Milk-based spice with natural nut flavor as well as preparation method and application of milk-based spice

Abstract

The invention relates to a milk-based spice with natural nut flavor as well as a preparation method and application thereof, and belongs to the technical field of preparation of dairy products. The milk-based spice with the natural nut flavor is obtained by adding the enzymolysis product and the anhydrous butter back into the protein solution; the enzymolysis product is obtained by performing enzymolysis on anhydrous cream and lipase. According to the method disclosed by the invention, a natural cream base material is taken as a starting point, and the natural cream flavor base which is high in strength, mellow in taste and free from foreign flavor is safely and controllably constructed through a biological enzymolysis technology. A natural nut precursor substance is ingeniously introduced, reaction conditions are accurately controlled, and pure and rich natural nut flavor is generated in situ in a milk-based system instead of simple addition. Finally, the cooperation and integration of a cream flavor forming way and a nut flavor forming way are realized, so that the two flavors are mutually infiltrated and promoted in the forming process, and a harmonious, full, lasting and high-grade composite flavor whole body is formed.

Description

Technical Field

[0001] This invention relates to the field of dairy product preparation technology, and in particular to a natural nut-flavored milk-based flavoring, its preparation method, and its application. Background Technology

[0002] Milk-based flavorings are a class of food flavorings with a rich, realistic milky aroma, obtained by processing dairy products or milk components (such as cream, butter, skim milk powder, whey powder, casein, etc.) through physical, chemical, or biological means. They are widely used in ice cream, chocolate, baked goods, candy, beverages, dairy products, desserts, and convenience foods, serving as key ingredients for enhancing product flavor and increasing consumer appetite and satisfaction. With the rapid development of the global food industry and consumers' pursuit of "clean label," natural, and healthy foods, the market demand for rich-flavored, natural, and high-quality milk-based flavorings is increasingly strong.

[0003] Among the many flavors favored by consumers, creamy and nutty flavors hold extremely important positions. Creamy flavors, with their rich, full-bodied, warm, and slightly sweet characteristics, are the soul of food; while nutty flavors (such as hazelnuts, almonds, walnuts, and macadamia nuts), with their unique roasted, caramelized, and oily aromas, give food a rich, elegant, and pleasant sensory experience. Organically combining these two top-tier flavors to create complex dairy-based flavorings that possess both a rich creamy base and typical nutty characteristics has become an important research and development direction in the food flavoring industry, with enormous market potential.

[0004] However, existing technologies face numerous technical challenges and bottlenecks in achieving pure, rich, and natural cream and nut flavors, especially in efficiently and stably blending them into the same milk-based system: the enhancement and realism of natural cream flavor, the acquisition and stability of natural nut flavor, and the synergistic effect and fusion of cream and nut flavors. The market urgently needs an innovative preparation method that can overcome these technical bottlenecks.

[0005] Existing dairy fat flavorings are mostly obtained through enzymatic hydrolysis of whey or butter, but these methods suffer from problems such as high substrate impurities, incomplete hydrolysis, and a lack of concentration of flavor compounds. Anhydrous butter (AMF), as a high-purity milk fat, contains almost no protein or lactose and is more suitable for lipase-directed catalysis in a low-water environment, potentially producing characteristic flavor compounds different from traditional systems. Furthermore, there are currently few systematic studies on lipase-directed reactions using anhydrous butter as a substrate, and there is a particular lack of publicly available reports on the characteristic aroma spectra and industrial applications of its products. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention provides a natural nut-flavored milk-based flavoring, its preparation method, and its application. This invention uses anhydrous butter as a substrate, and obtains a milk-based flavoring with nut, butter, and caramel flavors through lipase action, which is then added back into an emulsion system. This invention aims to provide a novel method for preparing a milk-based flavoring that perfectly blends the flavors of natural butter and nuts, filling a gap in existing technology and meeting the urgent demand of the high-end food manufacturing industry for top-quality natural compound flavor ingredients.

[0007] This invention is achieved through the following technical solution:

[0008] The first objective of this invention is to provide a natural nut-flavored milk-based flavoring, obtained by adding an enzymatic hydrolysate and anhydrous butter back into a protein solution; the enzymatic hydrolysate is obtained by enzymatic hydrolysis of anhydrous butter and lipase.

