Abalone umami peptide prepared from abalone cooking waste liquid and preparation method of abalone umami peptide

Abstract

The invention relates to the field of functional nutritional seasonings, and particularly discloses an abalone umami peptide prepared from an abalone cooking waste liquid and a preparation method thereof, and the preparation method comprises the following steps: (1) removing solid impurities from the abalone cooking waste liquid, and removing heavy metals by using electrodialysis; (2) adding the material obtained in the step (1) into specific protease accounting for 0.1-1% of the weight of the solid in the material obtained in the step (1), and carrying out an enzymolysis reaction under the conditions that the enzymolysis pH value is 6-7, the reaction temperature is 40-60 DEG C and the reaction time is 0.5-2 hours; and 3) removing impurities by using a nanofiltration membrane, and carrying out spray drying to obtain the abalone umami peptide. According to the abalone flavor peptide prepared by the method, the problem of environmental pollution caused by a large amount of abalone cooking waste liquid is solved, waste is turned into wealth, and a new way of resource utilization of the abalone cooking waste liquid is realized. The ratio gt of the molecular weight of the abalone umami peptide is 1-2KD; the seafood seasoning not only is high in antioxidant biological activity and low in heavy metal content, but also has rich seafood flavor, and can be widely applied to the field of functional nutritional seasonings.

Description

Technical Field

[0001] The invention relates to the field of functional nutritional seasonings, and in particular to an abalone umami peptide prepared by taking abalone cooking waste liquid as a raw material and a preparation method thereof. Background Art

[0002] In recent years, with the development of science and technology and the improvement of people's living standards, a large number of umami substances such as free amino acids and flavor nucleotides have been developed and put on the market, which are deeply loved by consumers. Although these umami substances are rich in umami, they do not have antioxidant effects. Therefore, there is a huge market demand for the development of umami peptides with antioxidant effects.

[0003] During the processing of abalone, a large amount of cooking waste liquid is generated, which pollutes the environment. Although these waste liquids are rich in peptides, water-soluble proteins, etc., they are high-quality raw materials for the development of umami peptides. At present, there is no patent for the development of abalone umami peptides using abalone cooking waste liquid as raw material. This is because the problem of excessive heavy metals in abalone cooking waste liquid has hindered its industrial production and promotion. Therefore, how to make high-value use of abalone cooking waste liquid resources, turn waste into treasure, develop abalone umami peptides with extremely high utilization value, extend the abalone industry chain, and alleviate environmental and corporate pressures is an urgent problem to be solved. Summary of the invention

[0004] A large amount of cooking waste liquid is generated during the development of abalone products, causing environmental pollution and other problems. The purpose of the present invention is to provide a method for preparing abalone umami peptides with mild reaction conditions, controllable quality, and suitable for large-scale production, so as to make full use of abalone cooking waste liquid, turn waste into treasure, and develop abalone umami peptides with high added value.

[0005] In order to solve the above technical problems, the first aspect of the present invention provides an abalone umami peptide prepared using abalone cooking waste liquid as raw material and a preparation method thereof, characterized in that: 1) Enzymatic hydrolysis reaction: adding protease to the abalone cooking waste liquid for enzymatic hydrolysis reaction; 2) Multi-stage membrane separation: Filter the solution after the enzymatic hydrolysis reaction in step 2) to remove peptides with molecular weight greater than 3KD and less than 800D; 3) Spray drying.

[0006] In some specific embodiments of the first aspect, in the step 1) enzymatic hydrolysis reaction, the pH value of the abalone cooking waste liquid is 6-7, in some embodiments, the pH value of the abalone cooking waste liquid is 6.3, in some embodiments, the pH value of the abalone cooking waste liquid is 6.5, and in some embodiments, the pH value of the abalone cooking waste liquid is 6.7.

