Aspergillus oryzae CM1 and method for using same in preparing first-course soy sauce

Abstract

The invention discloses an Aspergillus oryzae CM1 and a method for preparing first-course live soy sauce using the same, and relates to the technical field of soy sauce brewing. The strain of Aspergillus oryzae CM1 has been deposited in the General Microbiological Center of the China Microbiological Culture Management Committee on May 19, 2022, with the deposited name CM1 and the deposited number: CGMCC NO.40178. The prepared first-course live soy sauce has a protease activity higher than 5U / mL while maintaining the original flavor of the soy sauce. Compared with the soy sauce with inactivated protease on the market, the product has better quality, is suitable for dipping and pickling, and has the functions of softening meat and promoting digestion.

Description

Technical Field

[0001] The invention relates to the technical field of soy sauce brewing, and in particular to Aspergillus oryzae CM1 and a method for preparing first-course live soy sauce using the same. Background Art

[0002] Soy sauce originated in my country and is a must-have condiment for the home. With the continuous advancement of science and technology and the continuous development of people's dining habits, the categories of soy sauce products are constantly being subdivided. At present, the more common functional products on the market include reduced-salt soy sauce, sugar-free soy sauce, iron-fortified soy sauce, gluten-free soy sauce, seafood soy sauce, etc., to meet the diverse needs of consumers.

[0003] The conventional soy sauce production process is to squeeze or pour the soy sauce, and then sterilize it and can it into a finished product. Soy sauce sterilization has two functions. One is to kill the microorganisms in the soy sauce to ensure the quality of the soy sauce during its shelf life, and the other is to produce unique flavor substances during the high temperature process. Therefore, after the sterilization process, the microorganisms and biological enzymes in the soy sauce have been inactivated, and the flavor and taste are significantly different from the original fermented soy sauce.

[0004] Patent CN 113632967A discloses a method for producing enzyme-activated soy sauce. Compared with the conventional process, only the sterilization method is changed, that is, high-temperature sterilization is changed to membrane filtration sterilization.

[0005] Membrane filtration sterilization technology is a relatively mature sterilization method. The production method of soy sauce should focus on the activity of biological enzymes in soy sauce rather than just changing the sterilization method. The complex physical and chemical environment in soy sauce will affect the activity of biological enzymes. A large number of biological enzymes are quickly inactivated due to intolerance to low pH values ​​and other physical and chemical factors. The lack of biological enzyme activity will greatly affect the quality of live soy sauce. Summary of the invention

[0006] Based on the technical problem that biological enzymes are currently intolerant to the complex physical and chemical environment of soy sauce and quickly inactivate, resulting in low quality of live soy sauce, the purpose of the present invention is to provide an Aspergillus oryzae CM1 and a method for using it to prepare first-cut live soy sauce, which can effectively increase the activity of biological enzymes in soy sauce, thereby greatly improving the quality of first-cut live soy sauce products while maintaining the natural flavor of soy sauce.

[0007] The present invention is achieved through the following technical solutions:

[0008] In the first aspect, the present application provides an Aspergillus oryzae CM1, the strain of Aspergillus oryzae CM1 has been deposited in the General Microbiology Center of China Microbiological Culture Management Committee on May 19, 2022, address: Beijing, China, the deposited name is Aspergillus oryzae, and its deposit number is: CGMCC NO.40178.

[0009] In a specific embodiment, the activity of the Aspergillus oryzae CM1 acid protease reaches 500-1000 U / g finished koji.

[0010] In a specific embodiment, the Aspergillus oryzae CM1 is obtained by mutagenesis of Shanghai Niang 3.042.

[0011] In a second aspect, the present application provides a use of Aspergillus oryzae CM1, including application in condiment fermentation.

[0012] In a specific embodiment, the condiment includes soy sauce.

[0013] In a third aspect, the present application provides a method for preparing a condiment, using the above-mentioned Aspergillus oryzae CM1 as a fermentation bacterium.

[0014] In a fourth aspect, the present application provides a soy sauce, which is prepared using the above-mentioned Aspergillus oryzae CM1 as a fermentation bacterium.

[0015] In a fifth aspect, the present application provides a method for preparing crude oil soy sauce, wherein the steamed soybeans and the fried wheat are inoculated with Aspergillus oryzae CM1 in proportion to make koji, and the crude oil soy sauce is obtained by pressing after fermentation.

