CN114946993A - Bitter-free bean pulp delicious peptide and preparation method and application thereof - Google Patents
Bitter-free bean pulp delicious peptide and preparation method and application thereof Download PDFInfo
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- CN114946993A CN114946993A CN202210527943.4A CN202210527943A CN114946993A CN 114946993 A CN114946993 A CN 114946993A CN 202210527943 A CN202210527943 A CN 202210527943A CN 114946993 A CN114946993 A CN 114946993A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
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- A23J1/148—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by treatment involving enzymes or microorganisms
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23J1/142—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by extracting with organic solvents
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- A23J1/142—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by extracting with organic solvents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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Abstract
The invention discloses a bitter-free bean pulp delicious peptide and a preparation method and application thereof, and the bitter-free bean pulp delicious peptide comprises the following steps: (1) cooking the soybean meal at high temperature, and performing dual-bacterium starter propagation by using aspergillus oryzae and aspergillus niger to obtain a starter material; (2) adding water into the yeast material for hydrolysis reaction, filtering the obtained hydrolysate, inactivating enzyme, centrifuging, and taking supernatant; (3) and (3) carrying out ethanol fractionation and extraction on the supernatant, then carrying out sephadex separation and purification, and collecting components under a second characteristic peak according to a sephadex separation pattern formed by an AKTA protein purifier at 214nm to obtain the bitter-free bean pulp fresh taste peptide. The invention uses the compound protease system secreted by aspergillus oryzae and aspergillus niger to replace commercial enzyme to carry out enzymolysis on the soybean meal yeast to prepare the delicious peptide, and the method has the advantages of low production cost and industrialized extraction. In addition, the umami peptide of the invention has almost no bitter taste and has good stability in soy sauce.
Description
Technical Field
The invention relates to a method for preparing bitter-free delicious peptide from soybean meal and application of the bitter-free delicious peptide in soy sauce.
Background
With the continuous improvement of living standard of people, the diet concept of people gradually develops towards the directions of natural safety, health care nutrition, reasonable collocation and the like, the market proportion of the third generation compound condiment and the fourth generation pure natural condiment is larger and larger, and the delicious peptide which meets the requirement of health nutrition is more and more popular with consumers. Traditional single flavor agents such as monosodium glutamate (sodium glutamate) are strong in delicate flavor, but are monotonous in delicate flavor and insufficient in aftertaste, and excessive use of the flavor agents can also be harmful to body health; the mouth is dry and the tongue is dry after the I + G is too much eaten; the delicious peptide has the advantages of strong delicious taste, rich essential amino acids, easy digestion and absorption, nutrition and health care function, and the like, is more and more favored by people, is also an important component of the compound delicious seasoning, and at present, people separate and purify various delicious peptides from various animals and plants.
The soybean meal is a byproduct of soybean after oil removal, contains abundant protein and amino acid, but the soybean meal is less developed and applied in the food field at present due to the reasons that the human body digestion and absorption rate is reduced after the protein is denatured at high temperature, various anti-nutritional factors are contained and the like, and the research on continuously separating and purifying the delicious peptide is less because the bitter amino acid originally folded in the soybean protein is exposed after the soybean protein is decomposed, the bitter threshold is low, the taste is greatly influenced, and the soybean meal is the biggest problem in preparing the bean-derived delicious peptide.
The fanhairu separates the umami peptide similar to I + G from the bean pulp by means of continuous enzymolysis of endoprotease neutral protease, exoprotease compound protease and Flavourzyme, which provides possibility for directly extracting the umami peptide from the bean pulp, but the commercial enzyme has higher cost and is not suitable for industrial production, and the extraction of the umami peptide from the bean pulp by utilizing microbial starter propagation is not reported at present.
Disclosure of Invention
The invention aims to provide a method for industrially extracting fresh peptide without bitter taste from soybean meal, which utilizes aspergillus oryzae and aspergillus niger to simulate a soy sauce koji making process, and extracts, separates, purifies and debitterizes the soybean peptide by hydrolysis, ethanol fractionation and sephadex chromatography to finally obtain the fresh peptide without bitter taste.