[0007] In some specific embodiments of the first aspect, the enzymatic hydrolysis temperature of the abalone cooking waste liquid is 40-60°C, in some embodiments, the enzymatic hydrolysis temperature of the abalone cooking waste liquid is 45°C, in some embodiments, the enzymatic hydrolysis temperature of the abalone cooking waste liquid is 50°C, and in some embodiments, the enzymatic hydrolysis temperature of the abalone cooking waste liquid is 55°C.

[0008] In some specific embodiments of the first aspect, the enzymatic hydrolysis time of the abalone cooking waste liquid is 0.5~2h, in some embodiments, the enzymatic hydrolysis time of the abalone cooking waste liquid is 1.0h, and in some embodiments, the enzymatic hydrolysis time of the abalone cooking waste liquid is 1.5h.

[0009] In some specific embodiments of the first aspect, the peptides with a molecular weight greater than 3KD and a molecular weight less than 800D are filtered out by a nanofiltration membrane.

[0010] In some specific embodiments of the first aspect, the abalone cooking waste liquid in step 1) is further subjected to electrodialysis desorption to remove heavy metals before adding protease, and the electrodialysis desorption is specifically: S1: removing solid impurities from abalone cooking waste liquid; S2: Electrodialysis is used to remove heavy metals. The electrodialysis current is 8~12A and the electrodialysis flow rate is 0.6~1.2m 3 / h; In some embodiments, the electrodialysis current is 8A, in some embodiments, the electrodialysis current is 9A, in some embodiments, the electrodialysis current is 10A, in some embodiments, the electrodialysis current is 11A, and in some embodiments, the electrodialysis current is 12A.

[0011] In some embodiments, the electrodialysis flow rate is 0.7 m 3 / h, in some embodiments the electrodialysis flow rate is 0.8m 3 / h, in some embodiments the electrodialysis flow rate is 0.9m 3 / h, in some embodiments the electrodialysis flow rate is 1.0m 3 / h, in some embodiments the electrodialysis flow rate is 1.1m 3 / h.

[0012] In some specific embodiments of the first aspect, the protease is selected from one of neutral protease, ficin and bromelain.

[0013] In some specific embodiments of the first aspect, the mass of the protease feed is 0.1 wt% to 1 wt% of the dry weight of the solid content of the abalone cooking waste liquid.

[0014] In some specific embodiments of the first aspect, the solid impurities are removed by high-speed centrifugation, and the speed of the high-speed centrifuge is >10000rpm. In some embodiments, the speed of the high-speed centrifuge is >11000rpm. In some embodiments, the speed of the high-speed centrifuge is >12000rpm. In some embodiments, the speed of the high-speed centrifuge is >13000rpm.

[0015] In some specific embodiments of the first aspect, the outlet air temperature of the spray drying is 70~85℃, and the inlet air temperature is 140~170℃. In some embodiments, the outlet air temperature of the spray drying is 74℃, the outlet air temperature of the spray drying is 78℃, the outlet air temperature of the spray drying is 82℃, in some embodiments, the inlet air temperature is 145℃, in some embodiments, the inlet air temperature is 150℃, in some embodiments, the inlet air temperature is 155℃, and in some embodiments, the inlet air temperature is 160℃.

[0016] The second aspect of the present invention provides an abalone umami peptide, which is prepared by the preparation method of any embodiment of the first aspect.

[0017] In some specific embodiments of the second aspect, the proportion of umami peptides with a molecular weight of 1-2KD is >70%, in some embodiments, the proportion of umami peptides with a molecular weight of 1-2KD is >70%, in some embodiments, the proportion of umami peptides with a molecular weight of 1-2KD is >80%, and in some embodiments, the proportion of umami peptides with a molecular weight of 1-2KD is >90%.

[0018] The reagents used in the present invention are all purchased from the open and legal market without further purification.

