Preparation method of abalone umami peptide using abalone cooking waste liquid as raw material

Abstract

The present application relates to the field of functional nutritional condiment, and specifically discloses abalone umami peptide prepared by using abalone cooking waste liquid as raw material and a preparation method thereof, which comprises the following steps: (1) removing solid impurities from the abalone cooking waste liquid and removing heavy metals by electrodialysis; (2) adding specific protease to the material obtained in step (1) at a dosage of 0.1-1% of the weight of the solid content in the material obtained in step (1), and performing enzymatic reaction at a pH value of 6-7 and a reaction temperature of 40-60 DEG C for 0.5-2 h; (3) removing impurities by using nanofiltration membrane and performing spray drying to obtain abalone umami peptide. The abalone umami peptide prepared by the method solves the problem of environmental pollution caused by a large amount of abalone cooking waste liquid, and realizes a new way of resource utilization of abalone cooking waste liquid. The abalone umami peptide has a molecular weight of 1-2 KD and accounts for more than 70%, has high antioxidant bioactivity, low heavy metal content, and rich seafood flavor, and can be widely applied in the field of functional nutritional condiment.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of functional nutritional condiments, in particular to a kind of abalone umami peptide prepared with abalone cooking waste liquid as raw material and a preparation method thereof. BACKGROUND

[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 into the market, which are deeply loved by consumers. Although these umami substances are rich in umami, they do not have antioxidant effect. Therefore, there is a huge market demand for developing umami peptides with antioxidant effect.

[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 polypeptides and water-soluble proteins, they are excellent raw materials for developing umami peptides. Currently, there is no patent for developing 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 hinders its industrial production and promotion. Therefore, how to utilize abalone cooking waste liquid resources in a high-value way, turn waste into treasure, develop abalone umami peptides with extremely high utilization value, extend the abalone industry chain, and relieve environmental and enterprise pressure is a problem to be solved. SUMMARY

[0004] In view of the problem of a large amount of cooking waste liquid generated during the development of abalone products, which causes environmental pollution, the purpose of the present application is to provide a preparation method of abalone umami peptide with mild reaction conditions, controllable quality and suitable for large-scale production, which fully utilizes abalone cooking waste liquid, turns waste into treasure and develops abalone umami peptide with high added value.

[0005] To solve the above technical problems, the first aspect of the present application provides a kind of abalone umami peptide prepared with abalone cooking waste liquid as raw material and a preparation method thereof, characterized in that:

[0006] 1) Enzymatic reaction: proteinase is added to the abalone cooking waste liquid for enzymatic reaction;

[0007] 2) Multi-stage membrane separation: the solution after step 2) enzymatic reaction is filtered to remove peptides with molecular weight greater than 3KD and molecular weight less than 800D;

[0008] 3) Spray drying.

[0009] In some specific embodiments of the first aspect, the pH value of the abalone cooking waste liquid in the step 1) enzymatic reaction 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.

[0010] In some 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.

[0011] In some 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.

[0012] In some embodiments of the first aspect, the peptides with a molecular weight greater than 3KD and a molecular weight less than 800D are removed by a nanofiltration membrane.

[0013] In some embodiments of the first aspect, the abalone cooking waste liquid of step 1) is subjected to electrodialysis before the protease is added, to remove heavy metals, and the electrodialysis is specifically:

[0014] S1: removing solid impurities from the abalone cooking waste liquid;

[0015] S2: removing heavy metals by electrodialysis, the electrodialysis current is 8-12A, and the electrodialysis flow rate is 0.6-1.2m 3 / h;

[0016] 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.

[0017] In some embodiments, the electrodialysis flow rate is 0.7m 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.

[0018] In some embodiments of the first aspect, the protease is selected from one of neutral protease, ficin, and bromelain.

[0019] In some embodiments of the first aspect, the protease is added in an amount of 0.1wt%-1wt% of the dry weight of the solid content of the abalone cooking waste liquid.

[0020] In some embodiments of the first aspect, the solid impurities are removed by high speed centrifugation, and the speed of the high speed centrifuge is >10,000 rpm, in some embodiments, the speed of the high speed centrifuge is >11,000 rpm, in some embodiments, the speed of the high speed centrifuge is >12,000 rpm, and in some embodiments, the speed of the high speed centrifuge is >13,000 rpm.

