Soy sauce koji making technology based on spice formula

Abstract

The invention relates to a soy sauce koji-making process based on a spice formula, which comprises the following steps: cooking soaked soybeans until the digestibility is greater than or equal to 70%, and cooling to 30-35 DEG C; stir-frying and crushing the wheat; crushing spices to 20-50 meshes, wherein the use amount of the spices is 0.2-2% of the total weight of the raw materials; soybean and wheat are mixed according to the weight ratio of 7: 3; inoculating aspergillus oryzae accounting for 0.3-0.8% of the total weight of the raw materials; the starter propagation temperature is 28-32 DEG C; wherein the spice formula comprises the following components in percentage by weight: 15-23% of liquorice, 14-28% of Chinese yam, 2-8% of anise, 1-5% of citronella, 2-7% of rosemary, 2-10% of cinnamon, 1-3% of clove, 2-8% of pepper, 5-15% of pericarpium citri reticulatae, 7-15% of radix angelicae, 3-12% of mint, 5-10% of astragalus membranaceus and 2-10% of rhizoma smilacis glabrae. According to the technical scheme, through synergistic metabolism regulation and control of the spice combination, the activity content of finished koji enzyme and the abundance of an enzyme system are remarkably increased, and on the basis of not changing the dosage of the raw materials, the amino acid generation of the soy sauce is improved, and the glutamic acid proportion is promoted to be increased. And triple flavor synergy of'umami excitation-fishy smell neutralization-aftertaste extension 'is formed when the fish is contacted with the raw fish / sashimi body.

Description

Technical Field

[0001] The invention belongs to the field of soy sauce brewing and relates to a soy sauce koji making process based on a spice formula. Background Art

[0002] As a traditional Chinese condiment, soy sauce can be traced back to the Western Zhou Dynasty. In the early days, soybeans, wheat and other raw materials were used to form a liquid seasoning system rich in amino acids, sugars and esters through microbial fermentation. With the spread of food culture, soy sauce technology has derived a sub-category of sashimi soy sauce, whose core function is to enhance the umami level of fresh ingredients (such as sashimi and sushi) while suppressing fishy smell. Although the brewing process of sashimi soy sauce is derived from traditional soy sauce, it needs to meet higher requirements for flavor coordination, including fresh and sweet balance, aroma complexity and soft taste.

[0003] According to a report released by the International Seasoning Society (IFS), the compound annual growth rate of the global sashimi soy sauce market reached 7% between 2020 and 2025. It is expected that by 2030, the global sashimi soy sauce market will grow from US$2.8 billion in 2019 to US$4.2 billion, of which the market size of some Asian countries is expected to account for nearly two-thirds of the total market share ("2025-2030 Sashimi Soy Sauce Project Investment Value Analysis Report"). With the increase in consumer demand for natural and complex flavors, the scientific application of spices has become an important direction for the improvement of soy sauce. In traditional processes, spices are mostly added through later blending (such as kelp, bonito flakes, miso, etc.). Although they can give soy sauce a specific flavor, there are problems such as aroma volatilization loss, high cost, and insufficient fusion with the soy sauce base. Therefore, exploring the process innovation of integrating spices in the koji making stage has become a key breakthrough in optimizing the flavor generation path of sashimi soy sauce.

[0004] In the prior art, the traditional sashimi soy sauce production uses soybeans and wheat as raw materials, which are steamed and inoculated with Aspergillus oryzae to make koji, and the protease and amylase decompose proteins and polysaccharides to generate flavor precursors such as glutamic acid and glucose. The fermentation stage relies on the synergistic effect of lactic acid bacteria and yeast to form alcohols, organic acids and ester compounds, which constitute the basic umami taste and aroma of soy sauce. However, the flavor generation of this process is highly dependent on the natural metabolism of raw materials and strains, resulting in a single flavor of the finished product, which is difficult to meet the complex umami requirements required for sashimi seasoning.

[0005] With the refinement and classification of soy sauce functions, the prior art often uses a secondary blending method to improve the flavor. For example, a Chinese patent with publication number CN108433086A discloses a sashimi sushi soy sauce, which contains 20-80 parts of brewed soy sauce, 2-30 parts of water, 0.01-10 parts of alcohol, 0.001-2 parts of flavor enhancer, 0.001-0.01 parts of sweetener, 0.001-0.1 parts of colorant, and 0.001-0.01 parts of acidity regulator. Another example: some products use traditional fermentation technology and add fish extracts to obtain a rich and natural taste.

[0006] Since foods such as sashimi and raw fish lack the process of high-temperature cooking to blend with seasonings, the synergy and counteraction of the taste of seasonings and raw food will occur on the tip of the tongue and be captured by diners at the first moment. Therefore, the synergistic effect between the flavor substances in the soy sauce that directly acts on the food, the diffusion path of the flavor substances and the timing of the flavor substances have become important factors affecting diners' perception of the salty, fragrant and sweet taste of food.