[0009] In one embodiment of the present invention, the lipase is lipase A12.

[0010] In one embodiment of the present invention, the amount of lipase added is 100 U / g-150 U / g.

[0011] In one embodiment of the present invention, the enzymatic hydrolysis conditions are: 39℃-41℃ for 15 min-90 min;

[0012] The enzymatic hydrolysis speed is 80 rpm-100 rpm.

[0013] In one embodiment of the present invention, the protein solution is obtained by dissolving protein powder in water; the mass concentration of the protein solution is 5%-5.5%.

[0014] In one embodiment of the present invention, the mass ratio of the enzymatic hydrolysis product, anhydrous butter and protein solution is 3:6:91.

[0015] In one embodiment of the present invention, the volatile flavor compounds of the milk-based flavoring include acid compounds, aldehyde compounds, ketone compounds, alcohol compounds, ester compounds and alkene compounds;

[0016] The acid compounds include nonanoic acid, octanoic acid, hexanoic acid, and n-decanoic acid; the aldehyde compounds include hexanal, E-2-heptanal, E-2-nonenal, Z-2-heptanal, (Z)-2-butenal, benzaldehyde, and 3-methylbutanal; the ketone compounds include 2,3-octanedione, acetone, 2-heptanone, 2-undecanone, 2-nonanone, and 4-nonanone; the alcohol compounds include ethanol, 1-octanol, and 1-octen-3-ol; the ester compounds include ethyl octanoate; and the alkene compounds include D-limonene.

[0017] In one embodiment of the present invention, the fat concentration in the milk-based flavoring is 9%-11%.

[0018] A second objective of this invention is to provide a method for preparing the aforementioned milk-based flavoring, comprising the following steps:

[0019] (1) Heat the reaction substrate to melt it, add water, cool and stir; then add lipase to hydrolyze and inactivate the enzyme; remove water after separation to obtain the hydrolysis product;

[0020] (2) Add the enzymatic hydrolysis product obtained in step (1) and anhydrous butter to the protein solution, add stabilizer and shear to obtain a natural nut flavored milk flavor.

[0021] In one embodiment of the present invention, the heating and melting temperature is 60°C-65°C;

[0022] The mass ratio of the reaction substrate to water is 4:1-5:1;

[0023] The enzymatic hydrolysis conditions are: 80 rpm-100 rpm, 39℃-41℃ for 15 min-90 min;

[0024] The lipase is lipase A12, and the amount added is 100 U / g-150 U / g;

[0025] The mass concentration of the protein aqueous solution is 5%-5.5%;

[0026] The enzyme inactivation conditions are: enzyme inactivation at 80℃-85℃ for 15 min-20 min.

[0027] The shearing conditions are: 9000 rpm-10000 rpm, 1 min-3 min.

[0028] The stabilizer is selected from xanthan gum and / or carboxymethyl cellulose; the mass concentration of the stabilizer is 0.4%-0.6%.

[0029] A third objective of this invention is to provide the application of the aforementioned milk-based flavoring in food flavorings, dairy products, or plant-based beverages.

[0030] This invention uses natural milk base materials as a starting point and employs bio-enzymatic hydrolysis technology to safely and controllably construct a natural cream flavor base that is high in strength, rich in taste, and free of off-flavors. It cleverly introduces natural nut precursors and precisely controls the reaction conditions to generate a pure and rich natural nut flavor in situ within the milk base system, rather than simply adding them externally. Ultimately, it achieves synergy and integration between the cream flavor formation pathway and the nut flavor formation pathway, allowing the two flavors to permeate and promote each other during the formation process, forming a harmonious, full-bodied, long-lasting, and sophisticated complex flavor whole.

[0031] The technical solution of the present invention has the following advantages compared with the prior art:

[0032] (1) This invention provides a natural nut-flavored milk-based flavoring, its preparation method, and its application. The milk-based flavoring obtained by this invention has the following characteristics: enhanced milk flavor, natural nut flavor, and improved aroma harmony.

