A modified immobilized lipase and its use in the preparation of sauces

Abstract

The application relates to the technical field of by-products processing, and particularly discloses a modified immobilized lipase and application thereof in preparation of sauce. The application uses salmon bone row processing by-products, i.e. bone residues, modifies the bone residues, and adopts an adsorption-embedding combined method to immobilize lipase by taking the modified bone residues as a carrier, so that the enzyme activity recovery rate, stability and repeatability of the immobilized lipase are effectively improved. The modified immobilized lipase is combined with a compounded protease, so that the enzymolysis time can be shortened, the amino acid nitrogen level can be improved, and the production cost can be significantly reduced.

Description

Technical Field

[0001] This application relates to the field of by-product processing technology, and in particular to a modified immobilized lipase and its application in the preparation of seasoning sauces. Background Technology

[0002] Currently, salmon is not processed to a high degree, resulting in a large amount of scraps, including fish heads and bones, which are rarely utilized as resources, leading to low economic benefits. Salmon bones are high in protein, fat, and inorganic salts. They are commonly used in the production of seasoning sauces with compound proteases. However, the high fat content negatively impacts the effectiveness of fish protein hydrolysis and the flavor of the product. For example, patent CN118307628A, "An Atlantic Salmon Bone Filling Umami Peptide and Its Preparation Method and Application," uses flavor proteases and compound proteases to hydrolyze Atlantic salmon bone filling mud to produce seasoning sauce. Because salmon bones are rich in oil, they are difficult to separate, resulting in a low yield and a high susceptibility to oxidative rancidity, severely affecting the flavor of the seasoning sauce. Similarly, patent CN106071037B, "A Fish Bone Peptide Pretreatment Process for Slag Removal and Degreasing," addresses the high fat content by using processes such as steaming, pressing, and rinsing to remove oil before enzymatic hydrolysis to produce fish bone peptides. However, after high-temperature steaming, the endogenous enzymes in the salmon bones are deactivated, requiring the addition of more proteases. Furthermore, the oil oxidizes after high-temperature treatment, which is also a waste of resources.

[0003] Direct enzymatic hydrolysis using free enzymes typically results in enzyme preparations that can only be used once, cannot be recycled, and suffer from poor stability and high difficulty in separation and purification. Immobilized enzymes effectively avoid these problems. Patent CN119775070A, "A Fermented Fish Protein Composition and Its Preparation Process," discloses a method for using immobilized proteases to hydrolyze fish proteins. This method uses heat denaturation and low-temperature refolding to expose the enzyme cleavage sites, and then uses mesoporous silica as a carrier to adsorb and immobilize the protease. Patent CN118652730A, "A Method for Low-Temperature Crystallization Enrichment of Fish Oil EPA and DHA and High-Activity Fish Oil," uses a collagen modifier to crosslink and immobilize lipases, and the recovered lipases are reusable. However, both of these immobilization methods have certain drawbacks. For physically adsorbed immobilized enzymes, the interaction between the carrier and the enzyme is weak, the enzyme is prone to detachment, and the stability is generally poor. The crosslinking method involves a violent reaction, which has a significant impact on enzyme activity, and the process is complex and the carrier is expensive, making it difficult to industrialize. Summary of the Invention

[0004] The purpose of this application is to overcome the shortcomings of the prior art and provide a modified immobilized lipase and its application in the preparation of seasoning sauce.

[0005] To achieve the above objectives, the technical solution adopted in this application is as follows: This application provides a method for preparing a modified immobilized lipase, comprising the following steps: S1. Add water to the bone residue produced after processing salmon bones and rinse it to filter out the bone residue. S2. Dry and pulverize the bone residue to obtain bone residue powder, and then treat it with microwave to obtain modified bone residue; S3. Dissolve the lipase in a buffer solution, then add the modified bone residue obtained in step S2 for adsorption to obtain a mixture; S4. Add the mixture obtained in step S3 to the embedding agent, filter and wash to form a modified immobilized lipase.

