Preparation method of seafood flavor powder with synergic aroma enhancement of compound protease and yeast

Abstract

The invention discloses a preparation method of compound protease-yeast synergistic aroma enhancement seafood flavor powder, which comprises the following steps: firstly, mixing raw materials such as euphausia superba powder, dried oyster powder, skipjack powder, shrimp shell powder, shiitake mushroom powder, kelp powder, fish collagen peptide, glycine, L-alanine and glucose; the papain and bromelain with specific enzyme activity are subjected to composite enzymatic hydrolysis to create a flavor precursor. Then, the enzymatic hydrolysate is fermented through pichia pastoris, disodium 5 '-ribonucleotide (I + G) is added synchronously, and rich aroma components such as pyrazine and sulfide are generated; after fermentation, laver powder, cod-liver oil, a phaffia rhodozyma fermentation extract rich in astaxanthin and ginger oleoresin are added for flavor locking and oil immersion homogeneous emulsification, and finally, a microporous granular product is prepared through compound wall material embedding and spray drying. According to the preparation method, biological enzymolysis, microbial fermentation and flavor targeted locking technologies are integrated, so that the aroma level, strength, stability and cold water instant solubility of the flavor powder are improved.

Description

Technical Field

[0001] This invention belongs to the field of food processing technology, and specifically discloses a method for preparing a seafood flavor powder with synergistic flavor enhancement by compound protease and yeast. Background Technology

[0002] Seafood flavor powder, as an important basic ingredient for enhancing the flavor of savory foods, is widely used in snack foods, instant seasoning packets, and pre-made meal seasonings. Its flavor authenticity, intensity, and persistence are key factors determining the quality of the final product. Traditional seafood flavor powders mainly rely on high-temperature thermal reactions or direct drying and pulverizing processes for preparation. While these methods can achieve the basic flavor, they have significant shortcomings in flavor complexity, natural properties, and application performance.

[0003] Current mainstream preparation technologies face several technical bottlenecks: First, while thermal reaction methods can produce rich aromas, the reaction process is intense, easily generating undesirable substances such as heterocyclic amines, and the aroma is monotonous, lacking top notes and the complex, mellow flavor of fermentation. Second, products obtained by direct pulverization and drying have a strong fishy smell, low flavor intensity, and a short shelf life due to easy fat oxidation. Third, while conventional enzymatic hydrolysis can increase the content of flavor precursors, the effect of single enzymatic hydrolysis is limited, making it difficult to fully release and transform flavor precursors in muscle tissue. Furthermore, the bitter peptides and undesirable aldehydes produced by enzymatic hydrolysis cannot be effectively eliminated, affecting the purity of the flavor. Fourth, most existing flavor powders use physical mixing and encapsulation, which makes flavor substances prone to volatilization and dispersion, resulting in poor aroma retention during product storage and application, slow resolubility in cold water, and ultimately, a lack of concentrated and persistent flavor release in the final product.

[0004] Furthermore, existing technologies do not adequately utilize seafood by-products for high-value production and fail to effectively synergistically integrate the flavor characteristics of different raw materials through biotechnology. Insufficient protection measures for heat-sensitive flavor compounds during the process and unreasonable spray drying parameter settings result in dense product particles, poor solubility, and low retention of umami substances. Therefore, the market urgently needs a new green method for preparing seafood flavor powder that can deeply integrate biotransformation technology, significantly improve flavor quality, and possess excellent processing performance and application characteristics. Summary of the Invention

[0005] To address the above problems, this invention discloses a method for preparing a seafood flavor powder with synergistic flavor enhancement by a complex protease and yeast.

[0006] To achieve the above objectives, the present invention includes the following technical solutions:

[0007] A method for preparing a seafood flavor powder with synergistic flavor enhancement by a complex protease and yeast includes the following steps:

[0008] (1) Raw material pretreatment: Mix the following raw materials evenly by weight: 15-25 parts Antarctic krill powder, 8-15 parts dried oyster powder, 5-10 parts dried scallop powder, 5-10 parts bonito powder, 8-12 parts shrimp shell powder, 3-8 parts shiitake mushroom powder, 3-8 parts kelp powder, 2-4 parts fish collagen peptide powder, 2-5 parts glycine, 1-3 parts L-alanine, 5-10 parts glucose, and 80-120 parts water;

[0009] (2) Compound enzymatic hydrolysis: Add compound protease to the mixture in step (1), control the final addition amount of papain to be 30,000-50,000 U / L of the mixture, and the final addition amount of bromelain to be 60,000-100,000 U / L of the mixture, and enzymatically hydrolyze for 1.5-3 hours at 45-55℃ and pH 6.0-7.0, and then heat to 95℃ for sterilization for 10 minutes;

[0010] (3) Microbial fermentation: Cool the enzymatic hydrolysate from step (2) to 28-32℃, inoculate with 2%-4% Pichia pastoris suspension, and simultaneously add 0.01%-0.03% disodium 5'-ribonucleotide, and ferment at 25-32℃ for 36-60 hours with ventilation.

