A production process of low heavy metal cadmium small molecule peptide for deep processing of sea cucumber

Abstract

A kind of sea cucumber deep processing low heavy metal cadmium small molecule peptide production process, belong to sea cucumber small molecule active peptide deep processing technical field, in order to solve the problem of removing cadmium in the process of preparing small molecule peptide of sea cucumber, the gist includes preparing sea cucumber enzymatic hydrolysate;Remove cadmium in sea cucumber enzymatic hydrolysate, obtain the feed liquid of the sea cucumber enzymatic hydrolysate of removing cadmium, effect is able to obtain the sea cucumber small molecule peptide product with little heavy metal cadmium residue.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of deep processing of sea cucumber small molecule active peptides, and particularly relates to a production process of a small molecule peptide with low heavy metal cadmium obtained through deep processing of sea cucumber. BACKGROUND

[0002] Sea cucumber is one of the "eight treasures of the sea", and has existed on the earth for more than 600 million years, and is a recognized tonic and health product. Protein is the highest active component in sea cucumber, but its molecular weight is usually as high as 300 kDa, which is difficult to be directly absorbed and utilized. Therefore, people use biological enzymatic technology to prepare small molecule, high activity sea cucumber peptides. Sea cucumber peptides have the advantages of easy absorption, high biological potency, and no antigenicity, and have good solubility and stability.

[0003] However, in the preparation process of sea cucumber peptides, if there is no related removal process, heavy metals will also be enriched in the final product, such as the pollutant cadmium. Cadmium is one of the non-essential and harmful heavy metal elements in the human body, which is quickly absorbed in the digestive tract but very slowly degraded, easily accumulated in the liver or kidneys to cause harm, especially to the kidneys, and also causes osteoporosis and softening. The "Itai-Itai disease" reported by Japan is a chronic cadmium poisoning caused by eating cadmium-contaminated water and rice.

[0004] At present, environmental pollution, substandard breeding conditions and other problems are increasingly prominent, and cadmium is a heavy metal that is very easy to accumulate in sea cucumber during breeding. At present, GB 2762-2017 "National Food Safety Standard Limit of Contaminants in Foods" limits the cadmium content in fresh and frozen sea cucumber to 2.0 mg / kg (without internal organs), but there is no relevant limit requirement for sea cucumber products, which leads to that many sea cucumber peptide products on the market do not pay attention to the cadmium content and do not remove cadmium in the process. Therefore, the development of a production process of sea cucumber small molecule peptides with low heavy metal cadmium has great significance and value for the active peptide industry and society. SUMMARY

[0005] The application provides a production process of a small molecule peptide with low heavy metal cadmium obtained through deep processing of sea cucumber, which has the advantages of simple process, good cadmium removal effect, easy production, and contribution to food safety, and great help to the popularization and application of sea cucumber peptides in the fields of nutritional food, functional food and health products.

[0006] According to the production process of the small molecule peptide with low heavy metal cadmium obtained through deep processing of sea cucumber according to some embodiments of the application, the production process comprises the following steps:

[0007] Preparation of sea cucumber enzymatic solution;

[0008] Removal of cadmium in the sea cucumber enzymatic solution to obtain a feed liquid of the sea cucumber enzymatic solution from which cadmium is removed.

[0009] The sea cucumber fine processing low heavy metal cadmium small molecule peptide production process according to some embodiments of the application further comprises preparing sea cucumber peptide powder from the feed liquid.

[0010] The step of removing cadmium from the sea cucumber enzymatic hydrolysate in the sea cucumber fine processing low heavy metal cadmium small molecule peptide production process according to some embodiments of the application comprises adding seaweed ferment to the sea cucumber enzymatic hydrolysate.

