A golden orange lemon maillard reaction product essence as well as a preparation method and application thereof

Abstract

The application provides a golden orange and lemon Maillard reactant essence, a preparation method and application thereof, and belongs to the technical field of essence. Egg white albumin and lysozyme in egg white are separated, mixed with butter of cow milk, subjected to enzymolysis and fermentation to obtain a desensitization fermentation hydrolysate; golden orange and lemon slices are subjected to steaming and boiling, enzymolysis and fermentation to obtain a golden orange and lemon enzyme-assisted fermentation product, which is mixed with the desensitization fermentation hydrolysate and an additive and heated to perform Maillard reaction to obtain a Maillard reactant essence, which is embedded to obtain the golden orange and lemon Maillard reactant essence. The essence has fruit aroma, milk aroma and meat aroma, has rich, realistic and mellow fragrance, maximally retains the nutritional value, is green, healthy, nutritious and safe, has unique flavor, improves appearance, color and other adverse factors, prolongs the shelf life of the essence, improves the storage stability of the essence, and has a wide application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of essence, in particular to a kind of gold orange lemon Maillard reactant essence and its preparation method and application. BACKGROUND

[0002] Gold orange fruit contains rich vitamin A and vitamin C, can prevent pigment deposition, enhance skin luster and elasticity, slow down aging, avoid skin relaxation and wrinkle. It can also prevent civilization diseases such as vascular lesions and cancer, and can regulate qi, relieve cough, invigorate the stomach, resolve phlegm, prevent asthma and bronchitis. Gold orange also contains vitamin E, which is an important nutrient for maintaining vascular health, can strengthen the elasticity of microvessels, and can be used as an auxiliary food for hypertension, vascular sclerosis and heart disease. As a health food, gold orange preserves can stimulate appetite, drinking gold orange juice can quench thirst, and drinking it with radish juice and pear juice can cure cough. 80% of the vitamin C in gold orange is concentrated in the peel, up to 200 mg per 100 g. In addition to fresh food, gold orange can also be used to make tea. One of the larger uses is to process white sugar gold orange cake, licorice gold orange cake, jam, orange peel wine, gold orange juice, etc. The peel can also be used to extract aromatic oil.

[0003] Lemon is rich in vitamin C, sugar, calcium, phosphorus, iron, vitamin B1, vitamin B2, niacin, quinine acid, citric acid, malic acid, orange glycosides, naringin, coumarin, high potassium and low sodium, etc. It is very beneficial to the human body. Lemon can quench thirst, relieve heat and stimulate appetite, prevent cardiovascular disease, and eating lemon regularly can clear heat and resolve phlegm. Lemon has antibacterial and anti-inflammatory effects, can delay aging and inhibit pigment deposition, and can eliminate morning vomiting. Lemon is a treasure all over the body and an important raw material for processing green products.

[0004] Gold orange lemon has a wide range of effects and rich nutritional value. It has the effects of invigorating the spleen and stimulating appetite, clearing heat and relieving summer-heat, and moistening the intestines and promoting defecation. Moderate intake can effectively prevent heatstroke, increase appetite, promote intestinal peristalsis, and thus assist defecation. From the perspective of nutrition, gold orange lemon is rich in fruit acid, fructose, vitamin C, cellulose, and a large amount of vitamin E, which can whiten the skin, increase the speed of defecation, and refresh the mind.

[0005] With the improvement of people's living standards, the pursuit of healthy diet and natural food is becoming more and more obvious. Making gold orange lemon reactant essence is very scientific and reasonable in terms of taste and health and nutrition.

[0006] Maillard reaction flavor has high safety, low price, natural, realistic, round and strong market demand, and becomes the hotspot of flavor research and development in recent years. With the development of food industry, the understanding and requirements of consumers on aroma are continuously improved, and safety, natural and realistic become the trend of essence development. The preparation and application of Maillard reaction flavor also face new challenges and new problems.

[0007] Firstly, color problem. Beautiful color can bring consumers happy visual enjoyment, thereby promoting people's appetite and strengthening the sensory effect of food, which is expected in food processing process, such as chocolate color, coffee color, bread color, etc. However, Maillard reaction flavor also produces the color that people do not expect due to the non-enzymatic browning black pigment. Black pigment is a mixture with structure of reducing ketone, enamine or pyrrole, containing yellow, black, brown black and orange red different color groups. These color groups make the flavor present brown black or dark black color, thereby becoming the obstacle of application in some food fields.