[0021] In some embodiments of the first aspect, the outlet temperature of the spray drying is 70-85°C, and the inlet temperature of the spray drying is 140-170°C, in some embodiments, the outlet temperature of the spray drying is 74°C, in some embodiments, the outlet temperature of the spray drying is 78°C, in some embodiments, the outlet temperature of the spray drying is 82°C, in some embodiments, the inlet temperature of the spray drying is 145°C, in some embodiments, the inlet temperature of the spray drying is 150°C, in some embodiments, the inlet temperature of the spray drying is 155°C, and in some embodiments, the inlet temperature of the spray drying is 160°C.

[0022] The second aspect of the present application provides a fresh-taste peptide of abalone, which is prepared by the method of any one of the embodiments of the first aspect.

[0023] In some embodiments of the second aspect, the proportion of the fresh-taste peptide with a molecular weight of 1-2KD is >70%, in some embodiments, the proportion of the fresh-taste peptide with a molecular weight of 1-2KD is >70%, in some embodiments, the proportion of the fresh-taste peptide with a molecular weight of 1-2KD is >80%, and in some embodiments, the proportion of the fresh-taste peptide with a molecular weight of 1-2KD is >90%.

[0024] The reagents used in the present application are all purchased from open and legal markets, and are not further purified.

[0025] Term Explanation

[0026] Certain embodiments of the present application are now described in detail. The application is intended to cover all alternatives, modifications and equivalents thereof which are included within the scope of the application as defined in the claims. One skilled in the art will readily recognize from the disclosure herein, numerous other methods and materials similar or equivalent to those described herein, and the application extends to each such method and material, and is intended to embrace each and every such modification and change. The application is not limited to the methods and materials described herein, but extends to all methods and materials within the scope of the claims. In the event that one or more of the incorporated literature, patents, and similar materials differs from the claims or the disclosure herein, including terminology, definitions, materials, and / or methodologies, such conflicting provisions are intended to be controlled by the present claims, which shall prevail.

[0027] It is further recognized that certain features of the application, for clarity, are described in relation to only one or a few embodiments while they can be used in combination with one or more of the other embodiments. Conversely, various features of the application, for brevity, are described in relation to only one or a few embodiments while they can be used in combination with one or more of the other embodiments.

[0028] Unless otherwise stated, all technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art. All patents and publications related to this invention are incorporated herein by reference in their entirety.

[0029] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0030] In the following content, all figures disclosed herein, whether or not they use words such as "approximately" or "about," are approximate values. The value of each figure may vary by 1%, 2%, 5%, 7%, 8%, 10%, 15%, or 20%. Whenever a figure with a value of N is disclosed, any figure 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 "+ / -" indicates addition or subtraction.

[0031] Abalone cooking waste liquid is the cooking waste liquid produced after abalone is cooked in hot water.

[0032] [Molecular weight determination]

[0033] The molecular weight of abalone umami peptides was determined by gel permeation chromatography (GPC) according to the national standard GB / T22729-2008. A TSKgel G200 SWxL column (300 mm x 7.8 mm, 5 μm) was used, and the mobile phase was acetonitrile-water-trifluoroacetic acid (volume ratio 45:55:0.1). The samples were measured at a detection wavelength of 220 nm under the conditions of column temperature 30℃, flow rate 0.5 mL / min. The standard sample was: acetic acid-acetic acid-acetic acid. Mr 189.17, Ethanol-Ethanol-Tyrosine-Arginine: Mr 451.48, Bacillus enzyme: Mr 1422.69, aprotinin: Mr 6511.44, Cytochrome C: Mr 12384.

[0034] [Umami sensory evaluation]

[0035] Please 10 industry personnel, according to table 1 umami sensory evaluation form to the freshness of abalone umami peptide prepared in this embodiment, adopt the ten system, score calculation take its average value.

[0036] Table 1. Umami sensory evaluation standard table

[0037]

[0038] [Heavy metal content detection]

[0039] 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 methyl mercury in the product.

