Yeast protein zymolyte capable of enhancing salty taste and promoting digestion and absorption and preparation method of yeast protein zymolyte

Abstract

The invention discloses a yeast protein zymolyte for enhancing salty taste and promoting digestion and absorption and a preparation method thereof, and belongs to the field of food additives. The preparation method of the yeast protein zymolyte for enhancing the salty taste and promoting digestion and absorption comprises the following steps: adding yeast protein powder into deionized water to obtain a yeast protein solution; adding flavourzyme into the yeast protein solution, carrying out oscillatory reaction, and boiling; cooling to room temperature, adding neutral protease, and carrying out oscillation reaction; after the reaction is finished, boiling and centrifuging, storing supernate, and carrying out ultrafiltration to obtain yeast protein zymolyte; the prepared yeast protein zymolyte can become a natural product partially replacing sodium chloride, can also improve the digestion and absorption capacity of sodium salt reducing food, has an ideal raw material source of functional food ingredients or medicines for reducing blood pressure and resisting oxidation, and provides theoretical basis and technical support for developing salt reducing and functional food.

Description

Technical Field

[0001] The invention belongs to the field of food additives, and particularly relates to a yeast protein hydrolysate capable of enhancing saltiness and promoting digestion and absorption, and a preparation method thereof. Background Art

[0002] Sodium chloride (NaCl) is the main salty substance. A high-salt diet is associated with various chronic diseases, such as hypertension and cardiovascular disease. Therefore, reducing the NaCl content is particularly important for the development of the food processing industry and the research and development of healthy products.

[0003] The current salt reduction strategies mainly include the following methods: optimization of processing methods, substitution of non-sodium salts, addition of flavor enhancers and quality improvers, and optimization of salt structure and food texture. Among these methods, non-sodium salt substitution is one of the most commonly used methods. However, non-sodium salt substitution can cause product off-flavor. With the increasing demand of consumers for food flavor and health, the development of natural salt-enhancing substances has become a hot topic in recent years. To this end, a yeast protein hydrolysate that enhances saltiness and promotes digestion and absorption and a preparation method thereof are proposed. Summary of the invention

[0004] In view of the deficiencies of the prior art, the purpose of the present invention is to provide a yeast protein hydrolysate with enhanced saltiness and promoted digestion and absorption and a preparation method thereof, thereby solving the problems in the prior art.

[0005] The purpose of the present invention can be achieved through the following technical solutions:

[0006] The method for preparing a yeast protein hydrolysate capable of enhancing saltiness and promoting digestion and absorption comprises the following steps:

[0007] Adding yeast protein powder to deionized water to obtain a yeast protein solution;

[0008] Add flavor protease to the yeast protein solution, perform shaking reaction and boil; then cool to room temperature, add neutral protease and perform shaking reaction;

[0009] After the reaction is completed, the yeast protein hydrolysate is obtained by boiling and centrifuging, and the supernatant is stored and ultrafiltered.

[0010] Furthermore, the concentration of the prepared yeast protein solution is 2 wt %.

[0011] Furthermore, after flavor protease was added to the yeast protein solution, the oscillation reaction temperature was 50° C., the oscillation speed was 150 rpm, and the oscillation time was 4 h.

[0012] Furthermore, after neutral protease is added to the yeast protein solution, the temperature of the oscillation reaction is 50°C.

[0013] Furthermore, the centrifugation temperature is 4°C, the time is 15 min, and the rotation speed is 10000 rpm.

[0014] Furthermore, the component with a molecular weight less than 3KDa obtained by ultrafiltration is the yeast protein hydrolysate.

[0015] The yeast protein hydrolysate with enhanced salty taste and promoted digestion and absorption is prepared by using the above-mentioned preparation method.

[0016] The yeast protein hydrolysate with enhanced saltiness and promoted digestion and absorption is used in the preparation of pork sausage.

[0017] The application of the yeast protein hydrolysate capable of enhancing saltiness and promoting digestion and absorption in the preparation of food seasonings.

[0018] A food seasoning comprises the yeast protein hydrolysate which enhances salty taste and promotes digestion and absorption.

[0019] Beneficial effects of the present invention:

[0020] 1. The present invention obtains yeast protein hydrolysate with salty taste enhancement effect by enzymolysis of yeast protein and electronic tongue analysis.

