Efficient deodorization method of abalone pleopod enzymatic soluble collagen

Abstract

The invention discloses an efficient fishy smell removing method of abalone pleopod enzymatic soluble collagen, which comprises the following steps: ultrasonically treating the abalone pleopod enzymatic soluble collagen and covalently binding the treated abalone pleopod enzymatic soluble collagen with litchi rind polyphenol extracted by pulse and ultrasonic treatment, so that the binding between polyphenol and protein is remarkably enhanced; according to the present invention, the non-covalent binding of the fishy smell substance and the protein hydrophobic amino acid is weakened so as to easily remove the fishy smell substance and improve the overall flavor, and the litchi rind polyphenol is added, such that the fishy smell can be effectively removed, the flavor quality can be effectively improved, and the benefits of aging delaying, disease prevention and the like can be provided for consumers.

Description

Technical field:

[0001] The invention relates to a method for efficiently removing fishy smell by using abalone gastropod enzyme-catalyzed soluble collagen. Background technology:

[0002] Abalone, as one of the high-value marine single-shelled mollusks, is popular for its delicious taste and rich nutrients. It is rich in protein, fat, unsaturated fatty acids and other nutrients that are beneficial to the human body. It not only has traditional effects such as clearing heat and nourishing yin, improving eyesight and replenishing deficiency, benefiting the stomach, liver and kidneys, but is also known as one of the "eight treasures of seafood" and is a popular delicacy in coastal areas. However, the unsaturated fatty acids in abalone will produce small molecular compounds such as hexanal, heptanal, octanal, nonanal, 1-octen-3-ol, etc. during the oxidative decomposition process. Excessive accumulation of these small molecular compounds will affect the overall flavor quality of abalone.

[0003] The invention with the publication number CN118044595A and the name of the invention "A processing method for salt-ground quick-frozen abalone with original quality" discloses the use of low-temperature orbital oscillation coupled with food-grade hydrogen peroxide to remove the fishy substances of abalone and effectively reduce bacteria. The invention with the publication number CN115141868A and the name of the invention "A method for decolorizing and removing fishy smell from abalone protein peptide" discloses a method for decolorizing and removing fishy smell from abalone protein peptide, including the following specific steps: Step 1: Take fresh abalone, remove the abalone shell, clean the abalone meat, crush the meat, and pulp it to obtain abalone slurry; Step 2: Add warm water to the abalone slurry and stir it, raise the water temperature to 55℃-65℃, adjust the pH to 8.0, and obtain neutral abalone slurry; Step 3: Add composite protease to the neutral abalone slurry in step 2 for 5 hours, heat to 70-90°C to inactivate the enzyme, and collect the filtrate by centrifugation after 20-40 minutes; step 4: cool to 40°C, add the filtrate to a reactor, add activated carbon and yeast powder, and stir for 30-60 minutes; heat to 50-80°C for refining; step 5: filter and decarbonize through a plate and frame filter press, and collect the filtrate; step 6: filter and dry the filtrate to obtain abalone peptide with deodorization.

[0004] However, the above method needs to be further improved. Summary of the invention:

[0005] The invention aims to provide a method for efficiently removing fishy smell from abalone gastropod enzyme-catalyzed soluble collagen.

[0006] The present invention is achieved through the following technical solutions:

[0007] An efficient method for removing fishy smell from abalone gastropod enzyme-catalyzed soluble collagen, the method comprising the following steps:

[0008] (1) Ultrasonic treatment of abalone gastropod enzyme-mediated soluble collagen: ultrasonic treatment of abalone gastropod enzyme-mediated soluble collagen, ultrasonic power 200-800 W, preferably 400-800 W, ultrasonic time 15-30 minutes, preferably 20-30 minutes, the sample solution temperature is maintained at 14-16 ° C;

[0009] (2) Treating abalone gastropodase-mediated soluble collagen with litchi peel polyphenol extract: Add 10-50 mg / g of litchi peel polyphenol extract to the abalone gastropodase-mediated soluble collagen treated in step (1), adjust the pH to 9, mix well, and stir magnetically at 4-8° C. for 2-4 hours to obtain deodorized abalone gastropodase-mediated soluble collagen.

[0010] Preferably, step (2) is: add 40-50 mg / g of litchi peel polyphenol extract to the abalone gastropod enzyme-mediated soluble collagen treated in step (1), adjust the pH to 9, mix evenly, and stir magnetically for 3-4 hours at 6-8° C. to obtain abalone gastropod enzyme-mediated soluble collagen with deodorized fishy taste.

