Fermentation agent for improving yield of amino acid nitrogen and application of fermentation agent

Abstract

The invention belongs to the technical field of fermentation engineering, and particularly relates to a leavening agent for increasing the yield of amino acid nitrogen and application of the leavening agent. The leavening agent comprises independently packaged phytic acid and a microbial agent, wherein the volume ratio of the microbial agent to the phytic acid is (296-498): (2-4); the concentration of microorganisms in the microbial agent is 1 * 10 < 8 >-3 * 10 < 8 > / mL; the microorganisms comprise any one or more of actinomucor elegans, monascus and bacillus subtilis. The phytic acid is combined with the microbial agent to prepare the leavening agent, and the phytic acid can be used as a fermentation accelerant of the microbial agent to play a synergistic effect with the microbial agent, so that the content of amino acid nitrogen in the flavor base material is increased, and the quality and the nutritional value of the flavor base material are improved.

Description

Technical Field

[0001] The invention belongs to the technical field of fermentation engineering, and particularly relates to a starter culture for increasing the yield of amino acid nitrogen and application thereof. Background Art

[0002] Plant protein hydrolysates contain numerous active peptides and free amino acids. Free amino acids and peptides can enhance freshness, improve taste, and mask off-flavors. They also serve as flavor precursors and participate in complex chemical reactions. Furthermore, plant protein can be degraded into amino acids and peptides. Compared to protein, plant protein hydrolysates are more easily digested and absorbed by the human body, making them an excellent source of protein for nutritional and physiological functions. Therefore, they are particularly suitable for use in food processing for the elderly, infants, and patients. In recent years, protein hydrolysates have found applications in a variety of food products, including soy sauce, snack food seasonings, dressings, chicken bouillon seasonings, and savory flavorings. Demand for hydrolyzed plant protein in the food processing industry is increasing. Therefore, protein hydrolysates with higher free amino acid content and richer flavor hold great potential for application.

[0003] Currently, methods for hydrolyzing vegetable protein to obtain flavorful amino acid nitrogen include acid hydrolysis, enzymatic hydrolysis, and microbial fermentation. However, the hydrolyzate obtained with acid hydrolysis has a low amino acid nitrogen content, and post-processing with acid hydrolysis is complex and environmentally polluting. Enzymatic hydrolysis is costly, making it unsuitable for large-scale production. Microbial fermentation, on the other hand, offers mild reaction conditions, safe and reliable products, and reduces costs. However, the amino acid nitrogen content of hydrolyzed vegetable protein produced by fermentation methods in the prior art is generally low, at around 1%. Therefore, how to increase the amino acid nitrogen content of hydrolyzed vegetable protein using microbial fermentation has become an urgent problem in the field. Summary of the Invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a fermentation agent for increasing the yield of amino acid nitrogen and its application. The fermentation product has a high amino acid nitrogen content, low preparation cost, mild product and no pollution. The present invention specifically includes the following technical solutions:

[0005] A fermentation agent comprising phytic acid and a microbial agent;

[0006] The volume ratio of the microbial agent to phytic acid is 296-498:2-4;

[0007] The concentration of microorganisms in the microbial agent is 1×10 8 ~3×10 8 / mL;

[0008] The microorganisms include any one or more of Actinobacillus elegans, Monascus purpureus and Bacillus subtilis.

[0009] Preferably, the independent concentration of phytic acid is 60 to 80 wt.%.

[0010] Preferably, when the microorganisms are Actinobacillus elegans and Monascus purpureus, the concentration ratio of Actinobacillus elegans to Monascus purpureus in the microbial inoculant is 1-3:1-3;

[0011] When the microorganisms are Actinobacillus elegans and Bacillus subtilis, the concentration ratio of Actinobacillus elegans to Bacillus subtilis in the microbial inoculant is 1-3:1-3;

[0012] When the microorganisms are Actinobacillus elegans, Monascus purpureus and Bacillus subtilis, the concentration ratio of Actinobacillus elegans, Monascus purpureus and Bacillus subtilis in the microbial inoculant is 1-3:1-3:1-3.

