Glycosylation modified whey protein isolate hydrolysate and application thereof

Abstract

The invention belongs to the technical field of protein glycosylation modification, and particularly relates to a glycosylation-modified whey protein isolate zymolyte and application of the glycosylation-modified whey protein isolate zymolyte. According to the technology, trypsin is adopted to hydrolyze whey protein isolate, and then chitosan oligosaccharide and TGase enzyme are adopted to carry out glycosylation modification. The glycosylation modified whey protein isolate zymolyte has significantly improved emulsibility, foamability and oxidation resistance, has improved surface hydrophobicity, and is suitable for being used as an antioxidant or a food processing auxiliary material, such as an emulsifier and a foaming agent.

Description

Technical Field

[0001] The invention belongs to the technical field of protein glycosylation modification, and particularly relates to a glycosylated whey protein hydrolysate and its application. Background Art

[0002] Whey protein isolate (WPI) contains abundant essential amino acids, is easily digested and absorbed by organisms, and is therefore used as an important raw material and component of functional foods, nutritional products, etc., and is widely used in food production. However, WPI itself is unstable in nature, and its high-level structure is easily destroyed during processing and production, causing its physical, chemical and biological functions to change, thereby limiting the application of WPI. This area is usually modified by WPI, such as ultrasound, heating, enzymolysis, glycosylation, high-pressure homogenization, etc., in order to change its hydrophobic group distribution, spatial arrangement conformation and amino acid composition, and then improve the functional properties of whey protein isolate. However, a single technology has limited effect on the modification of whey protein.

[0003] Enzymatic hydrolysis is a common treatment method for reducing protein molecular weight, fragment size, and amino acid exposure characteristics. Glycosylation modification of some proteins after enzymatic hydrolysis may enhance their properties such as emulsification and antioxidant properties. However, since the enzymatic hydrolysis products of large molecular proteins are complex mixtures, and the products change with changes in enzymatic hydrolysis conditions, they are full of uncertainty, resulting in their glycosylation products being full of variation. There is also a lack of universal and accurate rules for the changes in functional properties such as emulsification and antioxidant properties of their products. Therefore, to date, this phenomenon has only been found in a few proteins. For example, CN119548613A discloses a glycosylated modified product of bird's nest enzymatic hydrolysate with good antioxidant properties, but it mainly relies on the antioxidant properties of the added resveratrol. This is significantly different from simply glycosylation modification of proteins.

[0004] Simple glycosylation of proteins without prior proteolysis has a widespread and predictable effect on improving functional properties such as emulsification, foaming, and film-forming properties. For example, CN119791278A discloses a chitosan-glycosylated casein derivative that can effectively load curcumin for use as a drug carrier. Zhang Xuan of Dalian University of Technology's academic paper, "Study on the Film-Forming Properties of Chitosan-Modified Whey Protein Modified by Transglutaminase," uses chitosan-glycosylated whey protein isolate to produce a glycosylated whey protein derivative with excellent film-forming properties.

[0005] As for whey protein isolate, its glycosylation modification is currently mainly used to load other active ingredients, such as curcumin. In the paper "Fabrication and characterization of novel TGase-mediated glycosylatedwhey protein isolatenanoparticles for curcumin delivery" published by Li Di et al. in Food Chemistry in 2024, the hydrophobicity, antioxidant properties and other characteristics of chitosan glycosylated WPI and its carrier nanoparticles loaded with curcumin were examined in detail. Compared with WPI, the hydrophobicity (H0) of chitosan glycosylated WPI was significantly improved, but significantly lower than that of non-glycosylated cross-linked WPI (that is, only glycosylation enzyme and WPI were reacted without adding chitosan). For details, please refer to Fig. 1G of the article. It can be seen that there is uncertainty in the glycosylation of WPI.

[0006] How to improve the functionality of WPI, such as hydrophobicity, and its activity, such as antioxidant activity, is one of the hot issues that still needs to be solved. Summary of the Invention

[0007] In response to the above technical problems, the present invention aims to provide a glycosylated whey protein isolate hydrolysate and its use, namely, first enzymatically hydrolyzing whey protein isolate (hereinafter referred to as WPI) into smaller peptides and possibly changing its amino acid exposure characteristics, and then glycosylation-modifying the hydrolysate. In the study of WPI glycosylation modification using this technology, the inventors unexpectedly obtained several glycosylated WPI hydrolysates with significantly improved antioxidant properties, as well as glycosylated WPI hydrolysates with significantly improved functional properties such as emulsification and foaming properties.