[0007] The existing soy sauce preparation technology for raw fish and sashimi has the following problems:

[0008] 1. Fragmentation of flavor generation pathways: The decomposition of raw materials in the koji-making stage is separated from the flavor generation process of spices, resulting in the failure of the two to form a synergistic metabolic network, which limits the natural formation of complex flavors. For example: the specificity conflict of enzymatic substrates and the lack of transformation pathways for secondary metabolites (such as glycyrrhetinic acid derivatives).

[0009] 2. Timing contradiction of adding spices: Although the later blending method can introduce specific flavors, its physical mixing mode makes it difficult to achieve molecular-level fusion of aroma components and soy sauce matrix, resulting in a single flavor level and insufficient durability. For example: spice flavor substances and soy sauce basic flavors (umami, sauce aroma) are only superimposed through physical mixing, rather than generating complex flavor molecules through the biochemical cascade reaction of enzymatic hydrolysis-fermentation; the "inert" addition of spices in the koji making stage results in the inability of lactic acid bacteria and yeast to use their active ingredients to build a synergistic metabolic network in subsequent fermentation.

[0010] 3. Disadvantages in process economy: Multi-stage processing (koji making → fermentation → blending) will increase energy consumption and raw material loss.

[0011] In addition, on the one hand, there are differences in understanding among those skilled in the art; on the other hand, the inventor studied a large number of documents and patents when making the present invention, but due to space limitations, not all details and contents are listed in detail. However, this does not mean that the present invention does not have the characteristics of these prior arts. On the contrary, the present invention already has all the characteristics of the prior art, and the applicant reserves the right to add relevant prior art to the background technology. Summary of the invention

[0012] Based on the above technical problems, one of the objects of the present invention is to provide a soy sauce koji making process based on a spice formula to overcome the deficiencies in the prior art.

[0013] To achieve the above object, the present invention adopts the following technical solution:

[0014] A soy sauce koji making process based on a spice formula comprises the following steps:

[0015] Soybeans and wheat are mixed and steamed until the digestibility is ≥70%, and the steamed raw materials are cooled to 30-35°C;

[0016] Mix the raw materials in the spice formula according to the proportion, and crush them into 20-50 meshes using a pulverizer, with the amount being 0.2-2% of the total weight of the raw materials;

[0017] Inoculate Aspergillus oryzae in an amount of 0.3-0.8‰ of the total weight of the raw materials;

[0018] The fermentation process takes 42 to 72 hours, during which:

[0019] The spice formula comprises 15-23% liquorice, 14-28% yam, 2-8% star anise, 1-5% citronella, 2-7% rosemary, 2-10% cinnamon, 1-3% clove, 2-8% prickly ash, 5-15% tangerine peel, 7-15% angelica, 3-12% mint, 5-10% astragalus and 2-10% smilax glabra.

[0020] According to a preferred embodiment, the pulverizer pulverizes to 20-50 meshes.

[0021] According to a preferred embodiment, soybeans and wheat are mixed in a mass ratio of 7:3.

[0022] According to a preferred embodiment, the temperature condition of the fermentation process is 28-32° C., and the humidity condition is 90-100%.

[0023] According to a preferred embodiment, the spice formula is 15% by weight of licorice, 14% by yam, 2% by star anise, 5% by lemongrass, 7% by rosemary, 10% by cinnamon, 1% by clove, 2% by pepper, 5% by tangerine peel, 15% by angelica, 12% by mint, 10% by astragalus, and 2% by wild smilax.

[0024] According to a preferred embodiment, the spice formula is 23% by weight of licorice, 16% by yam, 4% by star anise, 2% by lemongrass, 4% by rosemary, 4% by cinnamon, 3% by cloves, 4% by pepper, 7% by tangerine peel, 13% by angelica, 7% by mint, 7% by astragalus, and 6% by wild smilax.

[0025] According to a preferred embodiment, the spice formula is 20% by weight of licorice, 18% of yam, 8% of star anise, 1% of lemongrass, 2% of rosemary, 2% of cinnamon, 1% of cloves, 8% of prickly ash, 15% of tangerine peel, 7% of angelica, 3% of mint, 5% of astragalus, and 10% of smilax glabra.

[0026] According to a preferred embodiment, the weight proportion of spices in the raw materials is 0.2-2%. Preferably, the weight proportion of spices in the raw materials is 0.2%. The weight proportion of spices in the raw materials is 1%. The weight proportion of spices in the raw materials is 2%.

[0027] According to a preferred embodiment, the inoculation amount is 0.3-0.5‰ of the total weight of the raw material. Preferably, the inoculation amount is 0.3‰ of the total weight of the raw material. The inoculation amount is 0.4‰ of the total weight of the raw material. The inoculation amount is 0.5‰ of the total weight of the raw material.

[0028] One of the purposes of the present invention is to provide a soy sauce, which is prepared based on the soy sauce koji making process.