[0040] [Security evaluation]

[0041] The Kunming mice were gavaged with different doses of abalone umami peptide prepared in this embodiment, and each dose group was 1g / kg, 5g / kg, 10g / kg, 15g / kg.

[0042] [Antioxidant biological activity evaluation]

[0043] Select 25-30g healthy adult mice, except for the blank control group, the rest of the animals are modeled by intraperitoneal injection of D-galactose, design 1 model control group and 3 test sample dose groups (low dose group 100mg.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 is given the same volume of solvent, the mice are given the test product by gavage, at the same time of giving the test sample, the model control group and each dose group continue to be given the same dose of D-galactose intraperitoneal injection, the animals are sacrificed at the end of the experiment, and the antioxidant activity of the sample is measured.

[0044] Advantages of the present application:

[0045] Compared with the prior art, one embodiment of the present application has at least one of the following beneficial effects:

[0046] The present application uses abalone cooking waste liquid as raw material to develop abalone umami peptide, solves the bottleneck problem of environmental pollution caused by waste liquid in the processing of abalone products, and realizes a new way of high value utilization of abalone cooking waste liquid.

[0047] 2. The abalone umami peptide produced by the present application has high safety, no toxic side effects, low heavy metal content, and the proportion of abalone umami peptide with a molecular weight of 1-2KD is >70%, the sensory score of the product is >9, and the product can be applied in the field of functional nutritional condiments. DETAILED DESCRIPTION

[0048] The technical solutions in the embodiments of the present application will be clearly and completely described below in combination with the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0049] Embodiment 1

[0050] (1) 100L of abalone cooking waste liquid, after removing solid impurities by a centrifuge with a speed of >10000rpm, heavy metals are removed by electrodialysis, the current of electrodialysis is 10A, and the flow rate of electrodialysis is 0.8m 3 / h.

[0051] (2) The material obtained in step (1) is placed in a reaction kettle, and 1% of the weight of the solid in the material obtained in step (1) of ficin is added for enzymatic reaction, the pH value of the reaction system is controlled to be 6, the temperature of the enzymatic reaction is 60℃, and the time of the enzymatic reaction is 2h. After the enzymatic reaction is completed, macromolecular impurities are removed by using a nanofiltration membrane with a molecular weight cut-off of 3KD, and small molecular impurities are removed by using a nanofiltration membrane with a molecular weight cut-off of 800Da, and the material solution after membrane separation is spray dried, the outlet air temperature of spray drying is 85℃, and the inlet air temperature is 170℃, to obtain abalone umami peptide.

[0052] [Detection of molecular weight] The detection results show that the proportion of abalone umami peptide with a molecular weight of 1-2KD is 73%.

[0053] [Umami sensory evaluation] The evaluation results show that the freshness score of the abalone umami peptide is 9.16, and the sample has a rich umami taste

[0054] [Detection of heavy metal content] The inorganic arsenic, lead and methyl mercury contents of the abalone umami peptide prepared in embodiment 1 are all less than 0.1, which shows that the abalone umami peptide prepared in the embodiment has high safety.

[0055] [Safety evaluation] The animals in each dose group have no death, no abnormal behavior, and no obvious weight gain, which shows that the abalone umami peptide prepared in the embodiment has no toxic side effects.

[0056] [Antioxidant biological activity evaluation]

[0057] From Table 2, compared with the normal group, the MDA content in serum and liver of the D-galactose model group was extremely significantly increased (P<0.01) p <0.01), indicating that the mouse aging model was successfully modeled. 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 extremely significantly reduced (P<0.01) p <0.01), indicating that the abalone delicious peptide prepared in this embodiment had antioxidant activity.

[0058] Table 2. Effect of abalone delicious peptide on MDA content in serum and liver of mice

[0059]

[0060] Note: compared with the blank, ** p <0.01, compared with the model group, ## p <0.01

[0061] From Table 3, compared with the normal group, the SOD content in serum and liver of the D-galactose model group was extremely significantly reduced (P<0.01) p <0.01), indicating that the mouse aging model was successfully modeled. Compared with the model group, the SOD content in serum and liver of the control group and the dose group was increased, and the SOD content in serum and liver of the high-dose group was extremely significantly increased (P<0.01) p <0.01), indicating that the abalone delicious peptide prepared in this embodiment had antioxidant activity.