[0033] (2) The advantages of this invention are that the milk-based flavoring is enhanced with a natural nut flavor, the aroma is improved, and the enzymatic hydrolysis conditions of Lipase A “Amano”12 lipase are optimized, resulting in the most intense milk-based flavoring with a nut flavor. Attached Figure Description

[0034] To make the content of this invention easier to understand, the invention will be further described in detail below with reference to specific embodiments and accompanying drawings, wherein:

[0035] Figure 1 These are the acid values ​​of different enzymatic hydrolysates in this invention;

[0036] Figure 2 These are the sensory evaluation results of the emulsions with different milk-based flavorings added in this invention;

[0037] Figure 3 This refers to the acid value of lipase A12 at different enzymatic hydrolysis times in this invention;

[0038] Figure 4 This is a sensory evaluation of emulsions with added milk flavoring at different enzymatic hydrolysis times of lipase A12 in this invention;

[0039] Figure 5-A This is the principal component analysis diagram of SPME-GC-MS PLS-DA analysis of volatile flavors in this invention;

[0040] Figure 5-B This is a VIP value graph of PLS-DA analysis of volatile flavors by SPME-GC-MS in this invention;

[0041] Figure 5-C This is a bipolar plot of PLS-DA analysis of volatile flavors using SPME-GC-MS in this invention;

[0042] Figure 5-D This is a graph showing the predictive and accuracy of PLS-DA analysis of volatile flavors using SPME-GC-MS in this invention;

[0043] Figure 5-EThis is a displacement test analysis diagram of PLS-DA analysis of volatile flavor SPME-GC-MS in this invention;

[0044] Figure 6-A This is the principal component analysis diagram of PLS-DA analysis of volatile flavors by HS-GC-IMS in this invention;

[0045] Figure 6-B This is a VIP value graph of PLS-DA analysis of volatile flavors by HS-GC-IMS in this invention;

[0046] Figure 6-C This is a bipolar plot of PLS-DA analysis of volatile flavors by HS-GC-IMS in this invention;

[0047] Figure 6-D This is a graph showing the predictive and accuracy of PLS-DA analysis of volatile flavors by HS-GC-IMS in this invention;

[0048] Figure 6-E This is a displacement test analysis diagram of PLS-DA analysis of volatile flavors by HS-GC-IMS in this invention. Detailed Implementation

[0049] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0050] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, and the materials and reagents used are commercially available.

[0051] The following food-grade lipases are from Amano and Novozymes.

[0052] The xanthan gum and carboxymethyl cellulose were sourced from Anhui Runtian Biotechnology Co., Ltd.

[0053] Enzyme activity is defined in units of U / g: one unit of enzyme activity is defined as 1 μg of oleic acid produced per minute per gram of protein from the breakdown of substrate.

[0054] Example 1:

[0055] This embodiment provides a method for preparing a natural nut-flavored milk-based flavoring, the specific steps of which are as follows:

[0056] 50 g of anhydrous butter was heated and melted at 65°C, then 10 g of deionized water was added, the temperature was lowered to 40°C, and the mixture was stirred at 100 rpm at 40°C for 10 min. 150 U / g Amano lipase A “Amano” 12 was added, and the mixture was enzymatically hydrolyzed at 100 rpm at 40°C for 60 min. The enzyme was then inactivated at 85°C for 15 min. After the butter was separated into layers, the enzymatic hydrolysate was obtained.

[0057] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and emulsion 4 with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min, which is a natural nut-flavored milk-based flavoring.

[0058] Example 2:

[0059] This embodiment provides a method for preparing a natural nut-flavored milk-based flavoring, the specific steps of which are as follows:

[0060] First, melt 50 g of anhydrous butter at 65°C, then add 10 g of deionized water, cool to 40°C, and stir at 100 rpm at 40°C for 10 min; add 150 U / g Amano lipase A “Amano” 12, and enzymatically hydrolyze at 100 rpm at 40°C for 15 min; inactivate the enzyme at 85°C for 15 min, and obtain the enzymatic hydrolysate after the butter separates into layers.

[0061] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min, which is a natural nut-flavored milk-based flavoring.