[0006] This application provides a method for preparing modified immobilized lipase using low-cost raw materials. The method uses salmon bone scraps, a by-product of salmon bone processing, which is modified and then immobilized using an adsorption-embedding method. This effectively improves the enzyme activity recovery rate, stability, and reproducibility of the immobilized lipase.

[0007] In this process, bone residue is washed, dried, and pulverized, then modified using microwaves. This process creates abundant micropores and an uneven structure on the surface of the modified bone residue, increasing its specific surface area and exposing more contact sites for lipase. When mixed with lipase, the pores in the modified bone residue can adsorb the lipase. Subsequently, the modified bone residue is used to adsorb the lipase, and then the lipase is encapsulated and immobilized, effectively improving the stability and recyclability of the lipase.

[0008] This application employs an adsorption-embedding method to immobilize lipase. The conditions are mild and do not require complex chemical cross-linking processes. This method can largely maintain the enzyme activity recovery rate and stability of lipase. The bone residue generated from enzymatic hydrolysis can be reused as a carrier raw material, thus realizing a recyclable production process system.

[0009] In a preferred embodiment of the preparation method of the modified immobilized lipase described in this application, in step S2, the bone residue powder has a mesh size of 300-800 mesh.

[0010] In a preferred embodiment of the preparation method of the modified immobilized lipase described in this application, the microwave treatment in step S2 includes: the microwave power is 450~600W and the microwave time is 3~5min.

[0011] The carrier used in this application is derived from a byproduct of salmon bone processing. By modifying the bone residue through microwave treatment, it has been developed into a low-cost and high-efficiency immobilized enzyme carrier, realizing the recycling of waste resources.

[0012] In a preferred embodiment of the method for preparing the modified immobilized lipase described in this application, in step S3, the mass ratio of lipase to modified bone residue is 1:(1~1.5).

[0013] In a preferred embodiment of the preparation method of the modified immobilized lipase described in this application, in step S4, the encapsulating agent includes a mixture of calcium salt solution, sodium alginate and gelatin.

[0014] The calcium salt solution includes CaCl2 solution; calcium ions form a gel with sodium alginate, which is then co-encapsulated with gelatin to encapsulate lipase.

[0015] This application also provides a modified immobilized lipase prepared by the above-described method.

[0016] This application also provides the application of the above-mentioned modified immobilized lipase in the preparation of seasoning sauces.

[0017] This application also provides a method for producing a seasoning sauce, comprising the following steps: 1) Weigh salmon paste, add the modified immobilized lipase, neutral protease and collagenase mentioned above for enzymatic hydrolysis, and then inactivate the enzymes to obtain the hydrolysate; 2) Centrifuge the enzymatic hydrolysate, take the middle water layer, and obtain the seasoning sauce.

[0018] When salmon bone fillets are used to produce seasoning sauce through enzymatic hydrolysis with compound proteases, it is difficult to avoid problems such as low yield and poor flavor caused by high fat content. This application uses modified immobilized lipase, which can effectively improve the above problems. Modified immobilized lipase is essentially a protein. The modified immobilized lipase is used in combination with neutral protease and collagenase to avoid the modified immobilized lipase being hydrolyzed by proteases, thus ensuring the stability of the lipase and enabling its recycling.

[0019] In the technical solution of this application, modified immobilized lipase is used in combination with a compound protease to enzymatically hydrolyze salmon bone fillets. The immobilized lipase can specifically degrade ester bonds, destroying the lipid-protein encapsulation structure and fully exposing the protein substrate. Salmon bone fillets contain a high proportion of bones and skin. The use of neutral protease and collagenase allows for synergistic action, breaking down proteins into small peptides and free amino acids, increasing the release rate of amino acid nitrogen. Centrifugation is used to collect the middle water layer, yielding a seasoning sauce that balances flavor and quality. The upper layer is the oil layer, which has not yet been destroyed by high temperatures and can be pressed to obtain high-quality fish oil. The lower solid residue mainly consists of immobilized lipase and bone residue. Washing and sieving can separate the two. The immobilized lipase can be recycled, and the bone residue can be processed and reused as a carrier, achieving continuous, cyclical, and sustainable resource utilization.

