Yeast extract containing arginine as well as preparation method and application of yeast extract

Abstract

The invention relates to the field of yeast extracts, in particular to a yeast extract containing arginine as well as a preparation method and application of the yeast extract. Wherein the content of arginine in the yeast extract containing arginine is greater than or equal to 6%, and the preparation method comprises the following steps: step 1, culturing a strain to obtain yeast milk; 2, extracting the yeast milk obtained in the step 1, and performing solid-liquid separation to obtain supernate; step 3, performing enzymolysis on the supernate obtained in the step 2, and performing solid-liquid separation to obtain a yeast extract containing arginine; wherein in the step 3, the enzyme used for enzymolysis comprises one or more than two selected from a substance group consisting of papain, alkaline protease and neutral protease. By adopting the preparation method disclosed by the invention, the arginine in the yeast can be more effectively enriched and can be up to 7.35% at most, and the method can be widely applied to the fields of preparing flavor modifiers of foods or human nutrition supplements and the like.

Description

Technical Field

[0001] This invention belongs to the field of microbial application technology, specifically relating to an arginine-containing yeast extract, its preparation method, and its application. Background Technology

[0002] Arginine is a conditionally essential amino acid for maintaining the growth of newborn mammals and the body's nitrogen balance. As one of the important nutrients for human growth and development, it has the functions of promoting fetal growth, infant growth and development, immune regulation, anti-oxidation, anti-inflammation, promoting cell proliferation, anti-apoptosis, and regulating lipid metabolism. Therefore, adequate intake of arginine is particularly important for human growth and development and the maintenance of normal physiological functions. In addition, arginine can produce special aroma substances by reacting with sugars by heating (amino-carbonyl reaction).

[0003] In recent years, with economic development, people's demand for natural nutrient supplements has been increasing, and their pursuit of food flavors has become more diversified. Therefore, yeast extract rich in arginine can not only be used as a nutritional supplement but also as a food flavor improver, and it will have a broad market in the fields of nutrient supplementation and seasoning nutrition. However, the extraction and dissolution of arginine from yeast is currently limited and inefficient, which is not conducive to its industrial production. Summary of the Invention

[0004] The technical problem solved by this invention is that, in the prior art, the extraction and dissolution of arginine from yeast is limited and inefficient, which is not conducive to its industrial production.

[0005] To address the aforementioned technical problems, this invention provides an arginine-containing yeast extract, its preparation method, and its applications.

[0006] Specifically, the present invention provides the following technical solution: In a first aspect, the present invention provides a yeast extract containing arginine, wherein, based on the mass of the dry matter of the yeast extract, the arginine content in the yeast extract is greater than or equal to 6% and the protein content in the yeast extract is greater than or equal to 60%.

[0007] Preferably, the arginine content in the yeast extract is 6%-8% based on the mass of the dry matter of the yeast extract; and / or, the protein content in the yeast extract is 60%-70% based on the mass of the dry matter of the yeast extract.

[0008] Secondly, the present invention provides a method for preparing a yeast extract containing arginine, comprising the following steps: Step 1: Cultivate the strain to obtain yeast milk; Step 2: Extract the yeast milk obtained in Step 1, separate the solid and liquid components, and obtain the supernatant; Step 3: Enzymatically digest the supernatant obtained in Step 2, and separate the solid and liquid components to obtain a yeast extract containing arginine; In step 3, the enzymes used for enzymatic hydrolysis include one or more selected from the group consisting of papain, alkaline protease, and neutral protease.

[0009] Preferably, in step 1, the strain includes one or more selected from the group consisting of baker's yeast, brewer's yeast, saccharomyces cerevisiae, candidiasis, and kluyveromyces multispora.

[0010] And / or, preferably, in step 1, the culture medium used for yeast culture, by weight, comprises 17-23 parts glucose, 0.5-4 parts yeast extract, 0.1-2 parts MgSO4, and 0.5-5 parts KH2PO4. And / or, the incubation temperature is 25~40℃; and / or, the incubation time is 10-15 hours.

[0011] And / or, preferably, in step 1, the dry matter concentration of the yeast milk is 10%~20%; And / or, the color of the yeast milk is 30~60.

[0012] And / or, preferably, in step 2, water is added to the yeast milk obtained in step 1 to extract it to a dry matter concentration of 10%-20%; And / or, the extraction is selected from thermal extraction or homogenization and cell wall disruption extraction.

[0013] More preferably, the pH of the hot extraction is 4 to 8; and / or the temperature of the hot extraction is 60 to 99°C; and / or the time of the hot extraction is 80 to 100 min. Alternatively, more preferably, the pH for homogenization and cell disruption extraction is 4 to 8; and / or the pressure for homogenization and cell disruption extraction is 5 to 100 MPa.

[0014] And / or, preferably, in step 3, the dry matter concentration of the supernatant is 3% to 15%.

[0015] And / or, preferably, in step 3, papain is added first for enzymatic hydrolysis, and then alkaline protease or neutral protease is added for enzymatic hydrolysis.

[0016] More preferably, the amount of papain added is 0.01% to 1.0% based on the dry matter mass of the supernatant; and / or, the amount of neutral protease or alkaline protease added is 0.01% to 1.0% based on the dry matter mass of the supernatant; And / or, more preferably, the temperature for papain hydrolysis is 40-70°C; and / or, the pH for papain hydrolysis is 4-8; and / or, the hydrolysis time for papain is 2-12 h. And / or, more preferably, the temperature for alkaline protease hydrolysis is 40-70°C; and / or, the pH for alkaline protease hydrolysis is 7-11; and / or, the time for alkaline protease hydrolysis is 2-12 h. And / or, more preferably, the temperature for neutral protease hydrolysis is 40-70°C; and / or, the pH for neutral protease hydrolysis is 6-7.5; and / or, the time for neutral protease hydrolysis is 2-12 h.

[0017] And / or, preferably, after the enzymatic hydrolysis in step 3 is completed, the hydrolysate is subjected to solid-liquid separation, and the liquid is concentrated and / or dried to obtain a yeast extract containing arginine.

[0018] Thirdly, the present invention provides a yeast extract containing arginine, which is prepared by the method for preparing the yeast extract containing arginine.