[0062] Example 3:

[0063] This embodiment provides a method for preparing a natural nut-flavored milk-based flavoring, the specific steps of which are as follows:

[0064] First, melt 50 g of anhydrous butter at 65°C, then add 10 g of deionized water, cool to 40°C, and stir at 100 rpm at 40°C for 10 min; add 150 U / g Amano lipase A “Amano” 12, and enzymatically hydrolyze at 100 rpm at 40°C for 30 min; inactivate the enzyme at 85°C for 15 min, and obtain the enzymatic hydrolysate after the butter separates into layers.

[0065] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min, which is a natural nut-flavored milk-based flavoring.

[0066] Example 4:

[0067] This embodiment provides a method for preparing a natural nut-flavored milk-based flavoring, the specific steps of which are as follows:

[0068] First, melt 50 g of anhydrous butter at 65°C, then add 10 g of deionized water, cool to 40°C, and stir at 100 rpm at 40°C for 10 min; add 150 U / g Amano lipase A “Amano” 12, and enzymatically hydrolyze at 100 rpm at 40°C for 45 min; inactivate the enzyme at 85°C for 15 min, and obtain the enzymatic hydrolysate after the butter separates into layers.

[0069] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min, which is a natural nut-flavored milk-based flavoring.

[0070] Example 5:

[0071] This embodiment provides a method for preparing a natural nut-flavored milk-based flavoring, the specific steps of which are as follows:

[0072] First, melt 50 g of anhydrous butter at 65°C, then add 10 g of deionized water, cool to 40°C, and stir at 100 rpm at 40°C for 10 min; add 150 U / g Amano lipase A “Amano” 12, and hydrolyze at 100 rpm at 40°C for 90 min; inactivate the enzyme at 85°C for 15 min, and obtain the hydrolysate after the butter separates into layers.

[0073] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min, which is a natural nut-flavored milk-based flavoring.

[0074] Comparative Example 1:

[0075] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0076] 50 g of anhydrous butter was heated and melted at 65°C, then 10 g of deionized water was added, the temperature was lowered to 40°C, and the mixture was stirred at 100 rpm at 40°C for 10 min; 150 U / g Novozymes lipase Palatase 20000L was added, and the mixture was enzymatically hydrolyzed at 100 rpm at 40°C for 60 min; the enzyme was then inactivated at 85°C for 15 min, and the hydrolysate was obtained after the butter was separated into layers.

[0077] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0078] Comparative Example 2:

[0079] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0080] 50 g of anhydrous butter was heated and melted at 65°C, then 10 g of deionized water was added, the temperature was lowered to 40°C, and the mixture was stirred at 100 rpm at 40°C for 10 min; 150 U / g Novozymes lipase Lipozyme TL100L was added, and the mixture was enzymatically hydrolyzed at 100 rpm at 40°C for 60 min; the enzyme was then inactivated at 85°C for 15 min, and the hydrolysate was obtained after the butter was separated into layers.

[0081] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0082] Comparative Example 3:

[0083] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0084] 50 g of anhydrous butter was heated and melted at 65°C, then 10 g of deionized water was added, the temperature was lowered to 40°C, and the mixture was stirred at 100 rpm at 40°C for 10 min; 150 U / g Novozymes lipase Lipozyme TLIM was added, and the mixture was enzymatically hydrolyzed at 100 rpm at 40°C for 60 min; the enzyme was then inactivated at 85°C for 15 min, and the hydrolysate was obtained after the butter was separated into layers.

[0085] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0086] Comparative Example 4:

[0087] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0088] First, melt 50 g of anhydrous butter at 65°C, then add 10 g of deionized water, cool to 40°C, and stir at 100 rpm at 40°C for 10 min; add 150 U / g Novozymes lipase Lipozyme RM, and hydrolyze at 100 rpm at 40°C for 60 min; inactivate the enzyme at 85°C for 15 min, and obtain the hydrolysate after the butter separates into layers.

