Enrichment method of tuna cooking liquor goose carnosine

Abstract

The invention provides a method for enriching tuna cooking liquor goose carnosine, and relates to the field of high-value utilization of aquatic product processing byproducts. The enrichment method of the tuna cooking liquor goose carnosine comprises the following steps: taking the tuna cooking liquor, and carrying out pretreatment to remove suspended solids so as to obtain a pretreatment solution; filtering the pretreatment liquid through a ceramic membrane with the molecular weight cut-off of 3000-5000Da, and collecting permeate liquid; adding the permeate into a phosphate buffer solution for dynamic dilution, then filtering through an organic ultrafiltration membrane with the molecular weight cutoff of 150-200Da, and collecting the trapped fluid, so as to obtain enriched goose carnosine; wherein the dynamic dilution is to repeatedly add a phosphate buffer solution in the membrane filtration process, so as to avoid coprecipitation of purine and goose carnosine under high concentration. According to the method, coprecipitation of purine and the goose carnosine under high concentration is effectively avoided through the synergistic effect of multi-stage membrane separation (the combination of the ceramic membrane and the organic ultrafiltration membrane) and the dynamic dilution process, the yield of the goose carnosine can reach 1.28 g / L or above, and the purity of the goose carnosine can reach 86% or above.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of high-value utilization of aquatic processing by-products, in particular to a method for enriching goose muscle peptide from tuna cooking liquid. BACKGROUND

[0002] Goose muscle peptide is a kind of dipeptide with important physiological activity, which has significant effects on antioxidant and uric acid reduction, and is widely used in health products and medicines. A large amount of cooking liquid is produced during tuna processing, which contains a certain amount of goose muscle peptide. If it is directly discharged, it not only causes resource waste, but also pollutes the environment. Therefore, it has important economic and environmental value to enrich goose muscle peptide from tuna cooking liquid.

[0003] The existing technology has obvious deficiencies in separating goose muscle peptide from aquatic processing wastewater. For example, the patent document with publication number CN120025284A discloses a method for extracting goose muscle peptide and muscle peptide from tuna, which includes low-salt water extraction and high-salt water extraction under alkaline and elevated temperature conditions. The goose muscle peptide and / or muscle peptide extraction method provided by the present application is simple to operate, low in cost, and easy to use in large-scale industrialization. However, the purity of goose muscle peptide is less than 70%, which cannot meet the demand of industrial production for high-purity goose muscle peptide.

[0004] Secondly, Chinese invention patent CN119707823A proposes a goose muscle peptide extraction and preparation process and its application. This invention is finally beneficial to improve the extraction rate of tuna goose muscle peptide, and can effectively reduce the content of purine by combining membrane filtration. Although this method reduces the content of purine in tuna, the yield of goose muscle peptide in the final product is not high, less than 0.8g / L.

[0005] Therefore, it is urgent to develop an efficient and high-purity method for enriching goose muscle peptide from tuna cooking liquid. SUMMARY

[0006] (I) Technical problems solved

[0007] In view of the deficiencies of the prior art, the present application provides a method for enriching goose muscle peptide from tuna cooking liquid to improve the yield and purity of goose muscle peptide, realize the high-value utilization of tuna processing by-products, and solve the problems in the prior art that the separation efficiency of goose muscle peptide and impurities (such as purine and small molecule peptides) is low, the flux decays quickly due to dynamic membrane pollution, and the target substance is easy to co-precipitate and lose, resulting in a yield of less than 0.8g / L and a purity of less than 70%, which cannot meet the demand of industrial production.

[0008] (II) Technical solutions

[0009] To achieve the above purpose, the present application realizes the following technical solutions: a method for enriching goose muscle peptide from tuna cooking liquid, comprising the following steps:

[0010] Step one: raw material pretreatment

[0011] Take tuna cooking liquid, remove suspended solids by pretreatment to obtain pretreated liquid;

[0012] Step two: multi-stage membrane separation-ceramic membrane filtration

[0013] The pretreated liquid of step one is filtered through a ceramic membrane with a molecular weight cut-off of 3000-5000 Da, and the permeate is collected;

[0014] Step three: dynamic dilution-ultrafiltration membrane filtration

[0015] The permeate of step two is added to a phosphate buffer for dynamic dilution, and then filtered through an organic ultrafiltration membrane with a molecular weight cut-off of 150-200 Da, and the retentate is collected, i.e. the enriched goose muscle peptide; wherein the dynamic dilution is repeated addition of phosphate buffer during membrane filtration to avoid co-precipitation of purines and goose muscle peptide at high concentrations.

