters, detecting test results after the tests are finished, detecting and recording the yield of the polypeptide of each group of test objects by taking the yield of the polypeptide as an index, determining factors influencing polypeptide fermentation, determining the optimal conditions with the highest polypeptide yield, and improving the polypeptide fermentation yield, wherein the optimal fermentation conditions are as follows: fermentation time is 3 days, particle size of raw materials is 40 meshes, ratio of materials to liquid is 1:1.25, inoculation amount is 5%, fermentation temperature is 28 ℃, and ratio of composite strains-lactobacillus plantarum: streptococcus thermophilus: bacillus subtilis is 1:1:1, and under the conditions, the polypeptide yield is the most.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

A fermentation preparation method of walnut micromolecule polypeptide comprises the following steps: firstly, preprocessing walnut kernels, namely squeezing the walnut kernels by using a low-temperature physical squeezing technology, obtaining two products of walnut oil and walnut meal after squeezing is finished, and separating the two products, wherein the separation mode is as follows: separating the walnut oil and the walnut meal by a filter screen in a shaking mode, and standing the filter screen for a period of time to promote the walnut oil not to adhere to the filter screen and improve the extraction rate of the walnut oil; collecting walnut oil for eating, collecting walnut meal, introducing the walnut meal into a grinder for grinding and crushing, sieving the walnut meal with a 40-mesh sieve after crushing, preparing a plurality of 250mL beakers, containing 40.00g of walnut meal in each beaker, placing the beakers under a sterilization condition for aseptic operation and adding sterile water, inoculating fermentation seed liquid according to the inoculation amount, stirring and stirring for 5 minutes by a glass rod, promoting the materials in the beakers to be uniformly mixed, sealing by gauze, and performing solid state fermentation by using lactobacillus in combination with a biological fermentation technology, wherein the walnut meal needs to be weighed by a precise small balance when being contained in the beakers; the sterilization conditions are controlled as follows: the sterilization temperature is 121 ℃, and the sterilization time is 20 minutes; the mouth of the beaker is sealed by a rubber band when the gauze is sealed.

The method for preparing the walnut micromolecule polypeptide by fermentation comprises the following specific steps:

the method comprises the following steps: the method comprises the following steps: taking out the sample after the fermentation in the claim 1, and placing the sample in an oven to dry for 48 hours at the temperature of 55 ℃;

step two: after drying, crushing the materials, and then sieving the materials by a 40-mesh sieve to be used as a basic raw material for a test for later use;

step three: taking fermentation time (1, 2, 3, 4, 5, 6, 7, 8 and 9 days) as a single research parameter, researching an optimal fermentation time parameter, detecting a test result after respective fermentation time, taking the yield of the polypeptide as an index of the detection result, respectively recording the yield of the polypeptide after the fermentation time is finished each time, obtaining the optimal fermentation time of 3 days through tests, taking the fermentation time of a single factor as a variable in the test process in the three steps, registering a table, obtaining nine groups of polypeptide content test data under different fermentation times, drawing a line graph on the data, and visually judging the optimal fermentation time;

step four: after the fermentation time is determined (3 days), the optimal raw material particle size fermentation parameter is researched by taking the raw material crushed particle size (not crushed, 20, 40 and 60 meshes) as a single research parameter under the optimal fermentation time condition, the test result is detected after the fermentation time of 3 days, the detection result takes the yield of the polypeptide as an index, the yield of the polypeptide is detected and respectively recorded for each group of test objects, the test shows that the optimal fermented raw material particle size is 40 meshes under the condition that the optimal fermentation time is 3 days, the raw material crushed particle size of a single factor is taken as a variable in the test process of the fourth step, a table is registered, four groups of polypeptide content test data under different raw material crushed particle sizes are obtained, a broken line graph is drawn on the data, and the optimal raw material crushed particle size is visually judged;

step five: after the fermentation time (3 days) and the particle size of the raw material (40-mesh sieve) are determined, grouping tests are carried out under the conditions by taking the fermentation temperatures (28, 31, 34 and 37 ℃) as single research parameters to research the optimal fermentation temperature parameters, test results are detected after the tests are finished, the test results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and respectively recorded for each group of test objects, the tests show that the optimal fermentation temperature is 28 ℃ under the conditions of the fermentation time (3 days) and the particle size of the raw material (40-mesh sieve), the fermentation temperature of a single factor is taken as a variable in the test process in the fifth step, a table is registered to obtain four groups of polypeptide content test data under different fermentation temperatures, a broken line graph is drawn on the data, and the optimal fermentation temperature is visually judged;