[0089] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0090] Comparative Example 5:

[0091] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0092] First, melt 50 g of anhydrous butter at 65°C, then add 10 g of deionized water, cool to 40°C, and stir at 100 rpm at 40°C for 10 min; add 150 U / g Novozym 435 lipase and hydrolyze at 100 rpm at 40°C for 60 min; inactivate the enzyme at 85°C for 15 min, and obtain the hydrolysate after the butter separates into layers.

[0093] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0094] Comparative Example 6:

[0095] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0096] First, melt 50 g of anhydrous butter at 65°C, then add 10 g of deionized water, cool to 40°C, and stir at 100 rpm at 40°C for 10 min; add 150 U / g Novozymes lipase Lipozyme 435, and hydrolyze at 100 rpm at 40°C for 60 min; inactivate the enzyme at 85°C for 15 min, and obtain the hydrolysate after the butter separates into layers.

[0097] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0098] Comparative Example 7:

[0099] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0100] 50 g of anhydrous butter was heated and melted at 65°C, then 10 g of deionized water was added, the temperature was lowered to 40°C, and the mixture was stirred at 100 rpm at 40°C for 10 min; 150 U / g Amano lipase AY “Amano” 30SD was added, and the mixture was enzymatically hydrolyzed at 100 rpm at 40°C for 60 min; the enzyme was then inactivated at 85°C for 15 min, and the hydrolysate was obtained after the butter was separated into layers.

[0101] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0102] Comparative Example 8:

[0103] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0104] First, melt 50 g of anhydrous butter at 65°C, then add 10 g of deionized water, cool to 40°C, and stir at 100 rpm at 40°C for 10 min; add 150 U / g Amano lipase DF “Amano”15, and enzymatically hydrolyze at 100 rpm at 40°C for 60 min; inactivate the enzyme at 85°C for 15 min, and obtain the enzymatic hydrolysate after the butter separates into layers.

[0105] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 3% enzymatic hydrolysate and 6% anhydrous butter were added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0106] Comparative Example 9:

[0107] This comparative example provides a method for preparing a milk-based flavoring agent, the specific steps of which are as follows:

[0108] Protein powder (73% (w / w) protein) was dissolved in distilled water to prepare a 5.5% (w / w) protein solution; 9% anhydrous butter was added to the protein solution; stabilizers (0.3% xanthan gum and 0.2% carboxymethyl cellulose) were added; and an emulsion with a fat content of 9% (w / w) was prepared under high-speed shearing at 10,000 rpm for 1 min.

[0109] Performance testing

[0110] (1) The acid value (AV) of the enzymatic hydrolysates obtained in Examples 1-5 and Comparative Examples 1-8 was determined according to the method specified in GB 5009.229-2016. The results are as follows: Figure 1 As shown, different lipases have varying efficiencies in breaking down triglycerides into free fatty acids, resulting in different acid values.

[0111] (2) The aroma of the emulsions obtained in Example 1 and Comparative Examples 1-9 was compared using sensory evaluation. The results are as follows: Figure 2 As shown in Table 1, the sensory evaluation scores were as follows. It can be seen that the three sensory indicators of milk sweetness, milk aroma, and creaminess were consistent in the evaluation of these nine enzyme hydrolysates added to the emulsion. In most samples, enzymatic hydrolysis did not improve their milk aroma, milk sweetness, or creaminess. N-435 and L-TL100L exhibited a noticeable putrid odor, and their aroma harmony and acceptability were significantly reduced. A12 enzyme was found to significantly enhance the milk aroma, milk sweetness, creaminess, and fatiness, and added a nutty aroma, while also exhibiting the highest aroma harmony and acceptability.

[0112] Table 1. Sensory evaluation scores of emulsions containing nine different lipase hydrolysates.

[0113]

[0114] Continued from Table 1

[0115] Continued from Table 1

[0116]

[0117] (3) The acid value of the enzymatic hydrolysates obtained in Examples 1-5 was measured, and the results are as follows: Figure 3 As shown, the acid value of the hydrolysate increases with the extension of the enzymatic hydrolysis time, and the hydrolysis rate increases almost linearly, indicating that the lipase maintains high activity during this process.

