Tuna bone essence as well as preparation method and application thereof

Abstract

The invention relates to the technical field of food additives, in particular to tuna bone essence as well as a preparation method and application thereof. The preparation method of the tuna bone essence comprises the following steps: crushing to obtain tuna bone liquid; performing high-pressure cooking: performing high-pressure cooking on the tuna aggregate liquid, discarding waste residues, and taking supernate; performing enzymolysis reaction, namely performing enzymolysis reaction on the supernate twice to obtain tuna bone enzymolysis liquid; fermenting and deodorizing: adding saccharomyces cerevisiae into the tuna bone enzymatic hydrolysate, and fermenting to obtain tuna bone fermentation liquor; and performing Maillard reaction: adding D-xylose, L-cysteine and taurine into the tuna bone fermentation liquid, adjusting the pH value to 7.5-8.5, heating to 110-130 DEG C, and keeping the temperature for 40-60 minutes to obtain the tuna bone essence. According to the preparation method of the tuna bone essence provided by the invention, the flavor of tuna bones cannot be shielded, and the flavor of the tuna bones is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of food additives, and particularly relates to a tuna bone essence and a preparation method and application thereof. Background Art

[0002] There are many inconveniences in the preservation and transportation of fresh seafood, which results in relatively high prices of seafood foods, and it is difficult for residents in non-coastal areas to consume seafood foods. Seafood essences can simulate the unique aromas and flavors of seafood foods. They not only retain the freshness and mellowness of seafood, but also enable these flavors to be widely applied in food processing. Among the currently existing research reports on the preparation of seafood essences, there are mainly three types: one is to prepare through the Maillard reaction of seafood materials; the second is to prepare through the Maillard reaction of seafood materials plus simple blending; the third is to prepare completely through the blending of different monomer flavor raw materials. With the increasing pursuit of food quality by people, the market prospect of seafood essences prepared from seafood materials will be broader.

[0003] Currently, the seafood raw materials of Maillard reaction-type seafood essences are mostly shellfish, fish, shrimps and crabs, etc., and the process is mainly to carry out the Maillard reaction after enzymatic hydrolysis of the seafood raw materials. In the processing of such a preparation process of seafood essences, it is often still necessary to add spices and food flavor enhancers to further cover up the fishy smell of the seafood raw materials and improve the product flavor, but this will cover up the flavor of the seafood raw materials themselves and affect the flavor of the seafood raw materials themselves. Summary of the Invention

[0004] To solve the problem that in the prior art, in the processing of the preparation process of seafood essences, it is often still necessary to add spices and food flavor enhancers to further cover up the fishy smell of the seafood raw materials and improve the product flavor, but this will cover up the flavor of the seafood raw materials themselves and affect the flavor of the seafood raw materials themselves, the present invention provides a tuna bone essence and a preparation method and application thereof. The preparation method uses tuna bones as raw materials, through double-enzyme complex hydrolysis and Saccharomyces cerevisiae fermentation, combined with the Maillard reaction, to develop the by-product tuna bones generated in the processing of tuna into a high-value-added and strong-seafood-flavored edible essence, which will not cover up the flavor of the tuna bones themselves and improves the flavor of the tuna bones themselves.

[0005] To achieve the above object, the present invention adopts the following technical solutions: The present invention provides a preparation method of a tuna bone essence, comprising the following steps: Crushing: Select fresh tuna bones, wash them, cut them into bone segments, and add water to crush the bone segments to obtain a tuna bone material liquid.

[0006] High-pressure cooking: Cook the tuna bone aggregate liquid at 110°C to 130°C and 0.11 Mpa to 0.15 Mpa for 10 min to 20 min to achieve the effects of sterilization, softening the bone quality of the tuna bone, and improving the enzymatic hydrolysis efficiency of subsequent enzymatic hydrolysis reactions. After cooking, discard the waste residue and take the supernatant.

[0007] Enzymatic hydrolysis reaction: Heat the supernatant in a water bath at 45°C to 55°C, add alkaline protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 8.5 to 9.5, and perform the first enzymatic hydrolysis for 1.5 h to 2.5 h to fully destroy the protein tissue structure in the tuna bone; after the solution after the first enzymatic hydrolysis is heated to 90°C to 100°C to inactivate the enzyme for 10 min to 15 min, then add flavor protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 6 to 7, and perform the second enzymatic hydrolysis for 1.5 h to 2.5 h to decompose the bitter peptides in the tuna bone and further hydrolyze them to produce flavor amino acids, enhancing the flavor of the tuna bone itself; after the solution after the second enzymatic hydrolysis is heated to 90°C to 100°C to inactivate the enzyme for 10 min to 15 min, a tuna bone enzymatic hydrolysate is obtained.

[0008] Fermentation for deodorization: Add Saccharomyces cerevisiae to the tuna bone enzymatic hydrolysate and ferment at a constant temperature of 30°C for 3 d to 4 d to remove the fishy smell and impart a wine fragrance to the product, while retaining the flavor of the tuna bone itself; after fermentation, a tuna bone fermentation liquid is obtained.