[0011] Preferably, the concentration of abalone gastropod enzyme-catalyzed soluble collagen in step (1) is 6-10 mg / mL, more preferably 8-10 mg / mL.

[0012] The extraction of abalone gastropod enzymatic soluble collagen was as follows: first, the second part of the gastropod muscle was isolated from the abalone, homogenized, washed with deionized cold water, centrifuged, the precipitate was collected, 0.9M KCl was added and stirred for 12 hours, centrifuged, and then the precipitate was dissolved in 0.6M NaCl, stirred for 16 hours, centrifuged to collect the precipitate, and the precipitate was mixed with 10 times the volume of 0.1M NaOH was stirred for 12 hours to remove non-collagen substances, the extract was washed with deionized water to a neutral pH, and then the crude collagen was collected; the crude collagen was dissolved in 0.5 M acetic acid, stirred for 48 hours to remove acid-soluble collagen, and then centrifuged for 30 minutes. The insoluble part was digested with pepsin for 72 hours in a 0.5 M acetic acid environment, pH = 2.0, and 4°C. The mass ratio of enzyme to substrate was 3 / 20. The product was centrifuged for 10 minutes, and NaCl was added to the supernatant to a final concentration of 0.8 M. Centrifuged for 10 minutes, the precipitate was collected, dissolved in 0.5 M acetic acid, and dialyzed with 0.02 M NaHPO4 and 0.1 M acetic acid for 2 days to remove salt, thereby obtaining abalone gastropod enzyme-soluble collagen.

[0013] The preparation of the litchi peel polyphenol extract is as follows: the litchi peel is dried and crushed, and then screened through an 80-mesh sieve to obtain litchi peel powder with uniform particle size; 4-6g of litchi peel powder is evenly mixed with distilled water, and treated for 1-5 minutes, preferably 3-5 minutes, under a pulse electric field strength of 0.25-1.25kV / cm, preferably 0.75-1.25kV / cm, and then 50wt% or more of ethanol, preferably anhydrous ethanol, is added to adjust the ethanol concentration in the system to a range of 40wt%-80wt%, preferably 60wt%-80wt%, and then the solution is ultrasonically treated for 15 minutes at a power of 150W-300W, preferably 220W-300W, the entire extraction process is carried out in a temperature range of 25°C-56°C, preferably 46°C-56°C, and magnetic stirring is maintained during the ultrasonic treatment. Finally, the supernatant is obtained by centrifugation at a speed of 3000-6000 rpm, preferably 5000-6000 rpm, and a centrifugation time of 3-6 minutes, preferably 4-6 minutes, and then freeze-dried to finally obtain litchi peel polyphenol bioactive substances.

[0014] The beneficial effects of the present invention are as follows:

[0015] 1). The shear force and microjet generated during the ultrasonic treatment in step (1) will significantly change the secondary structure of the abalone gastropod enzyme-soluble collagen, making the protein molecules more flexible and loose, resulting in the hydrophobic amino acids and fishy substances originally hidden inside the protein being more easily exposed to the protein surface, which not only increases the hydrophobicity of the protein surface, but also provides more binding sites for molecules such as litchi peel polyphenols.

[0016] 2). The lychee peel polyphenols extracted by pulse and ultrasonic treatment have strong reactivity because they are rich in phenolic hydroxyl groups. Under alkaline conditions, these phenolic hydroxyl groups can react with nucleophilic groups (such as amino and proline residues) in abalone gastropod enzyme-soluble collagen to form covalent bonds. This covalent binding method has higher binding force and stability than non-covalent interactions (such as hydrogen bonds, hydrophobic interactions, etc.), thereby significantly enhancing the binding between polyphenols and proteins and weakening the non-covalent binding between fishy substances and protein hydrophobic amino acids, which is beneficial to the removal of fishy substances and the improvement of overall flavor.

[0017] 3). As a natural antioxidant, litchi peel polyphenols can remove excess free radicals in the body, inhibit lipid peroxidation, and protect biological macromolecules from damage induced by free radicals. Adding litchi peel polyphenols to abalone pre-cooked dishes can not only effectively remove the fishy smell and improve the flavor quality, but also provide consumers with benefits such as delaying aging and preventing diseases.