[0062] Table 3. Effect of delicious peptide on SOD content in serum and liver of mice

[0063]

[0064] Note: compared with the blank, ** p <0.01, compared with the model group, ## p <0.01

[0065] Example 2

[0066] (1) 500L of abalone cooking waste liquid, after removing solid impurities by a centrifuge with a speed of >10000rpm, heavy metals were removed by electrodialysis, the electrodialysis current was 11A, and the electrodialysis flow rate was 1m 3 / h.

[0067] (3) Place the material obtained in step (1) into a reaction vessel, add 0.5% of bromelain (based on the weight of solids in the material obtained in step (1)) for enzymatic hydrolysis, control the pH of the reaction system to 6.5, the temperature of the enzymatic hydrolysis reaction to 40℃, and the time of the enzymatic hydrolysis reaction to 1h. After the enzymatic hydrolysis reaction is completed, use a nanofiltration membrane with a molecular weight cutoff of 3KD to remove macromolecular impurities, use a nanofiltration membrane with a molecular weight cutoff of 800Da to remove small molecule impurities, and spray dry the material solution after membrane separation. The outlet air temperature of the spray dryer is 75℃ and the inlet air temperature is 150℃ to obtain abalone umami peptide.

[0068] [Molecular weight detection] The detection results showed that 79% of the abalone umami peptides had a molecular weight of 1-2 KD.

[0069] [Sensory Evaluation of Umami] The evaluation results showed that the umami peptides of abalone scored 9.37 points, indicating that the sample had a rich umami flavor.

[0070] [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 has high safety.

[0071] [Safety Evaluation] No animals in any dosage group died, exhibited no behavioral abnormalities, or showed significant weight gain, indicating that the abalone umami peptide prepared in this embodiment has no toxic side effects.

[0072] [Evaluation of Antioxidant Bioactivity]

[0073] As shown in Table 4, compared with the normal group, the serum and liver MDA levels in the D-galactose model group were significantly increased. p The result < 0.01 indicates that the mouse aging model was successfully established.

[0074] Compared with the model group, the serum and liver MDA levels in both the control and dosage groups were decreased, and the serum and liver MDA levels in the high-dose group were significantly decreased. p The value of < 0.01 indicates that the abalone umami peptide prepared in this embodiment has antioxidant activity.

[0075] Table 4. Effects of abalone umami peptides on MDA levels in mouse serum and liver

[0076]

[0077] Note: Compared with blank, ** p < 0.01, compared to the model group, ## p < 0.01

[0078] 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 extremely significantly reduced (P<0.01) p <0.01), indicating that the mouse aging model was successfully modeled. 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 was extremely significantly increased (P<0.01), indicating that the abalone umami peptide prepared in this embodiment had antioxidant activity. p

[0079] Table 5. Effect of umami peptide on SOD content in serum and liver of mice

[0080]

[0081] Note: compared with the blank, ** p <0.01, compared with the model group, ## p <0.01

[0082] Example 3

[0083] (1) 1000L of abalone cooking waste liquid, after removing solid impurities by a centrifuge with a speed of >10000rpm, heavy metals were removed by electrodialysis, the electrodialysis current was 12A, and the electrodialysis flow rate was 1.5m 3 / h.

[0084] (4) The material obtained in step (1) was placed in a reaction kettle, and 0.1% of neutral protease based on the weight of the solid content in the material obtained in step (1) was added for enzymatic reaction, the pH value of the reaction system was controlled at 7, the enzymatic reaction temperature was 50°C, and the enzymatic reaction time was 0.5h. After the enzymatic reaction was completed, macromolecular impurities were removed by using a nanofiltration membrane with a molecular weight cutoff of 2KD, small molecular impurities were removed by using a nanofiltration membrane with a molecular weight cutoff of 1KD, and the material solution after membrane separation was spray dried, with an air outlet temperature of 70°C and an air inlet temperature of 140°C, to obtain abalone umami peptide.

[0085] [Detection of molecular weight] The detection results showed that the proportion of abalone umami peptide with a molecular weight of 1-2KD was 76%.