[0009] Maillard reaction: Add D-xylose, L-cysteine, and taurine to the tuna bone enzymatic hydrolysate, adjust the pH to 7.5 to 8.5, heat to 110°C to 130°C and maintain for 40 min to 60 min to obtain tuna bone essence. Among them, D-xylose, as a reducing sugar, is one of the reaction substrates of the Maillard reaction. The addition of L-cysteine and taurine can improve the flavor and color of the product.

[0010] The present invention selects fresh tuna bones as raw materials. After washing them, they are cut into small sections of 2 cm to 3 cm, and then ground with water to obtain tuna bone slurry. Then, the tuna bone slurry is steamed at 110°C to 130°C and 0.11 Mpa to 0.15 Mpa for 10 min to 20 min to achieve sterilization, softening of the bone quality, and improvement of the enzymatic hydrolysis efficiency of subsequent enzymatic hydrolysis reactions. The supernatant liquid after discarding the bone residues is heated to 45°C to 55°C, alkaline protease is added, and the pH is adjusted to 8.5 to 9.5 with sodium bicarbonate and hydrochloric acid, and enzymatic hydrolysis is carried out for 1.5 h to 2.5 h. Then it is heated to 90°C to 100°C to inactivate the enzyme for 10 min to 15 min. Next, flavor protease is added, the pH is adjusted to 6 to 7 with sodium bicarbonate and hydrochloric acid, and enzymatic hydrolysis is carried out for 1.5 h to 2.5 h. Then it is heated to 90°C to 100°C to inactivate the enzyme for 10 min to 15 min. First, alkaline protease is used, and then flavor protease. The double-enzyme complex hydrolysis can first fully destroy the protein tissue structure in tuna bones, complete deep hydrolysis, then decompose bitter peptides, further hydrolyze to produce a large amount of flavor amino acids, and enhance the flavor of tuna bones themselves. After the hydrolysis of tuna bone enzymatic hydrolysate is completed, Saccharomyces cerevisiae is added and fermented at a constant temperature of 28°C to 30°C for 3 d to 4 d. During this process, yeast can ferment using aldehydes with fishy smells as substrates, and the alcohol produced during the fermentation process can also cover up the fishy smell and enhance the flavor of tuna bones themselves, without the need to add spices and food flavor enhancers subsequently. After the fermentation is completed, D-xylose, L-cysteine, and taurine are added to the tuna bone fermentation broth, the pH is adjusted to 7.5 to 8.5, and it is heated to 110°C to 130°C and maintained for 40 min to 60 min for Maillard reaction. Among them, D-xylose, as a reducing pentose, has low cost and high reaction activity, and is one of the commonly used Maillard reaction substrates in meat processing. L-cysteine and taurine are sulfur-containing compounds, which are indispensable for forming meat flavor, and can supplement the situation where the amino acid source capable of forming seafood flavor in tuna bone enzymatic hydrolysate is incomplete.

[0011] The tuna bone essence product prepared by the preparation method provided by the present invention fills the blank in the market that there is no edible essence prepared from tuna bones as raw materials. And through double-enzyme complex hydrolysis and Saccharomyces cerevisiae fermentation, combined with Maillard reaction, the waste tuna bones generated during the tuna processing are developed into a high-value-added edible essence with a strong seafood flavor, without the need to add spices and food flavor enhancers to further cover up the fishy smell of tuna bones and improve the product flavor. Moreover, it will not block the flavor of tuna bones themselves, and enhances the flavor of tuna bones themselves.

[0012] Further, the alkaline protease is a food-grade alkaline protease, and the enzyme activity is 199,000 U / g to 201,000 U / g.

[0013] Further, the flavor protease is a food-grade flavor protease, and the enzyme activity is 49,000 U / g to 51,000 U / g.

[0014] Further, the Saccharomyces cerevisiae is Angel brewing yeast.

[0015] Further, the mass ratio of the supernatant to the alkaline protease is 1: 0.01 to 0.012.

[0016] Further, the mass ratio of the solution after the first enzymatic hydrolysis to the flavor protease is 1: 0.012.

[0017] Further, the mass ratio of the Saccharomyces cerevisiae added to the tuna bone enzymatic hydrolysate is 1: 0.001 to 0.0016.

[0018] The present invention also provides a tuna bone essence prepared by the preparation method of the above-mentioned tuna bone essence.

[0019] Further, the tuna bone essence is made of the following materials in parts by weight: 10 to 15 parts of tuna bone, 100 to 150 parts of water, 1 to 1.5 parts of alkaline protease, 1 to 1.5 parts of flavor protease, 0.1 to 0.2 parts of Saccharomyces cerevisiae, 1.5 to 2.5 parts of D-xylose, 0.1 to 0.3 parts of L-cysteine, and 0.1 to 0.3 parts of taurine.