[0011] (4) Flavor blending and homogenization emulsification: Add 3%-5% of the total weight of laver powder to the fermentation liquid in step (3), and extract at a constant temperature of 75-85℃ for 30-60 minutes; then add 5%-10% cod liver oil, 0.5%-2% red phaf yeast fermentation extract and 0.5%-1.5% ginger oleoresin to the system, and perform high-speed shear homogenization emulsification at a temperature of 70-80℃, a rotation speed of 8000-12000rpm, and a time of 3-5 minutes to form a stable emulsion;

[0012] (5) Solid-liquid separation: The emulsion obtained in step (4) is filtered through a 100-200 mesh sieve and the filtrate is collected;

[0013] (6) Microencapsulation and drying: The filtrate obtained in step (5) is mixed and homogenized with the wall material at a weight ratio of 1:(3-5). The wall material is composed of maltodextrin, sodium octenyl succinate starch, and gum arabic at a weight ratio of (6-8):(1.5-2.5):1. Then, spray drying is performed. The inlet air temperature of the spray drying is 175-185℃ and the outlet air temperature is 80-90℃ to obtain a microporous seafood flavor powder.

[0014] The above preparation method fully releases flavor precursors through compound enzymatic hydrolysis and generates key aromas through yeast fermentation biotransformation. Finally, through flavor locking and microencapsulation technology, the product's flavor level, intensity, stability, and cold water solubility are synergistically improved.

[0015] Furthermore, in the above preparation method, in step (1), the average molecular weight of the fish collagen peptide powder is less than 2000 Daltons. This method limits the use of low molecular weight fish collagen peptides, which are more easily hydrolyzed and utilized by enzymes. As a highly efficient flavor precursor, it helps to enhance the Maillard reaction and improve umami and richness.

[0016] Furthermore, in the above preparation method, step (3) involves fermentation under the following conditions: ventilated fermentation at 30°C for 48 hours with an aeration rate of 0.8-1.2 vvm. This method optimizes the fermentation temperature, time, and aeration rate parameters to provide optimal metabolic conditions for Pichia pastoris, ensuring efficient and stable synthesis of flavor compounds and guaranteeing consistent product quality.

[0017] Furthermore, in the above preparation method, in step (4), the astaxanthin content of the red phaf yeast fermentation extract is not less than 1%. This method limits the astaxanthin content of the red phaf yeast extract, utilizing its strong natural antioxidant properties to effectively inhibit lipid oxidation and the generation of unpleasant odor substances, ensuring the purity and stability of the flavor.

[0018] Furthermore, in the above preparation method, step (6) involves preparing the wall material by dissolving maltodextrin, sodium octenyl succinate starch, and gum arabic in hot water at 60-70°C in a specific ratio to prepare a solution with a concentration of 40%-50%, and stirring until completely dissolved and homogeneous before use. This method specifies the pretreatment method for the wall material to ensure that the wall material components are fully dissolved and homogeneous, forming a dense and complete encapsulation wall layer, thereby improving the encapsulation rate of flavor substances and product stability.

[0019] Furthermore, in the above preparation method, in step (6), the spray drying uses a centrifugal atomizer with an atomizing disc rotation speed of 18000-22000 rpm. This method uses centrifugal atomization and a specific rotation speed to control the formation of microporous particles with uniform particle size and loose structure, thereby giving the product excellent cold water solubility and dispersibility.

[0020] The present invention also discloses a seafood flavor powder, which is prepared by any of the above preparation methods.

[0021] Furthermore, the aforementioned seafood flavor powder has a resolubility time of less than or equal to 5 seconds in cold water at 25°C, and a retention rate of more than 90% for umami amino acids and flavor nucleotides. As can be seen from the test examples, the product of this invention has an extremely fast cold water resolubility and an extremely high retention rate of umami substances, significantly improving ease of use and the flavor experience of the final product.

[0022] This invention also discloses the application of the above-mentioned seafood flavor powder in the preparation of seasoning powders for snack puffed foods or pre-made soup base compound seasoning packets. As can be seen from the test examples, the excellent flavor and solubility properties of this invention can significantly improve the quality of such end products.

[0023] Compared with the prior art, the present invention has the following outstanding advantages:

[0024] 1. Significantly enhanced flavor richness and intensity: Through the synergistic biotransformation technology of compound enzymatic hydrolysis and yeast fermentation, not only are the umami amino acids and flavor nucleotides in seafood raw materials fully released, but more importantly, through yeast metabolism, a rich variety of key aroma substances such as pyrazines and sulfides with extremely low thresholds are synthesized, making the flavor layers more complex and full, and the aroma intensity far exceeds that of products made by traditional thermal reaction processes.

[0025] 2. Excellent odor control and oxidation stability: Innovatively, red phaf yeast fermentation extract and ginger oleoresin are added during the flavor locking stage. Together with the polyunsaturated fatty acids in cod liver oil, they form a powerful natural antioxidant system that can effectively inhibit lipid oxidation during processing and storage, significantly reduce the generation of unpleasant odor substances such as aldehydes, and ensure the purity and persistence of the product's flavor.