[0086] [Umami sensory evaluation] The evaluation results showed that the freshness score of the abalone umami peptide was 9.71 points, and the sample had a rich umami taste

[0087] [Heavy metal content detection] The experimental results showed that the inorganic arsenic, lead and methyl mercury contents of the abalone umami peptide prepared in this embodiment were all less than 0.1, indicating that the abalone umami peptide prepared in this embodiment had high safety.

[0088] ​[Safety Evaluation] No animals in any dosage group died, exhibited no behavioral abnormalities, or showed significant weight gain, indicating that the abalone umami peptide prepared in this embodiment has no toxic side effects.

[0089] [Evaluation of Antioxidant Bioactivity]

[0090] As shown in Table 6, compared with the normal group, the MDA content in serum and liver of the D-galactose model group was significantly increased. p < 0.01 indicates successful establishment of the mouse aging model. Compared with the model group, the serum and liver MDA levels in both the control and dosage groups were decreased, and the serum and liver MDA levels in the high-dose group were significantly decreased ( p The value of < 0.01 indicates that the abalone umami peptide prepared in this embodiment has antioxidant activity.

[0091] Table 6. Effects of abalone umami peptides on MDA levels in mouse serum and liver

[0092]

[0093] Note: Compared with blank, ** p < 0.01, compared to the model group, ## p < 0.01

[0094] As shown in Table 7, compared with the normal group, the serum and liver SOD levels in the D-galactose model group were significantly reduced. p < 0.01 indicates successful establishment of the mouse aging model. Compared with the model group, the serum and liver SOD levels in both the control and dosage groups were increased, and the serum and liver SOD levels in the high-dose group were significantly increased ( p The value of < 0.01 indicates that the abalone umami peptide prepared in this embodiment has antioxidant activity.

[0095] Table 7. Effects of umami peptides on SOD levels in mouse serum and liver

[0096]

[0097] Note: Compared with blank, ** p < 0.01, compared to the model group, ## p < 0.01

[0098] Comparative Example 1

[0099] (1) 100L of abalone cooking waste liquid was centrifuged at a speed of >10000rpm to remove solid impurities, and then electrodialysis was used to remove heavy metals. The electrodialysis current was 10A and the electrodialysis flow rate was 0.8m³. 3 / h.

[0100] (2) Place the material obtained in step (1) into a reaction vessel, add 1% of A5 animal protein hydrolase (based on the weight of solids in the material obtained in step (1)) for enzymatic hydrolysis, control the pH of the reaction system to 6, the temperature of the enzymatic hydrolysis reaction to 60℃, and the time of the enzymatic hydrolysis reaction to 2h. After the enzymatic hydrolysis reaction is completed, use a nanofiltration membrane with a molecular weight cutoff of 3KD to remove macromolecular impurities, use a nanofiltration membrane with a molecular weight cutoff of 800Da to remove small molecule impurities, and spray dry the material solution after membrane separation. The outlet air temperature of the spray dryer is 85℃ and the inlet air temperature is 170℃ to obtain abalone umami peptides.

[0101] [Molecular weight detection] The detection results showed that 57% of the abalone umami peptides had a molecular weight of 1-2 kDa.

[0102] [Sensory Evaluation of Umami] The evaluation results showed that the umami peptide of abalone scored 7.34 points, indicating that the umami flavor of the sample was not strong.

[0103] [Heavy Metal Content Detection] The inorganic arsenic, lead and methylmercury contents of the abalone umami peptide prepared in Comparative Example 1 were all less than 0.1, indicating that the abalone umami peptide prepared in this example has high safety.

[0104] [Safety Evaluation] No animals in any dosage group died, exhibited no behavioral abnormalities, or showed significant weight gain, indicating that the abalone umami peptide prepared in this comparative study has no toxic side effects.

[0105] [Evaluation of Antioxidant Bioactivity]

[0106] As shown in Table 8, compared with the normal group, the MDA content in serum and liver of the D-galactose model group was significantly increased. p < 0.05 indicates successful establishment of the mouse aging model. Compared with the model group, the serum and liver MDA levels in both the control and dosage groups were decreased, and the serum and liver MDA levels in the high-dose group were significantly decreased ( p The value of < 0.05 indicates that the abalone umami peptide prepared in this comparative example has antioxidant activity.