[0018] In summary, the ultrasonic treatment of abalone gastropod enzyme-mediated soluble collagen in the present invention is covalently bound to litchi peel polyphenols extracted by pulse and ultrasonic treatment, which significantly enhances the binding between polyphenols and proteins and weakens the non-covalent binding between fishy substances and hydrophobic amino acids of proteins, thereby facilitating the removal of fishy substances and the improvement of the overall flavor. In addition, the addition of litchi peel polyphenols can not only effectively remove the fishy smell and improve the flavor quality, but also provide consumers with benefits such as delaying aging and preventing diseases. Specific implementation method:

[0019] The following is a further description of the present invention, rather than a limitation of the present invention.

[0020] References: Yuan Qixin, Zhu Beiwei, Dong Xiuping, et al. Extraction and property study of abalone gastropod collagen [J]. Journal of Dalian Polytechnic University, 2012, 31(01): 35-39. The steps for extracting enzymatic soluble collagen from abalone gastropod in the following examples and comparative examples are as follows: First, the second part of the gastropod muscle is separated from the abalone, then homogenized and washed with 10 times the volume of deionized cold water. Centrifuge for 15 minutes at a speed of 5000r / min to collect the precipitate. Add 10 times the volume of 0.9M KCl to the precipitate, stir for 12 hours, and centrifuge at 4000g / min for 10 minutes. Subsequently, dissolve the precipitate in 10 times the volume of 0.6M NaCl, stir for 16 hours, and centrifuge for 10 minutes at a speed of 5000r / min to collect the precipitate. Stir the precipitate with 10 times the volume of 0.1M NaOH for 12 hours to remove non-collagen substances. The extract was washed with deionized water to a neutral pH, and then the crude collagen was collected. The crude collagen was added to 10 times the volume of 0.5M acetic acid and stirred for 48 hours to remove acid-soluble collagen. After centrifugation at 10000r / min for 30 minutes, the insoluble part was digested with pepsin for 72 hours in a 0.5M acetic acid environment, pH=2.0, and 4°C. The mass ratio of enzyme to substrate was 3 / 20. The product was centrifuged at 6000r / min for 10 minutes, and NaCl was added to the supernatant to a final concentration of 0.8M, and centrifuged at 12000r / min for 10 minutes. The precipitate was collected, dissolved in 0.5M acetic acid, dialyzed with 0.02M NaHPO4 and 0.1M acetic acid for 2 days to remove salt, and finally freeze-dried to obtain abalone gastropod enzyme-soluble collagen.

[0021] Embodiment 1:

[0022] (1) Extraction of litchi peel polyphenols: The litchi peel was dried and crushed, and then screened through an 80-mesh sieve to obtain litchi peel powder with uniform particle size. 5 g of litchi peel powder was evenly mixed with 10 mL of distilled water, treated at a pulse electric field strength of 0.25 kV / cm for 1 minute, and then 50 wt% anhydrous ethanol was added to adjust the ethanol concentration in the system to within 40 wt%, and then the solution was ultrasonically treated at a power of 150 W for 15 minutes. The entire extraction process was carried out at a temperature range of 25°C, and magnetic stirring was maintained during the ultrasonic treatment. Finally, the supernatant was obtained by centrifugation at a speed of 3000 rpm and a centrifugation time of 3 minutes, and freeze-dried to finally obtain litchi peel polyphenol bioactive substances;

[0023] (2) Ultrasonic treatment of abalone gastropod enzyme-mediated soluble collagen: The abalone gastropod enzyme-mediated soluble collagen was dissolved in water to prepare an abalone gastropod enzyme-mediated soluble collagen aqueous solution with a concentration of 6 mg / mL, and then ultrasonically treated, wherein the ultrasonic power was 200 W, the ultrasonic time was 15 minutes, and the sample solution temperature was maintained at 14°C.

[0024] (3) Treatment of abalone gastropod enzyme-mediated soluble collagen with litchi peel polyphenols: Add 10 mg / g of litchi peel polyphenol extract to the ultrasonically treated abalone gastropod enzyme-mediated soluble collagen, adjust the pH to 9, mix well, and stir magnetically at 4°C for 2 hours to obtain deodorized abalone gastropod enzyme-mediated soluble collagen.

[0025] It was determined that the contents of the main odor substances hexanal, heptaldehyde, nonanal, 2-octen-1-ol and decanal in the abalone gastropod enzyme-soluble collagen of this embodiment decreased by 40.8%, 32.8%, 17.6%, 13.2% and 10.6% respectively compared with the untreated protein.