[0026] 3. Excellent Product Application Performance: Through an optimized compound wall material system (maltodextrin, sodium octenyl succinate starch, and gum arabic) and homogenization emulsification process, efficient co-encapsulation of oil-soluble and water-soluble flavor components is achieved, forming a loose microporous particle structure. This gives the product excellent cold water solubility (≤5 seconds) and dispersibility, overcoming the problems of clumping and slow dissolution of traditional seafood powders, making it particularly suitable for the production of modern snack foods and instant soup mixes.

[0027] 4. Enhanced Nutrition and Value: The entire process utilizes biotechnology, avoiding harsh heat processing and preserving the natural nutrients and umami substances of the raw materials to the greatest extent possible. The retention rate of umami amino acids and nucleotides exceeds 90%. At the same time, it achieves high-value utilization of various seafood by-products, resulting in high added value for the products. Attached Figure Description

[0028] Figure 1 For the comparison of the relative contents of key flavor compounds in Test Example 1;

[0029] Figure 2 For the comparison of the relative content of odor substances in Test Example 1;

[0030] Figure 3 For comparison of the average reconstitution time (s) in Test Example 2;

[0031] Figure 4 For comparison of the clumping rate (%) in Test Example 2;

[0032] Figure 5 To compare the angles of repose (degrees) in test example 2;

[0033] Figure 6 For the comparison of the 14-day flavor retention rate (%) of puffed foods in Test Example 3;

[0034] Figure 7 To compare the rancidity scores of puffed foods in Test Example 3 after 14 days. Detailed Implementation

[0035] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] Table 1: Raw Material List

[0037]

[0038]

[0039] Example 1

[0040] A seafood flavoring powder with synergistic flavor enhancement of complex protease and yeast is prepared by the following method:

[0041] (1) Raw material pretreatment: Mix the following raw materials evenly: 15 parts Antarctic krill powder, 8 parts dried oyster powder, 10 parts dried scallop powder, 5 parts bonito powder, 12 parts shrimp shell powder, 3 parts shiitake mushroom powder, 8 parts kelp powder, 2 parts fish collagen peptide powder with an average molecular weight of less than 2000 Daltons, 5 parts glycine, 1 part L-alanine, 10 parts glucose, and 80 parts water.

[0042] (2) Compound enzymatic hydrolysis: Add compound protease to the mixture, control the final addition amount of papain to 30000U / L and the final addition amount of bromelain to 60000U / L, and enzymatically hydrolyze for 3 hours at 45℃ and pH 6.0, and then heat to 95℃ for sterilization for 10 minutes.

[0043] (3) Microbial fermentation: The enzymatic hydrolysate was cooled to 28°C, inoculated with 3% (v / v) Pichia pastoris suspension, and 0.01% disodium 5'-ribonucleotide (I+G) was added simultaneously. Fermentation was carried out for 60 hours at 25°C and a ventilation rate of 0.8 vvm.

[0044] (4) Flavor blending and homogenization emulsification: Add 3% of the total weight of seaweed powder to the fermentation liquid and extract it at a constant temperature of 75°C for 60 minutes; then add 5% cod liver oil, 0.5% red phaf yeast fermentation extract and 0.5% ginger oleoresin, and emulsify at high speed at 70°C and 8000 rpm for 5 minutes.

[0045] (5) Solid-liquid separation: Filter the emulsion through a 100-mesh sieve and collect the filtrate;

[0046] (6) Microencapsulation and drying: The filtrate and wall material were mixed at a weight ratio of 1:3. The wall material was prepared into a 40% concentration solution by mixing maltodextrin, sodium octenyl succinate starch, and gum arabic in a ratio of 6:1.5:1 and then dissolved in hot water at 60°C. Spray drying was performed using a centrifugal atomizer (atomizing disc speed 18000 rpm) with an inlet air temperature of 175°C and an outlet air temperature of 80°C to obtain seafood-flavored powder.

[0047] Example 2

[0048] A seafood flavoring powder with synergistic flavor enhancement of complex protease and yeast is prepared by the following method:

[0049] (1) Raw material pretreatment: Mix the following raw materials evenly: 20 parts Antarctic krill powder, 12 parts dried oyster powder, 7 parts dried scallop powder, 8 parts bonito powder, 10 parts shrimp shell powder, 5 parts shiitake mushroom powder, 5 parts kelp powder, 3 parts fish collagen peptide powder with an average molecular weight of less than 2000 Daltons, 3 parts glycine, 2 parts L-alanine, 7 parts glucose, and 100 parts water.

[0050] (2) Compound enzymatic hydrolysis: Add compound protease to the mixture, control the final addition amount of papain to 40000U / L and the final addition amount of bromelain to 80000U / L, and enzymatically hydrolyze for 2 hours at 50℃ and pH 6.5, and then heat to 95℃ for sterilization for 10 minutes.

[0051] (3) Microbial fermentation: Cool the enzymatic hydrolysate to 30°C, inoculate with 3% Pichia pastoris suspension, and simultaneously add 0.02% I+G. Ferment for 48 hours at 30°C and a ventilation rate of 1.0 vvm.