[0020] In a preferred embodiment of the production method of the seasoning sauce described in this application, the mass ratio of the modified immobilized lipase, neutral protease and collagenase is (0.1~0.2):(0.25~0.5):(0.15~0.3), preferably 0.2:0.5:0.3.

[0021] In the technical solution of this application, modified immobilized lipase, neutral protease, and collagenase are used in combination, taking into account the characteristics of salmon bone fillets. This effectively increases the amino acid nitrogen value and reduces bitterness, resulting in a flavorful seasoning sauce. Furthermore, the combined use of these enzymes can shorten the enzymatic hydrolysis time and significantly reduce production costs.

[0022] This application employs modified immobilized lipase, neutral protease, and collagenase in the above-mentioned mass ratio. The modified immobilized lipase can specifically enzymatically break down ester bonds, disrupting the lipid-protein encapsulation structure and fully exposing the protein substrate. Salmon fillets contain a high proportion of bones and skin; the use of neutral protease and collagenase allows for synergistic action, breaking down proteins into small peptides and free amino acids, increasing the release rate of amino acid nitrogen, and thus improving the quality of the seasoning sauce.

[0023] This application also provides a seasoning sauce prepared by the above-mentioned method for producing seasoning sauce.

[0024] The flavor and quality of the seasoning sauce provided in this application are significantly improved. Through the optimization of the enzymatic hydrolysis process, the amino acid nitrogen and yield of the salmon seasoning sauce are significantly improved, the bitterness is effectively reduced, and the resulting seasoning sauce has a light fishy smell and a good seafood flavor. Due to the treatment of modified immobilized lipase, the residual oil in the seasoning sauce is effectively reduced, avoiding the layering caused by oil and the off-flavors caused by oil oxidation, and the seasoning sauce is stable.

[0025] Furthermore, the modified immobilized lipase treatment disrupts the lipid barrier surrounding the protein, effectively promoting amino acid release while maintaining stability. Experimental results show that after 20 repeated uses, the enzyme activity recovery rate of the modified immobilized lipase is still greater than 80%. The amino acid nitrogen value of the seasoning sauce obtained by the modified immobilized lipase and protease synergistic hydrolysis is greater than 0.69 g / 100 mL, the yield is greater than 70%, the turbidity is less than 31 NTU, the sensory evaluation is good, and the flavor and body shape remain stable after 180 days of storage at 4℃.

[0026] Compared with the prior art, this application has the following beneficial effects: This application provides a modified immobilized lipase and its application in the preparation of seasoning sauces. The application uses salmon bone scraps, a byproduct of salmon bone processing, which is modified and then immobilized using an adsorption-embedding method. This effectively improves the enzyme activity recovery rate, stability, and reproducibility of the immobilized lipase. Combining the modified immobilized lipase with a compound protease shortens the enzymatic hydrolysis time, increases the amino acid nitrogen level, and significantly reduces production costs. Specifically, the modified salmon bone scraps are used to immobilize the lipase, utilizing the byproduct as a carrier for high-value utilization rather than introducing additional exogenous carriers, thus reducing carrier raw material costs and potential safety hazards. The immobilized lipase has strong recyclability and can be recycled and reused multiple times. The carrier raw material can also be continuously obtained from the enzymatic hydrolysis waste, forming a recyclable resource utilization system and reducing production costs. Attached Figure Description

[0027] Figure 1 Flowchart for the preparation of modified immobilized lipase and seasoning sauce. Detailed Implementation

[0028] To better illustrate the purpose, technical solution, and advantages of this application, the following description will be provided in conjunction with the accompanying drawings and specific embodiments.

[0029] In the following examples and comparative examples, unless otherwise specified, the experimental methods used are conventional methods, and the materials and reagents used are commercially available unless otherwise specified. Furthermore, the raw materials used in each parallel experiment are the same.