[0020] Further, the tuna bone essence is made of the following materials in parts by weight: 12.5 parts of tuna bone, 125 parts of water, 1.25 parts of alkaline protease, 1.25 parts of flavor protease, 0.15 parts of Saccharomyces cerevisiae, 2.0 parts of D-xylose, 0.2 parts of L-cysteine, and 0.2 parts of taurine.

[0021] The present invention also provides the application of the tuna bone essence in the preparation of seafood products, and the tuna bone essence is used as a food additive.

[0022] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The present invention provides a preparation method of a tuna bone essence. The preparation method of the tuna bone essence provided by the present invention is obtained by double-enzyme composite hydrolysis, fermentation deodorization, and Maillard reaction of seafood-based ingredients. In the preparation process of the tuna bone essence, there is no need for blending, and there is no need to add spices and food flavor enhancers to further cover up the fishy smell of tuna bones and improve the product flavor. Moreover, it will not block the original flavor of tuna bones, but enhances the original flavor of tuna bones. The tuna bone essence product prepared by the preparation method provided by the present invention fills the blank in the market of preparing edible essence with tuna bones as raw materials. It is natural, safe, rich in seafood flavor, has a mellow taste, almost no fishy smell, has extremely high commercial value in the seafood food processing market, and has a simple production process and low cost, and can solve the environmental pollution caused by tuna waste.

[0023] 2. The main raw material used in the present invention is the fish bone, a by-product in the tuna processing. Tuna bones are rich in nutrients such as protein, unsaturated fatty acids, vitamins, and minerals. The tuna essence produced from it improves the utilization rate and added value of tuna bones, and also solves the problem of environmental pollution.

[0024] As a deep-sea fish, tuna not only has delicious meat and high nutritional value, but also is far from offshore pollution. Tuna is generally processed into sashimi or canned products. During the processing, about 60% of by-products are generated, including fish heads, fish bones, fish skins, internal organs, etc. These by-products are mostly used to produce fish meal as animal feed, and the fish meal processing will emit odors and a large amount of sewage, causing environmental pollution.

[0025] Tuna bones account for about 30% of the fish body weight and are the by-products with the largest proportion in the tuna processing. They are rich in nutrients such as protein, unsaturated fatty acids, vitamins, and minerals. Among the existing research reports on the development and utilization of tuna bones, they mainly include extracting bioactive peptides with tuna bones as raw materials and preparing tuna bone calcium powder. There is no relevant research on developing edible essence with tuna bones as raw materials. The preparation method of tuna bone essence provided by the present invention is a method for developing edible essence with tuna bones as raw materials, expanding the variety of seafood essences.

[0026] 3. The present invention uses tuna bones, a seafood ingredient, as the raw material to prepare Maillard-type seafood essence, without adding monomer flavor raw materials for blending, which conforms to the concept of natural safety.

[0027] 4. The present invention uses the method of fermenting and enzymolyzing the enzyme solution with Saccharomyces cerevisiae to remove fishy smell. Compared with the seafood essences on the market that rely on adding food flavor enhancers and spices to cover up the fishy smell, it can better retain the original flavor of seafood.

[0028] 5. Currently, the development of tuna bones on the market mainly focuses on extracting bioactive peptides with tuna bones as raw materials and preparing tuna bone calcium powder. The present invention can fill the market gap of preparing edible essence with tuna bones as raw materials.

[0029] 6. The present invention uses alkaline protease and flavor protease for double-enzyme complex hydrolysis to fully destroy the protein tissue structure in tuna bones, complete deep hydrolysis, then decompose bitter peptides, further hydrolyze to produce a large amount of flavor amino acids, and at the same time add two sulfur-containing compounds, L-cysteine and taurine, to enrich the amino acid source for Maillard reaction, enhancing the product flavor. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is the preparation process flow of the tuna bone essence in the embodiment of the present invention.

[0031] Figure 2This is the liquid chromatogram of sweet amino acids (serine, threonine, glycine, alanine, histidine, and proline) and umami amino acids (glutamic acid, aspartic acid, and lysine) in the tuna bone essence of the embodiment in the present invention; among them, 1 is aspartic acid, 2 is glutamic acid, 3 is serine, 4 is glycine, 5 is histidine, 6 is threonine, 7 is alanine, 8 is proline, and 9 is lysine. Detailed implementation manners

[0032] The following will describe the detailed implementation manners of the present invention in conjunction with the accompanying drawings. However, it should be understood that the protection scope of the present invention is not limited by the detailed implementation manners. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the protection scope of the present invention. The experimental methods described in the embodiments of the present invention are all conventional methods unless otherwise specified. The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0033] Embodiment 1: This embodiment provides a preparation method of tuna bone essence. The tuna bone essence includes the following raw materials in parts by weight: 12.5 parts of tuna bone, 125 parts of water, 1.25 parts of alkaline protease, 1.25 parts of flavor protease, 0.15 part of Saccharomyces cerevisiae, 2 parts of D-xylose, 0.2 part of L-cysteine, and 0.2 part of taurine.