step six: after the fermentation time (3 days), the raw material particle size (40-mesh sieve) and the fermentation temperature (28 ℃) are determined, grouping tests are carried out under the conditions by taking the inoculation amount (5, 10, 15, 20 and 25%) as a single research parameter to research the optimal fermentation inoculation amount, test results are detected after the tests are finished, the detection results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and respectively recorded for each group of test objects, the test shows that the optimal fermentation inoculation amount is 5% under the conditions of the fermentation time (3 days), the raw material particle size (40-mesh sieve) and the fermentation temperature (28 ℃), the inoculation amount of a single factor is taken as a variable in the test process in the sixth step, a table is registered to obtain five groups of polypeptide content test data under different inoculation amounts, a broken line graph is drawn on the data, and the optimal inoculation amount is visually judged;

step seven: after determining the fermentation time (3 days), the raw material particle size (40 mesh screen), the fermentation temperature (28 degrees) and the fermentation inoculum size (5%), the composite strain ratio was determined under these conditions-lactobacillus plantarum: streptococcus thermophilus: the bacillus subtilis (2:2:1, 2:1:1, 1:1:2 and 1:2:2) is a single research parameter grouping test to research the optimal fermentation inoculation amount, test results are detected after the tests are finished, the detection results take the yield of the polypeptide as an index, the yield of the polypeptide is detected and recorded for each group of test objects, the test shows that the optimal lactobacillus ratio is 1:1:1 under the conditions of fermentation time (3 days), raw material particle size (40 meshes), fermentation temperature (28 ℃) and fermentation inoculation amount (5%), and the compound strain ratio of a single factor-lactobacillus plantarum: streptococcus thermophilus: the bacillus subtilis is used as a variable, a table is registered, five groups of polypeptide content experimental data under different composite strain proportions are obtained, a line drawing is carried out on the data, and the optimal composite strain proportion is visually judged;

step eight: after the fermentation time (3 days), the raw material grain size (40 meshes), the fermentation temperature (28 degrees), the fermentation inoculation amount (5 percent) and the lactic acid bacteria ratio (1:1:1) are determined, under the conditions, the feed-liquid ratio (1:0.5, 1:0.75, 1:1, 1:1.25 and 1:1.5) are taken as single research parameters to carry out grouping tests to research the optimal fermentation inoculation amount, after the tests are finished, test results are detected, the test results take the yield of the polypeptide as an index, the yield of the polypeptide of each group of test objects is detected and respectively recorded, the test shows that the optimal feed-liquid ratio is 1:1.25 under the conditions of the fermentation time (3 days), the raw material grain size (40 meshes), the fermentation temperature (28 degrees), the fermentation inoculation amount (5 percent) and the lactic acid bacteria ratio (1:1: 1:1), the feed-liquid ratio of a single factor is taken as a variable in the test process in the step eight, registering a table to obtain five groups of polypeptide content experimental data under different fermentation times, drawing a line drawing on the data, and visually judging the optimal feed-liquid ratio;

step nine: when the walnut micromolecule polypeptide is prepared by fermentation, the optimal fermentation conditions are as follows: the fermentation time is 3 days, the grain size of the raw material is 40 meshes, the fermentation temperature is 28 ℃, the inoculation amount is 5%, the ratio of lactic acid bacteria is 1:1:1 and the ratio of material to liquid is 1:1.25, and under the condition, the polypeptide fermentation amount is the most.

The walnut kernels are squeezed by a low-temperature physical squeezing technology, two products of walnut oil and walnut dregs can be obtained after squeezing is finished, the walnut oil is separated, the walnut oil is collected and eaten, the walnut dregs are collected and then are led into a grinder to be ground and smashed, the walnut dregs are sieved by a 40-mesh sieve after being smashed, then a plurality of 250mL beakers are prepared, each beaker is filled with 40.00g of walnut dregs, then the beakers are placed under a sterilization condition to be subjected to aseptic operation and added with aseptic water, fermentation seed liquid is inoculated according to the inoculation amount and stirred for 5 minutes by a glass rod to promote materials in the beakers to be mixed and uniformly mixed, then gauze is used for sealing, solid state fermentation is carried out by utilizing lactic acid bacteria and combining with a biological fermentation technology, optimization treatment is carried out according to the influence condition of polypeptide fermentation, and fermentation time (1, 2, 3, 4, 5, 6, 7, 8 and 9 days) can be carried out according to fermentation time, Raw material crushed particle size (not crushed, 20, 40 and 60 mesh), fermentation temperature (28, 31, 34 and 37 degrees), inoculum size (5, 10, 15, 20 and 25%), composite strain ratio-lactobacillus plantarum: streptococcus thermophilus: the method comprises the steps of carrying out single research parameter grouping tests on bacillus subtilis (2:2:1, 2:1:1, 1:1:2 and 1:2:2) and material-liquid ratios (1:0.5, 1:0.75, 1:1, 1:1.25 and 1:1.5) to research optimal fermentation parameters, detecting test results after the tests are finished, detecting and recording the yield of the polypeptide of each group of test objects by taking the yield of the polypeptide as an index according to the detection results, determining factors influencing polypeptide fermentation, determining the optimal conditions with the highest polypeptide yield, and improving the polypeptide fermentation yield.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.