[0013] The present invention also provides the use of the above-mentioned fermentation agent in increasing the amino acid nitrogen content in the fermentation product.

[0014] The present invention also provides a method for increasing the amino acid nitrogen content in a fermentation product, comprising the following steps:

[0015] The starter and fermentation material as described above are mixed, and the resulting mixture is fermented to obtain a fermentation product.

[0016] Preferably, the fermentation temperature is 27-33°C, the fermentation speed is 150-250 r / min, and the fermentation time is 60-84 h;

[0017] The volume ratio of the starter to the fermentation material is 3-5:100.

[0018] Preferably, the fermentation material includes fermentation medium;

[0019] The fermentation medium comprises the following components in the following concentrations: 2-4% soy protein isolate, 1-2% glucose, 0.1-0.3% magnesium sulfate, 0.1-0.3% potassium dihydrogen phosphate, 0.05-0.15% yeast extract, 0.3-0.5% sodium nitrate, 0.01-0.03% zinc sulfate, 0.05-0.15 g / L calcium chloride, 0.03-0.07 g / L MnSO4, and 0.005-0.015 g / L FeSO4.

[0020] The percentages of the soy protein isolate, glucose, magnesium sulfate, potassium dihydrogen phosphate, yeast extract, sodium nitrate and zinc sulfate are all percentages by volume.

[0021] The present invention also provides application of the above method in preparing a flavor base.

[0022] Preferably, the fermentation product obtained by the method is subjected to solid-liquid separation, and the liquid portion is collected to obtain a flavor base.

[0023] Preferably, the solid-liquid separation method includes centrifugation, the centrifugal speed is 10000-14000 r / min, and the centrifugal time is 10 minutes.

[0024] The beneficial effects of the present invention are:

[0025] The present invention provides a starter, which comprises separately packaged phytic acid and a microbial agent, wherein the volume ratio of the microbial agent to the phytic acid is 296-498:2-4; the concentration of the microorganisms in the microbial agent is 1×10 8 ~3×10 8 The microorganisms include any one or more of Actinomucor elegans, Monascus purpureus, and Bacillus subtilis. The raw materials of the starter provided by the present invention can synergize with each other, and the raw materials are inexpensive and readily available. Fermenting the fermentation material using the starter of the present invention can increase the amino acid nitrogen content of the fermentation product, thereby enhancing the quality and nutritional value of the flavor base. Furthermore, the method provided by the present invention is low-cost, produces mild products, and is pollution-free. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments are briefly introduced below.

[0027] Figure 1 Schematic diagram of the results of amino acid nitrogen content in Experimental Example 1, Example 12 and Comparative Example 1;

[0028] Figure 2 Schematic diagram of the results of amino acid nitrogen content in Experimental Example 1, Example 13 and Comparative Example 2;

[0029] Figure 3 Schematic diagram of the results of amino acid nitrogen content in Experimental Example 1, Example 14 and Comparative Example 3;

[0030] Figure 4 Schematic diagram of the results of amino acid nitrogen content in Experimental Example 1, Example 15 and Comparative Example 4;

[0031] Figure 5 Schematic diagram of the results of amino acid nitrogen content in Experimental Example 1, Example 16 and Comparative Example 5;

[0032] Figure 6 Schematic diagram of the results of amino acid nitrogen content in Experimental Example 1, Example 17 and Comparative Example 6;

[0033] Figure 7Schematic diagram of the results of amino acid nitrogen content in Experimental Example 1, Example 18 and Comparative Example 7. DETAILED DESCRIPTION

[0034] The present invention provides a fermentation agent, comprising phytic acid and a microbial agent; the volume ratio of the microbial agent to the phytic acid is 296-498:2-4; the concentration of the microorganisms in the microbial agent is 1×10 8 ~3×10 8 / mL; the microorganisms include any one or more of Actinobacillus elegans, Monascus purpureus and Bacillus subtilis.