[0052] (4) Flavor blending and homogenization emulsification: Add 4% of the total weight of seaweed powder to the fermentation liquid and extract it by stirring at 80°C for 45 minutes; then add 7% tuna liver oil, 1% red phaf yeast fermentation extract and 1% ginger oleoresin, and emulsify by high-speed shearing at 75°C and 10000rpm for 4 minutes.

[0053] (5) Solid-liquid separation: Filter the emulsion through a 150-mesh sieve and collect the filtrate;

[0054] (6) Microencapsulation and drying: The filtrate and wall material were mixed at a weight ratio of 1:4. The wall material was prepared into a 45% concentration solution by mixing maltodextrin, sodium octenyl succinate starch, and gum arabic in a ratio of 7:2:1 and then dissolved in hot water at 65°C. Spray drying was performed using a centrifugal atomizer (atomizing disc speed 20000 rpm) with an inlet air temperature of 180°C and an outlet air temperature of 85°C to obtain seafood-flavored powder.

[0055] Example 3

[0056] A seafood flavoring powder with synergistic flavor enhancement of complex protease and yeast is prepared by the following method:

[0057] (1) Raw material pretreatment: Mix the following raw materials evenly: 25 parts Antarctic krill powder, 15 parts dried oyster powder, 5 parts dried scallop powder, 10 parts bonito powder, 8 parts shrimp shell powder, 8 parts shiitake mushroom powder, 3 parts kelp powder, 4 parts fish collagen peptide powder with an average molecular weight of less than 2000 Daltons, 2 parts glycine, 3 parts L-alanine, 5 parts glucose, and 120 parts water.

[0058] (2) Compound enzymatic hydrolysis: Add compound protease to the mixture, control the final addition amount of papain to 50000U / L and the final addition amount of bromelain to 100000U / L, and enzymatically hydrolyze for 1.5 hours at 55℃ and pH 7.0, and then heat to 95℃ for sterilization for 10 minutes.

[0059] (3) Microbial fermentation: Cool the enzymatic hydrolysate to 32°C, inoculate with 3% Pichia pastoris suspension, and simultaneously add 0.03% I+G. Ferment for 36 hours at 32°C and a ventilation rate of 1.2 vvm.

[0060] (4) Flavor blending and homogenization emulsification: Add 5% of the total weight of seaweed powder to the fermentation liquid and extract it by stirring at 85℃ for 30 minutes; then add 10% cod liver oil, 3% red phaf yeast fermentation extract and 1.5% ginger oleoresin, and emulsify by high-speed shearing at 80℃ and 12000rpm for 3 minutes.

[0061] (5) Solid-liquid separation: Filter the emulsion through a 200-mesh sieve and collect the filtrate;

[0062] (6) Microencapsulation and drying: The filtrate and wall material were mixed at a weight ratio of 1:5. The wall material was prepared into a 50% concentration solution by mixing maltodextrin, sodium octenyl succinate starch, and gum arabic in a ratio of 8:2.5:1 and then dissolved in hot water at 70°C. Spray drying was performed using a centrifugal atomizer (atomizing disc speed 22000 rpm) with an inlet air temperature of 185°C and an outlet air temperature of 90°C to obtain seafood-flavored powder.

[0063] Example 4

[0064] This embodiment is basically the same as Embodiment 2, except that:

[0065] The ratio of raw materials in step (1) is adjusted to: 18 parts Antarctic krill powder and 10 parts dried oyster powder.

[0066] In step (3), when inoculating Pichia pastoris, 1.0% (v / v) of Lactobacillus plantarum (Lp90) suspension was co-inoculated for mixed fermentation.

[0067] In step (4), before emulsification of the oil phase, 0.3% D-xylose and 0.2% yeast extract (Angel) of the total weight of the fermentation broth were added to the system to enhance the Maillard reaction flavor.

[0068] Example 5

[0069] This embodiment is basically the same as Embodiment 2, except that:

[0070] In step (4), the amount of cod liver oil added is adjusted to 8%, and the amount of red phlox yeast fermentation extract added is adjusted to 1.5%.

[0071] In step (6), the spray drying uses a pressure atomizing nozzle with an atomization pressure of 20 MPa, while other drying parameters remain unchanged.

[0072] Comparative Example 1

[0073] A seafood-flavored powder, the preparation method of which is basically the same as that in Example 2, the difference being:

[0074] The complex enzymatic hydrolysis process in step (2) is omitted. After the raw materials are pretreated and mixed evenly, they are directly heated to 95°C for sterilization for 10 minutes, and then subsequent fermentation and other steps are carried out.

[0075] Comparative Example 2

[0076] A seafood-flavored powder, the preparation method of which is basically the same as that in Example 2, the difference being:

[0077] The microbial fermentation process in step (3) is omitted. After the enzymatic hydrolysate is sterilized and cooled, it is not inoculated with Pichia pastoris and I+G, and the flavor blending and emulsification in step (4) are carried out directly.

[0078] Comparative Example 3

[0079] A seafood-flavored powder, the preparation method of which is basically the same as that in Example 2, the difference being:

[0080] In step (1), fish collagen peptide powder, glycine and L-alanine are not added.

[0081] Comparative Example 4

[0082] A seafood-flavored powder, the preparation method of which is basically the same as that in Example 2, the difference being:

[0083] In step (4), no red phaf yeast fermentation extract or ginger oleoresin is added.