[0016] Preferably, in step one, the pretreatment is centrifugation or coarse filtration; when using centrifugation, the centrifugation speed is 3000-5000 r / min, and the time is 10-20 min.

[0017] Preferably, in step two, the molecular weight cut-off of the ceramic membrane is 3500-4500 Da.

[0018] Preferably, in step three, the molecular weight cut-off of the organic ultrafiltration membrane is 160-180 Da.

[0019] Preferably, in step three, the concentration of the phosphate buffer is 0.01-0.05 mol / L, and the pH is 6.5-7.5.

[0020] Preferably, in step three, the number of dynamic dilutions is 2-4, and the volume of phosphate buffer added each time is 1 / 3-1 / 2 of the volume of the permeate in step two.

[0021] Preferably, in steps two and three, the operating pressure for membrane filtration is 0.1-0.3 MPa, and the temperature is 25-35°C.

[0022] Preferably, it further includes multi-stage adsorption between step one and step two:

[0023] First-stage adsorption: pump the pretreated liquid into a modified mesoporous silica adsorption column, the modified mesoporous silica adsorption column is MCM-41 type mesoporous silica modified with γ-aminopropyl triethoxysilane; the adsorption temperature of the modified mesoporous silica adsorption column is 25-30°C, and the flow rate is 1-2 BV / h;

[0024] Secondary adsorption: the breakthrough liquid after the first adsorption is pumped into a weak cation exchange resin column; the weak cation exchange resin column is of type D113, the adsorption temperature is 30-35 DEG C, and the flow rate is 0.5-1 BV / h.

[0025] Preferably, it also comprises low-temperature concentration after step three:

[0026] The carnosine retentate is vacuum concentrated to 1 / 10-1 / 15 of the original volume under a vacuum of 0.08-0.09 MPa and a temperature of 40-45 DEG C to obtain a carnosine enriched liquid.

[0027] (Three) beneficial effects

[0028] The application provides a method for enriching carnosine in tuna cooking liquid, and has the following beneficial effects:

[0029] 1. High yield: through the synergistic effect of multi-stage membrane separation (combination of ceramic membrane and organic ultrafiltration membrane) and dynamic dilution process, the coprecipitation of purines and carnosine at high concentrations is effectively avoided, and the yield of carnosine can reach more than 1.28 g / L, which is more than 50.6% higher than that of the conventional method (0.85 g / L), and the yield is ≥85%.

[0030] 2. High purity: the ceramic membrane (3000-5000 Da) can effectively retain macromolecular impurities, and the organic ultrafiltration membrane (150-200 Da) can accurately retain carnosine and remove salts and heavy metals, significantly improving the purity of the product. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 It is a method step schematic diagram of the embodiment one, the embodiment two and the embodiment three of the application;

[0032] Figure 2 It is a method step schematic diagram of the embodiment four of the application;

[0033] Figure 3 It is a method step schematic diagram of the embodiment five of the application. DETAILED DESCRIPTION

[0034] The technical solutions in the embodiments of the application will be clearly and completely described below with reference to the drawings in the embodiments of the application. Obviously, the described embodiments are only part of the embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the application.

[0035] Embodiment one:

[0036] As Figure 1As shown, this embodiment of the invention provides a method for enriching angiotensin in tuna cooking liquid, comprising the following steps:

[0037] Step 1: Raw material pretreatment:

[0038] Take the tuna cooking liquid, and pretreat it to remove suspended solids to obtain a pretreated liquid; wherein the pretreatment is centrifugation at a speed of 3000 r / min for 10 min;

[0039] Step 2: Multi-stage membrane separation - ceramic membrane filtration:

[0040] The pretreated solution from step one was filtered through a ceramic membrane with a molecular weight cutoff of 3500 Da, and the permeate was collected.

[0041] Step 3: Dynamic dilution - ultrafiltration membrane filtration:

[0042] The permeate from step two was dynamically diluted by adding phosphate buffer solution with a concentration of 0.01 mol / L and a pH of 6.5. The dynamic dilution was performed twice, with each addition of phosphate buffer solution being 1 / 3 of the volume of the permeate from step two. The solution was then filtered through an organic ultrafiltration membrane with a molecular weight cutoff of 160 Da, and the retentate was collected to obtain the enriched anserine peptide. The dynamic dilution involved repeatedly adding phosphate buffer solution during membrane filtration to prevent co-precipitation of high-concentration purines with anserine peptide.