[0035] As an embodiment, the volume ratio of the microbial agent of the present invention to phytic acid is 296-498:2-4; as an optional embodiment, the volume ratio of the microbial agent of the present invention to phytic acid is any one of 296:4, 297:3, 298:2, 396:4, 397:3, 398:2, 496:4, 497:3 and 498:2. As an embodiment, the concentration of phytic acid is 60-80wt.%; as an optional embodiment, the mass concentration of phytic acid can be any one of 60wt.%, 61wt.%, 62wt.%, 63wt.%, 64wt.%, 65wt.%, 66wt.%, 67wt.%, 68wt.%, 69wt.%, 70wt.%, 71wt.%, 72wt.%, 73wt.%, 74wt.%, 75wt.%, 76wt.%, 77wt.%, 78wt.%, 79wt.% and 80wt.%. As an embodiment, the concentration of microorganisms in the microbial inoculant is 1×10 8 ~3×10 8 As an optional embodiment, the concentration of microorganisms in the microbial agent can be 1×10 8 / mL, 2×10 8 / mL and 3×10 8 In a specific embodiment, the volume ratio of the microbial agent to phytic acid is 396:4, the mass concentration of phytic acid is 70wt.%, and the concentration of microorganisms in the microbial agent is 1×10 8 pieces / mL.

[0036] As an embodiment, when the microorganisms are Actinomucor elegans and Monascus purpureus, the concentration ratio of the Actinomucor elegans to Monascus purpureus is 1-3:1-3; as an optional embodiment, when the microorganisms are Actinomucor elegans and Monascus purpureus, the concentration ratio of the Actinomucor elegans to Monascus purpureus can be any one of 1:1, 1:2, 1:3, 2:1, 2:2, 2:3, 3:1, 3:2, and 3:3. In a specific embodiment, when the microorganisms are Actinomucor elegans and Monascus purpureus, the concentration ratio of the Actinomucor elegans to Monascus purpureus is 1:1. As an embodiment, when the microorganisms are Actinobacillus elegans and Bacillus subtilis, the concentration ratio of the Actinobacillus elegans to Bacillus subtilis is 1-3:1-3; as an optional embodiment, when the microorganisms are Actinobacillus elegans and Bacillus subtilis, the concentration ratio of the Actinobacillus elegans to Bacillus subtilis can be any one of 1:1, 1:2, 1:3, 2:1, 2:2, 2:3, 3:1, 3:2, and 3:3. In a specific embodiment, when the microorganisms are Actinobacillus elegans and Bacillus subtilis, the concentration ratio of the Actinobacillus elegans to Bacillus subtilis is 1:1.

[0037] As an embodiment, when the microbial agent includes Actinobacillus elegans, Monascus and Bacillus subtilis, the concentration ratio of Actinobacillus elegans, Monascus and Bacillus subtilis is 1-3:1-3:1-3; as an optional embodiment, when the microbial agent includes Actinobacillus elegans, Monascus and Bacillus subtilis, the concentration ratio of Actinobacillus elegans, Monascus and Bacillus subtilis can be 1:1:1, 1 : Any one of 1:2, 1:2:1, 1:2:2, 1:3:1, 1:1:3, 1:2:3, 1:3:2, 1:3:3, 2:1:1, 2:1:2, 2:2:1, 2:3:1, 2:1:3, 2:2:3, 2:3:2, 2:3:3, 3:1:1, 3:1:2, 3:2:1, 3:2:2, 3:3:1, 3:1:3, 3:2:3 and 3:3:2.

[0038] The present invention also provides the use of the above-described starter to increase the content of amino acid nitrogen in a fermentation product. The product obtained by fermentation using the starter of the present invention has an amino acid nitrogen content of 0.44% to 2.18%. When the microbial inoculum in the starter includes two or more microorganisms, the content of amino acid nitrogen in the fermentation product is above 1.64%. The starter of the present invention has simple ingredients, natural and pollution-free raw materials, and has a good fermentation-promoting effect, which can improve the fermentation effect of the fermentation base.