In order to achieve the above purpose, the operation steps of the invention are as follows:
a preparation method of bitter-free bean pulp delicious peptide comprises the following steps:
(1) preparing yeast: cooking the soybean meal at high temperature, and then making koji by using Aspergillus oryzae (Aspergillus oryzae) and Aspergillus niger (Aspergillus niger) to obtain a koji material;
(2) hydrolysis: adding water into the yeast material for hydrolysis reaction, filtering the obtained hydrolysate, inactivating enzyme, centrifuging, and taking supernatant;
(3) ethanol fractionation and extraction: concentrating the supernatant, adding edible anhydrous ethanol in several times to gradually increase alcohol content, gradually precipitating, adding water to redissolve the precipitate, and rotary evaporating to remove residual ethanol to obtain ethanol fractionation sample;
(4) and (3) separating and purifying the glucan gel: preparing an ethanol fractionation sample into an aqueous solution, connecting a chromatographic column filled with sephadex particles with an AKTA protein purifier for separation and purification, eluting with water, and collecting components under a second characteristic peak according to a sephadex separation pattern formed by the AKTA protein purifier under 214nm to obtain the bitter-free soybean meal delicious peptide.
Preferably, the inoculation ratio of the aspergillus oryzae to the aspergillus niger in the step (1) is 1: 1-6: 1(v/v), the inoculation amount is 0.1-0.5%, the culture temperature for preparing the koji is 25-38 ℃, the culture time is 48-72h, and the turning interval time is 6-10 h.
Preferably, the conditions of the hydrolysis reaction in step (2): the temperature is 30-50 ℃; the ratio of the feed to the liquid (g/g) is 1: 1-1: 6; pH 5-7 (including pH not adjusted in natural state); the hydrolysis time is 5-30 days.
Preferably, the filtration in the step (2) is warp cloth filtration and filter paper suction filtration in sequence; the enzyme deactivation condition is that the enzyme is deactivated in water bath at 90-95 ℃ for 15-20 min; the centrifugation condition is 5000-.
Preferably, the solid content after concentration in the step (3) is controlled to be 30-60%; adding edible absolute ethyl alcohol twice, wherein the content of the ethyl alcohol in a first precipitation system is controlled to be 50-70%; the ethanol content in the second precipitation system is controlled to be 75-90%.
Preferably, the ethanol fractionation extraction of step (3): concentrating the supernatant to solid content of 50%, adding anhydrous ethanol to make ethanol volume concentration 65%, stirring, centrifuging to remove precipitate, adding anhydrous ethanol to the supernatant to make ethanol volume concentration reach 85%, stirring, centrifuging, adding water to the precipitate for redissolution, and rotary steaming to remove residual ethanol.
Preferably, the conditions of the two times of stirring and centrifuging in the step (3) are as follows: stirring at 1500rpm of 1000-.
Preferably, the Sephadex G-15, Sephadex G-25 or Sephadex G-50 particles are used as the Sephadex gel particles in step (4).
Preferably, the elution conditions are: the flow rate is 1-2mL/min, the pump pressure is 0.40-0.45MPa, each 4mL tube is collected by tube.
Preferably, in step (4), the fraction under the second characteristic peak is concentrated into a paste by a rotary evaporator or into a powder by freeze-drying or spray-drying according to a separation pattern of Sephadex formed by AKTA protein purification apparatus.
The bitter-free bean pulp delicate flavor peptide powder is applied to soy sauce.
The invention carries out aspergillus niger and aspergillus oryzae dual-bacterium starter propagation and enzymolysis on the soybean meal, not only utilizes a compound protease system secreted by the aspergillus oryzae and the aspergillus niger to replace commercial enzyme to carry out enzymolysis on the soybean meal, but also can reduce the bitter taste of soybean peptide in an enzymolysis solution, and then carries out continuous separation and purification through ethanol classification and glucan gel chromatography to further remove the bitter taste of the soybean peptide, thereby preparing the delicious peptide taking the soybean meal as a raw material.