[0011] The seaweed ferment in the sea cucumber fine processing low heavy metal cadmium small molecule peptide production process according to some embodiments of the application is prepared by taking edible seaweed, adding 3-7 times the volume of pure water, homogenizing, then adding 5%-20% of the volume of the feed liquid of malt dextrin, adjusting the pH to 5.0-6.5, inoculating activated Lactobacillus plantarum, and fermenting at 28-35°C for 24-48 hours, and then concentrating and freeze-drying the feed liquid to obtain seaweed ferment.

[0012] The Lactobacillus plantarum in the sea cucumber fine processing low heavy metal cadmium small molecule peptide production process according to some embodiments of the application is Lactobacillus plantarum with the preservation number CGMCC No. 1258.

[0013] The edible seaweed in the sea cucumber fine processing low heavy metal cadmium small molecule peptide production process according to some embodiments of the application is Laminaria.

[0014] The sea cucumber fine processing low heavy metal cadmium small molecule peptide production process according to some embodiments of the application comprises adding 1%-2% of the seaweed ferment to the sea cucumber protein enzymatic hydrolysate, stirring and reacting at 50°C±2°C for 1-2 hours, centrifuging to obtain a clear liquid, and then introducing the clear liquid into an electrodialysis device.

[0015] First, start the first-stage electrodialysis, and when the feed liquid in the feed barrel is about 50%, start the second-stage electrodialysis, adjust the voltage to 65-75V, and control the first-stage conductivity at 1000-1100 μS / cm, and when the second-stage conductivity is less than 300 μS / cm, stop, and then introduce the feed liquid into a storage tank through a pipeline.

[0016] Raise the temperature of the feed liquid in the storage tank to 60-70°C, add 0.5-2% of the volume of the feed liquid of activated carbon, stir for 45-80 minutes for adsorption, and then filter the feed liquid to remove the activated carbon.

[0017] The sea cucumber fine processing low heavy metal cadmium small molecule peptide production process according to some embodiments of the application comprises separating the feed liquid after the activated carbon removal treatment, and spraying and drying the membrane permeate liquid to obtain sea cucumber peptide powder.

[0018] According to the sea cucumber deep processing low heavy metal cadmium small molecule peptide production process of some embodiments of the application, the preparation of the sea cucumber enzymatic hydrolysate includes crushing the cleaned sea cucumber raw material, placing it in an enzymatic hydrolysis tank, adding 8-20 times mass volume of pure water, stirring uniformly, then adding 2-5% of the mass of the sea cucumber of complex protease, enzymatic hydrolysis at 40-60 DEG C for 3-5 hours, the enzyme reaction pH value is controlled at 8.0-10.0, after the enzymatic hydrolysis is completed, the temperature is raised to 100 DEG C to inactivate the enzyme for 10 minutes, and then centrifugation is performed to obtain the sea cucumber enzymatic hydrolysate.

[0019] According to the sea cucumber deep processing low heavy metal cadmium small molecule peptide production process of some embodiments of the application, the mass ratio of the complex protease is: neutral protease: alkaline protease: flavor protease = (1-3): (2-5): (3-5).

[0020] The beneficial effects of the application are:

[0021] (1) The application provides a new sea cucumber peptide cadmium removal process. In the process, organic cadmium is converted into cadmium ions by using microbial fermentation products + electrodialysis + activated carbon, and then removed by cadmium ion migration under the action of an electric field. This process can safely remove cadmium from sea cucumber enzymatic hydrolysate, and has a very high removal rate. The experimental data of this effect are further illustrated by the examples and comparative examples of the application.