[0118] (4) Sensory evaluation was performed on the emulsions obtained in Examples 1-5 and Comparative Example 9, and the results are as follows: Figure 4 As shown in Table 2, the sensory evaluation scores were obtained from the samples. After A12 enzymatic hydrolysis, all aroma indicators were significantly improved overall. At a hydrolysis time of 30 min, the milk, cream, butter, and fat aromas of the emulsion were the highest, with improved aroma harmony and acceptability compared to the unhydrolyzed sample. Cheese and whey flavors decreased. Furthermore, a nutty flavor was developed after enzymatic hydrolysis, reaching its peak intensity at 45 min.

[0119] Table 2 Sensory evaluation scores of the added emulsions of hydrolysates at different enzymatic hydrolysis times

[0120]

[0121] Continued from Table 2

[0122] Continued from Table 2

[0123]

[0124] (5) SPME-GC-MS results were analyzed for the emulsions obtained in Examples 1-5 and Comparative Example 9.

[0125] The results of the identification of volatile flavor compounds are shown in Tables 3-8. A total of 50 volatile flavor compounds were detected, divided into 6 groups: 7 alcohols, 9 ketones, 8 acids, 12 esters, 10 aldehydes, and 5 alkenes. Among them, 18 substances were detected in the undigested samples, and 21 (15 min), 23 (30 min), 18 (45 min), 19 (60 min), and 25 (90 min) substances were detected after enzymatic hydrolysis. Enzymatic hydrolysis increased the variety of flavor compounds.

[0126] Table 3. Alcohols in volatile flavor compounds identified by SPME-GC-MS

[0127]

[0128] Table 4. Ketones of volatile flavor compounds identified by SPME-GC-MS

[0129]

[0130] Table 5. Acids of volatile flavor compounds identified by SPME-GC-MS

[0131]

[0132] Table 6 Aldehydes of volatile flavor compounds identified by SPME-GC-MS

[0133]

[0134] Table 7 Aldehydes of volatile flavor compounds identified by SPME-GC-MS

[0135]

[0136] Table 8. Olefins of volatile flavor compounds identified by SPME-GC-MS

[0137]

[0138] (6) PLS-DA analysis was performed on the volatile flavor (SPME-GC-MS) data. Figure 5-A The samples were found to be significantly different from those that were not enzymatically hydrolyzed, indicating that the flavor of the hydrolyzed samples changed significantly. Furthermore, 15 flavor compounds with a VIP value > 1 were identified. Figure 5-B ),Depend on Figure 5-C The bipolar plot shows that after adding the milk-based flavoring after A12 enzymatic hydrolysis, the levels of 2-heptanone and lactones in the emulsion increased significantly, resulting in nutty and milky aromas. Figure 5-D and Figure 5-E This indicates that the PLS-DA model has high confidence and high reliability and predictability. After OVA calculations, as shown in Table 9, a total of 22 volatile flavor compounds were identified, including 4 acids, 7 aldehydes, 6 ketones, 3 alcohols, 1 ester, and 1 alkene. Nonanoic acid, benzaldehyde, 4-nonanone, ethanol, 1-octanol, 1-octen-3-ol, and D-limonene were not previously identified in dairy products.

[0139] Nonanoic acid, octanoic acid, hexanoic acid, and decanoic acid constitute typical cheese, oily, and waxy aromas; hexanal, E-2-heptanal, E-2-nonenal, Z-2-heptanal, (Z)-2-butenal, benzaldehyde, and 3-methylbutanal constitute grassy, ​​fatty, and nutty aromas; 2,3-octanedione, acetone, 2-heptanone, 2-undecaneone, 2-nonenone, and 4-nonenone have creamy and fruity aromas; ethanol, 1-octanol, 1-octen-3-ol, ethyl octanoate, and D-limonene have fruity and citrus aromas.

[0140] Table 9 OVA values ​​of flavor compounds

[0141]

[0142] HS-GC-IMS performed well in identifying small molecules in butter, including compounds with carbon chains ranging from C2 to C9. As shown in Table 10, a total of 77 volatile flavor compounds were detected, categorized into 7 groups: 17 esters, 23 alcohols, 19 aldehydes, 14 ketones, 2 acids, 1 alkene, and acetoin. HS-GC-IMS is more suitable for detecting small molecules and trace compounds, detecting more alcohols, esters, and aldehydes compared to SPME-GC-MS, but not fewer acids.