[0026] Embodiment 2:

[0027] (1) Extraction of litchi peel polyphenols: The litchi peel was dried and crushed, and then screened through an 80-mesh sieve to obtain litchi peel powder with uniform particle size. 5 g of litchi peel powder was evenly mixed with 10 mL of distilled water, treated at a pulse electric field strength of 0.75 kV / cm for 3 minutes, and then anhydrous ethanol was added to adjust the ethanol concentration in the system to within the range of 60 wt%. The solution was then ultrasonically treated at a power of 220 W for 15 minutes. The entire extraction process was carried out within the temperature range of 46 ° C, and magnetic stirring was maintained during the ultrasonic treatment. Finally, the supernatant was obtained by centrifugation at a speed of 5000 rpm and a centrifugation time of 4 minutes, and was freeze-dried to finally obtain litchi peel polyphenol bioactive substances.

[0028] (2) Ultrasonic treatment of abalone gastropod enzyme-mediated soluble collagen: The abalone gastropod enzyme-mediated soluble collagen was dissolved in water to prepare an abalone gastropod enzyme-mediated soluble collagen aqueous solution with a concentration of 8 mg / mL, and then ultrasonically treated, wherein the ultrasonic power was 400 W, the ultrasonic time was 20 minutes, and the sample solution temperature was maintained at 15°C.

[0029] (3) Litchi peel polyphenol-treated abalone gastropodase-soluble collagen: Add 40 mg / g of lychee peel polyphenol extract to the ultrasonically treated abalone gastropodase-soluble collagen, adjust the pH to 9, mix evenly, and stir magnetically for 3 hours at 6°C to obtain the deodorized abalone gastropodase-soluble collagen. It was determined that the abalone gastropodase-soluble collagen of this embodiment, compared with the untreated protein, had a decrease in the contents of the main fishy substances hexanal, heptanal, nonanal, 2-octen-1-ol, and decanal by 43.9%, 38.7%, 23.6%, 20.5%, and 13.1%, respectively.

[0030] Embodiment 3:

[0031] (1) Extraction of litchi peel polyphenols: The litchi peel was dried and crushed, and then screened through an 80-mesh sieve to obtain litchi peel powder with uniform particle size. 5 g of litchi peel powder was mixed evenly with 10 mL of distilled water, treated at a pulse electric field strength of 1.25 kV / cm for 5 minutes, and then 80 wt% anhydrous ethanol was added to adjust the ethanol concentration in the system to within the range of 80 wt%. The solution was then ultrasonically treated at a power of 300 W for 15 minutes. The entire extraction process was carried out at a temperature range of 56°C, and magnetic stirring was maintained during the ultrasonic treatment. Finally, the supernatant was obtained by centrifugation at a speed of 6000 rpm and a centrifugation time of 6 minutes, and freeze-dried to finally obtain litchi peel polyphenol bioactive substances.

[0032] (2) Ultrasonic treatment of abalone gastropod enzyme-mediated soluble collagen: The abalone gastropod enzyme-mediated soluble collagen was dissolved in water to prepare an abalone gastropod enzyme-mediated soluble collagen aqueous solution with a concentration of 10 mg / mL, and then ultrasonically treated with an ultrasonic power of 800 W, an ultrasonic time of 30 minutes, and the sample solution temperature was maintained at 16°C.

[0033] (3) Litchi peel polyphenol-treated abalone gastropodase-soluble collagen: Add 50 mg / g of litchi peel polyphenol extract to the abalone gastropodase-soluble collagen treated in step (2), adjust the pH to 9, mix evenly, and stir magnetically for 4 hours at 8° C. to obtain abalone gastropodase-soluble collagen with deodorized fishy smell. It was determined that the abalone gastropodase-soluble collagen of this embodiment, compared with the untreated protein, had a decrease in the contents of the main fishy smell substances hexanal, heptanal, nonanal, 2-octen-1-ol, and decanal by 52.4%, 42.3%, 30.7%, 26.4%, and 16.8%, respectively.

[0034] Comparative Example 1

[0035] Reference Example 3, except that step (3) is not performed. It was determined that the contents of the main odor substances hexanal, heptaldehyde, nonanal, 2-octen-1-ol and decanal in the abalone gastropod enzyme-soluble collagen protein of this comparative example decreased by 17.2%, 8.3%, 4.5%, 3.2% and 1.2%, respectively, compared with the untreated protein.