[0039] The present invention also provides a method for increasing the amino acid nitrogen content in a fermentation product using the starter culture, which specifically comprises the following steps:

[0040] The starter and fermentation material as described above are mixed, and the resulting mixture is fermented to obtain a fermentation product.

[0041] The present invention mixes the starter agent with a fermentation material, and ferments and cultures the resulting mixture to obtain a fermentation product. In one embodiment, the starter agent includes a microbial agent and phytic acid; in an alternative embodiment, the volume ratio of the microbial agent to the phytic acid is 296-498:2-4; in an alternative embodiment, the volume ratio of the microbial agent to the phytic acid can be any one of 296:4, 297:3, 298:2, 396:4, 397:3, 398:2, 496:4, 497:3, and 498:2. In one embodiment, the volume ratio of the starter agent to the fermentation material is 3-5:100; in an alternative embodiment, the volume ratio of the starter agent to the fermentation material can be any one of 3:100, 4:100, and 5:100. In one embodiment, the fermentation material is a fermentation medium; in an alternative embodiment, the fermentation medium comprises the following components at the following concentrations: 2-4% soy protein isolate, 1-2% glucose, 0.1-0.3% magnesium sulfate, 0.1-0.3% potassium dihydrogen phosphate, 0.05-0.15% yeast extract, 0.3-0.5% sodium nitrate, 0.01-0.03% zinc sulfate, 0.05-0.15 g / L calcium chloride, 0.03-0.07 g / L MnSO4, and 0.005-0.015 g / L FeSO4; the percentages of soy protein isolate, glucose, magnesium sulfate, potassium dihydrogen phosphate, yeast extract, sodium nitrate, and zinc sulfate are all expressed in volume percentages. In one embodiment, the fermentation culture temperature is 27-33°C; in an alternative embodiment, the fermentation culture temperature can be any one of 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, and 33°C. As an embodiment, the fermentation culture speed is 150-250r / min. As an optional embodiment, the fermentation culture speed can be any one of 150r / min, 160r / min, 170r / min, 180r / min, 190r / min, 200r / min, 210r / min, 220r / min, 230r / min240r / min and 250r / min. As an embodiment, the fermentation culture time is 60-78h. As an optional embodiment, the fermentation culture time can be any one of 60h, 61h, 62h, 63h, 64h, 65h, 66h, 67h, 68h, 69h, 70h, 71h, 72h, 73h, 74h, 75h, 76h, 77h and 78h. In a specific embodiment, the present invention mixes the starter agent and the fermentation medium in a volume ratio of 4:100, the volume ratio of the microbial agent to phytic acid in the starter agent is 396:4, and the resulting mixture is fermented at 30°C and 200r / min for 72h to obtain a fermentation product.

[0042] The present invention also provides application of the above method in preparing a flavor base.

[0043] In one embodiment, the application comprises the steps of performing solid-liquid separation on the fermentation product obtained by the method, collecting the liquid portion, and obtaining a flavor base. In one embodiment, the solid-liquid separation comprises centrifugation at a speed of 10,000 to 14,000 rpm for 10 minutes. In another embodiment, the centrifugation speed may be 10,000 rpm, 11,000 rpm, 12,000 rpm, 13,000 rpm, or 14,000 rpm.

[0044] In order to further illustrate the present invention, the technical solution of the present invention is described in detail below with reference to the accompanying drawings and embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0045] Example 1 Preparation method of microbial spore suspension

[0046] In this example, a microbial spore suspension was prepared using Actinobacillus elegans, Monascus purpureus, and Bacillus subtilis. The deposit number of Actinobacillus elegans is CGMCC3.4389; the deposit number of Monascus purpureus is CGMCC3.7183; and the deposit number of Bacillus subtilis is CGMCC1.821. The specific preparation method is as follows:

[0047] 1) Strain activation

[0048] Activation of Actinobacillus elegans strain: Spread Actinobacillus elegans stored at -80°C on a PDA culture medium plate and place it in a constant temperature incubator at 30°C for activation. The activation process lasts for three generations, with each generation lasting 2 days.