[0107] Table 8. Effects of abalone umami peptides on MDA levels in mouse serum and liver

[0108]

[0109] Note: Compared with blank, ** p< 0.01, compared with the model group, ## p < 0.01, # p < 0.05

[0110] 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 extremely significantly reduced (P < 0.01), p < 0.01), indicating that the mouse aging model was successfully modeled. 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 was extremely significantly increased (P < 0.01), p < 0.01), indicating that the abalone umami peptide prepared in the comparative example had antioxidant activity.

[0111] Table 9. Effect of umami peptide on SOD content in serum and liver of mice

[0112]

[0113] Note: compared with the blank, ** p < 0.01, compared with the model group, ## p < 0.01

[0114] Comparative Example 2

[0115] 10 kg of abalone meat was added to the reaction kettle, 100 L of water was added, 1% of ficin was added based on the weight of the abalone meat for enzymatic hydrolysis reaction, the pH value of the reaction system was controlled at 6, the enzymatic hydrolysis reaction temperature was 60℃, and the enzymatic hydrolysis reaction time was 2 h. After the enzymatic hydrolysis reaction was completed, the macromolecular impurities were removed by using a nanofiltration membrane with a molecular weight cut-off of 3KD, the small molecular impurities were removed by using a nanofiltration membrane with a molecular weight of 800 Da, and the material solution after membrane separation was spray dried. The outflow temperature of the spray dryer was 85℃, and the inlet temperature was 170℃, to obtain abalone umami peptide.

[0116] The detection results of [molecular weight detection] showed that the proportion of abalone umami peptide with a molecular weight of 1-2KD was 43%.

[0117] The evaluation results of [umami sensory evaluation] showed that the freshness score of the abalone umami peptide was 5.23 points, and the sample had a not rich umami taste.

[0118] [Safety evaluation] The animals in each dose group had no death, no abnormal behavior, and no significant weight gain, indicating that the abalone umami peptide prepared in the comparative example had no toxic and side effects.

[0119] [Antioxidant biological activity evaluation]

[0120] From Table 10, compared with the normal group, the MDA content in the serum and liver of the D-galactose model group was significantly increased p < 0.05), indicating that the mouse aging model was successfully modeled. 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 was significantly reduced p < 0.05), indicating that the abalone umami peptide prepared in the comparative example had antioxidant activity.

[0121] Table 10. Effect of abalone umami peptide on MDA content in serum and liver of mice

[0122]

[0123] Note: compared with the blank, ** p < 0.01, compared with the model group, ## p < 0.01, # p < 0.05

[0124] From Table 11, compared with the normal group, the SOD content in the serum and liver of the D-galactose model group was extremely significantly reduced p < 0.01), indicating that the mouse aging model was successfully modeled. 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 was extremely significantly increased p < 0.01), indicating that the abalone umami peptide prepared in the comparative example had antioxidant activity.

[0125] Table 11. Effect of umami peptide on SOD content in serum and liver of mice

[0126]

[0127] Note: compared with the blank, ** p < 0.01, compared with the model group, ## p < 0.01

[0128] Comparative examples 4-9:

[0129] (1) 100 L of abalone cooking waste liquid, after removing solid impurities by a centrifuge with a speed of > 10,000 rpm, heavy metals were removed by electrodialysis, the electrodialysis current was 10 A, and the electrodialysis flow rate was 0.8 m 3 / h.

[0130] (2) The material obtained in step (1) is placed in a reaction kettle, and 1% of the weight of the solid in the material obtained in step (1) of ficin is added for enzymatic reaction, the pH value of the reaction system is controlled at 3-9, the enzymatic reaction temperature is 30-70°C, and the enzymatic reaction time is 2h. After the enzymatic reaction is completed, the impurities with the molecular weight shown in Table 13 are removed by nanofiltration membrane, and the material solution after membrane separation is spray dried, the air outlet temperature of spray drying is 85°C, and the air inlet temperature is 170°C, to obtain abalone umami peptide.

[0131] Table 12 Preparation conditions of Comparative Examples 4-9

[0132]

[0133] In Table 12, “-” indicates the same as Example 1; Comparative Example 8 is to remove macromolecular impurities by using a 3KD nanofiltration membrane without filtering out small molecular impurities, and Comparative Example 9 is to remove small molecular impurities by using an 800Da nanofiltration membrane without filtering out macromolecular impurities.