[0036] Comparative Example 2

[0037] Reference Example 3, except that step (2) is not performed. It was determined that the contents of the main odor substances hexanal, heptaldehyde, nonanal, 2-octen-1-ol and decanal in the abalone gastropod enzyme-mediated soluble collagen protein of this comparative example decreased by 31.7%, 19.4%, 13.8%, 7.5% and 2.3%, respectively, compared with the untreated protein.

[0038] By comparing Example 3 with Comparative Examples 1 and 2, it can be seen that the synergistic effect of step (2) and step (3) in Example 3 can more thoroughly remove the fishy smell of abalone. Compared with the untreated protein, the contents of the main fishy substances hexanal, heptaldehyde, nonanal, 2-octen-1-ol and decanal decreased by 40.8%, 32.8%, 17.6%, 13.2% and 10.6%, respectively.

[0039] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principles of the present invention shall be equivalent replacement methods and shall be included in the protection scope of the present invention.

Claims

1. An efficient method for removing fishy smell by catalyzing soluble collagen from abalone gastropods, characterized in that: The method comprises the following steps: (1) Ultrasonic treatment of abalone gastropod enzyme-mediated soluble collagen: ultrasonic treatment of abalone gastropod enzyme-mediated soluble collagen, with an ultrasonic power of 200-800 W, an ultrasonic time of 15-30 minutes, and a sample solution temperature maintained at 14-16° C.; (2) Treating abalone gastropodase-mediated soluble collagen with litchi peel polyphenol extract: Add 10-50 mg / g of litchi peel polyphenol extract to the abalone gastropodase-mediated soluble collagen treated in step (1), adjust the pH to 9, mix well, and stir magnetically at 4-8° C. for 2-4 hours to obtain deodorized abalone gastropodase-mediated soluble collagen.

2. The method according to claim 1, characterized in that In step (1), the ultrasonic power is 400-800W, and the ultrasonic time is 20-30 minutes.

3. The method according to claim 1, characterized in that Step (2) is: adding 40-50 mg / g of litchi peel polyphenol extract to the abalone gastropod enzyme-mediated soluble collagen processed in step (1), adjusting the pH to 9, mixing evenly, and magnetically stirring for 3-4 hours at 6-8° C. to obtain abalone gastropod enzyme-mediated soluble collagen with deodorized fishy taste.

4. The method according to claim 1, characterized in that: In step (1), the concentration of abalone gastropod enzyme-catalyzed soluble collagen is 6-10 mg / mL.

5. The method according to claim 1, characterized in that In step (1), the concentration of abalone gastropod enzyme-catalyzed soluble collagen is 8-10 mg / mL.

6. The method according to claim 1, characterized in that The preparation of litchi peel polyphenol extract is as follows: the litchi peel is dried and crushed, and then screened through an 80-mesh sieve to obtain litchi peel powder with uniform particle size; 4-6g of litchi peel powder is evenly mixed with distilled water, and treated at a pulse electric field strength of 0.25-1.25kV / cm for 1-5 minutes, and then more than 50wt% of ethanol is added to adjust the ethanol concentration in the system to a range of 40wt%-80wt%, and then the solution is ultrasonically treated for 15 minutes at a power of 150W-300W, the entire extraction process is carried out at a temperature range of 25°C-56°C, and magnetic stirring is maintained during the ultrasonic treatment; finally, the supernatant is obtained by centrifugation, and freeze-dried to finally obtain the litchi peel polyphenol bioactive substance.

7. The method according to claim 1, characterized in that The preparation of litchi peel polyphenol extract is as follows: the litchi peel is dried and crushed, and then screened through an 80-mesh sieve to obtain litchi peel powder with uniform particle size; 4-6g of litchi peel powder is evenly mixed with distilled water, and treated at a pulse electric field strength of 0.75-1.25kV / cm for 3-5 minutes, and then anhydrous ethanol is added to adjust the ethanol concentration in the system to a range of 60wt%-80wt%, and then the solution is ultrasonically treated for 15 minutes at a power of 220W-300W, the entire extraction process is carried out at a temperature range of 46°C-56°C, and magnetic stirring is maintained during the ultrasonic treatment; finally, a supernatant is obtained by centrifugation at a speed of 3000-6000rpm and a centrifugation time of 3-6 minutes, and the supernatant is freeze-dried to finally obtain litchi peel polyphenol bioactive substances.

8. The method according to claim 7, characterized in that Finally, the supernatant is obtained by centrifugation at a speed of 5000-6000 rpm and a centrifugation time of 4-6 minutes, and is freeze-dried to finally obtain litchi peel polyphenol bioactive substances.