[0019] Terminology Certain embodiments of the present invention are now described in detail. The present invention is intended to cover all substitutions, modifications and equivalent technical solutions, which are all included in the scope of the present invention as defined in the claims. It should be appreciated by those skilled in the art that many methods and materials similar or equivalent to those described herein can be used to practice the present invention. The present invention is by no means limited to the methods and materials described herein. In the event that one or more of the combined documents, patents and similar materials are different from or contradictory to the present application (including but not limited to defined terms, term applications, described technologies, etc.), the present application shall prevail.

[0020] It should be further appreciated that certain features of the invention, which for clarity are described in multiple separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which for brevity are described in a single embodiment, may also be provided separately or in any suitable sub-combination.

[0021] Unless otherwise specified, all technical terms used in the present invention have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. All patents and publications related to the present invention are incorporated herein by reference in their entirety.

[0022] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0023] In the following content, all numbers disclosed herein are approximate values, regardless of whether the words "about" or "approximately" are used. The value of each number may vary by 1%, 2%, 5%, 7%, 8%, 10%, 15% or 20%. Whenever a number with a value of N is disclosed, any number with a value of N+ / -1%, N+ / -2%, N+ / -3%, N+ / -5%, N+ / -7%, N+ / -8%, N+ / -10%, N+ / -15% or N+ / -20% will be explicitly disclosed, where "+ / -" means plus or minus.

[0024] Abalone cooking waste liquid is the cooking waste liquid produced after abalone is cooked in hot water.

[0025] [Molecular weight detection] With reference to the national standard GB / T22729-2008, the molecular weight of abalone umami peptide was determined by gel permeation chromatography. The chromatographic column was TSKgel G200 SWxL (300 mm X 7.8 mm, 5μm), the mobile phase was acetonitrile-water-trifluoroacetic acid (volume ratio of 45:55:0.1), and the sample was measured at a column temperature of 30°C and a flow rate of 0.5mL / min with a detection wavelength of 220nm. The standard samples were: tyrosine-tyrosine-tyrosine: Mr189.17, tyrosine-tyrosine-tyrosine-arginine: Mr451.48, bacillus enzyme: Mr1422.69, aprotinin: Mr6511.44, cytochrome C: Mr12384.

[0026] [Sensory evaluation of umami taste] Ten industry professionals were asked to conduct a sensory evaluation of the freshness of the abalone umami peptide prepared in this example according to the umami sensory scoring table in Table 1, using a ten-point system, and the scoring was calculated to take the average value.

[0027] Table 1. Umami sensory evaluation criteria [Heavy metal content detection] GB 500.11-2014 detects the content of inorganic arsenic in the abalone umami peptide prepared in this embodiment, GB 500.12-2017 detects the content of lead in the product, and GB 500.17-2021 detects the content of methylmercury in the product.

[0028] [Safety Evaluation] Different doses of the abalone umami peptide prepared in this example were intragastrically administered to Kunming mice, and the dose groups were 1 g / kg, 5 g / kg, 10 g / kg, and 15 g / kg, respectively.

[0029] [Evaluation of antioxidant biological activity] Healthy adult mice weighing 25-30 g were selected. Except for the blank control group, the other animals were injected intraperitoneally with D-galactose to establish the model. One model control group and three test sample dose groups (low dose group 100 mg.kg -1 .d -1 , medium dose group 500mg.kg -1 .d -1 and high dose group 1000mg.kg -1 .d -1 ), the model control group was given the same volume of solvent, and the test samples were given to mice by oral gavage. At the same time as the test samples, the model control group and each dose group continued to be given the same dose of D-galactose by intraperitoneal injection. At the end of the experiment, the animals were killed and the antioxidant activity of the samples was measured.

[0030] Advantages of the present invention: Compared with the prior art, a certain embodiment of the present invention has at least one of the following beneficial effects: The invention uses abalone cooking waste liquid as raw material to develop abalone umami peptides, solves the bottleneck problem of waste liquid polluting the environment in the abalone product processing process, and realizes a new way to high-value utilization of abalone cooking waste liquid.