[0049] Activation of Monascus: Spread the Monascus stored at -80°C on a PDA culture medium plate and place it in a constant temperature incubator at 30°C for activation. Activate for 3 generations, with each generation culturing for 3 days.

[0050] Activation of Bacillus subtilis: Bacillus subtilis stored at -80°C was spread on LB broth culture plates and placed in a constant temperature incubator at 30°C for activation. The activation process lasted for 3 generations, with each generation lasting 3 days.

[0051] 2) Preparation of spore suspension

[0052] The activated microbial strains were prepared into spore suspensions with sterile water. The concentration of the microbial spore suspensions was 1×10 8 pieces / mL.

[0053] Example 2 A fermentation medium

[0054] The volume percentages of the raw materials of the fermentation medium are as follows: 3% soy protein isolate, 1.5% glucose, 0.2% magnesium sulfate, 0.2% potassium dihydrogen phosphate, 0.1% yeast extract, 0.4% sodium nitrate, 0.02% zinc sulfate, 0.1 g / L calcium chloride, 0.05 g / L MnSO4 and 0.01 g / L FeSO4.

[0055] Example 3 A leavening agent

[0056] The fermentation agent is phytic acid and microbial agent packaged separately;

[0057] The volume ratio of the microbial agent to phytic acid is 296:4;

[0058] The microbial agent is a microbial spore suspension, and the concentration of the microbial spore suspension is 2×10 8 / mL;

[0059] The microbial spore suspension is Actinobacillus elegans spore suspension.

[0060] Example 4 A leavening agent

[0061] The fermentation agent is phytic acid and microbial agent packaged separately;

[0062] The volume ratio of the microbial agent to phytic acid is 396:4;

[0063] The microbial agent is a microbial spore suspension, and the concentration of microbial spores in the microbial agent is 1×10 8 / mL;

[0064] The microbial spore suspension is Actinobacillus elegans spore suspension.

[0065] Example 5 A leavening agent

[0066] The fermentation agent is phytic acid and microbial agent packaged separately;

[0067] The volume ratio of the microbial agent to phytic acid is 497:3;

[0068] The microbial agent is a microbial spore suspension, and the concentration of microbial spores in the microbial agent is 3×10 8 / mL;

[0069] The microbial spore suspension is Actinobacillus elegans spore suspension.

[0070] Example 6 A leavening agent

[0071] The components of the fermentation agent are the same as those in Example 4, except that the microbial spore suspension is a Monascus spore suspension.

[0072] Example 7 A leavening agent

[0073] The components of the fermentation agent are the same as those in Example 4, except that the microbial spore suspension is a Bacillus subtilis spore suspension.

[0074] Example 8 A leavening agent

[0075] The components of the fermentation agent are the same as those in Example 4, except that the microbial agent is composed of a concentration of 2×10 8 The spore suspension of Monascus purpureus and Actinomucor elegans at a volume ratio of 1:1 were mixed.

[0076] Example 9 A leavening agent

[0077] The components of the fermentation agent are the same as those in Example 4, except that the microbial agent is composed of a concentration of 2×10 8 The spore suspension of Actinobacillus elegans and the spore suspension of Bacillus subtilis were mixed at a volume ratio of 1:1.

[0078] Example 10 A leavening agent

[0079] The components of the fermentation agent are the same as those in Example 4, except that the microbial agent is composed of a concentration of 2×10 8 The Monascus spore suspension with a concentration of 100 spores / mL and the Bacillus subtilis spore suspension were mixed in a volume ratio of 1:1.

[0080] Example 11 A leavening agent

[0081] The components of the fermentation agent are the same as those in Example 4, except that the microbial agent is composed of a concentration of 3×10 8 The spore suspensions of Actinobacillus elegans, Monascus purpureus and Bacillus subtilis with the concentrations of spores / mL were mixed in a volume ratio of 1:1:1.