[0029] According to a preferred embodiment, the soy sauce is used for preparing sashimi or sashimi.

[0030] One of the objects of the present invention is also to provide a method for preparing soy sauce, comprising the following steps:

[0031] Mix soybeans and wheat, cook until the digestibility is ≥70%, and cool the cooked raw materials to 30-35°C;

[0032] Mix the raw materials in the spice formula according to the proportion, and crush them into 20-50 meshes using a pulverizer, with the amount being 0.2-2% of the total weight of the raw materials;

[0033] Inoculate Aspergillus oryzae in an amount of 0.3-0.8‰ of the total weight of the raw materials;

[0034] The fermentation process takes 42 to 72 hours;

[0035] Koji material: brine = 1:1.3~1.7 (W / V), fermentation time is more than one year,

[0036] The spice formula comprises 15-23% liquorice, 14-28% yam, 2-8% star anise, 1-5% citronella, 2-7% rosemary, 2-10% cinnamon, 1-3% clove, 2-8% prickly ash, 5-15% tangerine peel, 7-15% angelica, 3-12% mint, 5-10% astragalus and 2-10% smilax glabra.

[0037] According to a preferred embodiment, the ratio of koji material to brine is 1:1.7 (W / V).

[0038] According to a preferred embodiment, the fermentation time is 1 year.

[0039] The technical solution of the present invention achieves the improvement of enzyme activity, enrichment of flavor precursors and directional release of characteristic aroma in the koji making stage through the coordinated metabolic regulation of the spice combination, and finally enables the soy sauce to form a triple flavor synergy of "fresh taste stimulation-fishy smell neutralization-aftertaste extension" when it comes into contact with sashimi / sashimi. The comprehensive scores of Examples 1 and 3 (90 points and 84 points) verify its technical advantages in the raw food cooking scene and provide a scientific basis for the development of high-end condiments. DETAILED DESCRIPTION

[0040] In the description of the present invention, the terms are used for descriptive purposes only and are not to be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. In the description of the present invention, "plurality" means two or more, unless otherwise clearly and specifically defined.

[0041] It should be noted that in the technical field described in the present invention, the ratio of koji material to brine and fermentation time as a common means of adjusting product quality and characteristics has been widely recognized and practiced. Specifically, "koji material: brine = 1: 1.3 ~ 1.7 (W / V), fermentation time exceeds 1 year" is a conventional technical means for preparing soy sauce. Any change within this ratio range (i.e. 1: 1.3 to 1: 1.7 W / V), combined with the condition that the fermentation time exceeds 8 months, can achieve the expected effects and goals of the present invention. Therefore, the scope of protection required by the present invention application should include but is not limited to all the above ratios and their related fermentation time parameters, and is not limited to the specific values ​​mentioned in the embodiments. In addition, the Aspergillus oryzae used in the following embodiments is the soy sauce koji essence (koji species) of Jining Yuyuan Biotechnology Co., Ltd. Aspergillus oryzae is a common fermentation intermediate in soy sauce koji, and its various types or categories of Aspergillus oryzae are widely used as fermentation agents in the process of brewing soy sauce. Therefore, the present invention should not be limited to a specific type of Aspergillus oryzae.

[0042] Example 1

[0043] This embodiment provides a soy sauce koji making process based on a spice recipe. The soy sauce koji making process comprises the following steps:

[0044] S1: Soybean and wheat are mixed in a mass ratio of 7:3, washed and soaked for 12 hours, and steamed until the digestibility is ≥70%, and the steamed raw materials are cooled to 30-35°C.

[0045] S2: Mix the raw materials in the spice formula according to the proportion, and crush them into 20 meshes using a pulverizer. The amount used is 0.2% of the total weight of the raw materials.

[0046] S3: Inoculate Aspergillus oryzae at an inoculation amount of 0.3‰ of the total weight of the raw materials, and mix well with 40 times the weight of wheat flour.

[0047] S4: During the koji making process, the spice mixture and Aspergillus oryzae are evenly spread on the surface of the koji.

[0048] S5: Place the koji material in a koji making room, control the temperature at 28-32°C, the humidity at 90-100%, and the koji making time for 42 hours.

[0049] The spice formula is composed of the following raw materials in parts by weight:

[0050] Licorice 15%, yam 14%, star anise 2%, lemongrass 5%, rosemary 7%, cinnamon 10%, clove 1%, pepper 2%, tangerine peel 5%, angelica 15%, mint 12%, astragalus 10%, and wild yam 2%.

[0051] Example 2

[0052] The soy sauce koji making process includes the following steps:

[0053] S1: Soybean and wheat are mixed in a mass ratio of 7:3, washed and soaked for 12 hours, and steamed until the digestibility is ≥70%, and the steamed raw materials are cooled to 30-35°C.

[0054] S2: Mix the raw materials in the spice formula according to the proportion, and crush them into 50 meshes using a pulverizer. The amount used is 2% of the total weight of the raw materials.