Compared with the prior art, the invention has the following beneficial effects:
(1) in the invention, aspergillus oryzae and aspergillus niger are added into the bean pulp for starter propagation, and a composite protease system secreted by aspergillus oryzae and aspergillus niger is used for replacing commercial enzyme for enzymolysis to prepare the delicious peptide.
(2) A large amount of acid protease secreted by aspergillus niger is utilized, and the acid protease is mainly exonuclease, so that bitter amino acid at the tail end of a peptide chain can be cut off, thereby removing the bitter taste of soybean peptide, and the acid protease can also improve the flavor of hydrolysate; aspergillus oryzae is a good strain in the traditional brewing industry in China, and has wide application in soy sauce brewing, fermented soybean making and other aspects.
(3) Compared with two kinds of bean-source fresh-taste peptides (soybean peptide paste and soybean extract) sold in the market, the bean pulp fresh-taste peptide prepared by the invention almost has no bitter taste, and the other two kinds of bean pulp fresh-taste peptides have bitter taste; in addition, the fresh peptide of the soybean meal has good stability in soy sauce.
Drawings
FIG. 1 is a graph of Sephadex chromatographic separation.
FIG. 2 is a graph showing the comparison of the precipitation rate of three bean-derived umami peptides in soy sauce at different addition levels.
Detailed Description
The raw materials in the examples specifically adopt: soybean meal, commercially available; soybean peptide paste (purchased from Biotech Co., Ltd.), soybean extract (purchased from Biotech Co., Ltd.), and said Aspergillus oryzae strain CICC2339 (Shanghai brewing 3.042); aspergillus niger ATCC 16404.
Example 1: preparation method of bean pulp delicate flavor peptide powder
(1) High-temperature cooking: adding 50g of soybean meal, 10g of bran and 90mL of distilled water (the feed-liquid ratio is 1: 1.5) into a 500mL triangular flask, uniformly mixing, and then steaming and boiling for 20min at high temperature in a steaming and boiling pot.
(2) Activating strains: 0.2g NaNO was added to a 250mL Erlenmeyer flask 3 、0.1g K 2 HPO 4 、0.05g KCl、0.05g MgSO 4 、0.001g FeSO 4 3.0g of sucrose and 100mL of distilled water, sterilizing at 121 ℃ and 0.25MPa for 20min to obtain an activated culture medium, and standing to room temperature; selecting 3-4 rings of Aspergillus oryzae from PDA slant culture medium with inoculating loop, activating in activating culture medium (the process is carried out on a sterile operating platform), and activating at 220rpm and 30 deg.C in water bath oscillator for 36 h; aspergillus niger strains were activated as described above.
(3) Preparing yeast by using double bacteria: cooling the cooked material to 45 deg.C, inoculating 0.3% Aspergillus oryzae and Aspergillus niger at a ratio of 3: 1, performing solid state fermentation at 28 deg.C in a constant temperature incubator for 72h (until mature), and turning over every 6 h.
(4) Hydrolysis: adding water according to the yeast material-liquid ratio of 1: 2.1(g/g), stirring uniformly, hydrolyzing at 44.5 deg.C for 5.1 days, and adding 10% edible anhydrous ethanol (to prevent contamination of mixed bacteria).
(5) Filtering and centrifuging: filtering the obtained hydrolysate with 4 layers of gauze, filtering with filter paper, inactivating enzyme at 90 deg.C for 20min, centrifuging at 8000r/min for 20min, and collecting supernatant.