[0021] 2. The preparation process of the present invention has the advantages of simple process operation and high safety.

[0022] 3. The yeast protein hydrolysate prepared by the present invention can be a natural product that partially replaces sodium chloride, and can also improve the digestion and absorption capacity of sodium-reduced foods. It is an ideal raw material source for food ingredients or drugs with both blood pressure lowering and antioxidant functions, and provides a theoretical basis and technical support for the development of salt-reducing and functional foods. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0024] Figure 1 A diagram showing the saltiness enhancement effect of the yeast protein hydrolysate prepared by the present invention.

[0025] Figure 2 Analysis of quality characteristics of salt-reduced pork sausage made from yeast protein hydrolysate.

[0026] Figure 3 Analysis of in vitro digestibility and polypeptide content of yeast protein hydrolysate in salt-reduced pork intestines.

[0027] Figure 4Analysis of polypeptide content in in vitro digestion products of yeast protease hydrolysate and salt-reduced pork intestines after absorption.

[0028] Figure 5 Analysis of antioxidant capacity of in vitro digestion and absorption products of yeast protease hydrolysate and salt-reduced pork intestines.

[0029] Figure 6 Analysis of the ACE inhibitory ability of in vitro digestion and absorption products of yeast protease hydrolysate and salt-reduced pork intestines. DETAILED DESCRIPTION

[0030] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0031] Example 1

[0032] The method for preparing a yeast protein hydrolysate capable of enhancing saltiness and promoting digestion and absorption comprises the following steps:

[0033] S1, adding yeast protein powder to deionized water, keeping the material / liquid ratio at 1:50 (w / v), to obtain a yeast protein solution.

[0034] S2, adding flavor protease to the yeast protein solution, shaking the reaction at 50°C, 150 rpm, shaking for 4 hours, and boiling for 10 minutes to end the reaction; then the sample temperature was lowered to room temperature, neutral protease was added, and shaking the reaction was carried out at 50°C;

[0035] S3, after the reaction is completed, boil for 10 minutes, centrifuge (4°C, 10000 rpm) for 15 minutes, store the supernatant at 4°C and perform ultrafiltration; the component with a molecular weight less than 3KDa obtained by ultrafiltration is the yeast protein hydrolysate.

[0036] In this example, the electronic tongue was used to analyze the yeast protein hydrolysate. Figure 1 As shown, it can be seen that the prepared yeast protein hydrolysate has the effect of enhancing the salty taste of NaCl.

[0037] Example 2

[0038] In this example, the yeast protein hydrolysate prepared in Example 1 was used to prepare a yeast protein hydrolysate reduced-salt pork sausage, and a comparative analysis and evaluation was performed;

[0039] (1) Preparation of Yeast Protein Hydrolysate-Reduced Salt Pork Sausage

[0040] The fat and connective tissue visible in the pork were removed, and minced into minced meat using a meat grinder (5 mm plate) at 4°C. The corresponding sodium chloride was added to the pork sausages, which were divided into normal group pork sausage (3% NaCl), reduced salt group pork sausage (2.1% NaCl), and yeast protease hydrolysate group pork sausage (2.1% NaCl + 0.9% yeast protease hydrolysate). After mixing evenly, they were stored at 4°C for 16 hours, and then loaded into polyethylene casings (20 mm in diameter, 20 g per sausage) sealed at one end with a button machine, and the other end of the casing was sealed with a button machine after degassing. The sausages were then placed in an 80°C water bath and cooked for 30 minutes, and then immediately cooled in ice water.

[0041] 2) Sensory evaluation of salt-reduced pork sausage made from yeast protein hydrolysate

[0042] The sensory evaluation team consisted of 5 males and 5 females, aged 22 to 26 years old. All members of the team received sensory training and were able to accurately identify six basic taste characteristics. The standard taste solution was 0.80g / L citric acid, 10.00g / L sucrose, 0.35g / L quinine sulfate, 3.5g / L sodium chloride, 3.5g / L monosodium glutamate, and 0.85g / L tannin, which were used to describe sour, sweet, bitter, salty, fresh, and astringent, respectively. The sensory experiment was carried out at 25±2℃ and 65% humidity.