[0055] S3: Inoculate Aspergillus oryzae at an inoculation amount of 0.5‰ of the total weight of the raw materials, and mix well with 40 times the weight of wheat flour.

[0056] S4: During the koji making process, the spice mixture and Aspergillus oryzae are evenly spread on the surface of the koji.

[0057] S5: Place the koji material in a koji making room, control the temperature at 28-32°C, the humidity at 90-100%, and the koji making time for 72 hours.

[0058] The spice formula is composed of the following raw materials in parts by weight:

[0059] Licorice 23%, yam 16%, star anise 4%, lemongrass 2%, rosemary 4%, cinnamon 4%, clove 3%, pepper 4%, tangerine peel 7%, angelica 13%, mint 7%, astragalus 7%, and wild yam 6%.

[0060] Example 3

[0061] The soy sauce koji making process includes the following steps:

[0062] S1: Soybean and wheat are mixed in a mass ratio of 7:3, washed and soaked for 12 hours, and steamed until the digestibility is ≥70%, and the steamed raw materials are cooled to 30-35°C.

[0063] S2: Mix the raw materials in the spice formula according to the proportion, and crush them into 50 meshes using a pulverizer. The amount used is 1.0% of the total weight of the raw materials.

[0064] S3: Inoculate Aspergillus oryzae at an inoculation amount of 0.4‰ of the total weight of the raw materials, and mix well with 40 times the weight of wheat flour.

[0065] S4: During the koji making process, the spice mixture and Aspergillus oryzae are evenly spread on the surface of the koji.

[0066] S5: Place the koji material in a koji making room, control the temperature at 28-32°C, the humidity at 90-100%, and the koji making time for 64 hours.

[0067] The spice formula is composed of the following raw materials in parts by weight:

[0068] Licorice 20%, yam 18%, star anise 8%, lemongrass 1%, rosemary 2%, cinnamon 2%, clove 1%, pepper 8%, tangerine peel 15%, angelica 7%, mint 3%, astragalus 5%, and wild yam 10%.

[0069] Example 4

[0070] The spice formula is composed of the following raw materials in parts by weight:

[0071] Licorice 23%, yam 10%, star anise 4%, lemongrass 2%, rosemary 3%, cinnamon 4%, clove 2%, pepper 4%, tangerine peel 9%, angelica 14%, mint 10%, astragalus 8%, and wild yam 7%.

[0072] Koji making process:

[0073] S1: Mix soybeans and wheat in a mass ratio of 7:3, wash and soak for 12 hours, steam until the digestibility is ≥70%, and cool the steamed raw materials to 30-35°C.

[0074] S2: Mix the raw materials in the spice formula according to the proportion, and crush them into 50 meshes using a pulverizer. The amount used is 4% of the total weight of the raw materials.

[0075] S3: Inoculate Aspergillus oryzae at an inoculation amount of 0.8‰ of the total weight of the raw materials, and mix well with 40 times the weight of wheat flour.

[0076] S4: During the koji making process, the spice mixture and Aspergillus oryzae are evenly spread on the surface of the koji.

[0077] S5: Place the koji material in the koji making room, control the temperature at 28-32°C, the humidity at 90-100%, and the koji making time for 44 hours.

[0078] Example 5

[0079] The spice formula is composed of the following raw materials in parts by weight:

[0080] Licorice 10%, yam 5%, star anise 4%, lemongrass 15%, rosemary 8%, cinnamon 4%, clove 12%, pepper 14%, tangerine peel 1%, angelica 4%, mint 15%, astragalus 1%, and wild yam 7%.

[0081] Koji making process:

[0082] S1: Mix soybeans and wheat in a mass ratio of 7:3, wash and soak for 12 hours, steam until the digestibility is ≥70%, and cool the steamed raw materials to 30-35°C.

[0083] S2: Mix the raw materials in the spice formula according to the proportion, and crush them into 50 meshes using a pulverizer. The amount used is 1% of the total weight of the raw materials.

[0084] S3: Inoculate Aspergillus oryzae at an inoculation amount of 0.5‰ of the total weight of the raw materials, and mix well with 40 times the weight of wheat flour.

[0085] S4: During the koji making process, the spice mixture and Aspergillus oryzae are evenly spread on the surface of the koji.

[0086] S5: Place the koji material in the koji making room, control the temperature at 28-32°C, the humidity at 90-100%, and the koji making time for 42 hours.

[0087] Example 6

[0088] The koji making process of soy sauce includes:

[0089] S1: Mix soybeans and wheat in a mass ratio of 7:3, wash and soak for 12 hours, steam until the digestibility is ≥70%, and cool the steamed raw materials to 30-35°C.

[0090] S2: Inoculate Aspergillus oryzae at an inoculation amount of 0.3‰ of the total weight of the raw materials, and mix well with 40 times the weight of wheat flour.

[0091] S3: During the koji making process, the spice mixture and Aspergillus oryzae are evenly spread on the surface of the koji.