[0034] Among them, the specific operation steps of the preparation method of the tuna bone essence are as follows: S1, comminution: Select fresh tuna bones, wash them, cut them into fish bone segments, and add water to comminute the fish bone segments to obtain tuna bone slurry.

[0035] S2, high-pressure steaming: Steam the obtained tuna bone slurry at 120 °C and 0.13 Mpa for 15 min, discard the waste residue, and take the supernatant.

[0036] S3, enzymatic hydrolysis reaction: Take the prepared supernatant, keep the temperature at 50 °C by water bath heating, add alkaline protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 9, hydrolyze for 2 h, inactivate the enzyme at 100 °C for 10 min, then add flavor protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 7, hydrolyze for 2 h, and inactivate the enzyme at 100 °C for 10 min to obtain tuna bone hydrolyzate.

[0037] S4, fermentation and deodorization: Add Saccharomyces cerevisiae to the tuna bone hydrolyzate, ferment at a constant temperature of 30 °C for 4 d to obtain tuna bone fermentation broth.

[0038] S5, Maillard reaction: Add D-xylose, L-cysteine, and taurine to the enzymatically hydrolyzed solution of tuna bone. Add sodium bicarbonate and hydrochloric acid to adjust the pH to 8, heat to 120 °C and maintain for 50 min to obtain tuna bone essence.

[0039] Example 2: This example provides a method for preparing tuna bone essence. The tuna bone essence comprises the following raw materials by weight share: 15 parts of tuna bone, 150 parts of water, 1.5 parts of alkaline protease, 1.5 parts of flavor protease, 0.2 part of Saccharomyces cerevisiae, 2.5 parts of D-xylose, 0.3 part of L-cysteine, and 0.3 part of taurine.

[0040] Among them, the specific operation steps of the method for preparing tuna bone essence are as follows: S1, Crushing: Select fresh tuna bone, wash it, cut it into bone segments, and add water to crush the bone segments to obtain tuna bone slurry.

[0041] S2, High-pressure cooking: Cook the obtained tuna bone slurry at 120 °C and 0.13 Mpa for 15 min, discard the waste residue, and take the supernatant.

[0042] S3, Enzymatic hydrolysis reaction: Take the prepared supernatant, keep the temperature at 50 °C by water bath heating, add alkaline protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 9, hydrolyze for 2 h, inactivate the enzyme at 100 °C for 10 min, then add flavor protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 7, hydrolyze for 2 h, and inactivate the enzyme at 100 °C for 10 min to obtain the enzymatically hydrolyzed solution of tuna bone.

[0043] S4, Fermentation and deodorization: Add Saccharomyces cerevisiae to the enzymatically hydrolyzed solution of tuna bone, ferment at a constant temperature of 30 °C for 4 d to obtain the fermented solution of tuna bone.

[0044] S5, Maillard reaction: Add D-xylose, L-cysteine, and taurine to the enzymatically hydrolyzed solution of tuna bone. Add sodium bicarbonate and hydrochloric acid to adjust the pH to 8, heat to 120 °C and maintain for 50 min to obtain tuna bone essence.

[0045] Example 3: This example provides a method for preparing tuna bone essence. The tuna bone essence comprises the following raw materials by weight share: 10 parts of tuna bone, 100 parts of water, 1 part of alkaline protease, 1 part of flavor protease, 0.1 part of Saccharomyces cerevisiae, 1.5 parts of D-xylose, 0.1 part of L-cysteine, and 0.1 part of taurine.

[0046] Among them, the specific operation steps of the method for preparing tuna bone essence are as follows: S1, Crushing: Select fresh tuna bones, wash them, cut them into bone segments, and add water to crush the bone segments to obtain tuna bone aggregate liquid.

[0047] S2, High-pressure steaming: Steam the obtained tuna bone aggregate liquid at 120 °C and 0.13 MPa for 15 min, discard the waste residue, and take the supernatant.

[0048] S3, Enzymatic hydrolysis reaction: Take the prepared supernatant, heat it in a water bath to keep the temperature at 50 °C, add alkaline protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 9, carry out enzymatic hydrolysis for 2 h, inactivate the enzyme at 100 °C for 10 min, then add flavor protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 7, carry out enzymatic hydrolysis for 2 h, inactivate the enzyme at 100 °C for 10 min, to obtain tuna bone enzymatic hydrolysate.

[0049] S4, Fermentation for deodorization: Add Saccharomyces cerevisiae to the tuna bone enzymatic hydrolysate, ferment at a constant temperature of 30 °C for 4 d to obtain tuna bone fermentation liquid.

[0050] S5, Maillard reaction: Add D-xylose, L-cysteine, and taurine to the tuna bone enzymatic hydrolysate, add sodium bicarbonate and hydrochloric acid to adjust the pH to 8, heat to 120 °C and maintain for 50 min to obtain tuna bone essence.