[0019] Fourthly, the present invention provides an application of the arginine-containing yeast extract in the preparation of food flavor improvers or nutritional supplements.

[0020] Fifthly, the present invention provides a food flavor improver or nutritional supplement composition comprising the arginine-containing yeast extract.

[0021] Beneficial effects of the present invention (1) In this invention, yeast milk raw material is obtained by culturing yeast rich in arginine. The yeast extract is enriched by heating extraction or homogenization, enzymatic hydrolysis, concentration and / or drying processes, and the content of arginine is greater than or equal to 6%.

[0022] (2) In this invention, by adjusting the parameters in the extraction process and the enzymatic hydrolysis process, such as adjusting the extraction temperature, time, pH or homogenization pressure, as well as adjusting the order of added enzymes, the type of enzymes and the conditions of enzymatic hydrolysis, the yeast extract contains a higher content of amino acids.

[0023] (3) The yeast extract rich in arginine obtained by the present invention can be widely used in the preparation of food flavor improvers or human nutritional supplements.

[0024] Microbial strain preservation information The *Saccharomyces cerevisiae* Hansen Z 2.4 described in this invention was deposited on October 25, 2005, at the China Center for Type Culture Collection (CCTCC), accession number CCTCC NO: M 205130, at Wuhan University, Wuhan, China, postal code 430072; telephone number: (027) 68754052. This strain is described in patent application CN200810189433.0. Detailed Implementation

[0025] As described above, the purpose of this invention is to provide a yeast extract rich in arginine, its preparation method, and its application.

[0026] Specifically, the present invention provides a technical solution, including the following: Technical Solution 1: A yeast extract containing arginine, characterized in that, based on the mass of the dry matter of the yeast extract, the arginine content in the yeast extract is greater than or equal to 6%; and the protein content in the yeast extract is greater than or equal to 60%.

[0027] Technical Solution 2: The yeast extract containing arginine according to Technical Solution 1, wherein, based on the mass of the dry matter of the yeast extract, the arginine content in the yeast extract is 6%-8%; preferably, the arginine content in the yeast extract is 6.4%-7.5%; more preferably, the arginine content in the yeast extract is 7.0-7.4%. And / or, based on the mass of the dry matter of the yeast extract, the protein content in the yeast extract is 60%-70%; preferably, the protein content in the yeast extract is 65%-70%; more preferably, the protein content in the yeast extract is 65%-68%.

[0028] Technical Solution 3: A method for preparing the arginine-containing yeast extract as described in Technical Solution 1 or 2, characterized in that it includes the following steps: Step 1: Cultivate the strain to obtain yeast milk; Step 2: Extract the yeast milk obtained in Step 1, separate the solid and liquid components, and obtain the supernatant; Step 3: Enzymatically digest the supernatant obtained in Step 2, and separate the solid and liquid components to obtain a yeast extract containing arginine; In step 3, the enzymes used for enzymatic hydrolysis include one or more selected from the group consisting of papain, alkaline protease, and neutral protease.

[0029] Technical Solution 4: The method for preparing arginine-containing yeast extract according to Technical Solution 3, wherein, in step 1, the strain includes one or more selected from the group consisting of baker's yeast, brewer's yeast, saccharomyces cerevisiae, candidiasis, and kluyveromyces multispora; wherein, there is no limitation on the source of the strain, and any commercially available or prepared strains of those skilled in the art can be used; Preferably, the strain includes baker's yeast and / or brewer's yeast; wherein, there is no limitation on the source of the baker's yeast and / or brewer's yeast, and any commercially available or prepared baker's yeast and / or brewer's yeast skilled in the art can be used; More preferably, the strain is a brewer's yeast strain ( saccharomyces cerevisiae The accession number is: CCTCC NO: M 205130.

[0030] Technical Solution 5: A method for preparing arginine-containing yeast extract according to Technical Solution 3 or 4, wherein, in step 1, the culture medium used for yeast culture, by weight, comprises 17-23 parts glucose, 0.5-4 parts yeast extract, 0.1-2 parts MgSO4, and 0.5-5 parts KH2PO4; preferably, by weight, the culture medium used for yeast culture comprises 19-21 parts glucose, 1-3 parts yeast extract, 0.1-2 parts MgSO4, and 0.2-4 parts KH2PO4; And / or, the culture temperature is 25~40℃; preferably, the culture temperature is 33-38℃; for example, in some embodiments, the culture temperature can be 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40℃, or include the culture temperature within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, the incubation time is 10-15 hours; preferably, the incubation time is 12.5-14.5 hours; for example, in some embodiments, the incubation time can be 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5 or 15 hours, or include incubation time within the numerical range formed by any two of the above specific values ​​as endpoints.

[0031] Technical Solution 6: A method for preparing arginine-containing yeast extract according to any one of technical solutions 3-5, wherein, in step 1, the dry matter concentration of the yeast milk is 10%~20%; preferably, the dry matter concentration of the yeast milk is 16%~20%; more preferably, the dry matter concentration of the yeast milk is 16%~18%; for example, in some embodiments, the dry matter concentration of the yeast milk can be 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5% or 20%, or a dry matter concentration of yeast milk within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, the color intensity of the yeast milk is 30-60; preferably, the color intensity of the yeast milk is 38-43; more preferably, the color intensity of the yeast milk is 39-42; for example, in some embodiments, the color intensity of the yeast milk can be 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60, or include the color intensity of the yeast milk within the numerical range formed by any two of the above specific values ​​as endpoints.