[0008] The technical solutions of the present invention are as follows:

[0009] A glycosylated whey protein hydrolysate, wherein the glycosylated whey protein hydrolysate is a whey protein hydrolysate glycosylated with chitosan. The preparation method of the glycosylated whey protein hydrolysate comprises the following steps:

[0010] S1: preparing whey protein isolate protein hydrolysate: hydrolyzing whey protein isolate with trypsin to obtain whey protein isolate protein hydrolysate;

[0011] S2 glycosylation modification of whey protein isolate hydrolysate: The whey protein isolate hydrolysate is glycosylated using chitosan oligosaccharide and TGase enzyme to obtain a chitosan glycosylated whey protein isolate hydrolysate.

[0012] Preferably, the preparation method of the whey protein isolate hydrolysate described in step S1 comprises the following steps: dissolving the whey protein isolate in water, adjusting the pH to 7-9, adding trypsin, wherein the amount of trypsin added is 0.01g-0.02g trypsin per gram of whey protein isolate, the enzymatic hydrolysis temperature is 32°C-40°C, and the enzymatic hydrolysis time is 20min-40min.

[0013] Further preferably, in step S1, the amount of trypsin added is 0.012 g trypsin per g whey protein isolate, the enzymatic hydrolysis temperature is 37° C., and the enzymatic hydrolysis time is 30 min.

[0014] Preferably, step S2 comprises the following steps: adding chitosan oligosaccharide and TGase enzyme to the whey protein hydrolysate solution, and performing enzymatic glycosylation reaction at 32°C to 40°C and pH 7 to 8; the amount of chitosan oligosaccharide added is 20% to 100% of the mass of the whey protein hydrolysate.

[0015] More preferably, in step S2, the whey protein isolate hydrolysate solution has a mass volume percentage concentration of 2 to 5%.

[0016] Further preferably, in step S2, the amount of TGase added is 8 to 12 U / g whey protein hydrolysate.

[0017] The present invention further provides three uses of the aforementioned glycosylated whey protein hydrolysate:

[0018] (1) The use of the aforementioned glycosylated whey protein hydrolysate in the preparation of an antioxidant, wherein the amount of chitosan oligosaccharide added is 25% to 50% of the weight of the whey protein hydrolysate; preferably, the amount of chitosan oligosaccharide added is 30% to 35% of the weight of the whey protein hydrolysate.

[0019] (2) The use of the aforementioned glycosylated whey protein isolate hydrolysate in the preparation of a food emulsifier, wherein the amount of chitosan oligosaccharide added is 25% to 50% of the weight of the whey protein isolate hydrolysate.

[0020] (3) The use of the aforementioned glycosylated whey protein hydrolysate in the preparation of a food foaming agent, wherein the amount of chitosan oligosaccharide added is 25% to 35% of the weight of the whey protein hydrolysate.

[0021] Beneficial effects:

[0022] (1) The present invention uses transglutaminase (also known as TGase) to catalyze the coupling of amino sugars (chitooligosaccharides) with Gln residues in enzymatically hydrolyzed WPI molecules to form glycosylated products. Compared to the Maillard reaction, the reaction conditions are milder, the specificity is stronger, and the safety is higher.

[0023] (2) The glycosylated WPI hydrolysate prepared by the present invention has higher emulsification and emulsion stability. Compared with WPI, the emulsification and emulsion stability of the glycosylated WPI hydrolysate are improved by 75.72% and 27.68%.

[0024] (3) The glycosylated WPI hydrolysate provided by the present invention can effectively improve the antioxidant and foaming properties of WPI.

[0025] The above beneficial effects make the glycosylated WPI hydrolysate of the present invention more suitable as an antioxidant health food or auxiliary materials such as a foaming agent and an emulsifier. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the surface hydrophobicity measurement result diagram;

[0027] Figure 2 This is the result of emulsification test;

[0028] Figure 3 This is the result of emulsion stability test;

[0029] Figure 4 This is the result of foaming test;

[0030] Figure 5 This is the result of foaming stability test;

[0031] Figure 6 This is the result of DPPH free radical elimination activity determination;

[0032] Figure 7 This is the result of ABTS+ free radical elimination activity determination; DETAILED DESCRIPTION

[0033] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments are only used to explain the present invention and are not used to limit the scope of protection of the present invention.