(6) Ethanol fractionation and extraction: concentrating the supernatant at 50 deg.C by rotary evaporator to solid content of 50%, adding edible anhydrous ethanol into 500mL of supernatant to make ethanol concentration 65% (v: v, the same below), stirring at room temperature for 30min, centrifuging at 8000r/min for 15min to remove precipitate, adding anhydrous ethanol into the supernatant to make ethanol concentration reach 85%, stirring at room temperature for 30min, centrifuging at 8000r/min for 15min, adding water to redissolve the precipitate, removing residual ethanol by rotary evaporator at 50 deg.C, repeating the above steps twice, lyophilizing at-80 deg.C to obtain powder, and storing at-20 deg.C.
(7) And (3) separating and purifying the glucan gel: dissolving the obtained lyophilized powder in water to obtain a solution with a concentration of 50mg/mL, filtering with 0.22 μm water system filter membrane, and separating and purifying by connecting a glass chromatographic column of phi 2.6 × 60cm filled with Sephadex G-15 Sephadex with AKTA protein purifier. The sample amount is 2mL each time, distilled water is used as a mobile phase, the flow rate is 1mL/min, the pump pressure is 0.40MPa, each 4mL tube is collected in tubes, then according to a sephadex separation diagram formed by an AKTA protein purification instrument under 214nm, components under the same peak are combined, and are freeze-dried into powder at-80 ℃ by a freeze dryer and stored at-20 ℃ for later use. After separation by Sephadex, 4 fractions were separated, and the results are shown in FIG. 1, in which the yields of fractions III-1, III-2, III-3, and III-4 were 0.94%, 3.25%, 0.47%, and 0.06%, respectively.
(8) Sensory evaluation: performing sensory evaluation on different components obtained by gel separation, wherein the result is shown in table 1, the second characteristic peak component III-2 has the strongest delicate flavor, and is concentrated, rotary-steamed and freeze-dried into powder for facilitating the storage of the component, so as to be used as the prepared bean pulp delicate flavor peptide powder; in order to save working procedures and energy, the fresh peptide paste can also be directly concentrated into the fresh peptide paste.
Example 2: comparison of the properties of the umami peptide powder and other components of the sephadex chromatography
(1) Sensory evaluation and comparison of samples: 4 components obtained in example 1 by sephadex chromatography are taken for bitter and umami sensory comparative analysis, 10 evaluators (5 men and 5 women, sharp taste) conduct sensory evaluation and scoring on a sample solution with the mass fraction of 1% and standard substances with different concentrations in a sensory evaluation room, and after one sample is evaluated, the next sample is tasted after gargling and cleaning and 1min of rest. 0.35% monosodium glutamate solution and 0.5% L-leucine solution are selected as organoleptic standard substances with delicate flavor and bitter taste respectively, the standard substance is regulated to be 5 points, the scoring range is 0-10 points, the average value of a plurality of panelists is taken as the result, and the evaluation result is shown in table 1.
Table 1 shows the sensory comparison of the gel chromatography components
(2) Amino acid composition comparison: respectively taking 200mg of sample and 50mg of sulfosalicylic acid, adding distilled water, uniformly mixing, fixing the volume to 2mL, standing in a refrigerator at 4 ℃ for 1h, centrifuging at 10000r/min for 15min, removing precipitates, centrifuging again under the same condition, filtering by using a 0.22-micron filter membrane, and loading on a machine, wherein the amino acid composition is shown in Table 2; the result shows that the component III-2 umami peptide powder has the content of the umami amino acid of 9.549g/100g, wherein the content of glutamic acid accounts for 91.54 percent of the content of the umami amino acid.
Table 2 shows the amino acid composition comparison of each component of gel chromatography
Example 3: freshness characteristics of umami peptide powder
(1) Measuring the taste threshold of the bean pulp delicate flavor peptide powder: preparing a series of diluents of 100-500mg/L and increasing by taking 50mg/L as a gradient, sequentially evaluating the diluents of the samples from low concentration to high concentration, and averaging the results of multiple evaluators according to the taste threshold value of the substance, wherein the concentration of the diluent is just different from that of purified water and is identified, and the results are shown in Table 3.