[0022] (2) The application solves the safety hazard of cadmium pollution in sea cucumber peptides, has high safety, strong market competitiveness, and wide application prospect in the fields of nutritional food, functional food, health food, etc. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 is a process flow chart. DETAILED DESCRIPTION

[0024] The application is further illustrated below in conjunction with examples, but is not limited to the specific examples. The application proposes a sea cucumber deep processing low heavy metal cadmium small molecule peptide production process, which includes the following steps:

[0025] S101. Preparation of sea cucumber enzymatic hydrolysate: crush the cleaned sea cucumber raw material, place it in an enzymatic hydrolysis tank, add 8-20 times mass volume of pure water, stir uniformly, then add 2-5% of the mass of the sea cucumber (dry weight) of complex protease, enzymatic hydrolysis at 40-60 DEG C for 3-5 hours, the enzyme reaction pH value is controlled at 8.0-10.0, after the enzymatic hydrolysis is completed, the temperature is raised to 100 DEG C to inactivate the enzyme for 10 minutes, and then centrifugation is performed to obtain the sea cucumber enzymatic hydrolysate. Alternatively, the mass ratio of the complex protease in step S101 is: neutral protease: alkaline protease: flavor protease = (1-3): (2-5): (3-5).

[0026] S102. Preparation of seaweed enzyme: edible seaweed is taken, 3-7 times volume of pure water is added, homogenized, then 5%-20% volume of malt dextrin is added, pH is adjusted to 5.0-6.5, inoculated with activated Lactobacillus plantarum, fermented at 28-35°C for 24-48 hours, the feed liquid is concentrated and freeze-dried to obtain seaweed enzyme. Alternatively, the seaweed in step S102 is Laminaria. Alternatively, the Lactobacillus plantarum can be used in the food industry, the strain is preserved, and the preservation number is CGMCC No. 1258. Lactobacillus plantarum LP-ONLLY was preserved in China General Microbiological Culture Collection Center on December 6, 2004, the address is No. 13, North Zhongguancun Street, Haidian District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, the biological preservation number is CGMCC No. 1258, and it is disclosed in Chinese patent application publication CN1660300A.

[0027] S103. Removal of cadmium in sea cucumber enzymatic hydrolysate: 1%-2% seaweed enzyme is added to the sea cucumber protein enzymatic hydrolysate of step S101, stirred at 50°C±2°C for 1-2 hours, centrifuged to obtain a clear liquid, and then the clear liquid is introduced into an electrodialysis device. First, open the primary electrodialysis, and when the feed tank liquid is about 50%, open the secondary electrodialysis. Adjust the voltage to 65-75V, control the primary conductivity at 1000-1100 μS / cm, and when the secondary conductivity is <300 μS / cm, stop, and the liquid is introduced into a storage tank through a pipeline. The liquid in the storage tank is heated to 60-70°C, 0.5-2% activated carbon is added, stirred for 45-80 minutes for adsorption, and then the liquid is filtered for carbon removal treatment.

[0028] S104. Preparation of sea cucumber peptide powder: the liquid in step S103 is separated by membrane separation technology, the molecular weight cut-off is 1000 Da, and the filtrate is spray dried to obtain sea cucumber peptide powder.

[0029] Example 1: The production process of low heavy metal cadmium small molecule peptide of sea cucumber deep processing in this embodiment includes the following steps:

[0030] S101. Preparation of sea cucumber enzymatic hydrolysate: 1 kg of dried sea cucumber is cleaned and crushed, placed in an enzymatic hydrolysis tank, 15 times mass volume of pure water is added, stirred uniformly, then 4% of compound protease of sea cucumber (dry weight) is added, the mass ratio of compound protease is: neutral protease: alkaline protease: flavor protease = 3:5:3, enzymolysis is carried out at 52°C for 5 hours, the pH value of enzyme reaction is controlled at 9.0, after the enzymolysis is completed, the temperature is raised to 100°C for 10 minutes for enzyme inactivation, and centrifugation is carried out to obtain sea cucumber enzymatic hydrolysate.

[0031] S102. Preparation of seaweed enzyme: take 1 kg of kelp, add 4 times the volume of pure water homogenate, then add malt dextrin with a volume of 10% of the feed liquid, adjust the pH to 6.0, inoculate the activated lactobacillus plantarum (preserved number CGMCC NO.1258), and ferment at 30℃ for 24h. Concentrate the feed liquid and freeze-dry to obtain seaweed enzyme.