[0034] In the following examples, chitosan oligosaccharide was purchased from Zhejiang Jinke Pharmaceutical Co., Ltd. (molecular weight ≤ 1000 Da); whey protein isolate (92.6%, w / w; purchased from Hilmar, USA); trypsin was purchased from Shanghai Yuanye Biotechnology Co., Ltd.; and TGase enzyme was purchased from Jiangsu Yiming Fine Chemical Co., Ltd.

[0035] Example 1 A glycosylated WPI hydrolysate and its preparation method

[0036] Step 1: Preparation of WPI hydrolysate

[0037] 5 g of WPI was dissolved in 100 mL of water, the pH was adjusted to 8, 0.06 g of trypsin was added, and the mixture was hydrolyzed in a shaker at 37°C for 30 min. The hydrolysis was then stopped by heating in a 100°C water bath for 5 min to obtain a WPI hydrolysate. The WPI hydrolysate was centrifuged at 5000 rpm for 15 min, and the supernatant was freeze-dried to obtain a WPI hydrolysate powder.

[0038] Step 2: Preparation of glycosylated WPI hydrolysate

[0039] The WPI hydrolysate powder was dissolved in water at a mass concentration of 3% (w / v, 3g / 100mL). Chitosan oligosaccharides (WPI hydrolysate powder to chitosan oligosaccharide mass ratio of 4:1) and TGase enzyme (10U / g WPI hydrolysate) were added to the solution and reacted at 37°C and pH 7.5 for 3 hours. After the reaction, the enzyme was inactivated by heating in an 85°C water bath for 5 minutes, cooled to room temperature, and the pH of the solution was adjusted to 4.5 and centrifuged at 4000 rpm for 10 minutes to collect the precipitate. The solution was then repeatedly washed with distilled water at pH 4.5. After the precipitate was fully dissolved, it was freeze-dried to obtain the chitosan oligosaccharide-modified WPI hydrolysate.

[0040] Example 2. A glycosylated WPI hydrolysate and its preparation method

[0041] The preparation method is the same as that of Example 1, except that in step 2, the mass ratio of WPI hydrolysate powder to chitosan oligosaccharide is 3:1.

[0042] Example 3. A glycosylated WPI hydrolysate and its preparation method

[0043] The preparation method is the same as that of Example 1, except that in step 2, the mass ratio of WPI hydrolysate powder to chitosan oligosaccharide is 2:1.

[0044] Example 4. A glycosylated WPI hydrolysate and its preparation method

[0045] The preparation method is the same as that of Example 1, except that in step 2, the mass ratio of WPI hydrolysate powder to chitosan oligosaccharide is 1:1.

[0046] Example 5 Performance and antioxidant properties of different WPI products

[0047] First, the products of Comparative Examples 1 to 3 below were prepared for comparative study.

[0048] Comparative Example 1 WPI powder and preparation

[0049] In this comparative example, a comparative WPI (non-glycosylated and non-enzymatically hydrolyzed WPI) was prepared in the following steps:

[0050] 5 g of WPI was dissolved in 100 mL of water, the pH was adjusted to 8, and the mixture was allowed to react at 37°C on a shaker for 30 minutes. The mixture was then heated in a 100°C water bath for 5 minutes. The product was centrifuged at 5000 rpm for 15 minutes, and the supernatant was freeze-dried to obtain WPI powder. This product is referred to as WPI in the results below.

[0051] Comparative Example 2 WPI hydrolysate and its preparation

[0052] In this comparative example, a WPI for comparison (WPI that was simply enzymatically hydrolyzed without glycosylation modification) was prepared in the following steps:

[0053] 5 g of WPI was dissolved in 100 mL of water, the pH was adjusted to 8, and 0.06 g of trypsin was added. The mixture was then hydrolyzed in a shaker at 37°C for 30 minutes. The hydrolysis was then terminated by heating in a 100°C water bath for 5 minutes to obtain a WPI hydrolysate. The WPI hydrolysate was then centrifuged at 5000 rpm for 15 minutes. The supernatant was freeze-dried to obtain a WPI hydrolysate powder. This product is referred to as YWPH in the results below.