[0092] S4: Place the koji material in the koji making room, control the temperature at 28-32°C, the humidity at 90-100%, and the koji making time for 42 hours.

[0093] Example 7

[0094] Soy sauce was prepared based on the koji prepared by the koji making process of Examples 1 to 6. After the koji making was completed, the koji material was placed in a jacketed fermentation tank, and the raw materials prepared in Examples 1 to 6 were added to the tank at a ratio of 1.7 (mass volume ratio; W / V). The fermentation time was 1 year, and the prepared soy sauce was tested.

[0095] 1. Physical and chemical index testing

[0096] (I) Fragrance component detection

[0097] Sample processing method: take 1 mL of soy sauce sample, add 5 mL of ethanol, and perform ultrasonic extraction for 10 minutes; centrifuge (4000 rpm, 10 minutes), and take the supernatant; filter the supernatant with a 0.22 μm organic filter membrane to remove particulate matter; add an appropriate amount of internal standard n-pentanol to a final concentration of 10 μg / mL.

[0098] a. GC-MS conditions

[0099] Chromatographic column: DB-WAX (30m×0.25mm×0.25μm).

[0100] Carrier gas: helium, flow rate 1.0 mL / min.

[0101] Inlet temperature: 250℃.

[0102] Injection method: splitless injection, injection volume 1 μL.

[0103] Temperature program: Initial temperature: 40°C, hold for 2 minutes. Heating rate: 10°C / min.

[0104] Final temperature: 240°C, hold for 10 minutes.

[0105] b. Mass spectrometry (MS) conditions

[0106] Ion source: electron impact ion source (EI), 70 eV.

[0107] Ion source temperature: 230°C. Transfer line temperature: 250°C.

[0108] Scan mode: Full Scan, mass range: 50-200 m / z.

[0109] Selected ion monitoring (SIM) - aroma components: ethyl acetate: m / z 61, 70, 88; isopropenyl acetate: m / z 43, 69, 86; 2-methylbutanal: m / z 58, 71, 86; limonene: m / z 68, 93, 136; eugenol: m / z 164, 149, 103; cinnamaldehyde: m / z 131, 132, 103.

[0110] c. Preparation of standard curve

[0111] Preparation of standard solutions: prepare standard solutions of isopropenyl acetate, ethyl hexanoate, 2-methylbutyraldehyde, limonene, eugenol, and cinnamaldehyde (with concentration gradients of 0.1 μg / mL, 1 μg / mL, 10 μg / mL, 50 μg / mL, and 100 μg / mL, respectively). Add the same concentration of internal standard to each standard solution.

[0112] Injection and analysis: Injection and analysis were performed according to the above GC-MS conditions and the peak areas were recorded.

[0113] Draw a standard curve: Draw a standard curve with the peak area ratio of the target compound to the internal standard as the ordinate and the concentration as the abscissa.

[0114] Sample analysis injection: The treated sample solution is injected and analyzed according to GC-MS conditions.

[0115] Qualitative analysis: Confirm the target compound by comparing the retention time and mass spectrum with the standard.

[0116] Quantitative analysis: Calculate isopropenyl acetate, ethyl hexanoate, 2-methylbutyraldehyde, limonene, eugenol and cinnamaldehyde in the sample according to the standard curve.

[0117] d. Data processing

[0118] Qualitative analysis: The target compound was confirmed by comparing the mass spectrum with the NIST spectral library.

[0119] Quantitative analysis: The content of the target compound was calculated using the internal standard method.

[0120] Calculation formula:

[0121] C sample = (A sample × C internal standard × R standard curve) / A internal standard

[0122] Among them, C sample represents the concentration of the target compound in the sample; A sample represents the peak area of ​​the target compound; C internal standard represents the concentration of the internal standard; A internal standard represents the peak area of ​​the internal standard; R standard curve represents the slope of the standard curve.

[0123] (ii) The activity of protease in the protease is determined according to the commercial industry standard "SB / T 10317-1999 Protease Activity Assay".

[0124] (III) The spore count test is based on LS / T 6132-2018 Inspection of grain and oil - Detection of stored grain fungi - Spore counting method.

[0125] (IV) Amino Acid Nitrogen The detection of amino acid nitrogen is based on GB 5009.235-2016 National Food Safety Standard Determination of Amino Acid Nitrogen in Food.

[0126] (V) Determination of glutamate content

[0127] 1. Sample Pretreatment

[0128] Accurately pipette 1 mL of soy sauce sample and dilute it to 10 mL with ultrapure water (adjust the dilution factor according to the glutamic acid content). Pass the diluted sample through a 0.22 μm microporous filter membrane for later use.

[0129] 2. Preparation of standard solution

[0130] Glutamic acid standard stock solution:

[0131] Accurately weigh the glutamate standard, dissolve it in ultrapure water, and prepare a 1 mg / mL stock solution. Dilute the stock solution into a series of standard solutions with different concentrations (such as 10, 20, 50, 100, 200 μg / mL).