[0032] S103. Removal of cadmium in sea cucumber enzymatic hydrolysate: add 2% seaweed enzyme to the sea cucumber protease hydrolysate, stir at 50℃±2℃ for 2h, centrifuge to obtain the supernatant, then introduce the supernatant into the electrodialysis equipment. First, open the primary electrodialysis, and when the feed tank liquid is left about 50%, open the secondary electrodialysis. Adjust the voltage to 70V, control the primary conductivity at 1000μS / cm, and stop when the secondary conductivity is <100μS / cm. The feed liquid is introduced into the storage tank through the pipeline. The feed liquid in the storage tank is heated to 70℃, 1% activated carbon is added to the feed liquid, and stirred for 50min for adsorption, then the feed liquid is filtered for decarburization treatment.

[0033] S104. Preparation of sea cucumber peptide powder: the decarburized feed liquid is separated by membrane separation technology with a molecular weight cutoff of 1000Da, and the permeate is spray dried to obtain sea cucumber peptide powder.

[0034] Example 2: The production process of low heavy metal cadmium small molecule peptide in the sea cucumber deep processing in this embodiment includes the following steps:

[0035] S101. Preparation of sea cucumber enzymatic hydrolysate: take 1 kg of dried sea cucumber, clean and crush, add 10 times the volume of pure water to the enzymolysis tank, stir evenly, then add 3% of the compound protease of the mass of sea cucumber (dry weight), and the mass ratio of the compound protease is: neutral protease: alkaline protease: flavor protease = 2:4:4. Enzymolysis at 50℃ for 4h, the pH value of enzyme reaction is controlled at 8.5, and the enzyme is inactivated at 100℃ for 10min after enzyme reaction. Centrifugation to obtain sea cucumber enzymatic hydrolysate.

[0036] S102. Preparation of seaweed enzyme: take 1 kg of kelp, add 5 times the volume of pure water homogenate, then add malt dextrin with a volume of 13% of the feed liquid, adjust the pH to 5.8, inoculate the activated lactobacillus plantarum (strain number CGMCC NO.1258), and ferment at 32℃ for 36h. Concentrate the feed liquid and freeze-dry to obtain seaweed enzyme.

[0037] S103. Removal of cadmium from the sea cucumber protease hydrolysate: 1.5% seaweed enzyme was added to the sea cucumber protease hydrolysate of step S101, and stirred at 50°C±2°C for 2h. The supernatant was obtained by centrifugation, and then introduced into an electrodialysis device. First, primary electrodialysis was started, and when the remaining liquid in the tank was about 50%, secondary electrodialysis was started. The voltage was adjusted to 75V, and the primary conductivity was controlled at 1000μS / cm. When the secondary conductivity was less than 100μS / cm, the process was stopped, and the liquid was introduced into a storage tank through a pipeline. The liquid in the storage tank was heated to 65°C, 1% activated carbon was added, and stirred for 60min for adsorption. Then the liquid was filtered to remove the carbon.

[0038] S104. Preparation of sea cucumber peptide powder: the liquid in step S103 was separated by membrane separation technology with a molecular weight cut-off of 1000Da, and the permeate was spray dried to obtain sea cucumber peptide powder.

[0039] Comparative Example 1: The sea cucumber deep processing low heavy metal cadmium small molecule peptide production process described in this comparative example is the same as that in Example 1, except for the following differences. The differences include:

[0040] (1) Step S102 is not performed.

[0041] (2) The removal of cadmium from the sea cucumber protease hydrolysate in step S103 is different: in this comparative example, the sea cucumber protease hydrolysate of step S101 was directly introduced into an electrodialysis device.