[0082] Example 12: A method for increasing the amino acid nitrogen content in a flavor base

[0083] This example uses the fermentation medium described in Example 2 and the fermentation agent described in Example 4 for processing, and the method steps are as follows:

[0084] To a 250 mL Erlenmeyer flask, 100 mL of fermentation medium was added, along with 3.96% by volume of a microbial agent and 0.04% by volume of phytic acid, relative to the fermentation medium, and mixed. The mixture in the Erlenmeyer flask was fermented at 30°C and 200 rpm for 72 hours to obtain a fermentation product.

[0085] Example 13: A method for increasing the amino acid nitrogen content in a flavor base

[0086] The method of this embodiment is the same as that of Example 12, except that the leavening agent described in Example 6 is used for processing.

[0087] Example 14: A method for increasing the amino acid nitrogen content in a flavor base

[0088] The method of this embodiment is the same as that of Example 12, except that the leavening agent described in Example 7 is used for processing.

[0089] Example 15 A method for increasing the amino acid nitrogen content in a flavor base

[0090] The method of this embodiment is the same as that of Example 12, except that the leavening agent described in Example 8 is used for processing.

[0091] Example 16 A method for increasing the amino acid nitrogen content in a flavor base

[0092] The method of this embodiment is the same as that of Example 12, except that the leavening agent described in Example 9 is used for processing.

[0093] Example 17 A method for increasing the amino acid nitrogen content in a flavor base

[0094] The method of this embodiment is the same as that of Example 12, except that the leavening agent described in Example 10 is used for processing.

[0095] Example 18 A method for increasing the amino acid nitrogen content in a flavor base

[0096] The method of this embodiment is the same as that of Example 12, except that the fermentation agent described in Example 11 is used for processing.

[0097] Example 19: A method for preparing a flavor base using a fermentation product

[0098] The fermentation product is separated into solid and liquid by a centrifuge, and the liquid portion is collected to obtain a flavor base.

[0099] The centrifugal speed is 12000 r / min, and the centrifugal time is 10 min.

[0100] Comparative Example 1: A method for increasing the amino acid nitrogen content in a flavor base

[0101] The method of this comparative example is the same as that of Example 12, except that phytic acid is not added.

[0102] Comparative Example 2: A method for increasing the amino acid nitrogen content in a flavor base

[0103] The method of this comparative example is the same as that of Example 13, except that phytic acid is not added.

[0104] Comparative Example 3: A method for increasing the amino acid nitrogen content in a flavor base

[0105] The method of this comparative example is the same as that of Example 14, except that phytic acid is not added.

[0106] Comparative Example 4: A method for increasing the amino acid nitrogen content in a flavor base

[0107] The method of this comparative example is the same as that of Example 15, except that phytic acid is not added.

[0108] Comparative Example 5: A method for increasing the amino acid nitrogen content in a flavor base

[0109] The method of this comparative example is the same as that of Example 16, except that phytic acid is not added.

[0110] Comparative Example 6: A method for increasing the amino acid nitrogen content in a flavor base

[0111] The method of this comparative example is the same as that of Example 17, except that phytic acid is not added.

[0112] Comparative Example 7: A method for increasing the amino acid nitrogen content in a flavor base

[0113] The method of this comparative example is the same as that of Example 18, except that phytic acid is not added.

[0114] Experimental Example 1 Effect Verification Experiment

[0115] The fermentation products obtained by the methods described in Examples 12 to 18 and Comparative Examples 1 to 7 were centrifuged at 12,000 r / min for 10 min, and the supernatants were collected to obtain test samples 1 to 7 and control test samples 1 to 7, which were used to determine the content of amino acid nitrogen as described below:

[0116] Preparation of ninhydrin dye: Weigh 0.5 g of ninhydrin, 10 g of Na2HPO4·12H2O, 6 g of KH2PO4, and 0.3 g of fructose into a conical flask and dilute to 100 ml with distilled water to obtain ninhydrin dye. The ninhydrin dye is then stored in a brown reagent bottle in the dark.