[0030] The preparation process of modified immobilized lipase and seasoning sauce is as follows: Figure 1 As shown.

[0031] Example 1: A modified immobilized lipase and its preparation method This embodiment provides a method for preparing modified immobilized lipase, including the following steps: S1. Add twice the weight of production water to the bone residue produced after processing salmon bones, stir and rinse twice at 90℃ to remove residual grease, and filter out the bone residue with a sieve. S2. The bone residue is dried with hot air and pulverized to obtain 300-mesh bone residue powder. It is then treated with microwave (microwave power is 600W, microwave time is 3min) to obtain modified bone residue. S3. Dissolve 1g of lipase in 30mL of H3PO4 buffer, then add 1g of the modified bone residue obtained in step S2 for adsorption. After adsorption at 30℃, filter to obtain a mixture. S4. Add the mixture obtained in step S3 to 30 mL of 4% sodium alginate-gelatin solution (sodium alginate to gelatin mass ratio of 3:1), mix thoroughly, and then inject together into 0.05 mol / mL CaCl2 solution. Filter and wash the gel to form modified immobilized lipase.

[0032] Example 2: A modified immobilized lipase and its preparation method Compared with Example 1, the difference in Example 2 is that the mesh size of the bone residue powder obtained in step S2 is different, and 800-mesh bone residue powder is obtained. The other steps are the same as in Example 1.

[0033] Example 3: A modified immobilized lipase and its preparation method Compared with Example 2, Example 3 differs in that the microwave processing parameters in step S2 are different. The microwave power is 450W and the microwave time is 3min. The remaining steps are the same as in Example 1.

[0034] Example 4: A modified immobilized lipase and its preparation method Compared with Example 2, Example 4 differs in that the microwave processing parameters in step S2 are different. The microwave power is 650W and the microwave time is 3min. The remaining steps are the same as in Example 1.

[0035] Example 5: A modified immobilized lipase and its preparation method Compared with Example 2, Example 5 differs in that the microwave processing parameters in step S2 are different. The microwave power is 400W and the microwave time is 3min. The remaining steps are the same as in Example 1.

[0036] Example 6: A modified immobilized lipase and its preparation method Compared with Example 2, Example 6 differs in that the microwave processing parameters in step S2 are different. The microwave power is 700W and the microwave time is 3min. The remaining steps are the same as in Example 1.

[0037] Comparative Example 1 Comparative Example 1 provides a method for preparing immobilized lipase, comprising the following steps: S1. Add twice the weight of production water to the bone residue produced after processing salmon bones, stir and rinse twice at 90℃ to remove residual grease, and filter out the bone residue with a sieve. S2. The bone residue is dried and pulverized by hot air to obtain 800-mesh bone residue powder; S3. Dissolve 1g of lipase in 30mL of H3PO4 buffer, then add 1g of bone meal powder obtained in step S2 for adsorption. After adsorption at 30℃, filter to obtain a mixture. S4. Add the mixture obtained in step S3 to 30 mL of 4% sodium alginate-gelatin solution (sodium alginate to gelatin mass ratio of 3:1), mix thoroughly, and then inject together into 0.05 mol / mL CaCl2 solution. Filter and wash the gel to form immobilized lipase.

[0038] Comparative Example 1 was not subjected to microwave modification treatment.

[0039] The modified immobilized lipases prepared in Examples 1-6 and the immobilized lipase of Comparative Example 1 were used to prepare the seasoning sauce.

[0040] Example 7: A method for preparing a seasoning sauce This embodiment provides a method for preparing a seasoning sauce, including the following steps: 1) Take salmon bones and add water to a crusher and stir to make salmon paste. Add 0.2% modified immobilized lipase (Example 2), 0.5% neutral protease and 0.3% collagenase by mass concentration for enzymatic hydrolysis. Adjust the temperature of the salmon paste to 50°C and stir at a constant temperature for 10 hours. Then add salt, heat to 90°C and stir at a constant temperature for 20 minutes to inactivate the enzyme and obtain the enzymatic hydrolysate. 2) Centrifuge the enzymatic hydrolysate at 3000 r / min for 20 min, and take the middle water layer to obtain the seasoning sauce (the clear salmon seasoning sauce in the middle layer).