[0032] Technical Solution 7: A method for preparing arginine-containing yeast extract according to any one of technical solutions 3-6, wherein, in step 2, water is added to the yeast milk obtained in step 1 until the dry matter concentration is 10%-20%; preferably, water is added to the yeast milk obtained in step 1 until the dry matter concentration is 12%-16%; more preferably, water is added to the yeast milk obtained in step 1 until the dry matter concentration is 13%-15%; for example, in some embodiments, the dry matter concentration of the yeast milk obtained in step 1 can be 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5% or 20%, or the dry matter concentration of the yeast milk is within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, the extraction is selected from thermal extraction or homogenization and cell wall disruption extraction; Preferably, the extraction is a thermal extraction, and the pH of the thermal extraction is 4-8; more preferably, the pH of the thermal extraction is 6-7.5; for example, in some embodiments, the pH of the thermal extraction can be 4, 4.2, 4.4, 4.6, 4.8, 5, 5.2, 5.4, 5.6, 5.8, 6, 6.2, 6.4, 6.6, 6.8, 7, 7.2, 7.4, 7.6, 7.8 or 8, or a pH of the thermal extraction within the range of any two of the above specific values ​​as endpoints; a pH lower or higher than this pH range (i.e., an overly acidic or overly alkaline environment) will cause denaturation of components such as proteins, peptides and amino acids in the yeast cell contents in a strongly acidic or strongly alkaline environment, which will affect the subsequent enzymatic hydrolysis of proteins or polypeptides into amino acids, that is, affect the enrichment of arginine in the yeast extract; And / or, preferably, the thermal extraction temperature is 60-99°C; more preferably, the thermal extraction temperature is 75-99°C, 85-99°C, 88-96°C, or 90-94°C; for example, in some embodiments, the thermal extraction temperature can be 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99°C, or include thermal extraction temperatures within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the hot extraction time is 80-100 min; more preferably, the hot extraction time is 85-95 min; for example, in some embodiments, the hot extraction time is 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 min, or includes the hot extraction time within the numerical range formed by any two of the above specific values ​​as endpoints; Alternatively, preferably, the extraction is a homogenized cell-wall disruption extraction, with a pH of 4-8; more preferably, the pH of the homogenized cell-wall disruption extraction is 6-7.5; even more preferably, the pH of the homogenized cell-wall disruption extraction is 6.5-7.5; for example, in some embodiments, the pH of the homogenized cell-wall disruption extraction can be 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 or 8, or pH values ​​within the range of any two of the above specific values ​​as endpoints for homogeneous cell disruption extraction; And / or, preferably, the pressure for homogenization and cell disruption extraction is 5~100 MPa; more preferably, the pressure for homogenization and cell disruption extraction is 25~95 MPa; even more preferably, the pressure for homogenization and cell disruption extraction is 55~95 MPa; even more preferably, the pressure for homogenization and cell disruption extraction is 85~95 MPa; for example, in some embodiments, the pressure for homogenization and cell disruption extraction can be 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 3 9, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 MPa, or the pressure of homogeneous cell disruption extraction within the numerical range formed by any two of the above specific values ​​as endpoints.

[0033] Technical Solution 8: A method for preparing arginine-containing yeast extract according to any one of technical solutions 3-7, wherein, in step 3, the dry matter concentration of the supernatant is 3%~15%; Preferably, the dry matter concentration of the supernatant is 3% to 6%; More preferably, the dry matter concentration of the supernatant is 4% to 6%; For example, in some embodiments, the dry matter concentration of the supernatant can be 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or 15%, or the dry matter concentration of the supernatant can be within the range of values ​​formed by any two of the above specific values ​​as endpoints.

[0034] Technical Solution 9: The method for preparing arginine-containing yeast extract according to any one of Technical Solutions 3-8, wherein in step 3, papain is first added for enzymatic hydrolysis, and then alkaline protease or neutral protease is added for enzymatic hydrolysis. Preferably, the amount of papain added is 0.01% to 1.0% based on the dry matter mass of the supernatant; more preferably, the amount of papain added is 0.15% to 0.25%; for example, in some embodiments, the amount of papain added can be 0.01%, 0.05%, 0.10%, 0.15%, 0.20%, 0.25%, 0.30%, 0.35%, 0.40%, 0.45%, 0.50%, 0.55%, 0.60%, 0.65%, 0.70%, 0.75%, 0.80%, 0.85%, 0.90%, 0.95% or 1.00%, or an amount of papain added within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the amount of neutral protease or alkaline protease added is 0.01% to 1.0% based on the dry matter mass of the supernatant; more preferably, the amount of neutral protease or alkaline protease added is 0.05% to 0.4%; even more preferably, the amount of neutral protease or alkaline protease added is 0.1% to 0.4%; even more preferably, the amount of neutral protease or alkaline protease added is 0.2% to 0.4%; most preferably, the amount of neutral protease or alkaline protease added is 0.3% to 0.4%; for example, in some embodiments The amount of neutral or alkaline protease added can be 0.01%, 0.05%, 0.10%, 0.15%, 0.20%, 0.25%, 0.30%, 0.35%, 0.40%, 0.45%, 0.50%, 0.55%, 0.60%, 0.65%, 0.70%, 0.75%, 0.80%, 0.85%, 0.90%, 0.95% or 1.00%, or the amount of neutral or alkaline protease added within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the papain hydrolysis temperature is 40-70°C; more preferably, the papain hydrolysis temperature is 40-60°C; even more preferably, the papain hydrolysis temperature is 53-57°C; for example, in some embodiments, the papain hydrolysis temperature can be 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69 or 70°C, or a papain hydrolysis temperature within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the pH of the papain hydrolysis is 4-8; more preferably, the pH of the papain hydrolysis is 5-6; for example, in some embodiments, the pH of the papain hydrolysis can be 4, 4.2, 4.4, 4.6, 4.8, 5, 5.2, 5.4, 5.6, 5.8, 6, 6.2, 6.4, 6.6, 6.8, 7, 7.2, 7.4, 7.6, 7.8 or 8, or a pH of the papain hydrolysis that falls within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the papain hydrolysis time is 2-12 h; more preferably, the papain hydrolysis time is 4-8 h; even more preferably, the papain hydrolysis time is 5-7 h; for example, in some embodiments, the papain hydrolysis time can be 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5 or 12 h, or include papain hydrolysis times within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the alkaline protease hydrolysis temperature is 40-70°C; more preferably, the alkaline protease hydrolysis temperature is 42-48°C; for example, in some embodiments, the alkaline protease hydrolysis temperature can be 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69 or 70°C, or include alkaline protease hydrolysis temperatures within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the pH of alkaline protease hydrolysis is 7-11; more preferably, the pH of alkaline protease hydrolysis is 7-8; for example, in some embodiments, the pH of alkaline protease hydrolysis can be 7, 7.2, 7.4, 7.6, 7.8, 8, 8.2, 8.4, 8.6, 8.8, 9, 9.2, 9.4, 9.6, 9.8, 10, 10.2, 10.4, 10.6, 10.8 or 11, or the pH of alkaline protease hydrolysis can be within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, the alkaline protease hydrolysis time is 2-12 hours; more preferably, the alkaline protease hydrolysis time is 9-11 hours; for example, in some embodiments, the alkaline protease hydrolysis time can be 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5 or 12 hours, or include alkaline protease hydrolysis time within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the temperature for neutral protease hydrolysis is 40-70°C; more preferably, the temperature for neutral protease hydrolysis is 48-52°C; for example, in some embodiments, the temperature for neutral protease hydrolysis can be 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69 or 70°C, or include the temperature for neutral protease hydrolysis within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, preferably, the pH of the neutral protease hydrolysis is 6-7.5; more preferably, the pH of the neutral protease hydrolysis is 6.3-7.2; for example, in some embodiments, the pH of the neutral protease hydrolysis can be 6, 6.2, 6.4, 6.6, 6.8, 7, 7.2 or 7.5, or a pH of the neutral protease hydrolysis that falls within the numerical range formed by any two of the above specific values ​​as endpoints; And / or, the time for neutral protease hydrolysis is 2-12 hours; more preferably, the time for neutral protease hydrolysis is 9-11 hours; for example, in some embodiments, the time for neutral protease hydrolysis can be 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5 or 12 hours, or include neutral protease hydrolysis time within the numerical range formed by any two of the above specific values ​​as endpoints; More preferably, enzyme inactivation is performed after enzymatic hydrolysis; even more preferably, the enzyme inactivation conditions are: a temperature of 60~100℃ and / or an enzyme inactivation time of 0.1-1h.