[0043] The evaluators scored the saltiness of each yeast protein hydrolysate reduced-salt pork sausage sample, and the saltiness of the 3% NaCl pork sausage sample was 10 points. The evaluation environment was natural light in the laboratory, and each evaluation was conducted by an individual evaluator without contact or communication with each other. The evaluators rinsed their mouths with clean water between sample evaluations.

[0044] (3) Electronic tongue analysis of salt-reduced pork sausage using yeast protease hydrolysate

[0045] The taste attributes of pork sausage were evaluated using an electronic tongue. 5 g of yeast protein hydrolysate-reduced salt pork sausage was mixed with 50 g of ultrapure water and homogenized (1 min, 10,000 rpm), centrifuged (30 min, 10,000 rpm), the supernatant was filtered with filter paper, water was added to 80 ml after filtration, and then analyzed using an electronic tongue.

[0046] (4) Yeast protein hydrolysate reduces cooking loss and water retention capacity of salt-reducing pork sausage

[0047] Cooking loss was assessed by comparing the weight before and after cooking. Water holding capacity was calculated as the percentage of the weight of yeast protease hydrolysate reduced salt pork sausage retained after centrifugation to the initial weight.

[0048] (5) Microstructure of Yeast Protein Hydrolysate-Reduced Salt Pork Sausage

[0049] The gel samples were cut into slices with a thickness of 5 μm, then dehydrated by gradient ethanol and freeze-dried using a freeze dryer. Subsequently, the freeze-dried gel samples were gold / palladium coated by spray coating (Balzer, SCD004) and scanned under an EM30+ scanning electron microscope (COXEM, Korea).

[0050] Depend on Figure 2 As shown in A, using yeast protease hydrolysate to replace part of NaCl can improve the saltiness and umami taste of reduced-salt pork sausage and reduce the bitterness of reduced-salt pork sausage. Figure 2 As shown in Figure B, yeast protein hydrolysate can reduce the cooking loss of reduced-salt pork sausage. Figure 2 As shown in Figure C, yeast protein hydrolysate can improve the water holding capacity of reduced-salt pork sausage. Figure 2 As shown in D, yeast protein hydrolysate improved the microstructure of reduced-salt pork sausage, and the structure of the yeast protein hydrolysate group was more uniform compared with the reduced-sodium salt group.

[0051] Example 3

[0052] In this example, the yeast protein hydrolysate was used to verify the ability of the yeast protein hydrolysate to improve the in vitro digestibility of reduced-salt pork sausage.

[0053] The fat and connective tissue visible in the pork were removed, and minced into minced meat using a meat grinder (5 mm plate) at 4°C. The corresponding sodium chloride was added and divided into normal group pork sausage (3% NaCl), reduced salt group pork sausage (2.1% NaCl), and yeast protease hydrolysate group pork sausage (2.1% NaCl + 0.9% yeast protease hydrolysate). After mixing evenly, the sausages were stored at 4°C for 16 hours, and then packed into polyethylene casings (20 mm in diameter, 20 g per sausage) sealed at one end with a button machine, and the other end of the casing was sealed with a button machine after degassing. The sausages were then placed in an 80°C water bath and cooked for 30 minutes, and then immediately cooled in ice water.

[0054] The pork intestines were crushed and mixed with 10 mL of PBS (pH 7.2). The pH was adjusted to 2.0 by adding 2M HCl solution, and pepsin (25000 U / g) was added and shaken in a constant temperature shaking water bath at 150 rpm and 37°C for 2 h. The gastric digestion was terminated by adjusting the pH to 7.5 with 2M NaOH, and then pancreatic enzyme (2200 U / g) was added to start intestinal digestion, and the mixture was placed in a constant temperature shaking water bath at 150 rpm and 37°C for 2 h. After the reaction was completed, the mixture was heated in a 95°C water bath for 15 min. Then centrifuged at 4°C for 20 minutes (12,000 rpm) to separate the supernatant. The supernatant and precipitate were freeze-dried. The in vitro digestibility was determined.

[0055] The digestion products were processed by C-18 solid phase extraction column, and the samples were analyzed by LC-MS / MS. The obtained data were analyzed based on pFIND software and Uniprot database. The number of peptides was determined by searching the proteins from Sus scrofa in Uniprot (https: / / www.uniprot.org / ), Swiss-Prot and TreMBL databases.