[0054] Comparative Example 3 Cross-linked WPI enzymatic hydrolysate and its preparation

[0055] In this comparative example, a WPI modification (non-chitosan-modified glycosylated enzymatically hydrolyzed WPI) was prepared for comparison, and the steps were as follows:

[0056] Step 1: Dissolve 5g of whey protein isolate in 100mL of water, adjust the pH to 8, add 0.06g of trypsin, and hydrolyze in a shaker at 37°C for 30 minutes. Then, heat in a 100°C water bath for 5 minutes to stop the hydrolysis. The product is centrifuged at 5000rpm for 15 minutes, and the supernatant is freeze-dried to obtain WPI enzymatic hydrolysate powder.

[0057] Step 2: Dissolve the WPI hydrolysate powder in water at a concentration of 3% (w / v, 3g / 100mL). Add TGase (10U / g WPI hydrolysate) to the solution and react at 37°C, pH 7.5 for 3 hours. After the reaction, inactivate the enzyme in an 85°C water bath for 5 minutes, cool to room temperature, and freeze-dry to obtain cross-linked WPI hydrolysate powder. This product, glycosylated enzymatically hydrolyzed WPI without chitosan modification, is designated CG-YWPH in the results below.

[0058] Then, the WPI-related products of Examples 1-4 and Comparative Examples 1-3 were tested for functionality and antioxidant properties, respectively. The test methods are as follows:

[0059] 1. Surface hydrophobicity (H0) determination and results

[0060] The surface hydrophobicity (H0) of the products obtained in Examples 1-4 and Comparative Examples 1-3 was measured using an F-7100 fluorescence spectrophotometer, and ANS (1-cyclohexyl-3-sulfonic acid acrylate) was used as a fluorescent probe for analysis. The concentration of all samples was diluted to 0.1 mg / mL to 0.5 mg / mL, 4 mL of each sample was taken, and 20 μL of ANS solution (8 mmol / L) was added. After the samples were mixed, they were statically reacted at room temperature for 5 minutes, and the fluorescence intensity was measured at wavelengths of 370 nm (excitation) and 490 nm (emission). Linear regression analysis was used to obtain a graph showing the relationship between fluorescence intensity and protein concentration, and the initial slope was used as an indicator of protein hydrophobicity. The experimental results are shown in Figure 1 .

[0061] like Figure 1 As shown, the bar graphs of the ratios 1: 1, 2: 1, 3: 1, and 4: 1 correspond to the glycosylated WPI hydrolysates of Examples 4, 3, 2, and 1, respectively. As can be seen from the results, Comparative Example 2 has a higher surface hydrophobicity than Comparative Example 1. This is because enzymatic hydrolysis destroys the structure of the protein, exposing the hydrophobic amino acids to a polar environment, thereby increasing the surface hydrophobicity. The reduction in surface hydrophobicity in Comparative Example 3 compared to Comparative Example 2 is due to the Tgase enzyme causing covalent cross-linking of protein amino acids, forming aggregates, and the hydrophobic amino acids are again restricted within the aggregates, thereby causing a reduction in surface hydrophobicity. Chitosan oligosaccharides are water-soluble molecules with hydrophilic groups (including -OH and -NH2), and the connection of these functional groups covers the exposed hydrophobic groups, thereby reducing the surface hydrophobicity of glycosylated WPI nanoparticles. Therefore, the surface hydrophobicity of Examples 1-4 is all reduced. Due to the different addition ratios of whey isolate protein hydrolysate and chitosan oligosaccharide, Example 3 has the lowest surface hydrophobicity.

[0062] 2. Emulsification test and results

[0063] The samples of Examples 1-4 and Comparative Examples 1-3 were prepared as aqueous solutions (all at a concentration of 1 mg / mL) using deionized water and mixed with soybean oil at a ratio of 3:1 (v / v). The mixture was homogenized using a high-speed blender at 12,000 rpm for 1 minute. Aliquots of 0.5 mL of the liquid were removed from the bottom and diluted into 20 mL of a 0.1% (w / v, g / 100 mL) SDS (sodium dodecyl sulfate) solution at 0 and 10 minutes. The absorbance was measured at 500 nm. The emulsifying ability (EAI) and emulsion stability (ESI) were calculated using the following formulas:

[0064]

[0065] Where N is the dilution factor (N=100), C is the concentration of the protein solution (g / mL), and L is the optical path (L=1 cm). Oil phase fraction A0 and A 10 are the absorbance of the sample at 0 min and 10 min, respectively.