(2) Determining the umami enhancing effect of the bean pulp umami peptide powder: 0.35% (mass fraction, the same below) monosodium glutamate solution, 1% soybean meal flavor peptide solution, 0.7% edible salt solution, 0.35% monosodium glutamate + 1% soybean meal flavor peptide composite solution and 0.7% edible salt solution + 1% soybean meal flavor peptide solution were prepared, respectively, and the results are shown in table 3, with 0.35% monosodium glutamate solution as the standard and set to 5 points, the above solutions were scored and the average value was taken.
Table 3 shows the umami threshold and the compounding effect of the umami peptide powder of the soybean meal
Example 4: physicochemical properties of bean pulp delicate flavor peptide powder
(1) And (3) total sugar content determination: taking 1mL of hydrolysate which is filtered by a filter membrane of 0.22 mu m and subjected to suction filtration, diluting to 1000mL of the hydrolysate, taking 1mL of the hydrolysate, adding 1.0mL of phenol solution with volume fraction of 6%, fully and uniformly mixing by using a vortex oscillator, immediately adding 5.0mL of concentrated sulfuric acid, shaking uniformly, standing at room temperature for 20min, and determining the absorbance at 490 nm; the regression equation according to the standard curve is: a ═ 0.0191C-0.0358, R 2 0.9929. The result shows that the total sugar content of the bean pulp umami peptide powder is 20 percent.
(2) Protein content determination and determination steps: according to the detection of the Kjeldahl method in the national standard GB 5009.5-2016, the result shows that the protein content of the bean pulp umami peptide powder is 63%.
Example 5: the ratio of soybean meal flavor peptide to the physical and chemical properties of soybean extract
(1) Uniformly fixing the three bean-source umami peptides under the solid content of 65%, and comparing the reducing sugar content, the amino nitrogen content and the sensory evaluation of the three bean-source umami peptides.
(2) And (3) determining the content of reducing sugar: diluting 0.05g of three bean-source umami peptides to 100mL, adding 0.4mL of diluent into 1.2mL of 3, 5-dinitrosalicylic acid (DNS) boiling water bath, heating for 6min, rapidly placing into a refrigerator for cooling, adding 8.4mL of distilled water, and measuring the light absorption value at the wavelength of 540 nm; the standard curve regression equation is: a is 0.25017C +0.0072, R 2 ＝0.9974。
(3) And (3) measuring the content of amino nitrogen: 0.05g of three bean-source umami peptide is diluted to 100mL, 5mL of diluent is taken, 20mL of water is added, 0.02mol/L NaOH is used for titration until the pH value is 8.2, 5mL of formaldehyde is added, the NaOH is used for titration until the pH value is 9.2, and meanwhile, distilled water is used as a blank control.
Table 4 shows comparison of physicochemical properties of three kinds of umami peptides derived from beans
(4) Sensory evaluation: sensory evaluation was performed when the solid content of the three kinds of umami peptides derived from beans was 65% at the same time, and the results are shown in table 5, in the same manner as in example 2.
Table 5 is a sensory evaluation comparison of three bean-derived umami peptides
(5) Stability assay in thin salt light soy sauce applied: taking a plurality of 50mL centrifuge tubes, drying to constant weight, and marking as m 1 Adding 40g of soy sauce and 0%, 0.6%, 1.25%, 2.5%, 3.75% and 5% (mass fraction) of three different bean-derived flavor peptides, mixing well by using a vortex oscillator, sealing, standing at room temperature for 30 days, centrifuging at 4000r/min for 10min, slightly pouring out supernatant, drying to constant weight, and marking as m 2 The difference between the two is the quality of the precipitate.
The precipitation rate is calculated by the formula:
the results are shown in figure 2, and show that the soybean meal flavor peptide has the lowest precipitation rate in soy sauce and has good stability in soy sauce.