[0035] Technical Solution 10: A method for preparing arginine-containing yeast extract according to any one of technical solutions 3-9, wherein after enzymatic hydrolysis in step 3, the hydrolysate is concentrated and / or dried to obtain arginine-containing yeast extract; Preferably, the enzymatic hydrolysate is concentrated to obtain a paste-like yeast extract containing arginine; and / or, the enzymatic hydrolysate is dried to obtain a powdered yeast extract containing arginine. And / or, preferably, the drying is spray drying, vacuum drying or drum drying; More preferably, the spray drying temperature is 70-100°C.

[0036] Technical Solution 11: According to the preparation method of yeast extract containing arginine described in Technical Solution 10, wherein after enzymatic hydrolysis in step 3, the pH of the hydrolysate is adjusted to 5-6.5, solid-liquid separation is performed, and the liquid is then concentrated and / or dried to obtain yeast extract containing arginine. Preferably, the solid-liquid separation is centrifugation; More preferably, the centrifugation speed is 4500-5500 rpm; even more preferably, the centrifugation speed is 4900-5100 rpm. And / or, more preferably, the centrifugation time is 2-8 min; even more preferably, the centrifugation time is 4-6 min.

[0037] Technical Solution 12: A method for preparing arginine-containing yeast extract according to any one of Technical Solutions 3-11, wherein the source of papain is not limited in any way, and any commercially available or prepared papain can be used in this invention; preferably, the inventors have found that any commercially available or prepared papain with an enzyme activity of 200,000 U / g to 1,200,000 U / g can be used in this invention; more preferably, the enzyme activity of papain is 400,000 U / g to 800,000 U / g; even more preferably, the enzyme activity of papain is 550,000 U / g to 650,000 U / g; and even more preferably, the enzyme activity of papain is 580,000 U / g to 620,000 U / g. And / or, there is no limitation on the source of the alkaline protease, and any commercially available or prepared alkaline protease can be used in this invention; preferably, the inventors have found that as long as the enzyme activity of the commercially available or prepared alkaline protease is 200,000 U / g to 1,200,000 U / g, it can be used in this invention; more preferably, the enzyme activity of the alkaline protease is 400,000 U / g to 500,000 U / g; even more preferably, the enzyme activity of the alkaline protease is 430,000 U / g to 470,000 U / g; And / or, there is no limitation on the source of the neutral protease, and any commercially available or prepared neutral protease can be used in this invention; preferably, the inventors have found through research that as long as the enzyme activity of the commercially available or prepared neutral protease is 20,000 U / g to 150,000 U / g, it can be used in this invention; more preferably, the enzyme activity of the neutral protease is 20,000 U / g to 100,000 U / g; even more preferably, the enzyme activity of the neutral protease is 40,000 U / g to 60,000 U / g.

[0038] Technical Solution 13: A method for preparing arginine-containing yeast extract according to any one of Technical Solutions 3-12, wherein the source of the yeast extract is not limited in any way, and any commercially available or prepared yeast extract can be used in this invention; preferably, the inventors have found that as long as the total nitrogen content of the commercially available or prepared yeast extract is ≥10.0% and / or the amino nitrogen content is ≥5.0%, it can be used in this invention; more preferably, the total nitrogen content of the yeast extract is 10%~15%, and / or the amino nitrogen content is 5%~8%; More preferably, the yeast extract contains 40,700-40,900 mg / kg of trace elements by weight; even more preferably, the trace elements contain 31,906-31,916 mg / kg potassium, 5,733-5,743 mg / kg sodium, 350-360 mg / kg calcium, 2,667-2,677 mg / kg magnesium, 75-85 mg / kg zinc and / or 75-85 mg / kg iron; still more preferably, the trace elements contain 31,911.66 mg / kg potassium, 5,738.91 mg / kg sodium, 355.25 mg / kg calcium, 2,673.59 mg / kg magnesium, 80.94 mg / kg zinc and / or 80.21 mg / kg iron. And / or, more preferably, the yeast extract contains 5100-5500 ppm of vitamins by weight; even more preferably, the yeast extract contains 2-3 ppm of vitamin B1, 35-42 ppm of vitamin B2, 110-120 ppm of vitamin B5, 15-21 ppm of vitamin B6, 5-11 ppm of vitamin B7, 24-30 ppm of vitamin B9, 0.17-0.27 μg / 100g of vitamin B12, 3150-3250 ppm of choline, and inositol content... The content of vitamin B1 is 1520~1620ppm, and / or the content of niacin is 300~350ppm; more preferably, the yeast extract powder includes: vitamin B1 content of 2.3ppm, vitamin B2 content of 38.8ppm, vitamin B5 content of 115.0ppm, vitamin B6 content of 18.0ppm, vitamin B7 content of 7.9ppm, vitamin B9 content of 26.7ppm, vitamin B12 content of 0.23μg / 100g, choline content of 3206ppm, inositol content of 1577.7ppm, and / or niacin content of 328ppm; And / or, more preferably, the yeast extract comprises 58% to 64% total hydrolyzed amino acids by weight; even more preferably, the yeast extract comprises 60% to 63% total hydrolyzed amino acids; and still more preferably, the yeast extract comprises 61.21% total hydrolyzed amino acids.