[0143] Table 10 Volatile flavor compounds identified by HS-GC-IMS

[0144]

[0145] Continued from Table 10

[0146]

[0147] Continued from Table 10

[0148]

[0149] PLS-DA analysis was performed on the data identified by HS-GC-IMS. Figure 6-A As shown, similar to SPME-GC-MS, the samples after enzymatic digestion are clearly distinguishable from those without, but the distinguishing effect is lower for different digestion times. Figure 6-B As shown, a total of 15 key substances were identified. Both SPME-GC-MS and PLS-DA predictions showed the presence of 2-heptanone. Combined predictions from both methods revealed acids including acetic acid, hexanoic acid, octanoic acid, decanoic acid, and alcohol-aldehyde-ether compounds. Figure 6-C It can be seen that the significant increase in 2-heptanone compounds gives the sample a distinct nutty and creamy aroma. Figure 6-D and Figure 6-E It can be seen that the model fit has high confidence and has a certain degree of interpretability and predictability.

[0150] Analysis of volatile flavor compounds by combining SPME-GC-MS and HS-GC-IMS showed that the enzymatically hydrolyzed milk-based flavoring had a stronger flavor, enhancing its milky and nutty aromas. This molecular explanation provided the reason for the flavor enhancement, and was consistent with the results of sensory evaluation.

[0151] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A natural nut-flavored milk-based flavoring, characterized in that, It is obtained by adding the enzymatic hydrolysis product and anhydrous butter back into the protein solution; The enzymatic hydrolysis product was obtained by enzymatic hydrolysis of anhydrous butter with lipase.

2. The milk-based flavoring according to claim 1, characterized in that, The lipase is lipase A12.

3. The milk-based flavoring according to claim 1, characterized in that, The amount of lipase added is 100 U / g-150 U / g.

4. The milk-based flavoring according to claim 1, characterized in that, The conditions for enzymatic hydrolysis are: 39℃-41℃ for 15min-90min; And / or, the enzymatic hydrolysis is performed at a rotation speed of 80 rpm-100 rpm.

5. The milk-based flavoring according to claim 1, characterized in that, The protein solution is obtained by dissolving protein powder in water; the mass concentration of the protein solution is 5%-5.5%.

6. The milk-based flavoring according to claim 1, characterized in that, The mass ratio of the enzymatic hydrolysis product, anhydrous butter, and protein solution is 3:6:

91.

7. The milk-based flavoring according to claim 1, characterized in that, The volatile flavor compounds of the emulsion-based flavoring include acid compounds, aldehyde compounds, ketone compounds, alcohol compounds, ester compounds, and alkene compounds; The acid compounds include nonanoic acid, octanoic acid, hexanoic acid, and n-decanoic acid; the aldehyde compounds include hexanal, E-2-heptanal, E-2-nonenal, Z-2-heptanal, (Z)-2-butenal, benzaldehyde, and 3-methylbutanal; the ketone compounds include 2,3-octanedione, acetone, 2-heptanone, 2-undecanone, 2-nonanone, and 4-nonanone; the alcohol compounds include ethanol, 1-octanol, and 1-octen-3-ol; the ester compounds include ethyl octanoate; and the alkene compounds include D-limonene.

8. The milk-based flavoring according to claim 1, characterized in that, The fat concentration in the milk-based flavoring is 9%-11% by mass.

9. The method for preparing the emulsion-based flavoring according to any one of claims 1-8, characterized in that, Includes the following steps: The reaction substrate was heated and melted, water was added, and the mixture was cooled and stirred. Then, lipase was added for enzymatic hydrolysis to inactivate the enzyme. After separation, the water was removed to obtain the enzymatic hydrolysis product. The enzymatic hydrolysate obtained in step (1) and anhydrous butter were added to the protein solution, and a stabilizer was added and sheared to obtain a natural nut-flavored milk-based flavoring.

10. The use of any one of claims 1-8 in food flavorings, dairy products or plant-based beverages.

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