[0051] Comparative Example 1: This comparative example provides a preparation method of tuna bone essence, which is different from Example 2 in that fermentation for deodorization is not carried out. The tuna bone essence includes the following raw materials by weight share: 15 parts of tuna bones, 150 parts of water, 1.5 parts of alkaline protease, 1.5 parts of flavor protease, 0.2 part of Saccharomyces cerevisiae, 2.5 parts of D-xylose, 0.3 part of L-cysteine, and 0.3 part of taurine.

[0052] Among them, the specific operation steps of the preparation method of the tuna bone essence are as follows: S1, Crushing: Select fresh tuna bones, wash them, cut them into bone segments, and add water to crush the bone segments to obtain tuna bone aggregate liquid.

[0053] S2, High-pressure steaming: Steam the obtained tuna bone aggregate liquid at 120 °C and 0.13 Mpa for 15 min, discard the waste residue, and take the supernatant.

[0054] S3, Enzymatic hydrolysis reaction: Take the prepared supernatant, heat it in a water bath to keep the temperature at 50 °C, add alkaline protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 9, carry out enzymatic hydrolysis for 2 h, inactivate the enzyme at 100 °C for 10 min, then add flavor protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 7, carry out enzymatic hydrolysis for 2 h, inactivate the enzyme at 100 °C for 10 min, to obtain tuna bone enzymatic hydrolysate.

[0055] S4, Maillard reaction: Add D-xylose, L-cysteine, and taurine to the enzymatically hydrolyzed solution of tuna bones. Add sodium bicarbonate and hydrochloric acid to adjust the pH to 8, heat to 120 °C and maintain for 50 min to obtain tuna bone essence.

[0056] Comparative Example 2: This comparative example provides a method for preparing tuna bone essence, which is different from Example 2 in that L-cysteine is not added. The tuna bone essence comprises the following raw materials by weight: 15 parts of tuna bones, 150 parts of water, 1.5 parts of alkaline protease, 1.5 parts of flavor protease, 0.2 part of Saccharomyces cerevisiae, 2.5 parts of D-xylose, 0 part of L-cysteine, and 0 part of taurine.

[0057] Among them, the specific operating steps of the method for preparing the tuna bone essence are as follows: S1, Crushing: Select fresh tuna bones, wash them, cut them into bone segments, and add water to crush the bone segments to obtain tuna bone slurry.

[0058] S2, High-pressure cooking: Cook the obtained tuna bone slurry at 120 °C and 0.13 Mpa for 15 min, discard the waste residue, and take the supernatant.

[0059] S3, Enzymatic hydrolysis reaction: Take the prepared supernatant and keep the temperature at 50 °C by water bath heating. Add alkaline protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 9, hydrolyze for 2 h, inactivate the enzyme at 100 °C for 10 min, then add flavor protease, add sodium bicarbonate and hydrochloric acid to adjust the pH to 7, hydrolyze for 2 h, and inactivate the enzyme at 100 °C for 10 min to obtain the enzymatically hydrolyzed solution of tuna bones.

[0060] S4, Fermentation and deodorization: Add Saccharomyces cerevisiae to the enzymatically hydrolyzed solution of tuna bones, and ferment at a constant temperature of 30 °C for 4 d to obtain the fermented solution of tuna bones.

[0061] S5, Maillard reaction: Add D-xylose to the enzymatically hydrolyzed solution of tuna bones, add sodium bicarbonate and hydrochloric acid to adjust the pH to 8, heat to 120 °C and maintain for 50 min to obtain tuna bone essence.

[0062] Comparative Example 3 This comparative example refers to the preparation process of seafood flavor essence using squid offal in the Chinese patent with the patent number 202410401229.X, "Preparation process of seafood flavor essence using squid offal", to prepare a tuna bone essence, including the following steps: S1, Stir and crush the tuna bones to form crushed materials, mix the crushed materials and pure water according to a material-liquid ratio of 1:2, and then perform ultrasonic treatment at a power of 180 W for 15 minutes to form a stock solution.

[0063] S2. Adjust the pH value of the stock solution to 9.0, add alkaline protease at a dosage of 4000 U / g, conduct the first enzymatic hydrolysis, maintain the temperature at 55 °C for 5 hours, and then inactivate the enzyme at a temperature of 90 °C for 10 minutes.

[0064] S3. Add flavor protease to the stock solution that has completed the first enzymatic hydrolysis at a dosage of 1400 U / g, conduct the second enzymatic hydrolysis, maintain the temperature at 55 °C for 4 hours, and then inactivate the enzyme at a temperature of 90 °C for 10 minutes.

[0065] S4. Centrifuge and filter the stock solution that has completed the second enzymatic hydrolysis at a centrifugal speed of 4000 rpm for 20 minutes to obtain the enzymatic hydrolysate.

[0066] S5. Add 5% reducing sugar (4% glucose and 1% xylose), 0.2% food flavor enhancer I+G, and an amino acid mixture composed of 1% arginine, 1% alanine, 0.2% glycine, 0.1% taurine, 0.3% methionine, and 0.2% cysteine to the enzymatic hydrolysate according to weight percentage, adjust the pH value to 7.0, and maintain the Maillard reaction at 105 °C for 20 minutes to form a liquid flavor.