[0039] Technical Solution 14: A yeast extract containing arginine, characterized in that the yeast extract is prepared by any one of the preparation methods of the yeast extract containing arginine described in Technical Solutions 3-13.

[0040] Technical Solution 15: Application of the arginine-containing yeast extract described in Technical Solutions 1, 2 or 14 in the preparation of food flavor improvers or nutritional supplements; Preferably, the food flavor improver includes soy sauce, seasoning packets, or nut products; And / or, preferably, the nutritional supplement includes compound amino acid protein powder, beverage, capsule, or oral liquid.

[0041] Technical Solution 16: A food flavor improver or nutritional supplement composition, characterized in that the food flavor improver or nutritional supplement composition comprises the arginine-containing yeast extract described in Technical Solutions 1, 2 or 14.

[0042] In this invention, the method for detecting total nitrogen content refers to the National Standard of the People's Republic of China GB 5009.5-2016, National Food Safety Standard, Determination of Protein in Food, Method I, Kjeldahl Method.

[0043] In this invention, the method for detecting amino acid nitrogen content refers to the People's Republic of China National Standard GB5009.235-2016 Food Safety National Standard - Determination of Amino Acid Nitrogen in Food - Method 1 - Acidity Meter Method.

[0044] In this invention, the method for detecting vitamin B1 content refers to the People's Republic of China National Standard GB5009.84-2016 Food Safety National Standard - Determination of Vitamin B1 in Food - Method I - High Performance Liquid Chromatography.

[0045] In this invention, the method for detecting vitamin B2 content refers to the People's Republic of China National Standard GB5009.85—2016 Food Safety National Standard - Determination of Vitamin B2 in Food - Method I - High Performance Liquid Chromatography.

[0046] In this invention, the method for detecting vitamin B5 content refers to the People's Republic of China National Standard GB 5009.210-2023 Food Safety National Standard Determination of Pantothenic Acid in Food, Method III, Microbiological Method.

[0047] In this invention, the method for detecting vitamin B6 content refers to the People's Republic of China National Standard GB 5009.154-2023 Food Safety National Standard - Determination of Vitamin B6 in Food - Method III - High Performance Liquid Chromatography-Fluorescence Detection.

[0048] In this invention, the method for detecting vitamin B7 content refers to the People's Republic of China National Standard GB 5009.259-2023 Food Safety National Standard - Determination of Biotin in Food - Method II - Microbiological Method.

[0049] In this invention, the method for detecting vitamin B9 content refers to the National Standard of the People's Republic of China GB 5009.211-2022, National Food Safety Standard, Determination of Folic Acid in Food.

[0050] In this invention, the method for detecting vitamin B12 content refers to the People's Republic of China National Standard GB5009.285-2022 Food Safety National Standard - Determination of Vitamin B12 in Food - Method I - Liquid Chromatography.

[0051] In this invention, the method for detecting choline content refers to the People's Republic of China National Standard GB 5413.20-2022 National Food Safety Standard for the Determination of Choline in Infant Foods and Dairy Products, Method III: Liquid Chromatography-Tandem Mass Spectrometry.

[0052] In this invention, the method for detecting inositol content refers to the People's Republic of China National Standard GB 5009.270-2023 Food Safety National Standard - Determination of Inositol in Food - Method I - Gas Chromatography.

[0053] In this invention, the method for detecting nicotinic acid content refers to the People's Republic of China National Standard GB 5009.89-2023 Food Safety National Standard - Determination of Nicotinic Acid and Nicotinamide in Food - Method I - High Performance Liquid Chromatography.

[0054] In this invention, the method for detecting the total hydrolyzed amino acid content refers to the National Standard of the People's Republic of China GB5009.124-2016, National Food Safety Standard, Determination of Amino Acids in Food; the sum of the measured amino acids is the total hydrolyzed amino acid content.

[0055] In this invention, the method for detecting ash content refers to the People's Republic of China National Standard GB 5009.4-2016 Food Safety National Standard - Determination of Ash in Food - Method I - Determination of Total Ash in Food.

[0056] In this invention, the method for detecting sodium chloride content refers to the People's Republic of China National Standard GB 5009.44-2016 Food Safety National Standard Determination of Chloride in Food - Method 1: Potentiometric Titration.

[0057] In this invention, the method for detecting moisture content refers to the People's Republic of China National Standard GB 5009.3-2016 National Food Safety Standard - Determination of Moisture in Food - Method I - Direct Drying Method.

[0058] In this invention, the method for detecting arginine content refers to GB 5009.124-2016 Determination of Amino Acids in Food.

[0059] In this invention, the method for detecting protein content refers to GB 5009.5-2016 National Food Safety Standard - Determination of Protein in Food - Method I - Kjeldahl Method.

[0060] In this invention, the color of yeast milk is measured directly using a Konica Minolta CR-410 colorimeter.

[0061] Unless otherwise stated, all reagents / instruments used in the embodiments and comparative examples of this invention are conventional commercially available products. Information on the experimental materials and instruments used in this invention is shown in the table below: Table 1 Experimental Materials / Instruments and Suppliers In this invention, the dry matter content in the yeast milk refers to the content of all solid components remaining after removing water.

[0062] In this invention, the dry matter concentration of the light phase obtained in step 2 refers to the content of all solid components remaining after removing water.