[0134] Test data of Comparative Examples 4-9:

[0135] In Table 13, compared with the normal group, the MDA content in the liver of the D-galactose model group was extremely 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 extremely significantly decreased (P<0.01), indicating that the mouse aging model was successfully established. p p < 0.01), indicating that the mouse aging model was successfully established.

[0136] Comparative Examples 4-9 use a high-dose group feeding scheme, and the SOD in the liver of Comparative Examples 4-9 is only slightly increased compared with the model group, and the decrease of liver MDA is not significant. The SOD value and MDA value of Comparative Examples 4-9 are significantly worse than the SOD value 8.21±0.35 and MDA value 4.12±0.26 of the high-dose group in the scheme of Example 1, indicating that the antioxidant capacity of the abalone umami peptide prepared in Comparative Examples 4-9 has been significantly weakened. From the umami sensory evaluation, the umami sensory evaluation score of Comparative Examples 4-9 is between 3.22 and 7.59, significantly reducing the sweetness, and even appearing part of the bitter taste, which should be caused by the change of the peptide composition affecting the taste.

[0137] Table 13 Test data of Comparative Examples 4-9

[0138]

[0139] #is a significant difference, ##is an extremely significant difference;

[0140] Note: compared with the blank, ** ​p < 0.01, compared with the model group, ## p < 0.05;

[0141] The detection result of molecular weight detection shows that the proportion of the molecular weight of the abalone umami peptide is 1-2KD.

[0142] The above merely describes the preferred embodiments of the present application, but is not used 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 shall be included in the protection scope of the present application.

Claims

1. A method for preparing abalone umami peptide from abalone cooking waste liquid, characterized in that: 1) enzyme hydrolysis reaction: abalone cooking waste liquid is added with protease for enzyme hydrolysis reaction; 2) multi-stage membrane separation: the solution after step 1) enzyme hydrolysis reaction is filtered to remove peptides with molecular weight greater than 3KD and molecular weight less than 800D; 3) spray drying; in the 1) enzyme hydrolysis reaction, the pH value of the abalone cooking waste liquid is 6-7; in the 1) enzyme hydrolysis reaction, the enzyme hydrolysis temperature of the abalone cooking waste liquid is 40-60℃; the protease is selected from one of ficin, bromelain and neutral protease; in the step 1) enzyme hydrolysis reaction, the enzyme hydrolysis time of the abalone cooking waste liquid is 0.5-2h; the peptides with molecular weight greater than 3KD and molecular weight less than 800D are filtered by nanofiltration membrane; the mass of the protease is 0.1wt%-1wt% of the dry weight of the solid content of the abalone cooking waste liquid; before the abalone cooking waste liquid of step 1) is added with protease, it is subjected to electrodialysis to remove heavy metals, and the electrodialysis is specifically: S1: removing solid impurities from the abalone cooking waste liquid; the solid impurities are removed by high-speed centrifugation, and the speed of the high-speed centrifuge is >10000rpm; the outflow temperature of the spray drying is 70-85℃, and the inflow temperature is 140-170℃; the abalone umami peptide is prepared by the method of any one of claims 1-7; the proportion of the umami peptide with molecular weight of 1-2KD is >70%. ​ ​ ​ ​ ​ ​ 2. The method for preparing abalone umami peptides using abalone cooking waste liquid as raw material according to claim 1, characterized in that, ​ 3. The method for preparing abalone umami peptides using abalone cooking waste liquid as raw material according to claim 1, characterized in that, ​ 4. The method of claim 1, wherein, ​ 5. The method according to claim 1, wherein the method is characterized by, ​ ​ S2: removing heavy metals by electrodialysis, the electrodialysis current is 8-12 A, the electrodialysis flow is 0.6-1.2 m 3 / h.

6. The preparation method according to claim 5, characterized in that, ​ 7. The preparation method according to claim 1, characterized in that, ​ 8. A scallop umami peptide, characterized in that, ​ 9. The abalone umami peptide according to claim 8, characterized in that, ​

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