[0067] Comparative Example 4 This comparative example refers to the Chinese patent "A Seafood Flavor Paste Flavor and Its Preparation Method" with the patent number 202211367368.2 to prepare a tuna bone flavor, including the following steps: S1. Weigh 25 parts of tuna bones, add 60 parts of pure water, and 0.2 parts of collagenase. Maintain the temperature at 60 °C and the pH value at 7, and conduct enzymatic hydrolysis for 30 minutes. Then raise the temperature to 100 °C and heat for 30 minutes to inactivate the enzyme, obtaining a tuna bone collagen polypeptide hydrolysate; the enzyme activity of the collagenase is 100,000 U / g.

[0068] S2. Weigh 30 parts of the tuna bone collagen polypeptide hydrolysate, add 4 parts of yeast, 5 parts of granulated sugar, 1 part of I+G, 4 parts of glucose, 4 parts of amino acids, and 0.5 parts of ginger powder, and mix them thoroughly in a blender; then conduct the Maillard reaction at a reaction temperature of 100 °C and a reaction time of 150 minutes; the amino acids are a mixture of methionine, leucine, alanine, and cysteine with a mass ratio of 0.5:0.5:1.8:3.7.

[0069] S3. After the reaction is completed, take out the material and cool it at room temperature, and add xanthan gum and stir evenly to obtain it, where the addition amount of xanthan gum is 0.2% of the mass of the material after the reaction is completed.

[0070] Among them, the above-mentioned parts refer to mass parts.

[0071] Comparative Example 5 This comparative example prepares a tuna bone essence by referring to the Chinese patent "A Preparation Method of a Seafood Flavor Essence" with the patent number 200810152815.6, including the following steps: Crush the tuna bones and pass through a 16-mesh sieve to obtain crushed tuna bones; take 20 parts of crushed tuna bones, 10 parts of glucose, 10 parts of methionine, 10 parts of leucine, 20 parts of corn acid-hydrolyzed vegetable protein, 5 parts of acetic acid, 5 parts of edible ethanol, 5 parts of minced ginger, 5 parts of minced garlic, 1 part of astaxanthin, and 70 parts of salad oil by mass. Put each component into a high-pressure reaction kettle, mix well, quickly raise the temperature to 105°C, start the heat preservation reaction timing, control the temperature at 120°C, and the reaction time is 45 minutes. After the reaction is completed, cool to room temperature and filter to obtain the filtrate, which is the oily seafood flavor essence.

[0072] Carry out sensory evaluation on the tuna bone essences in Examples 1 to 3 and Comparative Examples 1 to 5. The evaluation group consists of 9 trained sensory assessors. The samples are randomly poured into paper cups, and the 9-person evaluation group evaluates the color, taste, smell, and acceptability of the samples respectively. The sensory quality evaluation criteria are shown in Table 1.

[0073] Table 1 Sensory Quality Evaluation Criteria for Tuna Bone Essence In the sensory score results, the scores for each sub-item of color, taste, smell, and acceptability are the arithmetic means of the sum of the corresponding sub-items after removing the highest and lowest scores. The total score is the sum of the scores for each sub-item of color, taste, smell, and acceptability, as shown in Table 2.

[0074] Table 2 Sensory Evaluation Score Table for Tuna Bone Essence As can be seen from Table 2, each index in Examples 1 to 3 of the present invention has a relatively high score. It can be seen therefrom that the tuna bone edible essence prepared by the present invention has a strong seafood flavor, an attractive color, and a mellow taste, and can be widely used in the preparation of seafood flavor foods.

[0075] In Comparative Example 1, compared with Example 2, without further fermentation and deodorization, and with the same mass of input materials, the score of Comparative Example 1 in terms of smell is lower than that of Example 2. This is because yeast can ferment with aldehydes with a fishy smell as the substrate, and the alcohol produced during the fermentation process can also cover up the fishy smell and enhance the flavor of the tuna bones themselves.

[0076] In Comparative Example 2, compared with Example 2, L-cysteine and taurine were not added, and when the other input materials were exactly the same in terms of mass, the scores of Comparative Example 2 in terms of taste and smell were lower than those of Example 2. This is because although the enzymatic hydrolysate contains amino acid sources for forming seafood flavors, it is still not comprehensive enough. L-cysteine and taurine are sulfur-containing compounds and are indispensable for forming meaty flavors.

[0077] In Comparative Example 3, the input materials, the mass of the input materials, and the preparation method refer to the preparation process for producing seafood flavor essence using squid offal in the Chinese patent with the patent number 202410401229.X. The scores of Comparative Example 3 in terms of taste and smell were lower than those of Example 2. This is because Comparative Example 3 did not perform yeast fermentation for deodorization, but relied on adding food flavor enhancer I+G to enhance umami and mask fishy smells, which would mask the original flavor of the tuna bones. In addition, the alcohol produced after yeast fermentation can further enhance the flavor of the essence, while Comparative Example 3 lacks this step of yeast fermentation, resulting in a decline in the overall taste and smell.