[0066] The experimental results are shown in Figure 2-Figure 3 .like Figure 2 and Figure 3 As shown, the bar charts for the 1:1, 2:1, 3:1, and 4:1 ratios correspond to the glycosylated WPI hydrolysates of Examples 4, 3, 2, and 1, respectively. The results show that compared to WPI, YWPH, and CG-YWP, the emulsifying properties of the samples of Examples 1-4 were significantly improved, and the emulsion stability of Examples 1-3 was significantly improved. Among them, Example 2 achieved the best emulsifying properties and emulsion stability, at 107.6% and 62.15%, respectively. Therefore, when the mass ratio of WPI hydrolysate powder to chitosan oligosaccharide is between 2 and 4:1, the glycosylated chitosan-modified WPI hydrolysate exhibits excellent emulsifying properties and emulsion stability, making it suitable as an emulsifier. However, it is clear that more chitosan content is not necessarily better, nor is less chitosan necessarily better. At a ratio of 4:1, the emulsifying properties and emulsion stability of the samples show a clear downward trend.

[0067] 3. Foaming test and results

[0068] The samples of Examples 1-4 and Comparative Examples 1-3 were prepared into aqueous solutions (all at a concentration of 1 mg / mL) using deionized water. 15 mL of the sample was placed in a 50 mL centrifuge tube and foamed at 10,000 rpm for 2 minutes using a high-speed homogenizer. The total volume (V1) at the dwell time was recorded. The foaming performance was calculated using the formula:

[0069]

[0070] Where: V0: initial volume of protein solution; V1: total volume of solution and foam after 2 min of homogenization; V2: total volume of solution and foam after 30 min of standing.

[0071] The experimental results are shown in Figure 4-Figure 5 .like Figure 4 and Figure 5 As shown, the bar graphs for the 1:1, 2:1, 3:1, and 4:1 ratios correspond to the glycosylated WPI hydrolysates of Examples 4, 3, 2, and 1, respectively. The results show that compared to WPI, YWPH, and CG-YWP, the foaming properties of the samples of Examples 1 and 2 were significantly improved, while the foaming stability of the samples of Examples 1-4 was significantly improved. Among them, the sample of Example 2 had the best foaming properties and foaming stability.

[0072] 4. DPPH free radical scavenging activity assay and results

[0073] The samples of Examples 1-4 and Comparative Examples 1-3 were prepared as aqueous solutions (all at a concentration of 1 mg / mL) using deionized water and mixed with a 0.2 mmol / L DPPH solution in anhydrous ethanol at a ratio of 1:1 (v / v). The mixture was reacted in the dark at 25°C for 30 minutes. The absorbance of the mixture at 517 nm was measured using a spectrophotometer. The DPPH free radical scavenging ability was calculated using the following formula:

[0074]

[0075] Among them, A a is the absorbance of the control group, which was prepared by using distilled water instead of the sample; A b is the absorbance of the sample.

[0076] The experimental results are shown in Figure 6 .like Figure 6 As shown, the bar graphs with ratios of 1:1, 2:1, 3:1, and 4:1 correspond to the glycosylated WPI hydrolysates of Examples 4, 3, 2, and 1, respectively. The results show that the DPPH radical scavenging activity of the samples of Examples 1-4 was significantly improved (relative to WPI), with only the sample of Example 2 significantly higher than that of WPI, YWPH, and CG-YWP. This also shows that not all chitosan-modified WPI hydrolysates can achieve DPPH radical scavenging activity higher than that of YWPH and CG-YWP.