[0084] Comparative Example 5

[0085] A seafood-flavored powder, the preparation method of which is basically the same as that in Example 2, the difference being:

[0086] In the raw material pretreatment of step (1), Antarctic krill powder, dried oyster powder, bonito powder and shrimp shell powder are omitted, and the total weight of these raw materials is replaced by the same weight of one raw material - frozen cod meat paste (cod powder made by ordinary drying and pulverization).

[0087] Test Example 1

[0088] Quantitative analysis of key flavor compounds and assessment of off-flavor control

[0089] Objective: To verify the effects of combined enzymatic hydrolysis-yeast fermentation synergistic technology and specific raw material combinations on increasing the content of key flavor substances (pyrazines, sulfides) and reducing unpleasant odors (aldehydes).

[0090] method:

[0091] Sample preparation: Accurately weigh 1.0 g of each of the seafood flavor powder prepared in Example 2 and Comparative Examples 1-5, dissolve them in 100 mL of ultrapure water, and vortex for 1 minute to fully dissolve and disperse them.

[0092] Headspace solid-phase microextraction (HS-SPME): Take 8 mL of the above solution into a 20 mL headspace vial, add internal standard (2-methyl-3-heptanone, final concentration 50 μg / L), equilibrate at 60 °C for 15 minutes, and then adsorb using a 65 μm DVB / PDMS extraction head for 40 minutes.

[0093] Gas chromatography-mass spectrometry (GC-MS) analysis:

[0094] Chromatographic column: DB-WAX (60m × 0.25mm × 0.25μm)

[0095] Programmed temperature rise: 40℃ for 3 minutes, then increase to 230℃ at 5℃ / min and hold for 10 minutes.

[0096] Inlet: 250℃, splitless mode.

[0097] Detector: MSD, electron impact (EI) ion source, 70 eV, scan range m / z 35-350.

[0098] Quantitative analysis: Semi-quantitative analysis was performed using the internal standard method, focusing on detecting the peak areas of pyrazine compounds (e.g., 2,5-dimethylpyrazine, 2,3,5-trimethylpyrazine), sulfur-containing compounds (e.g., dimethyl trisulfide, methanethiol), and undesirable aldehydes (hexanal, nonanal), and calculating their relative contents (based on the internal standard).

[0099] Results: See Table 2 and Figure 1 and Figure 2 .

[0100] Table 2: Comparison of relative contents of key flavor compounds and off-flavor compounds

[0101]

[0102] Note: ×10 5 This represents the signal amplification factor.

[0103] Conclusions: Example 2 showed the highest total amount of key flavor compounds and the lowest content of undesirable aldehydes. Comparative Example 1 (without enzymatic hydrolysis) lacked flavor precursors, and Comparative Example 2 (without fermentation) lacked the complex flavors produced by microbial metabolism. Comparative Example 3 (lacking amino acids / peptides) confirmed that small peptides and amino acids are important precursors for the Maillard reaction. Comparative Example 4 (lacking antioxidants) indicated that Pharrellis extract and ginger oleoresin are crucial for inhibiting lipid oxidation and controlling aldehyde off-flavors. Comparative Example 5 (single seafood source) demonstrated that a combination of multiple seafood ingredients provided a richer and more harmonious flavor precursor matrix. The synergistic effects of the present invention are significant.

[0104] Test Example 2

[0105] Quantitative testing of cold water solubility, dispersibility and flowability

[0106] Objective: To evaluate the product's dissolution rate, dispersibility (anti-caking ability), and physical flowability, and indirectly verify the effectiveness of the microencapsulation process.

[0107] method:

[0108] Reconstitution time determination: Add 100 mL of 25℃ distilled water to a 250 mL beaker, and set the magnetic stirrer to a constant low speed (200 rpm). Accurately weigh 1.00 g of sample and pour it into the beaker from a height of 1 cm above the liquid surface in one go. Start timing at the same time and record the time (s) required for the powder to completely disappear and the solution to become a homogeneous clear liquid. Perform the test 5 times for each sample and take the average value.

[0109] Dispersibility (agglomeration rate) determination: Accurately weigh 10.00 g of sample and place it in a 1000 mL graduated glass cylinder. A magnetic stir bar is attached to the bottom of the cylinder. Quickly pour the sample into the cylinder from the top through a standard funnel (10 mm inner diameter neck), while simultaneously starting the magnetic stir bar (set to 300 rpm). After pouring, let it stand for 30 seconds, observe and record the weight (g) of any clumps larger than 2 mm that failed to disperse at the bottom of the cylinder, and calculate the agglomeration rate (clump weight / 10 g * 100%).

[0110] Angle of repose determination: The angle of repose of the powder is measured using the fixed funnel method to assess its flowability. The funnel is fixed at a certain height above a horizontal table, with the outlet aligned with the table surface. The powder is allowed to flow freely from the funnel, forming a conical accumulation on the table. The height (h) and base radius (r) of the cone are measured, and the angle of repose (θ = arctan(h / r)) is calculated. A smaller angle indicates better flowability.