[0041] Example 8: A method for preparing a seasoning sauce This embodiment provides a method for preparing a seasoning sauce, including the following steps: 1) Take salmon bones and add water to a crusher and stir to make salmon paste. Add 0.1% modified immobilized lipase (Example 2), 0.25% neutral protease and 0.15% collagenase by mass concentration for enzymatic hydrolysis. Adjust the temperature of the salmon paste to 50°C and stir at a constant temperature for 10 hours. Then add salt, heat to 90°C and stir at a constant temperature for 20 minutes to inactivate the enzyme and obtain the enzymatic hydrolysate. 2) Centrifuge the enzymatic hydrolysate at 3000 r / min for 20 min, and take the middle water layer to obtain the seasoning sauce (the clear salmon seasoning sauce in the middle layer).

[0042] Example 9: A method for preparing a seasoning sauce Compared with Example 7, the difference in Example 9 is that the modified immobilized lipase of Example 1 is used, while the rest of the steps are the same as in Example 7.

[0043] Example 10: A method for preparing a seasoning sauce Compared with Example 7, the difference in Example 10 is that the modified immobilized lipase of Example 3 is used, while the rest of the steps are the same as in Example 7.

[0044] Example 11: A method for preparing a seasoning sauce Compared with Example 7, the difference in Example 11 is that the modified immobilized lipase of Example 4 is used, while the rest of the steps are the same as in Example 7.

[0045] Example 12: A method for preparing a seasoning sauce Compared with Example 7, the difference in Example 12 is that the modified immobilized lipase of Example 5 is used, while the rest of the steps are the same as in Example 7.

[0046] Example 3: A method for preparing a seasoning sauce Compared with Example 7, the difference in Example 13 is that the modified immobilized lipase of Example 6 is used, while the rest of the steps are the same as in Example 7.

[0047] Comparative Example 2: A method for preparing a seasoning sauce This comparative example provides a method for preparing a seasoning sauce, including the following steps: 1) Take salmon bones and add water to a crusher and stir to make salmon paste. Add 0.5% neutral protease and 0.3% collagenase by mass for enzymatic hydrolysis. Adjust the temperature of the salmon paste to 50℃ and stir at a constant temperature for 10 hours. Then add salt, heat to 90℃ and stir at a constant temperature for 20 minutes to inactivate the enzyme and obtain the enzymatic hydrolysate. 2) Centrifuge the enzymatic hydrolysate at 3000 r / min for 20 min, and take the middle water layer to obtain the seasoning sauce (the clear salmon seasoning sauce in the middle layer).

[0048] Comparative Example 2 did not use modified immobilized lipase.

[0049] Comparative Example 3: A method for preparing a seasoning sauce Compared with Example 7, the difference in Comparative Example 3 is that it uses the immobilized lipase of Comparative Example 1, while the rest of the steps are the same as in Example 7.

[0050] Comparative Example 4: A method for preparing a seasoning sauce Compared with Example 7, the difference in Comparative Example 4 is that a lipase with a mass concentration of 0.2% was used to directly replace the modified immobilized lipase with a mass concentration of 0.2%, while the other steps were the same as in Example 7.

[0051] Comparative Example 5: A method for preparing a seasoning sauce Compared with Example 7, the difference in Comparative Example 5 is that trypsin with a mass concentration of 0.3% is used instead of collagenase with a mass concentration of 0.3%, while the other steps are the same as in Example 7.

[0052] Experimental Example 1: Enzyme Activity Recovery Experiment (1) Using the modified immobilized lipases obtained in Examples 1-6 and the immobilized lipases obtained in Comparative Example 1, the proteins in the immobilization solution were immobilized. The immobilization rate was calculated using the Coomassie brilliant blue staining method. The results are shown in Table 1.