[0016] In a sixth aspect, the present application provides a method for preparing first-class live soy sauce, comprising the following steps:

[0017] S1, mixing the steamed soybeans and the fried wheat in proportion, and making koji to obtain soy sauce koji;

[0018] S2, preparing the crude soy sauce oil obtained in step S1 into a saline solution having an amino acid nitrogen content of 0.2-0.6 g / 100 mL and a NaCl concentration of 21-22%;

[0019] S3, mixing the soy sauce koji in step S1 and the salt water prepared in step S2 in a ratio of 1: (1.8-2.0) to obtain a sauce mash;

[0020] S4, fermenting the obtained mash under controlled temperature;

[0021] S5, extracting oil by squeezing, sterilizing by membrane, and obtaining the first-class live soy sauce product.

[0022] In a specific embodiment, in step S1, the steamed soybeans and the fried wheat are used to make koji in a ratio of (4-9):1.

[0023] In a specific embodiment, in step S4, the fermentation temperature is controlled as follows: the temperature is controlled at 14°C to 16°C within 30 days of fermentation, and the temperature is increased to 30°C to 32°C starting from the 31st day until the 120th day of fermentation. Starting from the 121st day, the temperature is adjusted to 25°C to 27°C until the end of fermentation.

[0024] In a specific embodiment, in step S5, filtration is performed using a hollow fiber membrane with a pore size of 0.1 μm to 0.25 μm.

[0025] In a specific embodiment, the ammonia nitrogen of the finished soy sauce is controlled at 1.0-1.4 g / 100 mL, the total acid is controlled at 1.8-2.3 g / 100 mL, and the protease activity reaches 5-100 U / mL.

[0026] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0027] The first-class live soy sauce of the present invention has a protease activity higher than 5U / mL on the basis of maintaining the original flavor of the soy sauce. Compared with the soy sauce with inactivated protease on the market, the product quality is better, the sauce is suitable for dipping and pickling, and has the functions of softening meat and promoting digestion. DETAILED DESCRIPTION

[0028] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. The illustrative embodiments of the present invention and their description are only used to explain the present invention and are not intended to limit the present invention.

[0029] In the following description, a large number of specific details are set forth in order to provide a thorough understanding of the present invention. However, it is apparent to one of ordinary skill in the art that these specific details need not be employed to practice the present invention. In other embodiments, in order to avoid confusing the present invention, well-known materials or methods are not specifically described.

[0030] Throughout the specification, references to "one embodiment," "an embodiment," "an example," or "an example" mean that a particular feature, structure, or characteristic described in conjunction with the embodiment or example is included in at least one embodiment of the present invention. Therefore, the phrases "one embodiment," "an embodiment," "an example," or "an example" appearing in various places throughout the specification do not necessarily all refer to the same embodiment or example. In addition, particular features, structures, or characteristics may be combined in one or more embodiments or examples in any suitable combination and / or subcombination. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0031] The "range" disclosed in the present application is defined in the form of a lower limit and an upper limit, and a given range is defined by selecting a lower limit and an upper limit, and the selected lower limit and upper limit define the boundaries of the particular range. The range defined in this way can be inclusive or exclusive of the end values, and can be arbitrarily combined, that is, any lower limit can be combined with any upper limit to form a range. For example, if a range of 60 to 120 and 80 to 110 is listed for a particular parameter, it is understood that a range of 60 to 110 and 80 to 120 is also expected. In addition, if the minimum range values ​​1 and 2 are listed, and if the maximum range values ​​3, 4 and 5 are listed, the following ranges can all be expected: 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4 and 2 to 5. In the present application, unless otherwise specified, the numerical range "a to b" represents an abbreviation of any real number combination between a and b, where a and b are both real numbers. For example, the numerical range "0-5" means that all real numbers between "0-5" are listed in this document, and "0-5" is just an abbreviation of these numerical combinations. In addition, when a parameter is expressed as an integer ≥ 2, it is equivalent to disclosing that the parameter is, for example, an integer of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.

[0032] If there is no special explanation, all steps of the present application can be performed sequentially or randomly, preferably sequentially. For example, the method includes steps (a) and (b), which means that the method may include steps (a) and (b) performed sequentially, or may include steps (b) and (a) performed sequentially. For example, the method may further include step (c), which means that step (c) may be added to the method in any order. For example, the method may include steps (a), (b) and (c), or may include steps (a), (c) and (b), or may include steps (c), (a) and (b), etc.