[0063] The dry matter content of the yeast milk and the dry matter content of the light phase obtained in step 2 were determined using the following drying and weighing method: (1) Place the cleaned evaporating dish into the oven and dry it at 105±2℃ for 1 hour. Then place it into the desiccator, cool and weigh it. Repeat the drying and weighing of the evaporating dish until a constant weight is reached. Record it as W1. (2) Take 10g (i.e. m) of sample and put it into an evaporating dish that has been weighed to constant weight. Place the evaporating dish in an oven and dry it at 105±2℃ for 1h. Then place it in a desiccator, cool and weigh it. Repeat drying and weighing the evaporating dish until constant weight is achieved. Record it as W2. (3) Calculate the dry matter content according to Formula 1: Dry matter content = ( W 2- W 1) / m×100% Formula 1 Wherein: the dry matter content is expressed in %; W1 is the weight of the evaporating dish, g; W2 is the weight of the evaporating dish and dry matter, g; and m is the weight of the sample, g.

[0064] To better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0065] Example 1 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The color value of the yeast milk was 40, and the dry matter content in the yeast milk was 16.5 w / w.

[0066] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0067] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the temperature was adjusted to 92℃ for thermal extraction for 90 min. Finally, the thermal extract was separated using a centrifuge to obtain a light phase (supernatant) with a dry matter concentration of 5.0 w / w.

[0068] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.05% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 85℃ to inactivate the enzyme for 0.5 hours, obtaining an enzymatic hydrolysate rich in arginine.

[0069] Step 4: Separation and Concentration / Drying The pH of the enzymatic hydrolysate obtained in step 3 was adjusted to 5.6-6.0 using citric acid solution or sodium hydroxide solution. The solution was then separated by centrifugation at 5000 rpm for 5 minutes to obtain a clear liquid. This clear liquid was then dried in a spray drying tower to prepare a powdered yeast extract rich in arginine at a temperature of 70-100℃. Based on the total dry matter mass of the yeast extract, the yeast extract powder contained 7.05% arginine, 3.5% moisture, and 68.0% protein.

[0070] Alternatively, the clear liquid can be concentrated using a rotary evaporator under the following conditions: temperature 85°C, time 50 min, and vacuum degree -0.1 MPa, until it becomes a paste-like yeast extract rich in arginine with a dry matter content of approximately 65 w / w%.

[0071] Example 2 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13.5 hours to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained with a color of 42 and a dry matter content of 17w / w.

[0072] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0073] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the temperature was adjusted to 92℃ for hot extraction for 90 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.8 w / w.

[0074] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Then, add 0.2% papain based on the dry matter content of the light phase (supernatant) and hydrolyze for 6 hours. Next, lower the temperature to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.1% alkaline protease based on the dry matter content of the light phase (supernatant) and hydrolyze for 10 hours. Finally, raise the temperature to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0075] Step 4: The steps for separating and concentrating / drying the enzymatic hydrolysate are the same as in Example 1.

[0076] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 7.10% arginine, 4.8% moisture, and 67.8% protein.

[0077] Example 3 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13.8 hours to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The color of the yeast milk was 39 and the dry matter content was 17.3 w / w.

[0078] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0079] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the temperature was set to 92℃ for 90 min of hot extraction. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 5.1 w / w.

[0080] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool down to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.2% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0081] Step 4: The separation and concentration / drying steps are the same as in Example 1.

[0082] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 7.20% arginine, 3.7% moisture, and 67.2% protein.

[0083] Example 4 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13.5 hours to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The color of the yeast milk was 39.5 and the dry matter content of the yeast milk was 16.8 w / w.

[0084] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0085] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the temperature was adjusted to 92℃ for thermal extraction for 90 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 5.0 w / w.

[0086] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.3% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, raise the temperature to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0087] Step 4: The separation and concentration / drying steps are the same as in Example 1.

[0088] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 7.25% arginine, 3.4% moisture, and 66% protein.

[0089] Example 5 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13.7 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The color of the yeast milk was 40 and the dry matter content was 17.3 w / w.

[0090] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0091] Step 2: Extraction of yeast milk by heating: First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the heat extraction temperature was set to 92℃ for 90 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.9 w / w.

[0092] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool down to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.4% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0093] Step 4: The separation and concentration / drying steps are the same as in Example 1.

[0094] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 7.35% arginine, 3.9% moisture, and 65% protein.

[0095] Example 6 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The color value of the yeast milk was 41.5 and the dry matter content of the yeast milk was 16.3 w / w.

[0096] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0097] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the heat extraction temperature was set to 85℃ for 90 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.8 w / w.

[0098] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, lower the temperature to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.4% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, raise the temperature to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0099] Step 4: The separation and concentration / drying steps are the same as in Example 1.

[0100] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.85% arginine, 3.9% moisture, and 60% protein.

[0101] Example 7 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The color value of the yeast milk was 42 and the dry matter content of the yeast milk was 16.2 w / w.

[0102] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0103] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the temperature for hot extraction was set to 75°C for 90 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.7 w / w.

[0104] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool down to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.4% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine. Step 4: The separation and concentration / drying steps are the same as in Example 1.

[0105] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.55% arginine, 3.85% moisture, and 61% protein.

[0106] Example 8 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13.6 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The color value of the yeast milk was 39.5 and the dry matter content of the yeast milk was 17 w / w.

[0107] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0108] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the heat extraction temperature was set to 65°C for 90 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.7 w / w.

[0109] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool down to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.4% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0110] Step 4: The separation and concentration / drying steps are the same as in Example 1.

[0111] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.50% arginine, 3.7% moisture, and 60% protein.

[0112] Example 9 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The yeast milk color was 41 and the dry matter content was 16.5 w / w.

[0113] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0114] Step 2: High-pressure homogenization extraction of yeast milk: First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and high-pressure homogenization extraction was performed at a pressure of 10 MPa. Finally, the homogenized extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.5 w / w.

[0115] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.1% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, raise the temperature to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0116] Step 4: The steps for separating and concentrating / drying the enzymatic hydrolysate are the same as in Example 1.

[0117] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.55% arginine, 3.5% moisture, and 61% protein.

[0118] Example 10 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13.5 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The yeast milk color was 40 and the dry matter content was 16.8 w / w.

[0119] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0120] Step 2: High-pressure homogenization extraction of yeast milk First, water is added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH is adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and high-pressure homogenization extraction is performed at a pressure of 30 MPa. The homogenized extract is then separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.45 w / w.