[0043] In steps two and three, the membrane filtration operation pressure is 0.1 MPa and the temperature is 25°C.

[0044] Example 2:

[0045] like Figure 1 As shown, this embodiment of the invention provides a method for enriching angiotensin in tuna cooking liquid, comprising the following steps:

[0046] Step 1: Raw material pretreatment:

[0047] Take the tuna cooking liquid, and pretreat it to remove suspended solids to obtain a pretreated liquid; wherein the pretreatment is centrifugation at a speed of 4000 r / min for 15 min;

[0048] Step 2: Multi-stage membrane separation - ceramic membrane filtration:

[0049] The pretreated solution from step one was filtered through a ceramic membrane with a molecular weight cutoff of 4000 Da, and the permeate was collected.

[0050] Step 3: Dynamic dilution - ultrafiltration membrane filtration:

[0051] The permeate of step two is added into phosphate buffer for dynamic dilution, the concentration of the phosphate buffer is 0.03 mol / L, the pH is 7.0, and the dynamic dilution is performed for 3 times, the volume of the phosphate buffer added each time is 1 / 3 of the volume of the permeate in step two, then filtration is performed through an organic ultrafiltration membrane with a molecular weight cut-off of 170 Da, and the cut-off liquid is collected, thereby obtaining the enriched carnosine; wherein, the dynamic dilution is repeatedly adding the phosphate buffer in the membrane filtration process to avoid the coprecipitation of purine and carnosine under high concentration.

[0052] In step two and step three, the operating pressure of the membrane filtration is 0.2 MPa, and the temperature is 30℃.

[0053] Example three:

[0054] As shown in Figure 1 , the embodiment of the present application provides a method for enriching carnosine in tuna cooking liquid, which comprises the following steps:

[0055] Step one: pretreatment of raw materials:

[0056] The tuna cooking liquid is taken and pretreated to remove suspended solids, thereby obtaining a pretreated liquid; wherein, the pretreatment is centrifugation, and the centrifugal speed is 5000 r / min, and the time is 20 min;

[0057] Step two: multi-stage membrane separation-ceramic membrane filtration:

[0058] The pretreated liquid of step one is filtered through a ceramic membrane with a molecular weight cut-off of 4500 Da, and the permeate is collected;

[0059] Step three: dynamic dilution-ultrafiltration membrane filtration:

[0060] The permeate of step two is added into phosphate buffer for dynamic dilution, the concentration of the phosphate buffer is 0.05 mol / L, the pH is 7.5, and the dynamic dilution is performed for 4 times, the volume of the phosphate buffer added each time is 1 / 2 of the volume of the permeate in step two, then filtration is performed through an organic ultrafiltration membrane with a molecular weight cut-off of 180 Da, and the cut-off liquid is collected, thereby obtaining the enriched carnosine; wherein, the dynamic dilution is repeatedly adding the phosphate buffer in the membrane filtration process to avoid the coprecipitation of purine and carnosine under high concentration.

[0061] In step two and step three, the operating pressure of the membrane filtration is 0.3 MPa, and the temperature is 35℃.

[0062] Example four:

[0063] As shown in Figure 2 , the embodiment of the present application provides a method for enriching carnosine in tuna cooking liquid, which comprises the following steps:

[0064] Step one: raw material pretreatment:

[0065] Take tuna cooking liquid, remove suspended solids by pretreatment to obtain pretreated liquid; wherein the pretreatment is centrifugation or coarse filtration; when using centrifugation, the centrifugation speed is 4000 r / min, and the time is 15 min;

[0066] Also includes multi-stage adsorption:

[0067] First-stage adsorption: pump the pretreated liquid into a modified mesoporous silica adsorption column, the modified mesoporous silica adsorption column is MCM-41 type mesoporous silica modified by γ-aminopropyl triethoxysilane; the adsorption temperature of the modified mesoporous silica adsorption column is 30℃, the flow rate is 2BV / h, and the diameter-height ratio of the modified mesoporous silica adsorption column is 1:10, and the bed volume (BV) to pretreated liquid volume ratio is 1:8; wherein the preparation method of the modified mesoporous silica adsorption column is: adding MCM-41 mesoporous silica into an ethanol solution (mass concentration 8%) of γ-aminopropyl triethoxysilane, refluxing at 70℃ for 6h, and then washing and drying to obtain;