[0132] 3. Chromatographic conditions

[0133] Chromatographic column: C18 reverse phase column (250 mm × 4.6 mm, 5 μm).

[0134] Mobile phase: Phase A is 0.05 M potassium dihydrogen phosphate buffer (pH 2.5-3.0); Phase B is methanol.

[0135] Gradient elution program (example): 0-5 minutes: 95% A, 5% B; 5-15 minutes: linear change to 70% A, 30% B; 15-20 minutes: return to initial conditions (95% A, 5% B); flow rate: 1.0 mL / min; detection wavelength: 210 nm (if using UV detector); column temperature: 30°C. Injection volume: 10-20 μL.

[0136] 4. Sample determination

[0137] Inject the standard series solutions in sequence, record the peak area, and draw a standard curve. Inject the sample to be tested, record the peak area of ​​glutamic acid. Calculate the glutamic acid content in the sample based on the standard curve.

[0138] 5. Data Analysis

[0139] Standard curve: Draw a standard curve with the concentration of glutamate standard as the horizontal axis and the peak area as the vertical axis to obtain the linear regression equation.

[0140] Sample calculation: Substitute the sample peak area into the standard curve equation to calculate the glutamate concentration.

[0141] Glutamate content = (C*V*D) / m,

[0142] C: glutamate concentration in the sample (μg / mL); V: sample volume (mL); D: dilution factor; m: sample mass (g)

[0143] (VI) Sensory evaluation

[0144] In the sensory evaluation, the body shape is evaluated by pouring soy sauce into a small white ceramic dish and shaking it slightly. The aroma is evaluated by smelling. The color is evaluated by visual observation and combined with the body shape test results. The taste is evaluated by taking the same weight of raw fish meat and dipping it in an appropriate amount of soy sauce.

[0145] 2. Experimental Results

[0146] (I) Physical and chemical index test results

[0147] Table 1

[0148]

[0149] (II) Sensory evaluation

[0150] Table 2

[0151]

[0152]

[0153] From the experimental data in Table 1, it can be seen that there are significant differences in the enzyme activity, spore count, amino acid nitrogen content and key flavor substances content of different examples. Among them, the release efficiency of flavor substances in different examples is significantly different:

[0154] (1) Enzyme activity in koji: The enzyme activities of Examples 1 and 3 were relatively high, 1380 U / g and 1480 U / g, respectively, while the enzyme activity of Example 2 was relatively high, 1180 U / g. The enzyme activities of Examples 4, 5, and 6 were significantly reduced, 870 U / g, 850 U / g, and 940 U / g, respectively. The results showed that Examples 1 and 3 had higher enzymatic hydrolysis efficiency in the koji-making stage.

[0155] (2) Spore count: The spore count of Example 3 was the highest, which was 6.9×108 CFU / g, followed by the spore count of Example 1, which was 5.8×108 CFU / g, and then the spore count of Example 2, which was 3.6×108 CFU / g. The spore counts of Examples 4, 5, and 6 were significantly lower, which were 1.3×108 CFU / g, 1.4×108 CFU / g, and 2×108 CFU / g, respectively, indicating that the microbial activity of Examples 1 and 3 was stronger.

[0156] (3) Amino acid nitrogen content: Example 3 had the highest amino acid nitrogen content (1.03 g / 100 mL), followed by Examples 1 and 2 (1.01 g / 100 mL and 0.98 g / 100 mL), while Examples 4, 5, and 6 had lower amino acid nitrogen contents (0.9 g / 100 mL, 0.91 g / 100 mL, and 0.94 g / 100 mL), indicating that the protein decomposition of Examples 1 and 3 was more complete.

[0157] (4) Ethyl acetate: Example 3 had the highest content (589 μg / L), followed by Example 1 (534 μg / L), followed by Example 2 (434 μg / L) and Example 5 (219 μg / L). Examples 4 and 6 had extremely low contents (32 μg / L and 40.21 μg / L).

[0158] (5) Isopropenyl acetate: Example 3 has the highest content (1097 μg / L), followed by Example 1 (1059 μg / L), followed by Example 2 (965 μg / L) and Example 5 (770 μg / L). Examples 4 and 6 have relatively low contents (206 μg / L and 239.28 μg / L).

[0159] (6) 2-Methylbutanal: The content in Example 1 is 329 μg / L, the content in Example 2 is 299 μg / L, the content in Example 3 is 374 μg / L, the content in Example 5 is 174 μg / L, the content in Example 4 is 32 μg / L, and the content in Example 6 is 59.89 μg / L.

[0160] (7) Limonene: the content in Example 1 is 57 μg / L; the content in Example 2 is 513 μg / L; the content in Example 3 is 239 μg / L; the content in Example 4 is 813 μg / L; the content in Example 5 is 239 μg / L; the content in Example 6 is 0 μg / L.