[0031] 2. The abalone umami peptide produced by the present invention has high safety, no toxic side effects, low heavy metal content, the proportion of umami peptides with a molecular weight of 1-2KD is greater than 70%, and the product umami sensory sensory score is greater than 9 points, and can be applied in the field of functional nutritional seasonings. DETAILED DESCRIPTION

[0032] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] Example 1 (1) After 100L of abalone cooking wastewater was centrifuged at a speed of >10,000rpm to remove solid impurities, heavy metals were removed by electrodialysis at an electrodialysis current of 10A and an electrodialysis flow rate of 0.8m 3 / h.

[0034] (2) The material obtained in step (1) is placed in a reactor, and 1% of the weight of the solid calculated by the solid content in the material obtained in the above step (1) of fig protease is added for enzymatic hydrolysis reaction, and the pH value of the reaction system is controlled to be 6, the temperature of the enzymatic hydrolysis reaction is 60°C, and the time of the enzymatic hydrolysis reaction is 2h. After the enzymatic hydrolysis reaction is completed, a nanofiltration membrane with a molecular weight cutoff of 3KD is used to remove macromolecular impurities, and a nanofiltration membrane with a molecular weight of 800Da is used to remove small molecular impurities. The material solution after membrane separation is spray dried, and the outlet air temperature of the spray drying is 85°C and the inlet air temperature is 170°C to obtain abalone umami peptide.

[0035] [Molecular weight detection] The test results show that the proportion of abalone umami peptide molecular weight in the range of 1-2KD is 73%.

[0036] [Umami sensory evaluation] The evaluation results showed that the freshness score of abalone umami peptide was 9.16 points, and the sample had a strong umami taste. [Heavy metal content detection] The inorganic arsenic, lead and methylmercury contents of the abalone umami peptide prepared in Example 1 are all less than 0.1, indicating that the abalone umami peptide prepared in this example is highly safe.

[0037] [Safety Evaluation] There was no death in the animals of each dosage group, and no abnormal behavior and no significant weight gain, indicating that the abalone umami peptide prepared in this example has no toxic side effects.

[0038] [Evaluation of antioxidant biological activity] As shown in Table 2, compared with the normal group, the MDA content in the serum and liver of the D-galactose model group was significantly increased (p<0.01), indicating that the mouse aging model was successfully established. Compared with the model group, the MDA content in the serum and liver of the control group and the dose group was reduced, and the MDA content in the serum and liver of the high-dose group mice was significantly reduced (p<0.01), indicating that the abalone umami peptide prepared in this example has antioxidant activity.

[0039] Table 2. Effects of abalone umami peptides on the MDA content in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01 As shown in Table 3, compared with the normal group, the SOD content in the serum and liver of the D-galactose model group was significantly reduced (p<0.01), indicating that the mouse aging model was successfully established. Compared with the model group, the SOD content in the serum and liver of the control group and the dose group was increased, and the SOD content in the serum and liver of the high-dose group mice was significantly increased (p<0.01), indicating that the abalone umami peptide prepared in this example has antioxidant activity.

[0040] Table 3. Effects of umami peptides on SOD levels in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01 Example 2 (1) After 500L of abalone cooking wastewater was centrifuged at a speed of >10,000rpm to remove solid impurities, heavy metals were removed by electrodialysis at an electrodialysis current of 11A and an electrodialysis flow rate of 1m 3 / h.

[0041] (3) The material obtained in step (1) is placed in a reactor, and 0.5% of the weight of bromelain calculated by the solid content in the material obtained in the above step (1) is added for enzymatic hydrolysis reaction, and the pH value of the reaction system is controlled to be 6.5, the temperature of the enzymatic hydrolysis reaction is 40° C., and the time of the enzymatic hydrolysis reaction is 1 hour. After the enzymatic hydrolysis reaction is completed, a nanofiltration membrane with a molecular weight cutoff of 3KD is used to remove macromolecular impurities, and a nanofiltration membrane with a molecular weight of 800Da is used to remove small molecular impurities. The material solution after membrane separation is spray dried, and the outlet air temperature of the spray drying is 75° C. and the inlet air temperature is 150° C., to obtain abalone umami peptide.