[0056] like Figure 3 As shown in Figure A, yeast protein hydrolysate can improve the in vitro digestibility of reduced sodium pork sausage. Figure 3 As shown in Figure B, yeast protease hydrolysate can increase the amount of polypeptides in the in vitro digestion products of reduced sodium pork sausage.

[0057] Example 4

[0058] In this example, the yeast protein hydrolysate was used to verify the ability of the yeast protein hydrolysate to improve the in vitro absorption capacity of the salt-reduced pork sausage.

[0059] The small intestinal tissue of mice was collected, and the small intestine was ligated with cotton thread. 100 μL of the digestion products of the normal group pork intestine, the reduced salt group pork intestine, and the yeast protease hydrolysate group were injected into the 3 cm small intestinal segment, respectively, and placed in 10 mL of Krebs Ringer buffer, and incubated in a shaker at 100 rpm and 37 ° C for 4 h. The buffer was collected and freeze-dried, and the absorbed peptides were analyzed by LC-MS / MS.

[0060] like Figure 4 As shown in the results, yeast protease hydrolysate can increase the amount of absorbed peptides in the in vitro digestion products of reduced sodium pork intestines.

[0061] Example 5

[0062] In this example, the yeast protein hydrolysate was used to verify the antioxidant capacity of reduced-salt pork sausage.

[0063] (1) Hydroxyl radical scavenging ability

[0064] 1 mL of 6 mM salicylic acid-ethanol solution and 1 mL of 6 mM FeSO4 solution were mixed with the digestion product solution and the absorption product solution, respectively. Then 1 mL of H2O2 (5 mM) was added to the reactants and vortexed for 10 s, incubated at 37°C for 60 min, and then the absorbance was measured at 520 nm. Each group was measured in parallel 3 times.

[0065] Hydroxyl radical scavenging rate (%) = [A 0-520 -(A 1-520 -A 2-520 )] / A 0-520 ×100%

[0066] In the formula, A 0-520 , A 1-520 and A 2-520 They are the absorbance values ​​of the mixture of water and H2O2, sample solution and H2O2, and sample solution and water respectively.

[0067] (2) DPPH free radical scavenging activity

[0068] 0.5 mL (1 mg / mL) of digestion product solution and absorption product solution were added to 0.5 mL 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH)-ethanol solution (0.1 mM ethanol), respectively, and stored at 25°C in the dark for 30 min, and then the absorbance was measured at 517 nm. Each group was measured in parallel 3 times.

[0069] DPPH free radical scavenging rate (%) = [A 0-517 -(A 1-517 -A 2-517 )] / A 0-517 ×100%

[0070] In the formula, A 0-517 , A 1-517 , A 2-517 They are the absorbance values ​​of the mixture of ethanol and DPPH solution, sample solution and DPPH solution, and polypeptide solution and ethanol.

[0071] (3) ABTS free radical scavenging ability

[0072] Equal volumes of 2,2-azino-di(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt (ABTS) solution (7.0 mM) and K2O8S2 solution (2.45 mM) were stored at 25°C in the dark for 16 h, and then diluted with PBS to an absorbance of 0.70 ± 0.02 at 734 nm. 3 mL of the diluted ABTS solution was added to 0.2 mL of the digestion product solution and the absorption product solution, respectively, and incubated at 25°C in the dark for 60 min, and then the absorbance was detected at 734 nm. Each group was measured in parallel 3 times.

[0073] ABTS free radical scavenging activity (%) = [A 0-734 -(A 1-734 -A 2-734 )] / A 0-734 ×100%

[0074] Among them, A 0-734 , A 1-734 , A 2-734 They are the absorbance values ​​of the mixture of water and ABTS solution, sample solution and ABTS solution, and sample solution and water, respectively.

[0075] like Figure 5As shown in the figure, among the digestion products and absorption products, the yeast protein hydrolysate group has the strongest ability to scavenge free radicals. Figure 5 As shown in A, the hydroxyl radical scavenging rate of the digestion product of the yeast protease hydrolysate group was 55.71%, and the hydroxyl radical scavenging rate of the absorption product was 61.06%; Figure 5 As shown in Figure B, the DPPH free radical scavenging rate of the digestion product of the yeast protease hydrolysate group was 63.84%, and the DPPH free radical scavenging rate of the absorption product was 77.26%; Figure 5 As shown in Figure C, the ABTS free radical scavenging rate of the digestion product of the yeast protein hydrolysate group is 77.42%, and the ABTS free radical scavenging rate of the absorption product is 87.29%. It can be analyzed that the yeast protein hydrolysate improves the in vitro antioxidant capacity of the digestion and absorption products of the reduced-salt pork sausage.