[0161] (8) Eugenol: the content in Example 1 is 91.2 μg / L; the content in Example 2 is 785 μg / L; the content in Example 3 is 356 μg / L; the content in Example 4 is 1536 μg / L; the content in Example 5 is 356 μg / L; the content in Example 6 is 0 μg / L.

[0162] (9) Cinnamaldehyde: the content in Example 1 is 13.8 μg / L; the content in Example 2 is 117 μg / L; the content in Example 3 is 101 μg / L; the content in Example 4 is 216 μg / L; the content in Example 5 is 101 μg / L; the content in Example 6 is 0 μg / L.

[0163] (10) Glutamic acid content: the content of Example 1 is 21.2 mg / mL; the content of Example 2 is 20.6 mg / mL; the content of Example 3 is 21.8 mg / mL; the content of Example 4 is 15.98 mg / mL; the content of Example 5 is 16.48 mg / mL; the content of Example 6 is 11.4 mg / mL.

[0164] From the experimental data in Table 2, it can be seen that there are significant differences in sensory quality and fragrance emphasis in different embodiments:

[0165] (1) The color score is as follows: Example 1 is 28 points (reddish brown or light reddish brown, bright color, glossy); Example 2 is 29 points (reddish brown or light reddish brown, bright color, glossy); Example 3 is 25 points (reddish brown or light reddish brown, bright color, glossy); Example 4 is 25 points (reddish brown or light reddish brown, dull); Example 5 is 26 points (reddish brown or light reddish brown, dull); Example 6 is 25 points (reddish brown or light reddish brown, dull).

[0166] (2) The aroma score was 29 points for Example 1 (strong sauce aroma, ester aroma and floral and fruity aroma); 29 points for Example 2 (strong sauce aroma, ester aroma and floral and fruity aroma); 26 points for Example 3 (strong sauce aroma, ester aroma and floral and fruity aroma); 15 points for Example 4 (with sauce aroma and ester aroma); 18 points for Example 5 (with sauce aroma and ester aroma); and 25 points for Example 6 (relatively strong sauce aroma and ester aroma).

[0167] (3) The taste score is as follows: Example 1 is 25 points (delicious, mellow, fresh, salty and sweet); Example 2 is 23 points (delicious, mellow, fresh, salty and sweet); Example 3 is 26 points (delicious, mellow, fresh, salty and sweet); Example 4 is 20 points (delicious, salty and sweet); Example 5 is 19 points (delicious, salty and sweet); Example 6 is 22 points (delicious, salty and sweet).

[0168] (4) The physical score was 8 points for Example 1 (clear, free of impurities); 8 points for Example 2 (clear, free of impurities); 7 points for Example 3 (clear, free of impurities); 8 points for Example 4 (clear, free of impurities); 8 points for Example 5 (clear, free of impurities); and 9 points for Example 6 (clear, free of impurities).

[0169] (5) The total score was 90 points for Example 1; 89 points for Example 2; 84 points for Example 3; 68 points for Example 4; 71 points for Example 5; and 81 points for Example 6.

[0170] In all examples, the enzyme activity and spore count of the finished product are at a good level in the industry, the amino acid nitrogen compound meets the national standard for premium soy sauce, and the overall flavor is harmonious, which is at an excellent level among soy sauce products.

[0171] Among these flavor substances, ethyl acetate brings the unique ester aroma of soy sauce, isopropenyl acetate brings floral and fruity aroma, and 2-methylbutanal brings sweet and floral and fruity aroma.

[0172] The present invention proposes a synergistic fermentation process integrating licorice and other spices in the koji-making stage, which has remarkable effects on the efficient generation of flavor precursor substances, the generation of umami substances and the directional release of characteristic flavor substances.

[0173] Specifically, the amino acid nitrogen content (1.03 g / 100 mL) and glutamic acid content (21.8 mg / mL) of Example 3 were significantly higher than those of the other groups, wherein the components of the main umami amino acids and glutamic acid were significantly increased, and the ratio of glutamic acid to amino acid nitrogen in Example 3 was also significantly higher than those in Comparative Example 4 and Comparative Example 5, indicating that the soy sauce prepared with the koji of Example 3 can increase the proportion of glutamic acid production, and the higher umami substances can significantly enhance the taste when combined with raw food, and have an inhibitory effect on fishy smell.

[0174] Eugenol and cinnamaldehyde produce spicy and sweet aromas, respectively. The soy sauce (Examples 1 to 3) of the present invention is rich in cinnamaldehyde and has a controlled limonene content, so that it can combine with amino acids during fermentation to produce fruity esters without exceeding the taste threshold to produce bitterness and irritation, thereby avoiding taste conflict between soy sauce and wasabi (or mustard).