[0117] The test samples 1 to 7 and the control group test samples 1 to 7 were respectively subjected to gradient dilution. The gradient dilution was as follows: the samples were diluted 10, 100, and 1000 times in sequence. 2 mL of the test samples of different concentrations were respectively taken into colorimetric tubes, 1 mL of ninhydrin color developer was added, and the mixture was heated in a 95°C water bath for 15 minutes. After cooling for 5 minutes, 7 mL of 45% ethanol solution was added, and the mixture was mixed again and allowed to stand for 15 minutes. After the sample color was developed, the absorbance of the colored test sample at 568 nm was measured using a UV-visible spectrophotometer. The amino acid concentration was obtained by comparing with the standard curve, and then the amino acid nitrogen value was calculated. The experiment was repeated 3 times and the average value was taken.

[0118] To create a standard curve: Prepare 100 mg of glycine and dilute to 100 mL with distilled water to obtain a 1000 mg / L glycine solution. This solution is then diluted to obtain sample solutions with concentrations of 0.1, 0.5, 1, 2, 4, 6, 8, 10, 20, 30, 50, 80, and 100 mg / L. Follow the above assay steps for different glycine concentrations. After completion of the assay, create a glycine standard curve and derive the corresponding equation.

[0119] The equation of the glycine standard curve is: y = 0.01672x + 0.04922, R 2 =0.99868.

[0120] The results of the determination of the amino acid nitrogen content in the test sample 1 (the fermentation product of Example 12) and the control group test sample 1 (the fermentation product of Comparative Example 1) are as follows: Figure 1 As shown in Table 1; the results of the determination of the amino acid nitrogen content in the test sample 2 (the fermentation product of Example 13) and the control group test sample 2 (the fermentation product of Comparative Example 2) are as follows Figure 2 As shown in Table 2; the results of the determination of the amino acid nitrogen content in the test sample 3 (the fermentation product of Example 14) and the control group test sample 3 (the fermentation product of Comparative Example 3) are as follows Figure 3 As shown in Table 3; the results of the determination of the amino acid nitrogen content in the test sample 4 (fermentation product of Example 15) and the control group test sample 4 (fermentation product of Comparative Example 4) are as follows Figure 4 As shown in Table 4; the results of the determination of the amino acid nitrogen content in the test sample 5 (the fermentation product of Example 16) and the control group test sample 5 (the fermentation product of Comparative Example 5) are as follows Figure 5 As shown in Table 5; the results of the determination of the amino acid nitrogen content in the test sample 6 (the fermentation product of Example 17) and the control group test sample 6 (the fermentation product of Comparative Example 6) are as follows Figure 6 As shown in Table 6; the results of the determination of the amino acid nitrogen content in the test sample 7 (the fermentation product of Example 18) and the control group test sample 7 (the fermentation product of Comparative Example 7) are as follows Figure 7and as shown in Table 7.

[0121] The times shown in Tables 1 to 7 are the fermentation times of different microorganisms.

[0122] Table 1 Effect of different fermentation methods on amino acid nitrogen production (g / 100ml)

[0123]

[0124] Table 2 Effect of different fermentation methods on amino acid nitrogen production (g / 100ml)

[0125]

[0126] Table 3 Effect of different fermentation methods on amino acid nitrogen production (g / 100ml)

[0127]

[0128]

[0129] Table 4 Effect of different fermentation methods on amino acid nitrogen production (g / 100ml)

[0130]

[0131] Table 5 Effect of different fermentation methods on amino acid nitrogen production (g / 100ml)

[0132]

[0133] Table 6 Effect of different fermentation methods on amino acid nitrogen production (g / 100ml)

[0134]

[0135]

[0136] Table 7 Effect of different fermentation methods on amino acid nitrogen production (g / 100ml)

[0137]

[0138] From Tables 1 to 7 and Figures 1 to 7 As can be seen, the present invention combines different microbial spore suspensions, either alone or in combination, with phytic acid for fermentation of flavor bases, producing unexpected technical benefits. Examples 12-16 and Example 18 significantly increased the amino acid nitrogen content in the flavor base, thereby enhancing the quality and nutritional value of the flavor base. Furthermore, the fermentation performance of the samples from Examples 12-16 and Example 18 was significantly different from that of Comparative Examples 1-7 and Example 17.