[0078] In Comparative Example 4, the input materials, the mass of the input materials, and the preparation method refer to a Chinese patent with the patent number 202211367368.2 for a seafood flavor paste essence and its preparation method. The scores of Comparative Example 4 in terms of color, taste, and smell were lower than those of Example 2. This is because Comparative Example 4 used collagenase to hydrolyze the tuna bones, and the hydrolysis effect on the bone collagen of the tuna bones was good, but the hydrolysis effect on other macromolecular proteins on the tuna bones was weak. This would lead to fewer types and amounts of amino acids in the enzymatic hydrolysate of the tuna bones than in Example 2, directly affecting the color, taste, and smell after the subsequent Maillard reaction. In addition, Comparative Example 4 did not perform yeast fermentation for deodorization, but relied on adding food flavor enhancer I+G and ginger powder as spices to enhance umami and mask fishy smells, which would mask the original flavor of the tuna bones. In addition, the alcohol produced after yeast fermentation can further enhance the flavor of the essence, while Comparative Example 4 lacks this step of yeast fermentation, resulting in a decline in the overall taste and smell.

[0079] In Comparative Example 5, the input materials, the mass of the input materials, and the preparation method refer to the preparation method of a seafood flavor essence in a Chinese patent with the patent number 200810152815.6. The score of Comparative Example 5 in terms of color, taste, and smell is lower than that of Example 2. This is because in Comparative Example 5, the tuna bones were not enzymatically hydrolyzed, which would result in the inability to release a large amount of flavor amino acids in the tuna bones, directly affecting the color, taste, and smell of the subsequent Maillard reaction. In addition, in Comparative Example 5, yeast fermentation was not carried out for deodorization, but instead, food flavor enhancer I+G, minced ginger and scallions as spices, and acetic acid were added to enhance the umami taste and mask the fishy smell. This would mask the original flavor of the tuna bones. In addition, during the process of yeast fermentation of the enzymatically hydrolyzed solution of tuna bones, in addition to producing alcohol, a small amount of flavor esters would also be produced, further enhancing the flavor of the essence. However, in Comparative Example 5, yeast fermentation was not carried out, but instead, edible ethanol was directly added, resulting in a decrease in the overall taste and smell compared to Example 2.

[0080] In summary, the tuna bone essence product prepared by the preparation method provided by the present invention fills the gap in the market where there is no edible essence prepared from tuna bones. Through double-enzyme complex hydrolysis and Saccharomyces cerevisiae fermentation, combined with the Maillard reaction, the tuna bones, which are by-products generated during the tuna processing, are developed into a high-value-added edible essence with a strong seafood flavor. It does not mask the original flavor of the tuna bones and enhances the original flavor of the tuna bones. Moreover, this application does not require the addition of spices and food flavor enhancers to further mask the fishy smell and improve the product flavor.

[0081] At the same time, the present invention also used the PITC pre-column derivation - reversed-phase high-performance liquid chromatography method to detect the sweet amino acids (serine, threonine, glycine, alanine, histidine, and proline) and umami amino acids (glutamic acid, aspartic acid, and lysine) in the tuna bone essence prepared in Example 1. The specific detection method is as follows:

[0082] 1. Reagents and instruments: Phenyl isothiocyanate (PITC, purity > 98%): Sigma Company, USA; Acetonitrile (chromatographic pure): Shanghai Zhanyun Chemical Co., Ltd.; Sodium acetate trihydrate (analytical pure): Hangzhou Chemical Reagent Co., Ltd.; Triethylamine (analytical pure): Shanghai Zhanyun Chemical Co., Ltd.; n-Hexane (analytical pure): Sinopharm Chemical Reagent Co., Ltd.

[0083] LC-20AT liquid chromatograph: Shimadzu Corporation, Japan.

[0084] 2. Experimental procedures 1) Sample derivatization: Accurately transfer 1 mL of the sample solution into a 10 mL stoppered test tube. Add 0.5 mL each of a 14% triethylamine - acetonitrile solution by volume and a 1.2% phenyl isothiocyanate - acetonitrile solution by volume. Mix well, let stand for 30 min at room temperature, then add 2 mL of n - hexane. After shaking for 60 s with a vortex mixer and letting stand for 10 min to extract the excess PITC in the reaction, take the lower clear liquid, filter it through a 0.45 μm organic filter membrane, and wait for reverse - phase high - performance liquid chromatography analysis.

[0085] Among them, the sample solution refers to the tuna bone essence prepared in Example 1.

[0086] 2) Chromatographic conditions: Chromatographic column: SHIMSEN Ankylo C18 chromatographic column (250 mm×4.6 mm, 5 μm); Mobile phase A: 0.1 mol / L sodium acetate buffer - acetonitrile (97:3, V / V); Mobile phase B: acetonitrile - water (4:1, V / V); Flow rate: 0.8 mL / min; Detection wavelength: 254 nm; Column temperature: 30 °C; Injection volume: 20 μL; Elution program: see Table 3.