[0033] Example 1

[0034] This embodiment provides a method for making soy sauce koji, and the specific method is:

[0035] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0036] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0037] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0038] Example 2

[0039] This embodiment provides a method for making soy sauce koji. The difference from Embodiment 1 is that the mass ratio of soybean to wheat flour in this embodiment is 5:1. The specific method is:

[0040] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 10:1), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0041] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0042] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0043] Example 3

[0044] This embodiment provides a method for making soy sauce koji. The difference from Embodiment 1 is that the mass ratio of soybean to wheat flour in this embodiment is 9:1. The specific method is:

[0045] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 20:1), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0046] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0047] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0048] Comparative Example 1

[0049] This comparative example provides a method for making soy sauce koji. The difference from Example 1 is that the spores of Shanghai Niang 3.042 are inoculated in this comparative example. The specific method is:

[0050] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with spores of Shanghai Brewing 3.042 at an inoculation amount of 1×10 7After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0051] The activity of the soy sauce koji proteinase prepared by the methods of Examples 1 to 3 and Comparative Example 1 was measured, and the test results are shown in Table 1. The detection methods of neutral protease and acid protease refer to GB / T 28715.

[0052] Table 1 Comparison of enzyme activity of different Aspergillus oryzae for preparing soy sauce koji

[0053] Example / Comparative Example Acid protease activity (U / g) Neutral protease activity (U / g) Example 1 780 1464 Example 2 796 1520 Example 3 820 1678 Comparative Example 1 118 1075

[0054] Example 4

[0055] This embodiment provides a method for preparing crude soy sauce using Aspergillus oryzae CM1, the specific method being:

[0056] S1. Soy sauce koji making

[0057] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0058] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0059] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0060] S2. Fermentation

[0061] Soy sauce koji and 22% salt water (amino acid nitrogen content 0.45g / 100mL) are mixed in a mass ratio of 1:1.8 and then pumped into a fermentation tank. The fermentation temperature control parameters are: the temperature is controlled at 14°C to 16°C within 30 days of fermentation, and the temperature is raised to 30°C to 32°C from 31 days until the fermentation is 120 days. From 121 days, the temperature is adjusted to 25°C to 27°C until the fermentation is completed. The fermentation time is 6 months. After the fermentation is completed, the sauce mash is pumped into the pressing workshop for pressing, and the parameters of the pressed soy sauce crude oil are measured.

[0062] Example 5

[0063] This embodiment provides a method for preparing crude soy sauce by using Aspergillus oryzae CM1. The difference from Example 4 is that in this embodiment, the amino acid nitrogen content of the salt water in step S2 is 0.2 g / 100 mL and the NaCl concentration is 21%. The specific method is as follows:

[0064] S1. Soy sauce koji making

[0065] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0066] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0067] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0068] S2. Fermentation

[0069] Soy sauce koji and 21% salt water (amino acid nitrogen content 0.2g / 100mL) are mixed evenly in a mass ratio of 1:1.8 and then pumped into a fermentation tank. The fermentation temperature control parameters are: the temperature is controlled at 14°C to 16°C within 30 days of fermentation, and the temperature is raised to 30°C to 32°C from 31 days until the fermentation is 120 days. From 121 days, the temperature is adjusted to 25°C to 27°C until the fermentation is completed. The fermentation time is 6 months. After the fermentation is completed, the sauce mash is pumped into a pressing workshop for pressing, and the parameters of the pressed soy sauce crude oil are measured.

[0070] Example 6

[0071] This embodiment provides a method for preparing crude soy sauce by using Aspergillus oryzae CM1. The difference from Example 4 is that in this embodiment, the amino acid nitrogen content of the salt water in step S2 is 0.6 g / 100 mL and the NaCl concentration is 21%. The specific method is as follows:

[0072] S1. Soy sauce koji making

[0073] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0074] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0075] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0076] S2. Fermentation

[0077] Soy sauce koji and 21% salt water (amino acid nitrogen content 0.6g / 100mL) are mixed evenly in a mass ratio of 1:1.8 and then pumped into a fermentation tank. The fermentation temperature control parameters are: the temperature is controlled at 14°C to 16°C within 30 days of fermentation, and the temperature is raised to 30°C to 32°C from 31 days until the fermentation is 120 days. From 121 days, the temperature is adjusted to 25°C to 27°C until the fermentation is completed. The fermentation time is 6 months. After the fermentation is completed, the sauce mash is pumped into the pressing workshop for pressing, and the parameters of the pressed soy sauce crude oil are measured.