[0042] Comparative Example 2: The sea cucumber deep processing low heavy metal cadmium small molecule peptide production process described in this comparative example is the same as that in Example 1, except for the following differences. The differences include:

[0043] (1) The removal of cadmium from the sea cucumber protease hydrolysate in step S103 is different: in this comparative example, 2% seaweed enzyme was added to the sea cucumber protease hydrolysate of step S101, and stirred at 50°C±2°C for 2h, and the supernatant was obtained by centrifugation.

[0044] (2) The preparation of sea cucumber peptide powder in step S104 is different: in this comparative example, the supernatant in step S103 was directly separated by membrane separation technology with a molecular weight cut-off of 1000Da, and the permeate was spray dried to obtain sea cucumber peptide powder.

[0045] Comparative Example 3: The sea cucumber deep processing low heavy metal cadmium small molecule peptide production process described in this comparative example is the same as that in Example 1, except for the following differences. The differences include:

[0046] (1) Steps S102 and S103 are not performed.

[0047] (2) The sea cucumber peptide powder in step S104 is prepared in different ways. In the present comparative example, the sea cucumber proteolytic liquid in S101 is directly separated by membrane separation technology with a molecular weight cut-off of 1000 Da, and the permeate is spray dried to obtain sea cucumber peptide powder.

[0048] Results verification: The sea cucumber raw material and the sea cucumber peptides prepared in Examples 1 and 2 and Comparative Examples 1, 2 and 3 of the present application were used as samples, and the cadmium content was detected to verify the cadmium removal effect of the production process.

[0049] Detection method: GB 5009.15-2014 "Determination of cadmium in food safety national standard"

[0050] Detection results: As shown in Table 1.

[0051] Table 1 Cadmium content in each sea cucumber raw material and sea cucumber peptide powder

[0052] Project Example 1 Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Cadmium content of sea cucumber (dry weight basis) (mg / kg) 0.563 0.563 0.563 0.563 0.563 Cadmium content of sea cucumber peptide (mg / kg) ND 0.004 0.172 0.146 0.484 Removal rate, % 100％ 99.29％ 69.45％ 74.07％ 14.03％

[0053] ND: not detected

[0054] Conclusion: 1. The results of Examples 1 and 2 show that the production process of the present application can effectively remove the pollutant cadmium in sea cucumber peptides, with a removal rate of more than 99%.

[0055] 2. The results of Comparative Examples 1 and 2 show that when a single operation is performed, the adsorption and removal effect of cadmium by sea algae enzyme fermented by Lactobacillus plantarum is better than that by electrodialysis + activated carbon adsorption.

[0056] 3. The results of Comparative Example 3 show that direct membrane separation and spray drying of sea cucumber proteolytic liquid cannot achieve the effect of removing cadmium.

[0057] 4. The results of Examples 1 and 2 and Comparative Examples 1, 2 and 3 show that the effect of using sea algae enzyme fermented by Lactobacillus plantarum + electrodialysis + activated carbon adsorption in combination is better than that of a single operation.

[0058] Sea cucumber is a water organism that is easily contaminated by cadmium. The cadmium content in each organ of sea cucumber from high to low is: respiratory tree > intestine > body wall > male gland > female gland. The market for sea cucumber peptides is currently in chaos, and sea cucumber peptides are prepared from various parts of sea cucumber, with inconsistent raw material sources and quality, large price differences, and no relevant national standards for the cadmium content limit of sea cucumber peptides, which poses a safety risk. The above experiments of the present application show that the sea algae enzyme prepared by fermentation of Lactobacillus plantarum can effectively adsorb and remove heavy metal cadmium in sea cucumber proteolytic liquid, and the removal rate of cadmium reaches more than 99% when combined with electrodialysis and activated carbon.

[0059] While the application has been described by way of example, it should be appreciated that modifications and additions can be made without departing from the scope of the application as recited in the claims.