[0068] Second-stage adsorption: pump the breakthrough liquid after the first-stage adsorption into a weak cation exchange resin column, and the diameter-height ratio of the weak cation exchange resin column is 1:12, and the bed volume to first-stage adsorption breakthrough liquid volume ratio is 1:10; the type of the weak cation exchange resin column is D113, the adsorption temperature is 35℃, and the flow rate is 1BV / h;

[0069] Step two: multi-stage membrane separation-ceramic membrane filtration:

[0070] Filter the pretreated liquid (after multi-stage adsorption) of step one through a ceramic membrane with a molecular weight cut-off of 4000Da to collect the permeate;

[0071] Step three: dynamic dilution-ultrafiltration membrane filtration:

[0072] Add phosphate buffer to the permeate of step two for dynamic dilution, the concentration of the phosphate buffer is 0.03mol / L, the pH is 7.0, and the number of dynamic dilution is 3 times, the volume of phosphate buffer added each time is 1 / 3 of the volume of the permeate in step two, and then filter through an organic ultrafiltration membrane with a molecular weight cut-off of 170Da to collect the retentate, i.e. the enriched goose muscle peptide; wherein the dynamic dilution is to repeatedly add phosphate buffer during membrane filtration to avoid co-precipitation of purines and goose muscle peptide at high concentrations.

[0073] In steps two and three, the operating pressure of membrane filtration is 0.2MPa, and the temperature is 30℃.

[0074] Example five:

[0075] As Figure 3As shown, this embodiment of the invention provides a method for enriching angiotensin in tuna cooking liquid, comprising the following steps:

[0076] Step 1: Raw material pretreatment:

[0077] Take the tuna cooking liquid, and pretreat it to remove suspended solids to obtain a pretreated liquid; wherein, the pretreatment is centrifugation or coarse filtration; when centrifugation is used, the centrifugation speed is 4000 r / min and the time is 15 min;

[0078] Step 2: Multi-stage membrane separation - ceramic membrane filtration:

[0079] The pretreated solution from step one was filtered through a ceramic membrane with a molecular weight cutoff of 4000 Da, and the permeate was collected.

[0080] Step 3: Dynamic dilution - ultrafiltration membrane filtration:

[0081] The permeate from step two was dynamically diluted by adding phosphate buffer solution with a concentration of 0.03 mol / L and a pH of 7.0. The dynamic dilution was performed three times, with each addition of phosphate buffer solution equal to one-third of the volume of the permeate from step two. The solution was then filtered through an organic ultrafiltration membrane with a molecular weight cutoff of 170 Da, and the retentate was collected to obtain the enriched anserine peptide. The dynamic dilution involved repeatedly adding phosphate buffer solution during membrane filtration to prevent co-precipitation of high-concentration purines with anserine peptide.

[0082] In steps two and three, the membrane filtration operation pressure is 0.2 MPa and the temperature is 30°C.

[0083] It also includes low-temperature concentration:

[0084] The retentate of goose muscle peptides was concentrated to 1 / 15 of its original volume under vacuum conditions of 0.09 MPa and 45°C to obtain a goose muscle peptide enrichment solution.

[0085] Comparative example:

[0086] The same batch of tuna was treated using the method published under CN120025284A to extract goose muscle peptides.

[0087] Experimental example:

[0088] The performance of anserine in Examples 1, 2, 3, 4, 5, and the comparative example was tested (the anserine content was determined by high performance liquid chromatography (HPLC): C18 column (250 mm × 4.6 mm, 5 μm), mobile phase was 0.1% phosphoric acid aqueous solution-acetonitrile (95:5), flow rate was 1.0 mL / min, detection wavelength was 210 nm, and column temperature was 30 °C). The results are shown in the table below:

[0089] Group Goose skin yield (g / L) Purity (%) Enrichment fold (times) Example 1 1.28 86 42 Example 2 1.32 88 48 Example 3 1.30 87 46 Example 4 1.32 88 50 Example 5 1.33 90 48 Comparative Example 0.85 75 35

[0090] In summary, the enrichment of anserine by the present invention can significantly improve the yield and purity of anserine in tuna cooking broth. Experimental data show that, compared to the method disclosed in CN120025284A used in the comparative example, the anserine yield in each embodiment reached over 1.28 g / L, a significant improvement over the comparative example, with a yield exceeding 85%. This demonstrates a significant improvement in both anserine adsorption rate and product purity in the embodiments of the present invention. In Examples 1 to 3, by rationally controlling the centrifugation parameters of the raw material pretreatment, the molecular weight cutoff of the ceramic membrane and ultrafiltration membrane, the concentration and pH value of the phosphate buffer, and the number and volume of dynamic dilutions, anserine can be more effectively enriched from the tuna cooking broth.