[0053] In the formula: m1 is the protein mass in the solution before fixation / μg; m2 is the protein mass in the solution after fixation / μg.

[0054] (2) Using the modified immobilized lipases obtained in Examples 1-6 and the immobilized lipases obtained in Comparative Example 1, olive oil was hydrolyzed and the enzyme activity was determined.

[0055] The specific method is as follows: Take 4 mL of 0.05 mol / L phosphate buffer and 4 mL of 0.225 g / mL olive oil emulsion, preheat at 37°C for 10 min, add 25 mg of the modified immobilized lipase obtained in Examples 1-6 (or the immobilized lipase obtained in Comparative Example 1), shake for 20 min, add 15 mL of 95% ethanol to stop the reaction, use phenolphthalein as an indicator, and titrate the produced fatty acids with 0.05 mol / L NaOH solution. Enzyme activity is defined as the amount of enzyme required to generate 1 μmol of fatty acid per minute as one enzyme activity unit.

[0056] (3) The enzyme activity was measured after the immobilized lipase was repeatedly recycled 20 times, and the enzyme activity recovery rate was calculated. The results are shown in Table 1.

[0057] Table 1 According to the data in Table 1, the modified immobilized lipases prepared in Examples 1-6 of this application still had an enzyme activity recovery rate of more than 80% after being repeatedly recycled and used 20 times. Among them, the enzyme activity recovery rate of the modified immobilized lipases in Examples 5-6 was lower than that of the modified immobilized lipases in Examples 2-4, indicating that the power of microwaves has an impact on the enzyme activity recovery rate of modified immobilized lipases.

[0058] The immobilized lipase in Comparative Example 1, without microwave modification, had a lower enzyme activity recovery rate than that in Examples 1-6. Salmon bone scraps, a byproduct of salmon bone processing, were used for microwave modification. This modified bone scraps developed abundant micropores and an uneven structure, increasing the specific surface area and exposing more contact sites for the lipase. When mixed with the lipase, the pores in the modified bone scraps could adsorb the lipase. Furthermore, an adsorption-embedding method was employed to immobilize the lipase using the modified bone scraps as a carrier, effectively improving the enzyme activity recovery rate, stability, and reproducibility of the immobilized lipase.

[0059] Test Example 2: Sensory Evaluation and Shelf Life Test Sensory evaluation tests were conducted on the seasoning sauces prepared in Examples 7-13 and Comparative Examples 2-5. The sensory evaluation criteria and results are shown in Tables 2 and 3.

[0060] Table 2 Table 3 The amino acid nitrogen value, yield, turbidity, etc. of the seasoning sauces prepared in Examples 7-13 and Comparative Examples 2-5 were determined, and the shelf life was tested at 4°C and 25°C. The results are shown in Table 4.

[0061] Table 4 As shown in Table 2, the seasoning sauces prepared in Examples 7-13 all had high amino acid nitrogen content and yield, low turbidity, and good sensory scores and shelf life. Among them, the seasoning sauce of Example 7 had the best overall performance. The amino acid nitrogen content of the seasoning sauce of Example 7 was 0.86 g / 00mL, the yield was 86.7%, the turbidity was 2.77 NTU, the sensory score was 9.2 points, and it still maintained stable flavor and body shape after being stored at 4℃ for 180 days and at 25℃ for 90 days.

[0062] The seasoning sauces prepared by immobilized lipase and co-protease hydrolysis in Examples 7-13 have an amino acid nitrogen value greater than 0.69 g / 100 mL, a yield greater than 70%, a turbidity less than 31 NTU, good sensory evaluation, and can still maintain flavor and shape stability after 120 days at 4℃ and 60 days at 25℃.

[0063] Comparative Example 2 did not contain modified immobilized lipase. The prepared sauce had lower amino acid nitrogen value and yield, higher turbidity, and poor sensory evaluation. It only lasted 15 days at 4°C. This indicates that adding modified immobilized lipase can improve the quality of the sauce.