[0076] Example 6

[0077] In this example, the yeast protease hydrolysate was used to verify the ACE inhibition ability of reduced-salt pork sausage.

[0078] ACE (20 μL, 0.1 U / mL) was added to the 96-well plate, 40 μL HEPES buffer (80 mmol / L, pH 8.3) was added to the blank well, and 15 mg / mL digestion product solution and absorption product solution were added to the sample wells. Finally, 50 μL of preheated 1 mmol / L FAPGG was added to each well and mixed thoroughly immediately. The initial absorbance of the blank well and sample well at a wavelength of 340 nm (A1, B1) was measured, and then the absorbance (A2, B2) was measured after incubation at 37 ° C in the dark for 30 minutes. The ACE inhibition rate was then calculated using the following equation:

[0079] ACE inhibition rate (%) = (AB) / A×100%

[0080] The absorbance difference A of the blank well is calculated as: A=A1-A2, and the absorbance difference B of the sample well is calculated as: B=B1-B2.

[0081] like Figure 6 As shown in the results, the yeast protease hydrolysate group had the strongest ability to inhibit ACE activity among the digested products and absorbed products. The ACE inhibition rate of the digested products of the yeast protease hydrolysate group was 40.36%; the ACE inhibition rate of the absorbed products was 55.07%. This indicates that yeast protease hydrolysate improves the ability of the digested and absorbed products of reduced-salt pork sausage to inhibit ACE activity.

[0082] In the description of this specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0083] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of the present invention to be protected.

Claims

1. A method for preparing a yeast protein hydrolysate with enhanced saltiness and improved digestion and absorption, characterized in that: The following steps are involved: Adding yeast protein powder to deionized water to obtain a yeast protein solution; Adding flavor protease to the yeast protein solution, shaking the solution and boiling the solution; Then, the temperature was lowered to room temperature, neutral protease was added, and an oscillating reaction was performed; After the reaction is completed, the yeast protein hydrolysate is obtained by boiling and centrifuging, and the supernatant is stored and ultrafiltered.

2. The method for preparing a yeast protein hydrolysate with enhanced saltiness and improved digestion and absorption according to claim 1, characterized in that: The concentration of the prepared yeast protein solution was 2 wt %.

3. The method for preparing a yeast protein hydrolysate with enhanced saltiness and improved digestion and absorption according to claim 1, characterized in that: After flavor protease was added to the yeast protein solution, the oscillation reaction temperature was 50° C., the oscillation speed was 150 rpm, and the oscillation time was 4 h.

4. The method for preparing a yeast protein hydrolysate with enhanced saltiness and improved digestion and absorption according to claim 1, characterized in that: After neutral protease was added to the yeast protein solution, the temperature of the oscillation reaction was 50°C.

5. The method for preparing a yeast protein hydrolysate with enhanced saltiness and improved digestion and absorption according to claim 1, characterized in that: The centrifugation temperature was 4°C, the time was 15 min, and the rotation speed was 10000 rpm.

6. The method for preparing a yeast protein hydrolysate with enhanced saltiness and improved digestion and absorption according to claim 1, characterized in that: The component with a molecular weight less than 3KDa obtained by ultrafiltration is the yeast protein hydrolysate.

7. A yeast protein hydrolysate with enhanced saltiness and improved digestion and absorption, characterized in that: It is prepared using the preparation method described in any one of claims 1 to 6.

8. Use of the yeast protein hydrolysate capable of enhancing saltiness and promoting digestion and absorption as claimed in claim 7 in the preparation of pork sausage.

9. Use of the yeast protein hydrolysate capable of enhancing saltiness and promoting digestion and absorption as claimed in claim 7 in preparing food seasonings.

10. A food seasoning, characterized in that: The yeast protein hydrolysate with enhanced salty taste and promoted digestion and absorption as claimed in claim 7.

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