[0091] Example 4 adds a multi-stage adsorption step, with the first-stage adsorption using a modified mesoporous silica adsorption column and the second-stage adsorption using a weak cation exchange resin column, further improving the enrichment effect. Example 5 adds a low-temperature concentration step after step 3, where the anserine retentate is concentrated under vacuum, improving the yield and purity of the anserine.

[0092] The method of this invention also enables the high-value utilization of tuna processing byproducts. Tuna cooking liquid, originally wastewater from seafood processing, can be used to extract high-value anserine peptides using this method, reducing waste emissions and aligning with the concept of sustainable development. Furthermore, the method is relatively simple to operate, and the parameters of each step are easy to control, meeting the demands of industrial production for high-yield, high-purity anserine peptides, thus possessing broad market application prospects and good economic benefits.

[0093] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for enriching goose muscle peptides in tuna cooking liquid, characterized in that: Includes the following steps: Step 1: Raw material pretreatment: Take the tuna cooking liquid, and pre-treat it to remove suspended solids to obtain the pre-treated liquid; Step 2: Multi-stage membrane separation - ceramic membrane filtration: The pretreated solution from step one is filtered through a ceramic membrane with a molecular weight cutoff of 3000-5000 Da, and the permeate is collected. Step 3: Dynamic dilution - ultrafiltration membrane filtration: The permeate from step two was dynamically diluted by adding phosphate buffer, and then filtered through an organic ultrafiltration membrane with a molecular weight cutoff of 150-200 Da. The retentate was collected to obtain enriched anserine peptides. The dynamic dilution involved repeatedly adding phosphate buffer during membrane filtration to avoid co-precipitation of high-concentration purines with anserine peptides.

2. The method for enriching goose muscle peptides in tuna cooking liquid according to claim 1, characterized in that: In step one, the pretreatment is centrifugation or coarse filtration; when centrifugation is used, the centrifugation speed is 3000-5000 r / min and the time is 10-20 min.

3. The method for enriching goose muscle peptides in tuna cooking liquid according to claim 1, characterized in that: In step two, the molecular weight cutoff of the ceramic membrane is 3500-4500 Da.

4. The method for enriching goose muscle peptides in tuna cooking liquid according to claim 1, characterized in that: In step three, the molecular weight cutoff of the organic ultrafiltration membrane is 160-180 Da.

5. The method for enriching goose muscle peptides in tuna cooking liquid according to claim 1, characterized in that: In step three, the concentration of the phosphate buffer solution is 0.01-0.05 mol / L, and the pH is 6.5-7.

5.

6. The method for enriching goose muscle peptides in tuna cooking liquid according to claim 1, characterized in that: In step three, the dynamic dilution is performed 2-4 times, and the volume of phosphate buffer added each time is 1 / 3-1 / 2 of the volume of permeate in step two.

7. The method for enriching goose muscle peptides in tuna cooking liquid according to claim 1, characterized in that: In steps two and three, the membrane filtration operation pressure is 0.1-0.3 MPa and the temperature is 25-35℃.

8. The method for enriching angiotensin in tuna cooking liquid according to any one of claims 1-7, characterized in that: It also includes multi-stage adsorption between step one and step two: Primary adsorption: The pretreatment liquid is pumped into a modified mesoporous silica adsorption column, wherein the modified mesoporous silica adsorption column is an MCM-41 type mesoporous silica modified with γ-aminopropyltriethoxysilane; the adsorption temperature of the modified mesoporous silica adsorption column is 25-30℃, and the flow rate is 1-2 BV / h. Secondary adsorption: The permeate after primary adsorption is pumped into a weak cation exchange resin column; the weak cation exchange resin column is model D113, with an adsorption temperature of 30-35℃ and a flow rate of 0.5-1 BV / h.

9. A method for enriching angiotensin in tuna cooking liquid according to any one of claims 1-7, characterized in that: This also includes low-temperature concentration following step three: The retentate of goose muscle peptides was concentrated under vacuum at a pressure of 0.08-0.09 MPa and a temperature of 40-45℃ to 1 / 10-1 / 15 of its original volume to obtain a goose muscle peptide enrichment solution.

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