Claims

1. A production process of a low heavy metal cadmium small molecule peptide of sea cucumber deep processing, characterized in that, The application relates to a method for preparing sea cucumber peptide powder. The method comprises the following steps: preparing sea cucumber enzymatic hydrolysate; removing cadmium in the sea cucumber enzymatic hydrolysate to obtain a sea cucumber enzymatic hydrolysate solution; and preparing sea cucumber peptide powder from the solution. The step of removing cadmium in the sea cucumber enzymatic hydrolysate comprises adding seaweed enzyme into the sea cucumber enzymatic hydrolysate. The seaweed enzyme is obtained by adding pure water and malt dextrin into seaweed, inoculating activated lactobacillus plantarum, and then performing fermentation, wherein the lactobacillus plantarum is lactobacillus plantarum with a preservation number of CGMCC No. 1258. The method further comprises preparing sea cucumber peptide powder from the solution.

2. The production process of low heavy metal cadmium small molecule peptide of sea cucumber deep processing according to claim 1, characterized in that, The seaweed enzyme is prepared in the following manner: edible seaweed is taken, 3-7 times the volume of pure water is added, homogenization is performed, then 5%-20% of the volume of malt dextrin is added, the pH value is adjusted to 5.0-6.5, activated lactobacillus plantarum is inoculated, and then fermentation is performed at 28-35 DEG C for 24-48 hours; and the solution is concentrated and freeze-dried to obtain the seaweed enzyme.

3. The production process of low heavy metal cadmium small molecule peptide of sea cucumber deep processing according to claim 1, characterized in that, The edible seaweed is kelp.

4. The production process of low heavy metal cadmium small molecule peptide of sea cucumber deep processing according to claim 3, characterized in that, 1%-2% of the seaweed enzyme is added into the sea cucumber enzymatic hydrolysate, stirring reaction is performed at 50 DEG C + / - 2 DEG C for 1-2 hours, and then centrifugation is performed to obtain a clear solution; and then the clear solution is introduced into an electrodialysis device.

5. The production process of low heavy metal cadmium small molecule peptide of sea cucumber deep processing according to claim 1 or 3, characterized in that, Firstly, primary electrodialysis is started, and when the solution in the barrel is about 50%, secondary electrodialysis is started, the voltage is adjusted to 65-75 V, the conductivity of the primary electrodialysis is controlled to be 1000-1100 mu S / cm, and when the conductivity of the secondary electrodialysis is less than 300 mu S / cm, the solution is stopped and introduced into a storage tank through a pipeline. The solution in the storage tank is heated to 60-70 DEG C, 0.5-2% of the volume of activated carbon is added, stirring treatment is performed for 45-80 minutes for adsorption, and then the solution is filtered for carbon removal treatment. The solution after the carbon removal treatment is separated, the molecular weight is cut off to be 1000 Da, and the membrane liquid is spray dried to obtain sea cucumber peptide powder.

6. The production process of low heavy metal cadmium small molecule peptide of sea cucumber deep processing according to claim 5, characterized in that, The sea cucumber enzymatic hydrolysate is prepared in the following manner: clean sea cucumber raw materials are crushed, put into an enzymatic hydrolysis tank, 8-20 times the mass volume of pure water is added, stirring is uniformly performed, then 2-5% of the mass of the sea cucumber is added into complex protease, enzymatic hydrolysis is performed at 40-60 DEG C for 3-5 hours, the pH value of the enzyme reaction is controlled to be 8.0-10.0, the temperature is increased to 100 DEG C for 10 minutes for enzyme inactivation after the enzymatic hydrolysis is completed, and then centrifugation is performed to obtain the sea cucumber enzymatic hydrolysate.

7. The production process of low heavy metal cadmium small molecule peptide of sea cucumber deep processing according to claim 1, characterized in that, The mass ratio of the complex protease is as follows: neutral protease: alkaline protease: flavor protease = 1-3: 2-5: 3-5. 8.The production process of low heavy metal cadmium small molecule peptide of sea cucumber deep processing according to claim 7, characterized in that, ​

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