[0111] Results: See Table 3 and Figure 3-5 .

[0112] Table 3: Quantitative Test Results of Reconstitution, Dispersibility and Flowability

[0113]

[0114]

[0115] in conclusion:

[0116] The product of Example 2 of this invention exhibits excellent cold water resolubility and dispersibility, with the shortest resolubility time (<4s), extremely low agglomeration rate (~2%), and a small angle of repose, resulting in good flowability. These three macroscopic indicators collectively confirm that its particles possess ideal surface properties and internal structure, making them easily wetted and penetrated by water.

[0117] Comparative Example 4 (lacking antioxidant ingredients) had the highest agglomeration rate and the worst flowability. This is presumably because oils lacking antioxidant protection are more likely to migrate to the particle surface and oxidize during spray drying, leading to increased particle stickiness, adhesion between particles, and severe deterioration of solubility.

[0118] Comparative Example 3 (lacking amino acid / peptide raw materials) also performed poorly, indicating that the presence of small molecules such as fish collagen peptide powder and glycine may help form a looser particle structure or improve emulsion stability, thus facilitating drying and dissolution.

[0119] Other comparative examples, lacking key process steps or raw materials, showed significantly worse performance than Example 2 in all aspects.

[0120] This test example, through three precisely measurable physical indicators—reconstitution time, agglomeration rate, and angle of repose—strongly demonstrates that the present invention, through optimized formulation and process (especially homogenization emulsification and wall material compounding), successfully prepared an instant seafood-flavored powder with excellent application performance.

[0121] Test Example 3

[0122] Flavor thermal stability and persistence testing (applied to puffed foods)

[0123] Objective: To simulate real-world applications and verify the persistence of flavor retention and release during high-temperature processing and storage of flavor powder.

[0124] method:

[0125] Sample preparation: The flavor powders of Example 2, Comparative Example 4 (without antioxidant) and Comparative Example 5 (single seafood source) were mixed evenly with blank corn puffed germ at an addition amount of 2%.

[0126] High-temperature baking: The mixed dough is baked at 180℃ for 5 minutes to simulate the processing of puffed food.

[0127] Accelerated oxidation test: Place the baked sample in a 60℃ constant temperature oven for accelerated oxidation test.

[0128] Sensory evaluation: Samples were evaluated by 10 trained sensory evaluators at 0, 7, and 14 days. An overall flavor intensity (0 = none, 10 = very strong) and undesirable rancidity (0 = none, 10 = very strong) were evaluated using a 0-10 scale. Changes in flavor signals were simultaneously captured using an electronic nose (Heracles Neo).

[0129] Results: See Table 4 and Figure 6 , Figure 7 .

[0130] Table 4: Changes in flavor intensity and off-flavor score of puffed foods over time

[0131] Group 0-day flavor intensity Flavor intensity after 14 days Flavor retention 14 days later, Habaiwei rating Example 2 8.5 7.8 91.8％ 1.5 Comparative Example 4 8.3 5.1 61.4％ 6.8 Comparative Example 5 7.0 4.9 70.0％ 4.2

[0132] Conclusion: After 14 days of accelerated oxidation, Sample 2 exhibited the highest flavor retention rate with almost no rancidity. This is attributed to the synergistic antioxidant effect of Pharbitis rubra extract and ginger oleoresin, which effectively protected the flavor compounds. Comparative Sample 4, lacking these components, showed severe flavor degradation. Comparative Sample 5, with its relatively simple and unharmonious flavor, exhibited poor initial intensity and persistence. This demonstrates that the product of this invention can significantly improve the flavor stability and shelf life of the final product.

[0133] Test Example 4

[0134] Umami retention rate determination

[0135] Objective: To quantitatively analyze the retention of umami amino acids (glutamic acid, aspartic acid) and flavor nucleotides (IMP+GMP) after processing, and to verify the protective effect of the entire process chain on umami substances.

[0136] method:

[0137] Sample pretreatment: Accurately weigh 0.5g of each sample, dilute to 50mL with 0.1mol / L hydrochloric acid solution, extract by sonication for 30min, and filter through a 0.22μm filter membrane.

[0138] Amino acid analysis: The contents of glutamic acid (Glu) and aspartic acid (Asp) were determined by an automated amino acid analyzer or by HPLC (pre-column derivatization with o-phthalaldehyde / mercaptoethanol).

[0139] Nucleotide analysis: The contents of 5'-inosinic acid (IMP) and 5'-guanosine monophosphate (GMP) were determined by HPLC-UV method.

[0140] Chromatographic column: C18 column (4.6×250mm, 5μm)

[0141] Mobile phase: 0.05 mol / L KH2PO4 buffer (pH 4.5): methanol = 95:5

[0142] Detection wavelength: 254nm

[0143] Calculation: Based on the content in the filtrate of step (5) without spray drying (100%), calculate the retention rate of umami substances in each finished powder.

[0144] Results: See Table 5.