[0042] [Molecular weight detection] The test results show that the proportion of abalone umami peptides with a molecular weight of 1~2KD is 79%.

[0043] [Umami sensory evaluation] The evaluation results showed that the freshness score of abalone umami peptide was 9.37 points, and the sample had a strong umami taste. [Heavy metal content detection] The experimental results show that the inorganic arsenic, lead and methylmercury contents of the abalone umami peptide prepared in this embodiment are all less than 0.1, indicating that the abalone umami peptide prepared in this embodiment is highly safe.

[0044] [Safety Evaluation] There was no death in the animals of each dosage group, and no abnormal behavior and no significant weight gain, indicating that the abalone umami peptide prepared in this example has no toxic side effects.

[0045] [Evaluation of antioxidant biological activity] As shown in Table 4, compared with the normal group, the MDA content in the serum and liver of the D-galactose model group was significantly increased (p<0.01), indicating that the mouse aging model was successfully established.

[0046] Compared with the model group, the MDA content in serum and liver of the control group and the dose group was reduced, and the MDA content in serum and liver of the high-dose group was significantly reduced (p<0.01), indicating that the abalone umami peptide prepared in this example has antioxidant activity.

[0047] Table 4. Effects of abalone umami peptides on the MDA content in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01 As shown in Table 5, compared with the normal group, the SOD content in the serum and liver of the D-galactose model group was significantly reduced (p<0.01), indicating that the mouse aging model was successfully established. Compared with the model group, the SOD content in the serum and liver of the control group and the dose group was increased, and the SOD content in the serum and liver of the high-dose group mice was significantly increased (p<0.01), indicating that the abalone umami peptide prepared in this example has antioxidant activity.

[0048] Table 5. Effects of umami peptides on SOD levels in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01 Example 3 (1) After 1000L of abalone cooking wastewater was centrifuged at a speed of >10,000rpm to remove solid impurities, heavy metals were removed by electrodialysis at an electrodialysis current of 12A and an electrodialysis flow rate of 1.5m 3 / h.

[0049] (4) The material obtained in step (1) is placed in a reactor, and 0.1% of the weight of the solid calculated by the solid content in the material obtained in the above step (1) is added with a neutral protease to carry out an enzymatic hydrolysis reaction, and the pH value of the reaction system is controlled to be 7, the temperature of the enzymatic hydrolysis reaction is 50° C., and the time of the enzymatic hydrolysis reaction is 0.5 h. After the enzymatic hydrolysis reaction is completed, a nanofiltration membrane with a molecular weight cutoff of 2KD is used to remove macromolecular impurities, and a nanofiltration membrane with a molecular weight cutoff of 1KD is used to remove small molecular impurities. The material solution after membrane separation is spray dried, and the outlet air temperature of the spray drying is 70° C. and the inlet air temperature is 140° C., to obtain abalone umami peptide.

[0050] [Molecular weight detection] The test results show that the proportion of abalone umami peptides with a molecular weight of 1~2KD is 76%.

[0051] [Umami sensory evaluation] The evaluation results showed that the freshness score of abalone umami peptide was 9.71 points, and the sample had a strong umami taste. [Heavy metal content detection] The experimental results show that the inorganic arsenic, lead and methylmercury contents of the abalone umami peptide prepared in this embodiment are all less than 0.1, indicating that the abalone umami peptide prepared in this embodiment is highly safe.

[0052] [Safety Evaluation] There was no death in the animals of each dosage group, and no abnormal behavior and no obvious weight gain, indicating that the abalone umami peptide prepared in this example has no toxic side effects.