[0121] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and hydrolyze for 6 hours. Then, cool to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.2% alkaline protease based on the dry matter content of the light phase (supernatant), and hydrolyze for 10 hours. Finally, heat to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0122] Step 4: The steps for separating and concentrating / drying the enzymatic hydrolysate are the same as in Example 1.

[0123] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.62% arginine, 3.45% moisture, and 62.5% protein.

[0124] Example 11 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The yeast milk color was 41 and the dry matter content was 16.20 w / w.

[0125] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0126] Step 2: High-pressure homogenization extraction of yeast milk First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and high-pressure homogenization extraction was performed at a pressure of 60 MPa. Finally, the homogenized extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.6 w / w.

[0127] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool down to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.3% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0128] Step 4: The steps for separating and concentrating / drying the enzymatic hydrolysate are the same as in Example 1.

[0129] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.80% arginine, 3.5% moisture, and 60.5% protein.

[0130] Example 12 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained with a color of 40.5 and a dry matter content of 16 w / w.

[0131] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0132] Step 2: High-pressure homogenization extraction of yeast milk First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the high-pressure homogenization pressure was 90 MPa. Finally, the homogenized extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.4 w / w.

[0133] Step 3: Enzymatic hydrolysis: First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool down to 45℃, adjust the pH to 7.3-7.5 using citric acid solution or sodium hydroxide solution, and add 0.4% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0134] Step 4: The steps for separating and concentrating / drying the enzymatic hydrolysate are the same as in Example 1.

[0135] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.80% arginine, 3.45% moisture, and 60.5% protein.

[0136] Example 13 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13.5 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The yeast milk color was 39.5 and the dry matter content was 16.5 w / w.

[0137] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0138] Step 2: High-pressure homogenization extraction of yeast milk First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the heat extraction temperature was set to 92℃ for 90 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.6 w / w.

[0139] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool to 50℃, adjust the pH to 6.5-7.0 using citric acid solution or sodium hydroxide solution, and add 0.03% neutral protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 65℃ for enzyme inactivation for 30 minutes to obtain an enzymatic hydrolysate rich in arginine.

[0140] Step 4: The steps for separating and concentrating / drying the enzymatic hydrolysate are the same as in Example 1.

[0141] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.45% arginine, 3.2% moisture, and 62.5% protein.

[0142] Example 14 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 35℃ for 13 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The yeast milk color was 40 and the dry matter content was 16.60 w / w.

[0143] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0144] Step 2: High-pressure homogenization extraction of yeast milk: First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 14 w / w%. Then, the pH was adjusted to 7.0-7.2 using citric acid solution or sodium hydroxide solution, and the heat extraction temperature was set to 92℃ for 90 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 4.52 w / w.

[0145] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 5.3-5.6 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 55℃. Add 0.2% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 6 hours. Then, cool down to 50℃, adjust the pH to 6.5-7.0 using citric acid solution or sodium hydroxide solution, and add 0.06% neutral protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 10 hours. Finally, heat to 65℃ for enzyme inactivation for 30 minutes to obtain an enzymatic hydrolysate rich in arginine.

[0146] Step 4: The steps for separating and concentrating or drying the enzymatic hydrolysate are the same as in Example 1.

[0147] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.65% arginine, 3.55% moisture, and 63% protein.

[0148] Example 15 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 25℃ for 15 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The yeast milk color was 30 and the dry matter content was 10.0 w / w.

[0149] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0150] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 10 w / w%. Then, the pH was adjusted to 4.0-4.2 using citric acid solution or sodium hydroxide solution, and the heat extraction temperature was set to 60°C for 100 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 3.0 w / w.

[0151] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 7.7-8.0 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 40℃. Add 0.05% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 12 hours. Then, cool to 70℃, adjust the pH to 10.8-11.0 using citric acid solution or sodium hydroxide solution, and add 1.0% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 2 hours. Finally, raise the temperature to 65℃ for enzyme inactivation, and the enzyme inactivation time is 30 minutes, yielding an enzymatic hydrolysate rich in arginine.

[0152] Step 4: The separation and concentration / drying steps are the same as in Example 1.

[0153] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.00% arginine, 3.5% moisture, and 60% protein.

[0154] Example 16 Step 1: Yeast fermentation culture The Saccharomyces cerevisiae strain (preservation number CCTCC NO: M 205130) was cultured on a culture medium at 40℃ for 10 h to obtain a fermentation broth. After centrifugation and multiple washings, yeast milk was obtained. The yeast milk color was 60 and the dry matter content was 10.0 w / w.

[0155] The culture medium used for cultivation, by weight, consists of 20 parts glucose, 2 parts yeast extract, 1 part MgSO4, and 3 parts KH2PO4.

[0156] Step 2: Extracting yeast milk by heating First, water was added to the yeast milk obtained in step 1 to adjust its dry matter content to 20 w / w%. Then, the pH was adjusted to 6.0-6.2 using citric acid solution or sodium hydroxide solution, and the heat extraction temperature was set to 95°C for 80 min. Finally, the heated extract was separated by centrifugation to obtain a light phase (supernatant) with a dry matter concentration of 5.5 w / w.

[0157] Step 3: Enzymatic hydrolysis First, adjust the pH of the light phase (supernatant) obtained in step 2 to 4.0-4.3 using citric acid solution or sodium hydroxide solution, and adjust the temperature to 50℃. Add 1.0% papain based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 2 hours. Then, cool down to 50℃, adjust the pH to 7.7-8.0 using citric acid solution or sodium hydroxide solution, and add 1.0% alkaline protease based on the dry matter content of the light phase (supernatant), and the enzymatic hydrolysis time is 8 hours. Finally, raise the temperature to 65℃ to inactivate the enzyme for 30 minutes, obtaining an enzymatic hydrolysate rich in arginine.

[0158] Step 4: The separation and concentration / drying steps are the same as in Example 1.

[0159] Based on the total dry matter mass of the yeast extract, the yeast extract powder contains 6.9% arginine, 3.2% moisture, and 60.5% protein.

[0160] While the specific embodiments of the present invention have been described above, they are not intended to limit the scope of protection of the present invention. Based on the technical solutions of the present invention, various modifications or variations that can be made by those skilled in the art without creative effort are still within the scope of protection of the present invention.