[0064] Comparative Example 3, using an immobilized lipase without modification, produced a sauce with lower amino acid nitrogen value and yield than the sauce in Example 7, higher turbidity, and poorer sensory evaluation. It also had a shelf life of 30 days at 4°C. Immobilized lipase treatment disrupts the lipid barrier surrounding the protein, effectively promoting amino acid release while maintaining stability. Experiments showed that after 20 repeated uses, the enzyme activity recovery rate of the immobilized lipase was still greater than 80%. The prepared sauce, obtained by enzymatic hydrolysis of the immobilized lipase and protease, had an amino acid nitrogen value greater than 0.8 g / 100 mL, a yield greater than 80%, turbidity less than 10 NTU, and good sensory evaluation. It maintained flavor and shape stability after 180 days at 4°C and 90 days at 25°C.

[0065] Comparative Example 4 used lipase with a mass concentration of 0.2% to directly replace modified immobilized lipase with a mass concentration of 0.2%, and Comparative Example 5 used trypsin with a mass concentration of 0.3% to replace collagenase with a mass concentration of 0.3%. The prepared sauces had lower amino acid nitrogen values ​​and yields than the sauces in Example 7, and also had higher turbidity, poorer sensory evaluation, and a shelf life of 30 days at 4°C.

[0066] The experimental data above show that the technology of using the modified immobilized lipase and compound protease combined to enzymatically hydrolyze salmon bones has significant advantages in improving the amino acid nitrogen and flavor of the seasoning sauce. In addition, since the immobilized lipase removes oil, it can reduce turbidity, effectively improve the stability of the seasoning sauce, and extend its shelf life.

[0067] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application and are not intended to limit the scope of protection of this application. Although this application has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of this application without departing from the substance and scope of the technical solutions of this application.

Claims

1. A method for preparing a modified immobilized lipase, characterized in that, Includes the following steps: S1. Add water to the bone residue produced after processing salmon bones and rinse it to filter out the bone residue. S2. Dry and pulverize the bone residue to obtain bone residue powder, and then treat it with microwave to obtain modified bone residue; S3. Dissolve the lipase in a buffer solution, then add the modified bone residue obtained in step S2 for adsorption to obtain a mixture; S4. Add the mixture obtained in step S3 to the embedding agent, filter and wash to form a modified immobilized lipase.

2. The method for preparing the modified immobilized lipase as described in claim 1, characterized in that, In step S2, the bone residue powder has a mesh size of 300-800 mesh.

3. The method for preparing the modified immobilized lipase as described in claim 1, characterized in that, In step S2, the microwave processing includes: the microwave power is 450~600W, and the microwave time is 3~5min.

4. The method for preparing the modified immobilized lipase as described in claim 1, characterized in that, In step S3, the mass ratio of lipase to modified bone residue is 1:(1~1.5).

5. The method for preparing the modified immobilized lipase as described in claim 1, characterized in that, In step S4, the encapsulating agent includes a mixture of calcium salt solution, sodium alginate, and gelatin.

6. The modified immobilized lipase prepared by the method according to any one of claims 1 to 5.

7. The application of the modified immobilized lipase as described in claim 6 in the preparation of seasoning sauce.

8. A method for producing a seasoning sauce, characterized in that, Includes the following steps: 1) Weigh salmon paste, add the modified immobilized lipase, neutral protease and collagenase as described in claim 6 for enzymatic hydrolysis, and then inactivate the enzymes to obtain the hydrolysate; 2) Centrifuge the enzymatic hydrolysate, take the middle water layer, and obtain the seasoning sauce.

9. The method for producing the seasoning sauce as described in claim 8, characterized in that, The mass ratio of the modified immobilized lipase, neutral protease and collagenase is (0.1~0.2):(0.25~0.5):(0.15~0.3).

10. A seasoning sauce prepared by the method for producing seasoning sauce as described in claim 8 or 9.

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