[0053] [Evaluation of antioxidant biological activity] As shown in Table 6, compared with the normal group, the MDA content in the serum and liver of the D-galactose model group was significantly increased (p<0.01), indicating that the mouse aging model was successfully established. Compared with the model group, the MDA content in the serum and liver of the control group and the dose group was reduced, and the MDA content in the serum and liver of the high-dose group mice was significantly reduced (p<0.01), indicating that the abalone umami peptide prepared in this example has antioxidant activity.

[0054] Table 6. Effects of abalone umami peptides on the MDA content in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01 As shown in Table 7, compared with the normal group, the SOD content in the serum and liver of the D-galactose model group was significantly reduced (p<0.01), indicating that the mouse aging model was successfully established. Compared with the model group, the SOD content in the serum and liver of the control group and the dose group was increased, and the SOD content in the serum and liver of the high-dose group mice was significantly increased (p<0.01), indicating that the abalone umami peptide prepared in this example has antioxidant activity.

[0055] Table 7. Effects of umami peptides on SOD levels in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01 Comparative Example 1 (1) After 100L of abalone cooking wastewater was centrifuged at a speed of >10,000rpm to remove solid impurities, heavy metals were removed by electrodialysis at an electrodialysis current of 10A and an electrodialysis flow rate of 0.8m 3 / h.

[0056] (2) The material obtained in step (1) is placed in a reactor, and 1% of the weight of the solid calculated by the solid content in the material obtained in the above step (1) is added to the A5 animal protein hydrolase for enzymatic hydrolysis reaction, and the pH value of the reaction system is controlled to be 6, the temperature of the enzymatic hydrolysis reaction is 60°C, and the time of the enzymatic hydrolysis reaction is 2h. After the enzymatic hydrolysis reaction is completed, the macromolecular impurities are removed by using a nanofiltration membrane with a molecular weight cutoff of 3KD, and the small molecular impurities are removed by using a nanofiltration membrane with a molecular weight of 800Da. The material solution after membrane separation is spray dried, and the outlet air temperature of the spray drying is 85°C and the inlet air temperature is 170°C to obtain abalone umami peptide.

[0057] [Molecular weight detection] The test results show that the proportion of abalone umami peptides with a molecular weight of 1~2KD is 57%.

[0058] [Umami sensory evaluation] The evaluation results showed that the freshness score of abalone umami peptide was 7.34 points, and the sample had a umami taste that was not strong.

[0059] [Heavy metal content detection] The inorganic arsenic, lead and methylmercury contents of the abalone umami peptide prepared in Comparative Example 1 are all less than 0.1, indicating that the abalone umami peptide prepared in this example is highly safe.

[0060] [Safety Evaluation] There was no death in the animals of each dosage group, and no abnormal behavior, and no obvious weight gain, indicating that the abalone umami peptide prepared in this comparative ratio had no toxic side effects.

[0061] [Evaluation of antioxidant biological activity] As shown in Table 8, compared with the normal group, the MDA content in the serum and liver of the D-galactose model group increased significantly (p<0.05), indicating that the mouse aging model was successfully established. Compared with the model group, the MDA content in the serum and liver of the control group and the dose group decreased, and the MDA content in the serum and liver of the high-dose group mice decreased significantly (p<0.05), indicating that the abalone umami peptide prepared in this comparative example has antioxidant activity.

[0062] Table 8. Effects of abalone umami peptides on MDA levels in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01, # p<0.05 As shown in Table 9, compared with the normal group, the SOD content in the serum and liver of the D-galactose model group was significantly reduced (p<0.01), indicating that the mouse aging model was successfully established. Compared with the model group, the SOD content in the serum and liver of the control group and the dose group was increased, and the SOD content in the serum and liver of the high-dose group mice was significantly increased (p<0.01), indicating that the abalone umami peptide prepared in this comparative example has antioxidant activity.