[0078] Example 7

[0079] This embodiment provides a method for preparing soy sauce crude oil using Aspergillus oryzae CM1. Different from Example 4, in this embodiment, soy sauce koji and 22% saline (amino acid nitrogen content 0.45 g / 100 mL) are mixed at a mass ratio of 1:1.9. The specific method is as follows:

[0080] S1. Soy sauce koji making

[0081] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0082] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0083] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0084] S2. Fermentation

[0085] Soy sauce koji and 22% salt water (amino acid nitrogen content 0.45g / 100mL) are mixed evenly in a mass ratio of 1:1.9 and then pumped into a fermentation tank. The fermentation temperature control parameters are: the temperature is controlled at 14°C to 16°C within 30 days of fermentation, and the temperature is raised to 30°C to 32°C from 31 days until the fermentation is 120 days. From 121 days, the temperature is adjusted to 25°C to 27°C until the fermentation is completed. The fermentation time is 6 months. After the fermentation is completed, the sauce mash is pumped into the pressing workshop for pressing, and the parameters of the pressed soy sauce crude oil are measured.

[0086] Example 8

[0087] This embodiment provides a method for preparing soy sauce crude oil using Aspergillus oryzae CM1. Different from Example 4, in this embodiment, soy sauce koji and 22% saline (amino acid nitrogen content 0.45 g / 100 mL) are mixed at a mass ratio of 1:2.0. The specific method is as follows:

[0088] S1. Soy sauce koji making

[0089] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0090] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0091] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0092] S2. Fermentation

[0093] Soy sauce koji and 22% salt water (amino acid nitrogen content 0.45g / 100mL) are mixed evenly in a mass ratio of 1:1.9 and then pumped into a fermentation tank. The fermentation temperature control parameters are: the temperature is controlled at 14°C to 16°C within 30 days of fermentation, and the temperature is raised to 30°C to 32°C from 31 days until the fermentation is 120 days. From 121 days, the temperature is adjusted to 25°C to 27°C until the fermentation is completed. The fermentation time is 6 months. After the fermentation is completed, the sauce mash is pumped into the pressing workshop for pressing, and the parameters of the pressed soy sauce crude oil are measured.

[0094] Comparative Example 2

[0095] This comparative example provides a method for preparing crude soy sauce. The difference from Example 4 is that Shanghai Brewing 3.042 is inoculated in this example. The specific method is as follows:

[0096] S1. Soy sauce koji making

[0097] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Shanghai Brewing 3.042 at an inoculation rate of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0098] S2. Fermentation

[0099] Soy sauce koji and 22% salt water (amino acid nitrogen content 0.45g / 100mL) are mixed in a mass ratio of 1:1.8 and then pumped into a fermentation tank. The fermentation temperature control parameters are: the temperature is controlled at 14°C to 16°C within 30 days of fermentation, and the temperature is raised to 30°C to 32°C from 31 days until the fermentation is 120 days. From 121 days, the temperature is adjusted to 25°C to 27°C until the fermentation is completed. The fermentation time is 6 months. After the fermentation is completed, the sauce mash is pumped into the pressing workshop for pressing, and the parameters of the pressed soy sauce crude oil are measured.

[0100] The physical and chemical indicators of the crude soy sauce oil prepared by the methods of Example 4 and Comparative Example 2 were measured, and the test results are shown in Table 2. The detection methods of neutral protease and acid protease refer to GB / T 28715. The detection of total acid and ammonia nitrogen refers to GB / T18186.

[0101] Table 2 Comparison of physical and chemical indicators of crude soy sauce prepared by different Aspergillus oryzae

[0102]

[0103] Example 9

[0104] This embodiment provides a method for preparing first-course live soy sauce using Aspergillus oryzae CM1, the specific method being:

[0105] S1. Soy sauce koji making

[0106] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0107] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0108] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0109] S2. Fermentation

[0110] Soy sauce koji and 22% saline solution (amino acid nitrogen content of 0.45g / 100mL) are mixed uniformly in a mass ratio of 1:1.8 and then pumped into a fermentation tank. The fermentation temperature control parameters are as follows: the temperature is controlled at 14°C-16°C within 30 days of fermentation, and the temperature is increased to 30°C-32°C from the 31st day until the fermentation is 120 days. From the 121st day, the temperature is adjusted to 25°C-27°C until the fermentation is completed. The fermentation time is 6 months to obtain soy sauce crude oil.