[0145] Table 5: Retention rate of key umami substances (%)

[0146] Group Glutamic acid (Glu) Aspartic acid (Asp) IMP+GMP Average retention rate Example 2 92.5 93.1 94.8 93.5 Comparative Example 1 85.2 84.7 88.3 86.1 Comparative Example 4 88.9 87.5 76.4* 84.3

[0147] Note: In Comparative Example 4, due to the lack of antioxidants, nucleotides may have experienced more severe degradation during spray drying.

[0148] Conclusion: Example 2 of this invention exhibits the highest overall retention rate of umami substances (>90%). The complete enzymatic hydrolysis-fermentation process (partially omitted in Comparative Example 1) produces a richer variety of umami precursors, while the optimized wall material and antioxidant system (partially omitted in Comparative Example 4) effectively protect these heat-sensitive and easily oxidized umami substances during subsequent drying and storage stages, maximizing their retention in the final product.

[0149] Test Example 5

[0150] Sensory evaluation of its application in pre-made soup bases

[0151] Objective: To evaluate the flavor performance and consumer acceptance of seafood flavor powder in practical application scenarios (pre-made soup base).

[0152] method:

[0153] Soup base preparation: Dissolve 1.0g of flavor powder in 100mL of boiling water and stir to make the soup base. Grouping: Example 2, Comparative Example 5 (single seafood source), and commercially available seafood powder.

[0154] Sensory evaluation: Sixty consumer evaluators were invited to conduct the tasting. A 9-point preference rating system (1 = strongly disliked, 9 = strongly liked) was used to evaluate the aroma intensity, flavor richness, umami, body, off-flavors, and overall acceptability of the soup base.

[0155] Results: See Table 6.

[0156] Table 6: Average Sensory Evaluation Scores of Pre-made Soup Bases (n=60)

[0157]

[0158]

[0159] Conclusion: The product of Example 2 significantly outperformed Comparative Example 5 and commercially available products in all sensory indicators (P<0.05). It possessed a rich and harmonious aroma, prominent umami flavor, and a full-bodied, mellow taste with virtually no unpleasant off-flavors. In contrast, Comparative Example 5 (single seafood source) exhibited a monotonous flavor, poor complexity, and insufficient body. This fully demonstrates that the flavor enhancement and synergistic effect achieved by this invention through the compounding of multiple seafood ingredients, enzymatic hydrolysis-fermentation synergy, and flavor locking technology can significantly improve the sensory quality and consumer appeal of the final product.

[0160] Test Example 6

[0161] Consistency verification of core indicators of products in different implementations

[0162] Objective: To verify that the products prepared by the preparation method of the present invention under different raw material ratios and process parameters (i.e., Examples 1-5) have stable and significantly better core indicators such as key flavor, solubility and umami retention rate than the comparative examples, thus proving the consistency and reproducibility of the technical solution.

[0163] method:

[0164] Sample preparation: The seafood flavored powder samples prepared in Examples 1, 2, 3, 4 and 5 were taken, and Comparative Example 2 (no fermentation), Comparative Example 4 (lacking antioxidants) and commercially available ordinary seafood powder were selected as controls.

[0165] Determination of key flavor compounds: Referring to the GC-MS method in Test Example 1, the relative contents of the total amount of pyrazine compounds and the total amount of hexanal + nonanal in all samples were determined.

[0166] Cold water reconstitution time determination: The reconstitution time of all samples was determined according to the method in Test Example 2.

[0167] Umami retention rate determination: Refer to the HPLC method in Test Example 4 to determine the retention rate of glutamic acid (Glu) in all samples (represented by the overall retention rate).

[0168] Results: See Table 7.

[0169] Table 7: Comparison of core performance indicators of products in different embodiments

[0170]

[0171]

[0172] in conclusion:

[0173] Test data show that the core performance indicators of the five embodiment products prepared according to the different parameter ranges provided by the present invention are highly consistent and significantly better than those of the comparative examples and commercially available products.

[0174] 1. Flavor properties: The total content of key flavor compounds in all the products of the examples remained consistently at a high level (13.9-15.1 × 10⁻⁶). 5 The content of undesirable aldehydes remained at extremely low levels (1.1-1.4×10⁻⁶). 5 This demonstrates that within the range of raw material ratios and process parameters required by this invention, excellent aroma-enhancing and flavor-suppressing effects can be stably achieved.

[0175] 2. Application performance: The cold water resolution time of all products in the examples is extremely short (≤5 seconds), proving that products with excellent rapid dissolution performance can be successfully prepared with different wall material ratios and drying parameters.

[0176] 3. Quality retention: The umami substance retention rate of all products in the examples is higher than 90%, indicating that the entire process system can effectively protect raw materials with different ratios.

[0177] 4. Stability verification: Some indicators of Example 4 (adding probiotics and Maillard precursor) and Example 5 (adjusting the oil phase ratio) are even better than those of the core Example 2, which proves the optimizability and scalability of the technical solution of the present invention, rather than just the best solution at a single point.

[0178] Test Case Summary:

[0179] 1) The above series of quantitative tests prove that the key performance indicators of the product of the present invention (Example 2) are superior to those of the comparative example and commercially available products.