[0063] Table 9. Effects of umami peptides on SOD levels in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01 Comparative Example 2 10kg of abalone meat of Haliotis discus was added to a reactor, 100L of water was added, and 1% of the weight of the abalone meat was added for enzymolysis reaction. The pH value of the reaction system was controlled to be 6, the temperature of the enzymolysis reaction was 60°C, and the time of the enzymolysis reaction was 2h. After the enzymolysis reaction, a nanofiltration membrane with a molecular weight cutoff of 3KD was used to remove macromolecular impurities, and a nanofiltration membrane with a molecular weight of 800Da was used to remove small molecular impurities. The material solution after membrane separation was spray dried, and the outlet temperature of the spray dried was 85°C and the inlet temperature was 170°C to obtain abalone umami peptide.

[0064] [Molecular weight detection] The test results show that the proportion of abalone umami peptides with a molecular weight of 1~2KD is 43%.

[0065] [Umami sensory evaluation] The evaluation results showed that the freshness score of abalone umami peptide was 5.23 points, and the sample had a umami taste that was not strong.

[0066] [Safety Evaluation] There was no death in the animals of each dosage group, and no abnormal behavior, and no obvious weight gain, indicating that the abalone umami peptide prepared in this comparative ratio had no toxic side effects.

[0067] [Evaluation of antioxidant biological activity] As shown in Table 10, compared with the normal group, the MDA content in the serum and liver of the D-galactose model group increased significantly (p<0.05), indicating that the mouse aging model was successfully established. Compared with the model group, the MDA content in the serum and liver of the control group and the dose group decreased, and the MDA content in the serum and liver of the high-dose group mice decreased significantly (p<0.05), indicating that the abalone umami peptide prepared in this comparative example has antioxidant activity.

[0068] Table 10. Effects of abalone umami peptides on the MDA content in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01, # p<0.05 As shown in Table 11, compared with the normal group, the SOD content in the serum and liver of the D-galactose model group was significantly reduced (p<0.01), indicating that the mouse aging model was successfully established. Compared with the model group, the SOD content in the serum and liver of the control group and the dose group was increased, and the SOD content in the serum and liver of the high-dose group mice was significantly increased (p<0.01), indicating that the abalone umami peptide prepared in this comparative example has antioxidant activity.

[0069] Table 11. Effects of umami peptides on SOD levels in serum and liver of mice Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.01 Comparative Examples 4 to 9: (1) After 100L of abalone cooking wastewater was centrifuged at a speed of >10,000rpm to remove solid impurities, heavy metals were removed by electrodialysis at an electrodialysis current of 10A and an electrodialysis flow rate of 0.8m 3 / h.

[0070] (2) The material obtained in step (1) is placed in a reactor, and 1% of the weight of the solid calculated by the solid content in the material obtained in the above step (1) is added to perform enzymatic hydrolysis reaction, and the pH value of the reaction system is controlled to be 3-9, the temperature of the enzymatic hydrolysis reaction is 30-70°C, and the time of the enzymatic hydrolysis reaction is 2h. After the enzymatic hydrolysis reaction is completed, the impurities with molecular weights shown in Table 13 are removed by nanofiltration membrane, and the material solution after membrane separation is spray dried, and the outlet air temperature of the spray drying is 85°C and the inlet air temperature is 170°C to obtain abalone umami peptide.

[0071] Table 12 Preparation conditions of comparative examples 4 to 9 In Table 12, “—” indicates the same as Example 1; Comparative Example 8 uses a 3KD nanofiltration membrane to remove large molecular impurities without filtering out small molecular impurities, and Comparative Example 9 uses an 800Da nanofiltration membrane to remove small molecular impurities without filtering out large molecular impurities.

[0072] Test data of comparative examples 4 to 9: In Table 13, compared with the normal group, the MDA content in the liver of the D-galactose model group was significantly increased (p<0.01), indicating that the mouse aging model was successfully established. Compared with the normal group, the SOD content in the liver of the D-galactose model group was significantly decreased (p<0.01), indicating that the mouse aging model was successfully established.