[0077] 5. ABTS+ free radical scavenging activity assay and results

[0078] ABTS solution (7 mmol / L) and K2S2O8 solution (2.45 mmol / L) were mixed in a 2:1 volume ratio and allowed to react in the dark for 12–16 hours to generate stable ABTS+ free radicals. The ABTS+ solution was then diluted to an absorbance of 0.7 ± 0.02 at 734 nm. The sample was dissolved in deionized water (0.1 mg / mL), and 0.2 mL of the sample solution was mixed with 3.8 mL of the ABTS+ solution and allowed to react in the dark for 5 minutes. The absorbance was measured at 734 nm using a spectrophotometer. The ABTS+ free radical scavenging activity was calculated as follows:

[0079]

[0080] Where Ao is the absorbance of the control group, which was prepared using distilled water instead of the sample; A is the absorbance of the sample.

[0081] The experimental results are shown in Figure 7 .like Figure 6As shown, the bar graphs with ratios of 1:1, 2:1, 3:1, and 4:1 correspond to the glycosylated WPI hydrolysates of Examples 4, 3, 2, and 1, respectively. As can be seen from the results, the samples of Examples 1-4 have improved ABTS+ free radical scavenging activity compared to WPI. However, only the samples of Examples 1-3 have significantly higher ABTS+ free radical scavenging activity than YWPH and CG-YWP; there is no significant difference in the ABTS+ free radical scavenging activity of the samples of Examples 1-3, but they still show a trend of first increasing and then decreasing. This is similar to the DPPH free radical scavenging activity, further proving that not all chitosan-modified WPI hydrolysates can achieve antioxidant properties higher than YWPH and CG-YWP.

[0082] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the technical principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A glycosylated whey protein hydrolysate, characterized in that: The glycosylated whey protein hydrolysate is a whey protein hydrolysate modified by glycosylation with chitosan. The preparation method of the glycosylated whey protein hydrolysate comprises the following steps: S1: preparing whey protein isolate protein hydrolysate: hydrolyzing whey protein isolate with trypsin to obtain whey protein isolate protein hydrolysate; S2 glycosylation modification of whey protein isolate hydrolysate: The whey protein isolate hydrolysate is glycosylated using chitosan oligosaccharide and TGase enzyme to obtain chitosan glycosylated whey protein isolate hydrolysate.

2. The glycosylated whey protein hydrolysate according to claim 1, characterized in that The preparation method of the whey isolated protein hydrolysate comprises the following steps: Dissolve whey protein isolate in water, adjust the pH to 7-9, add trypsin, the amount of trypsin added is 0.01g-0.02g per gram of whey protein isolate, the enzymatic hydrolysis temperature is 32℃-40℃, and the enzymatic hydrolysis time is 20min-40min.

3. The glycosylated whey protein hydrolysate according to claim 2, wherein The amount of trypsin added was 0.012 g of trypsin per g of whey protein isolate, the enzymolysis temperature was 37° C., and the enzymolysis time was 30 min.

4. The glycosylated whey protein hydrolysate according to claim 1, wherein The step S2 comprises the following steps: adding chitosan oligosaccharide and TGase enzyme to the whey protein isolate hydrolysate solution, and performing an enzymatic glycosylation reaction at 32° C. to 40° C. and pH 7 to 8; the amount of chitosan oligosaccharide added is 20% to 100% of the mass of the whey protein isolate hydrolysate.

5. The glycosylated whey protein hydrolysate according to claim 4, characterized in that The mass volume percentage concentration of the whey isolate protein hydrolysate solution is 2% to 5%.

6. The glycosylated whey protein hydrolysate according to claim 5, characterized in that The amount of TGase added is 8-12 U / g whey protein hydrolysate.

7. Use of the glycosylated whey protein hydrolysate according to claim 6 in the preparation of an antioxidant, characterized in that: The added amount of the chitosan oligosaccharide is 25% to 50% of the mass of the whey isolate protein hydrolysate.

8. The use according to claim 7, characterized in that The added amount of the chitosan oligosaccharide is 30% to 35% of the mass of the whey isolate protein hydrolysate.

9. Use of the glycosylated whey protein hydrolysate according to claim 6 in the preparation of a food emulsifier, characterized in that: The added amount of the chitosan oligosaccharide is 25% to 50% of the mass of the whey isolate protein hydrolysate.

10. Use of the glycosylated whey protein hydrolysate according to claim 6 in preparing a food foaming agent, characterized in that: The added amount of the chitosan oligosaccharide is 25% to 35% of the mass of the whey isolate protein hydrolysate.

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