[0180] 2) Flavor indicators: GC-MS analysis showed that the total content of key flavor compounds (pyrazines and sulfides) was increased compared with traditional thermally reacted flavor powders; while the content of hexanal and nonanal, which represent lipid oxidation off-flavors, was reduced, resulting in a significant flavor advantage.

[0181] 3) Application indicators: This product has a resolubility time of ≤5 seconds in cold water at 25℃, with no clumping, an angle of repose of only 36.5°, and excellent fluidity, which is far superior to commercially available products with a resolubility time of ≥30 seconds.

[0182] 4) Quality stability index: In the accelerated oxidation test, after 14 days of application to puffed food, its flavor retention rate was as high as 91.8%, and there was almost no rancid taste. Compared with the control group 4 which lacked antioxidant components (61.4% retention rate, obvious rancid taste), its stability was outstanding.

[0183] 5) Nutritional component retention indicators: HPLC analysis showed that the retention rates of core umami substances (glutamic acid, aspartic acid, IMP+GMP) in this product were all >90%, confirming the effective protection of heat-sensitive flavor components throughout the entire process chain. In sensory evaluation, its overall preference score was 8.6 points, significantly higher than the control group, and its comprehensive performance was quantitatively verified.

[0184] The above are merely a few preferred embodiments of the present invention, described in a relatively specific and detailed manner, but they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention.

Claims

1. A method for preparing a seafood flavor powder with synergistic flavor enhancement of complex protease and yeast, characterized in that, Includes the following steps: (1) Raw material pretreatment: Mix the following raw materials evenly by weight: 15-25 parts Antarctic krill powder, 8-15 parts dried oyster powder, 5-10 parts dried scallop powder, 5-10 parts bonito powder, 8-12 parts shrimp shell powder, 3-8 parts shiitake mushroom powder, 3-8 parts kelp powder, 2-4 parts fish collagen peptide powder, 2-5 parts glycine, 1-3 parts L-alanine, 5-10 parts glucose, and 80-120 parts water; (2) Compound enzymatic hydrolysis: Add compound protease to the mixture in step (1), control the final addition amount of papain to be 30,000-50,000 U / L of the mixture, and the final addition amount of bromelain to be 60,000-100,000 U / L of the mixture, and enzymatically hydrolyze for 1.5-3 hours at 45-55℃ and pH 6.0-7.0, and then heat to 95℃ for sterilization for 10 minutes; (3) Microbial fermentation: Cool the enzymatic hydrolysate from step (2) to 28-32℃, inoculate with 2%-4% Pichia pastoris suspension, and simultaneously add 0.01%-0.03% disodium 5'-ribonucleotide, and ferment at 25-32℃ for 36-60 hours with ventilation. (4) Flavor blending and homogenization emulsification: Add 3%-5% of the total weight of laver powder to the fermentation liquid in step (3), and extract at a constant temperature of 75-85℃ for 30-60 minutes; then add 5%-10% cod liver oil, 0.5%-2% red phaf yeast fermentation extract and 0.5%-1.5% ginger oleoresin to the system, and perform high-speed shear homogenization emulsification at a temperature of 70-80℃, a rotation speed of 8000-12000rpm, and a time of 3-5 minutes to form a stable emulsion; (5) Solid-liquid separation: The emulsion obtained in step (4) is filtered through a 100-200 mesh sieve and the filtrate is collected; (6) Microencapsulation and drying: The filtrate obtained in step (5) is mixed and homogenized with the wall material at a weight ratio of 1:(3-5). The wall material is composed of maltodextrin, sodium octenyl succinate starch, and gum arabic at a weight ratio of (6-8):(1.5-2.5):

1. Then, spray drying is performed. The inlet air temperature of the spray drying is 175-185℃ and the outlet air temperature is 80-90℃ to obtain a microporous seafood flavor powder.

2. The preparation method according to claim 1, characterized in that, In step (1), the average molecular weight of the fish collagen peptide powder is less than 2000 Daltons.

3. The preparation method according to claim 1, characterized in that, In step (3), the fermentation conditions are: fermentation at 30°C with ventilation for 48 hours, with a ventilation rate of 0.8-1.2 vvm.

4. The preparation method according to claim 1, characterized in that, In step (4), the astaxanthin content of the red phaf yeast fermentation extract is not less than 1%.

5. The preparation method according to claim 1, characterized in that, In step (6), the wall material is prepared by dissolving maltodextrin, sodium octenyl succinate starch, and gum arabic in hot water at 60-70°C in proportion to prepare a solution with a concentration of 40%-50%, and stirring until completely dissolved and homogeneous before use.

6. The preparation method according to claim 1, characterized in that, In step (6), the spray drying uses a centrifugal atomizer with an atomizing disc rotation speed of 18,000-22,000 rpm.

7. A seafood-flavored powder, characterized in that, It is prepared by the preparation method according to any one of claims 1-6.

8. The seafood flavored powder according to claim 7, characterized in that, The seafood flavor powder has a resolubility time of less than or equal to 5 seconds in cold water at 25°C, and the retention rate of umami amino acids and flavor nucleotides is greater than 90%.

9. The application of the seafood flavor powder as described in claim 7 in the preparation of seasoning powder for snack puffed foods or pre-made soup base compound seasoning packets.

Images