[0139] Secondly, from Table 6 and Figure 6 As can be seen, there is an antagonistic effect between Bacillus subtilis and Monascus, while a synergistic effect is observed between Bacillus subtilis, Monascus, and Actinomucor elegans. This may be due to the complementary metabolites produced by the individual strains, resulting in a better overall effect. This may induce a stress response between Bacillus subtilis and Monascus, prompting each to enhance certain physiological functions or produce specific metabolites to cope with the competitive pressure from the other, thus having a positive impact on the overall combination of the three. This may also indirectly have a positive effect on Actinomucor elegans.

[0140] In summary, the present invention utilizes phytic acid as a fermentation promoter, combined with one or more of Actinomucor elegans and Bacillus subtilis and Monascus purpureus, and fermentation conditions to significantly increase the amino acid nitrogen content in the flavor base, thereby enhancing the quality and nutritional value of the flavor base. Furthermore, the experimental strains used in the method of the present invention are of clear and readily available origin, and the composition of the fermentation broth culture medium can be adjusted within a certain range to accommodate different production needs and conditions, thus offering good flexibility and operability.

[0141] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments like this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A leavening agent, characterized in that Including phytic acid and microbial agents; The volume ratio of the microbial agent to phytic acid is 296-498:2-4; The concentration of microorganisms in the microbial agent is 1×10 8 ~3×10 8 / mL; The microorganisms include any one or more of Actinobacillus elegans, Monascus purpureus and Bacillus subtilis.

2. The leavening agent according to claim 1, wherein The independent concentration of the phytic acid is 60 to 80 wt.%.

3. The leavening agent according to claim 1, wherein When the microorganisms are Actinobacillus elegans and Monascus purpureus, the concentration ratio of Actinobacillus elegans to Monascus purpureus in the microbial inoculant is 1-3:1-3; When the microorganisms are Actinobacillus elegans and Bacillus subtilis, the concentration ratio of Actinobacillus elegans to Bacillus subtilis in the microbial inoculant is 1-3:1-3; When the microorganisms are Actinobacillus elegans, Monascus purpureus and Bacillus subtilis, the concentration ratio of Actinobacillus elegans, Monascus purpureus and Bacillus subtilis in the microbial inoculant is 1-3:1-3:1-3.

4. Use of the starter culture according to any one of claims 1 to 3 in increasing the amino acid nitrogen content in a fermentation product.

5. A method for increasing the content of amino acid nitrogen in a fermentation product, characterized in that: The steps include: The starter according to any one of claims 1 to 3 is mixed with a fermentation material, and the obtained mixture is fermented and cultured to obtain a fermentation product.

6. The method according to claim 5, wherein The fermentation temperature is 27-33°C, the fermentation speed is 150-250 r / min, and the fermentation time is 60-84 h; The volume ratio of the starter to the fermentation material is 3-5:

100.

7. The method according to claim 6, wherein The fermentation material includes fermentation medium; The fermentation medium comprises the following components in the following concentrations: 2-4% soy protein isolate, 1-2% glucose, 0.1-0.3% magnesium sulfate, 0.1-0.3% potassium dihydrogen phosphate, 0.05-0.15% yeast extract, 0.3-0.5% sodium nitrate, 0.01-0.03% zinc sulfate, 0.05-0.15 g / L calcium chloride, 0.03-0.07 g / L MnSO4, and 0.005-0.015 g / L FeSO4. The percentages of the soy protein isolate, glucose, magnesium sulfate, potassium dihydrogen phosphate, yeast extract, sodium nitrate and zinc sulfate are all percentages by volume.

8. Use of the method according to any one of claims 5 to 7 in the preparation of a flavor base.

9. The use according to claim 8, characterized in that The fermentation product obtained by the method is subjected to solid-liquid separation, and the liquid portion is collected to obtain a flavor base.

10. The use according to claim 9, characterized in that The solid-liquid separation method includes centrifugation, the centrifugal speed is 10000-14000 r / min, and the centrifugal time is 10 minutes.

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