[0073] Comparative Examples 4 to 9 adopted a high-dose group feeding scheme. Compared with the model group, the SOD in the liver of Comparative Examples 4 to 9 was only slightly increased, and the liver MDA was not significantly reduced. The SOD value and MDA value of Comparative Examples 4 to 9 were significantly deteriorated from the SOD value of 8.21±0.35 and the MDA value of 4.12±0.26 of the high-dose group in Example 1, indicating that the antioxidant capacity of the abalone umami peptide prepared in Comparative Examples 4 to 9 was significantly weakened. From the perspective of umami sensory evaluation, the umami sensory evaluation scores of Comparative Examples 4 to 9 were between 3.22 and 7.59, which significantly reduced the sweetness and even had some bitterness, which should be caused by the change in the composition of the peptides that affect the taste.

[0074] Table 13 Test data of comparative examples 4 to 9

[0075] # is a significant difference, ## is a very significant difference; Note: Compared with blank, ** p<0.01, compared with the model group, ## p<0.05; [Molecular weight detection] The test results show that the molecular weight of abalone umami peptides is between 1 and 2 KD.

[0076] The above description is only a preferred embodiment of the present application and is not intended to limit the protection scope of the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application are included in the protection scope of the present application.

Claims

1. A method for preparing abalone umami peptides using abalone cooking waste liquid as raw material, characterized in that: 1) Enzymatic hydrolysis reaction: adding protease to the abalone cooking waste liquid for enzymatic hydrolysis reaction; 2) Multi-stage membrane separation: Filter the solution after the enzymatic hydrolysis reaction in step 2) to remove peptides with molecular weight greater than 3KD and less than 800D; 3) Spray drying.

2. The method for preparing abalone umami peptide using abalone cooking waste liquid as raw material according to claim 1, characterized in that: In the enzymolysis reaction of step 1), the pH value of the abalone cooking waste liquid is 6-7, and / or the enzymolysis temperature of the abalone cooking waste liquid is 40-60° C., and / or the enzymolysis time of the abalone cooking waste liquid is 0.5-2 h.

3. The method for preparing abalone umami peptides using abalone cooking waste liquid as raw material according to any one of claims 1 and 2, characterized in that: The peptides with a molecular weight greater than 3KD and a molecular weight less than 800D are filtered out by the nanofiltration membrane.

4. The preparation method according to any one of claims 1 to 3, characterized in that: The protease is selected from one of neutral protease, ficin and bromelain.

5. The preparation method according to any one of claims 1 to 4, characterized in that: The mass of the protease feed is 0.1wt%~1wt% of the dry weight of the solid content of the abalone cooking waste liquid.

6. Abalone umami peptide prepared from abalone cooking waste liquid as raw material and a preparation method thereof according to any one of claims 1 to 5, characterized in that: The abalone cooking waste liquid in step 1) is further subjected to electrodialysis desorption before adding protease to remove heavy metals. The electrodialysis desorption is specifically as follows: S1: removing solid impurities from abalone cooking waste liquid; S2: Electrodialysis is used to remove heavy metals. The electrodialysis current is 8~12A and the electrodialysis flow rate is 0.6~1.2m 3 / h.

7. The preparation method according to claim 6, characterized in that: The solid impurities are removed by high-speed centrifugation, and the rotation speed of the high-speed centrifuge is >10000rpm.

8. The preparation method according to any one of claims 6 or 7, characterized in that: The outlet air temperature of the spray drying is 70-85°C, and the inlet air temperature is 140-170°C.

9. An abalone umami peptide, characterized in that: The method is prepared by any one of claims 1 to 8.

10. The abalone umami peptide according to claim 9, characterized in that The proportion of umami peptides with a molecular weight of 1~2KD is >70%.

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