Although the above examples describe the operation flow of the product of the present invention and the properties of the product in the greatest possible detail, it is only a part of the examples of the present invention, but not all examples, and one can still prepare the umami peptide (powder) from other materials according to the extraction method of the present invention, and these examples are all within the scope of the present invention.
Claims (10)
1. A preparation method of bitter-free bean pulp delicious peptide is characterized by comprising the following steps:
(1) preparing yeast: cooking the soybean meal at high temperature, and then making koji by using Aspergillus oryzae (Aspergillus oryzae) and Aspergillus niger (Aspergillus niger) to obtain a koji material;
(2) hydrolysis: adding water into the yeast material for hydrolysis reaction, filtering the obtained hydrolysate, inactivating enzyme, centrifuging, and taking supernatant;
(3) ethanol fractionation and extraction: concentrating the supernatant, adding edible anhydrous ethanol in several times to gradually increase alcohol content, precipitating step by step, adding water to the precipitate for redissolving, and rotary evaporating to remove residual ethanol to obtain ethanol fractionation sample;
(4) and (3) separating and purifying the glucan gel: preparing a sample subjected to ethanol fractionation into an aqueous solution, connecting a chromatographic column filled with sephadex particles with an AKTA protein purifier for separation and purification, eluting with water, and collecting components under a second characteristic peak according to a sephadex separation pattern formed by the AKTA protein purifier at 214nm to obtain the bitter-free soybean meal delicious peptide.
2. The process according to claim 1, wherein the Aspergillus oryzae and Aspergillus niger are inoculated in the step (1) at a ratio of 1: 1 to 6: 1(v/v), the amount of the inoculum is 0.1 to 0.5%, the temperature for the koji preparation is 25 to 38 ℃, the time for the culture is 48 to 72 hours, and the interval between the koji turnover is 6 to 10 hours.
3. The method according to claim 1, wherein the conditions of the hydrolysis reaction in step (2): the temperature is 30-50 ℃; the ratio of the feed to the liquid (g/g) is 1: 1-1: 6; the pH value is 5-7; the time is 5-30 days.
4. The method according to claim 1, wherein the filtration conditions in step (2) are warp cloth filtration followed by filter paper suction filtration; the enzyme deactivation condition is that the enzyme is deactivated in water bath at 90-95 ℃ for 15-20 min; the centrifugation condition is 5000-.
5. The method according to claim 1 or 2 or 3 or 4, wherein the concentration in step (3) is controlled to a solid content of 30-60%; firstly, adding absolute ethyl alcohol into the supernatant to enable the volume concentration of the ethyl alcohol to be 50-70%, stirring and centrifuging to remove precipitates, then continuously adding the absolute ethyl alcohol into the supernatant to enable the volume concentration of the ethyl alcohol to reach 75-90%, stirring and centrifuging, adding water into precipitates obtained by secondary centrifugation for redissolving, and performing rotary evaporation to remove residual ethyl alcohol.
6. The method according to claim 1 or 2 or 3 or 4, wherein the conditions of the two times of stirring and centrifuging in the step (3) are as follows: stirring at 1500rpm of 1000-.
7. The method according to claim 1 or 2 or 3 or 4, wherein the sample fractionated with ethanol in the step (4) is prepared into an aqueous solution having a concentration of 30 to 70 mg/mL; the Sephadex G-15, Sephadex G-25 or Sephadex G-50 particles are used as the Sephadex gel particles.
8. The production method according to claim 1 or 2 or 3 or 4, wherein the elution conditions in the step (4) are: the flow rate is 1-2mL/min, and the pump pressure is 0.40-0.45 MPa; concentrating the second characteristic peak component into paste by rotary evaporator or freeze drying or spray drying to obtain powder.
9. The bitter-free bean pulp umami peptide prepared by the method of any one of claims 1 to 8.
10. The use of the non-bitter soybean meal umami peptide of claim 9 in soy sauce.
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