[0175] Combined with the sensory taste evaluation, it can be known that the spice combination co-fermented with the koji provided by the present invention can not only neutralize the raw fishy taste of raw food, but also combine with fish fat through esters (such as isopropenyl acetate) to form a floral and fruity aftertaste, and can also form a synergistic and complementary taste through a small amount of limonene, eugenol and rich cinnamaldehyde, forming a cool top note, a warm middle note and a floral and sweet base note in the mouth, which complements the oily feeling of, for example, salmon belly.

[0176] In terms of visual sense, since the sashimi soy sauce is directly poured on the food by the diners, the color and shape are also the key to the evaluation. Thanks to the influence of the spices on the oxidation inhibition and colloidal stability of the soy sauce of the present invention, the soy sauce is reddish brown and shiny, and has a certain wall-hanging property.

[0177] In addition, in the process of exploring the flavor of sashimi soy sauce, the present invention unexpectedly discovered that a spice combination including licorice, yam, star anise, lemongrass, rosemary, cinnamon, cloves, pepper, tangerine peel, angelica dahurica, mint, astragalus, and smilax glabra has a positive promoting effect on the enzyme activity and the number of spores in the soy sauce koji making process at a specific ratio (see blank group Example 6 and experimental group Examples 1 to 3), and after changing the spice ratio, the enzyme activity and the number of spores in the soy sauce koji making process are inhibited (see control group Examples 4 and 5).

[0178] It should be noted that the above specific embodiments are exemplary, and those skilled in the art can come up with various solutions inspired by the disclosure of the present invention, and these solutions also belong to the disclosure scope of the present invention and fall within the protection scope of the present invention. Those skilled in the art should understand that the description of the present invention is illustrative and does not constitute a limitation of the claims. The protection scope of the present invention is defined by the claims and their equivalents.

Claims

1. A soy sauce koji making process based on a spice formula, characterized in that: The following steps are involved: Soybeans and wheat are mixed and steamed until the digestibility is ≥70%, and the steamed raw materials are cooled to 30-35°C; Mix the raw materials in the spice formula according to the proportion, and crush them into 20-50 meshes using a pulverizer, with the amount being 0.2-2% of the total weight of the raw materials; Inoculate Aspergillus oryzae at an inoculation rate of 0.3-0.8‰ of the total weight of the raw materials; The fermentation process takes 42 to 72 hours, during which: The spice formula is 15-23% by weight of liquorice, 14-28% of yam, 2-8% of star anise, 1-5% of citronella, 2-7% of rosemary, 2-10% of cinnamon, 1-3% of clove, 2-8% of prickly ash, 5-15% of tangerine peel, 7-15% of angelica dahurica, 3-12% of mint, 5-10% of astragalus and 2-10% of smilax glabra.

2. The soy sauce koji making process according to claim 2, characterized in that: The soybeans and wheat are mixed in a mass ratio of 7:

3.

3. The soy sauce koji making process according to claim 2 or 3, characterized in that: The temperature condition of the koji making process is 28-32° C., and the humidity condition is 90-100%.

4. The soy sauce koji making process according to any one of claims 1 to 3, characterized in that: The spice formula is 15% by weight of licorice, 14% of yam, 2% of star anise, 5% of lemongrass, 7% of rosemary, 10% of cinnamon, 1% of clove, 2% of prickly ash, 5% of tangerine peel, 15% of angelica dahurica, 12% of mint, 10% of astragalus, and 2% of smilax glabra.

5. The soy sauce koji making process according to any one of claims 2 to 4, characterized in that: The weight proportion of the spices in the raw materials is 0.2-2%.

6. The soy sauce koji making process according to any one of claims 2 to 5, characterized in that: The inoculation amount is 0.3 to 0.8‰ of the total weight of the raw materials.

7. A method for preparing soy sauce, comprising the following steps: Mix soybeans and wheat, cook until the digestibility is ≥70%, and cool the cooked raw materials to 30-35°C; Mix the raw materials in the spice formula according to the proportion, and crush them into 20-50 meshes using a pulverizer, with the amount being 0.2-2% of the total weight of the raw materials; Inoculate Aspergillus oryzae at an inoculation rate of 0.3-0.8‰ of the total weight of the raw materials; The koji making process takes 42 to 72 hours; Koji material: brine = 1:1.3~1.7 (W / V), fermentation time is more than one year, The spice formula comprises, by weight, 15-23% liquorice, 14-28% yam, 2-8% star anise, 1-5% citronella, 2-7% rosemary, 2-10% cinnamon, 1-3% clove, 2-8% prickly ash, 5-15% tangerine peel, 7-15% angelica, 3-12% mint, 5-10% astragalus and 2-10% smilax glabra.

8. The method for preparing soy sauce according to claim 7, characterized in that: The koji material: brine = 1:1.7 (W / V).

9. A soy sauce, characterized in that The soy sauce is prepared based on the soy sauce koji making process described in claims 1 to 7 or the soy sauce preparation method described in claim 8.

10. The soy sauce according to claim 9, characterized in that The soy sauce is used for preparing sashimi or sashimi.

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