[0087] Table 3 Analytical gradient elution program The detection results are shown in Figure 2 .

[0088] Taking the peak area as the detection result, the amino acid content is calculated by the external standard method. As can be seen from Figure 2 , the tuna bone essence contains sweet amino acids (serine, threonine, glycine, alanine, histidine, and proline) and umami amino acids (glutamic acid, aspartic acid, and lysine). The contents of 9 amino acids are shown in Table 4.

[0089] Table 4 Contents of sweet amino acids and umami amino acids in tuna bone essence From the above results, it can be seen that the tuna bone essence prepared in the examples of the present invention is rich in various sweet amino acids, significantly improving the utilization rate and added value of tuna bones, and also solving the environmental pollution.

[0090] Although the preferred embodiments of the present invention have been described, those skilled in the art can make additional changes and modifications to these embodiments once they know the basic creative concepts.

[0091] Obviously, those skilled in the art can make various changes and deformations to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and deformations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention also intends to include these modifications and deformations.

Claims

1. A method for preparing tuna bone flavor, characterized in that: The steps include: Select fresh tuna bones, wash them, cut them into fish bone segments, add water to crush the fish bone segments, and obtain tuna bone liquid; High-pressure cooking: cooking the tuna bone liquid at 110° C. to 130° C. and 0.11 MPa to 0.15 MPa for 10 min to 20 min to soften the tuna bone; after the cooking is completed, discard the waste residue and take the supernatant; Enzymatic hydrolysis reaction: heating the supernatant at 45° C. to 55° C., adding alkaline protease, adjusting the pH to 8.5 to 9.5, and performing the first enzymatic hydrolysis for 1.5 h to 2.5 h to fully destroy the organizational structure of the protein in the tuna bone; The solution after the first enzymatic hydrolysis is heated to 90°C-100°C, the enzyme is inactivated, and then flavor protease is added to adjust the pH to 6-7. The second enzymatic hydrolysis is performed for 1.5h-2.5h to decompose the bitter peptides in the tuna bones, and further hydrolyze them to produce flavor amino acids; the solution after the second enzymatic hydrolysis is heated to inactivate the enzyme to obtain a tuna bone enzymatic hydrolyzate; Fermentation to remove the fishy smell: adding brewer's yeast to the tuna bone enzymatic hydrolysate for fermentation to remove the fishy smell and impart wine aroma, while retaining the flavor of the tuna bone itself; After fermentation, a tuna bone fermented liquid is obtained; Maillard reaction: add D-xylose, L-cysteine ​​and taurine to the tuna bone fermentation liquid, adjust the pH to 7.5-8.5, heat to 110° C.-130° C. and keep for 40 min-60 min to obtain the tuna bone flavor.

2. The method for preparing tuna bone flavor according to claim 1, characterized in that: The alkaline protease is a food-grade alkaline protease with an enzyme activity of 199,000 U / g to 201,000 U / g.

3. The method for preparing tuna bone flavor according to claim 1, characterized in that: The flavor protease is a food-grade flavor protease with an enzyme activity of 49,000 U / g to 51,000 U / g.

4. The method for preparing tuna bone flavor according to claim 1, characterized in that: The mass ratio of the supernatant to the alkaline protease is 1:0.01-0.

012.

5. The method for preparing tuna bone flavor according to claim 1, characterized in that: The mass ratio of the solution after the first enzymatic hydrolysis to the flavor protease is 1:0.01-0.

012.

6. The method for preparing tuna bone flavor according to claim 1, characterized in that: The mass ratio of adding the brewer's yeast to the tuna bone enzymatic hydrolysate is 1:0.001-0.0016.

7. The tuna bone flavor prepared by the preparation method of the tuna bone flavor according to claim 1.

8. The tuna bone flavor according to claim 7, characterized in that: The invention is prepared from the following materials in parts by weight: 10 to 15 parts of tuna bones, 100 to 150 parts of water, 1 to 1.5 parts of alkaline protease, 1 to 1.5 parts of flavor protease, 0.1 to 0.2 parts of brewer's yeast, 1.5 to 2.5 parts of D-xylose, 0.1 to 0.3 parts of L-cysteine ​​and 0.1 to 0.3 parts of taurine.

9. The tuna bone flavor according to claim 8, characterized in that: The invention is prepared from the following materials in parts by weight: 12.5 parts of tuna bones, 125 parts of water, 1.25 parts of alkaline protease, 1.25 parts of flavor protease, 0.15 parts of brewer's yeast, 2.0 parts of D-xylose, 0.2 parts of L-cysteine ​​and 0.2 parts of taurine.

10. Use of the tuna bone flavor according to claim 7 in the preparation of seafood products, wherein the tuna bone flavor is used as a food additive.

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