Claims

1. A yeast extract containing arginine, characterized in that, Based on the mass of the dry matter of the yeast extract, the yeast extract contains arginine at a rate of 6% or higher and protein at a rate of 60% or higher.

2. The yeast extract containing arginine according to claim 1, wherein, Based on the mass of the dry matter of the yeast extract, the arginine content in the yeast extract is 6%-8%; preferably, the arginine content in the yeast extract is 6.4%-7.5%; more preferably, the arginine content in the yeast extract is 7.0-7.4%. And / or, based on the mass of the dry matter of the yeast extract, the protein content in the yeast extract is 60%-70%; preferably, the protein content in the yeast extract is 65%-70%; more preferably, the protein content in the yeast extract is 65%-68%.

3. A method for preparing the arginine-containing yeast extract according to claim 1 or 2, characterized in that, It includes the following steps: Step 1: Cultivate the strain to obtain yeast milk; Step 2: Extract the yeast milk obtained in Step 1, separate the solid and liquid components, and obtain the supernatant; Step 3: Enzymatically digest the supernatant obtained in Step 2, and separate the solid and liquid components to obtain a yeast extract containing arginine; In step 3, the enzymes used for enzymatic hydrolysis include one or more selected from the group consisting of papain, alkaline protease, and neutral protease.

4. The method for preparing the arginine-containing yeast extract according to claim 3, wherein, In step 1, the strain includes one or more selected from the group consisting of baker's yeast, brewer's yeast, saccharomyces cerevisiae, candidiasis, and kluyveromyces multispora. Preferably, the strain is a brewer's yeast strain (Saccharomyces cerevisiae). saccharomyces cerevisiae The accession number is: CCTCCNO: M 205130.

5. The method for preparing the arginine-containing yeast extract according to claim 3 or 4, wherein, In step 1, the culture medium used for yeast culture, by weight, comprises 17-23 parts glucose, 0.5-4 parts yeast extract, 0.1-2 parts MgSO4, and 0.5-5 parts KH2PO4; preferably, by weight, the culture medium used for yeast culture comprises 19-21 parts glucose, 1-3 parts yeast extract, 0.1-2 parts MgSO4, and 0.2-4 parts KH2PO4. And / or, the incubation temperature is 25~40℃; and / or, the incubation time is 10-15 hours.

6. The method for preparing the arginine-containing yeast extract according to any one of claims 3-5, wherein, In step 1, the dry matter concentration of the yeast milk is 10% to 20%; preferably, the dry matter concentration of the yeast milk is 16% to 20%. And / or, the color intensity of the yeast milk is 30-60; preferably, the color intensity of the yeast milk is 38-43; more preferably, the color intensity of the yeast milk is 39-42.

7. The method for preparing the arginine-containing yeast extract according to any one of claims 3-6, wherein, In step 2, water is added to the yeast milk obtained in step 1 until the dry matter concentration reaches 10%-20% for extraction; preferably, water is added to the yeast milk obtained in step 1 until the dry matter concentration is 12%-16%. And / or, the extraction is selected from thermal extraction or homogenization and cell wall disruption extraction.

8. The method for preparing the arginine-containing yeast extract according to claim 7, wherein, The pH of the hot extraction is 4-8; and / or the temperature of the hot extraction is 60-99°C; and / or the time of the hot extraction is 80-100 min; preferably, the pH of the hot extraction is 6-7.5; and / or the temperature of the hot extraction is 75-99°C; and / or the time of the hot extraction is 85-95 min. Alternatively, the pH of the homogenized cell wall disruption extraction is 4-8; preferably, the pH of the homogenized cell wall disruption extraction is 6-7.5; and / or, the pressure of the homogenized cell wall disruption extraction is 5-100 MPa; preferably, the pressure of the homogenized cell wall disruption extraction is 25-95 MPa.

9. The method for preparing the arginine-containing yeast extract according to any one of claims 3-8, wherein, In step 3, the dry matter concentration of the supernatant is 3% to 15%; preferably, the dry matter concentration of the supernatant is 3% to 6%; more preferably, the dry matter concentration of the supernatant is 4% to 6%.

10. The method for preparing the arginine-containing yeast extract according to any one of claims 3-9, wherein, In step 3, papain is added first for enzymatic hydrolysis, followed by alkaline or neutral protease for further enzymatic hydrolysis.

11. The method for preparing the arginine-containing yeast extract according to claim 10, wherein, Based on the dry matter mass of the above supernatant, the amount of papain added is 0.01%~1.0%; preferably, the amount of papain added is 0.15%~0.25%; and / or, based on the dry matter mass of the above supernatant, the amount of neutral protease or alkaline protease added is 0.01%~1.0%; preferably, the amount of neutral protease or alkaline protease added is 0.05%~0.4%. And / or, the temperature for papain hydrolysis is 40-70℃; and / or, the pH for papain hydrolysis is 4-8; and / or, the hydrolysis time for papain is 2-12h. And / or, the temperature for alkaline protease hydrolysis is 40-70℃; and / or, the pH for alkaline protease hydrolysis is 7-11; and / or, the time for alkaline protease hydrolysis is 2-12h. And / or, the temperature for neutral protease hydrolysis is 40-70℃; and / or, the pH for neutral protease hydrolysis is 6-7.5; and / or, the time for neutral protease hydrolysis is 2-12h.

12. The method for preparing arginine-containing yeast extract according to any one of claims 3-11, wherein, After the enzymatic hydrolysis in step 3 is completed, the hydrolysate is subjected to solid-liquid separation, and the liquid is concentrated and / or dried to obtain a yeast extract containing arginine.

13. A yeast extract containing arginine, characterized in that, The yeast extract is prepared by the method for preparing arginine-containing yeast extract according to any one of claims 3-12.

14. The use of the yeast extract containing arginine as described in claim 1 or 2, or the yeast extract containing arginine as described in claim 13, in the field of preparing food flavor improvers or nutritional supplements.

15. A food flavor improver or nutritional supplement composition, characterized in that, The food flavor improver or nutritional supplement composition comprises the arginine-containing yeast extract of claim 1 or 2, or the arginine-containing yeast extract of claim 13.

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