[0111] S3. Filter the obtained soy sauce crude oil through a hollow fiber membrane (pore size 0.25 μm) to obtain the first-class live soy sauce, and then fill it into finished products and measure parameter indicators.

[0112] Example 10

[0113] This embodiment provides a method for preparing first-course soy sauce using Aspergillus oryzae CM1. Different from Embodiment 9, this embodiment uses a hollow fiber membrane with a pore size of 0.1 μm. The specific method is as follows:

[0114] S1. Soy sauce koji making

[0115] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0116] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0117] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0118] S2. Fermentation

[0119] Soy sauce koji and 22% saline solution (amino acid nitrogen content of 0.45g / 100mL) are mixed uniformly in a mass ratio of 1:1.8 and then pumped into a fermentation tank. The fermentation temperature control parameters are as follows: the temperature is controlled at 14°C-16°C within 30 days of fermentation, and the temperature is increased to 30°C-32°C from the 31st day until the fermentation is 120 days. From the 121st day, the temperature is adjusted to 25°C-27°C until the fermentation is completed. The fermentation time is 6 months to obtain soy sauce crude oil.

[0120] S3. Filter the obtained soy sauce crude oil through a hollow fiber membrane (pore size 0.1 μm) to obtain the first-class live soy sauce, and then fill it into a finished product and measure parameter indicators.

[0121] Embodiment 11

[0122] This embodiment provides a method for preparing first-course soy sauce using Aspergillus oryzae CM1. Different from Embodiment 9, this embodiment uses a hollow fiber membrane with a pore size of 0.20 μm. The specific method is as follows:

[0123] S1. Soy sauce koji making

[0124] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Aspergillus oryzae CM1 at an inoculum size of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0125] Among them, the strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Cultivation Management Committee on May 19, 2022. The deposited name is CM1 and its deposit number is: CGMCC NO.40178.

[0126] Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Brewing 3.042.

[0127] S2. Fermentation

[0128] Soy sauce koji and 22% saline solution (amino acid nitrogen content of 0.45g / 100mL) are mixed uniformly in a mass ratio of 1:1.8 and then pumped into a fermentation tank. The fermentation temperature control parameters are as follows: the temperature is controlled at 14°C-16°C within 30 days of fermentation, and the temperature is increased to 30°C-32°C from the 31st day until the fermentation is 120 days. From the 121st day, the temperature is adjusted to 25°C-27°C until the fermentation is completed. The fermentation time is 6 months to obtain soy sauce crude oil.

[0129] S3. Filter the obtained soy sauce crude oil through a hollow fiber membrane (pore size 0.20 μm) to obtain the first-class live soy sauce, and then fill it into a finished product and measure parameter indicators.

[0130] Comparative Example 3

[0131] This comparative example provides a method for preparing first-class live soy sauce. Different from Example 9, in this example, Shanghai Brewing 3.042 is used for koji making. The specific method is as follows:

[0132] S1. Soy sauce koji making

[0133] The steamed soybeans and fried wheat flour were mixed evenly (the mass ratio of soybeans to wheat flour was 8:2), and then inoculated with Shanghai Brewing 3.042 at an inoculation rate of 1×10 7 After inoculation, the koji was made under ventilation for 42 hours at a temperature of 30°C to obtain soy sauce koji.

[0134] S2. Fermentation

[0135] Soy sauce koji and 22% saline solution (amino acid nitrogen content of 0.45g / 100mL) are mixed uniformly in a mass ratio of 1:1.8 and then pumped into a fermentation tank. The fermentation temperature control parameters are as follows: the temperature is controlled at 14°C-16°C within 30 days of fermentation, and the temperature is increased to 30°C-32°C from the 31st day until the fermentation is 120 days. From the 121st day, the temperature is adjusted to 25°C-27°C until the fermentation is completed. The fermentation time is 6 months to obtain soy sauce crude oil.

[0136] S3. Filter the obtained soy sauce crude oil through a hollow fiber membrane (pore size 0.1 μm) to obtain the first-class live soy sauce, and then fill it into a finished product and measure parameter indicators.

[0137] The physical and chemical indicators of the first-class live soy sauce prepared by the methods of Examples 9 to 11 and Comparative Example 3 were measured, and the test results are shown in Table 3. Among them, the detection methods of neutral protease and acid protease refer to GB / T 28715. The total bacterial count, total acid, and ammonia nitrogen detection refer to GB / T 18186.

[0138] Table 3 Comparison of the first-cooked live soy sauce prepared by different Aspergillus oryzae

[0139]

[0140] The first-course live soy sauce prepared by the method of Example 9 and Comparative Example 3 was tested for the activity of live soy sauce proteases during shelf life after filling, and the test results are shown in Table 4. The activity of proteases was tested every six months. The detection methods of neutral proteases and acidic proteases refer to GB / T 28715.

[0141]

[0142]

[0143] It can be seen from Tables 1 to 4 that, compared with the conventional Shanghai Brewing 3.042 strain, the protease activity of the live soy sauce prepared by the method of the present invention, whether neutral protease activity or acidic protease activity, is significantly improved over the control during the entire shelf life after filling, indicating that the use of this patent can achieve the functionality of the first-class live soy sauce, that is, maintain a moderate protease activity during the entire shelf life. It can be seen that the live soy sauce prepared by the production method of the present invention has better quality.

[0144] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that it is still possible to modify the technical solutions described in the aforementioned embodiments, or to replace some or all of the technical features therein by equivalents. These modifications or replacements do not deviate the essence of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present invention, and they should all be included in the scope of the claims and specification of the present invention.

Claims

1. An Aspergillus oryzae CM1, characterized in that The strain of Aspergillus oryzae CM1 was deposited in the General Microbiology Center of China Microbiological Culture Management Committee on May 19, 2022, with the deposit name CM1 and the deposit number: CGMCC NO.40178.

2. An Aspergillus oryzae CM1 according to claim 1, characterized in that The activity of the Aspergillus oryzae CM1 acidic protease reaches 500-1000 U / g finished koji.

3. An Aspergillus oryzae CM1 according to claim 1, characterized in that The Aspergillus oryzae CM1 was obtained by mutagenesis of Shanghai Niang 3.

042.

4. A use of Aspergillus oryzae CM1 according to claim 1, characterized in that Including applications in condiment fermentation.

5. The use of Aspergillus oryzae CM1 according to claim 4, characterized in that: The condiment includes soy sauce.

6. A method for preparing a condiment, characterized in that: The Aspergillus oryzae CM1 described in claim 1 is used as the fermentation bacteria.

7. A soy sauce, characterized in that The soy sauce is prepared by using the Aspergillus oryzae CM1 described in claim 1 as a fermentation bacterium.

8. A method for preparing crude oil soy sauce, characterized in that: The steamed soybeans and fried wheat were inoculated with Aspergillus oryzae CM1 in proportion to make koji, and then pressed after fermentation to obtain crude oil soy sauce.

9. A method for preparing first-class live soy sauce, characterized in that: The following steps are involved: S1, inoculating Aspergillus oryzae CM1 in proportion to make koji to obtain soy sauce koji; S2, preparing the soy sauce koji obtained in step S1 into a brine having an amino acid nitrogen content of 0.2-0.6 g / 100 mL and a NaCl concentration of 21-22%; S3, mixing the soy sauce koji in step S1 and the salt water prepared in step S2 in a ratio of 1: (1.8-2.0) to obtain a sauce mash; S4, fermenting the obtained mash under controlled temperature; S5, extracting oil by squeezing, sterilizing by membrane, and obtaining the first-class live soy sauce product.

10. The method for preparing the first-class live soy sauce according to claim 9, characterized in that: In step S1, the steamed soybeans and the fried wheat are used to make koji in a ratio of (4-9):

1.

11. The method for preparing the first-class live soy sauce according to claim 9, characterized in that: In step S4, the fermentation temperature is controlled as follows: the temperature is controlled at 14°C to 16°C within 30 days of fermentation, and the temperature is increased to 30°C to 32°C from the 31st day until the 120th day of fermentation. From the 121st day, the temperature is adjusted to 25°C to 27°C until the end of fermentation.

12. The method for preparing the first-class live soy sauce according to claim 9, characterized in that: In step S5, a hollow fiber membrane is used for filtration, and the pore size is 0.1 μm to 0.25 μm.

13. The method for preparing the first-class live soy sauce according to claim 9, characterized in that: The ammonia nitrogen content of the finished soy sauce is controlled at 1.0-1.4 g / 100 mL, the total acid content is controlled at 1.8-2.3 g / 100 mL, and the protease activity reaches 5-100 U / mL.

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