[0008] Secondly, viscosity problem. The material of Maillard reaction flavor contains hydrolyzed protein of animals and plants and natural food raw materials. These materials form a mixed system with poor flowability due to protein denaturation, sugar and amino acid reaction and solvent during the thermal reaction process. In product application, high viscosity food flavor has poor dispersibility due to gelation or emulsification, thereby leading to instability of the whole food system.

[0009] Finally, the preparation process is difficult due to the problem of turbidity and precipitation. Similarly, due to the inclusion of hydrolyzed protein of animals and plants and natural food raw materials in the reaction material, these materials finally precipitate in the product system, causing turbidity in appearance. Therefore, it is necessary to remove these precipitates to obtain a system uniform product. However, due to the high viscosity of the product, the process of filtration, centrifugation and other process means is bound to face process difficulties.

[0010] In summary, Maillard reaction flavor is affected by three problems of color, viscosity and precipitation. These three problems become the application obstacle of this kind of flavor. Therefore, how to obtain a natural food flavor with rich and pure flavor, safety, certain efficacy and high appearance acceptance will solve the technical problems of the present application. SUMMARY

[0011] The present application aims to provide a kind of gold orange lemon Maillard reaction flavor and its preparation method and application, it has fruit fragrance, milk fragrance and meat flavor simultaneously, fragrance is rich, realistic, mellow, fragrance attracts consumer, maximum retention its nutritional value, green and healthy, nutrition safety, flavor is unique, improve the appearance, color and other adverse factors of Maillard reaction flavor, avoid volatile flavor volatile loss, prolong the shelf life of flavor, improve its storage stability, with broad application prospect.

[0012] The technical scheme of the present application is realized as follows:

[0013] The present application provides a kind of gold orange lemon Maillard reaction flavor preparation method, egg white in egg white is separated from lysozyme, mixed with milk cream to add compound protease, enzymolysis, fermentation, obtain desensitization fermentation hydrolysate;Gold orange dry, lemon slice is cooked, enzymolysis, fermentation, obtain gold orange lemon enzyme assisted fermentation product, mix with desensitization fermentation hydrolysate, additive heating is carried out Maillard reaction, obtain Maillard reaction flavor, embedding, obtain gold orange lemon Maillard reaction flavor.

[0014] As a further improvement of the present application, the following steps are included:

[0015] S1. chicken egg white protein and lysozyme separation: separate egg white from yolk, add water and stir to mix evenly, centrifuge, take supernatant, collect dialysate for second ultrafiltration, cut off filtrate and freeze-dry, obtain chicken egg white protein, dialysate freeze-drying, obtain lysozyme;

[0016] S2. enzyme assisted hydrolysis and conformation change reaction of exogenous enzyme: mix milk cream, chicken egg white protein prepared in step S1, lysozyme prepared in step S1 evenly, add water, add compound protease, enzymolysis, temperature reaction, obtain desensitization hydrolysate;

[0017] S3. fermentation: inoculate the first fermentation bacteria into the desensitization hydrolysate prepared in step S2, and culture by fermentation, filter, freeze-dry, to prepare desensitization fermentation hydrolysate;

[0018] S4. gold orange lemon enzyme assisted fermentation: wash gold orange dry and lemon slice, dry, add water, heat and cook, cool to room temperature, obtain gold orange lemon mixed pulp, add compound enzyme, enzymolysis, inoculate the second fermentation bacteria, culture by fermentation, filter, dry, to prepare gold orange lemon enzyme assisted fermentation product;

[0019] S5. preparation of additive: mix compound amino acid, D-xylose and glucose evenly to prepare additive;

[0020] S6. Maillard reaction: the desensitization fermentation hydrolysate prepared in step S3, the gold orange lemon enzyme assisted fermentation product prepared in step S4, and the additive prepared in step S5 are mixed, water is added, and the mixture is heated and stirred to react, thereby obtaining a Maillard reaction product essence;

[0021] S7. Embedding: chitosan is dissolved in an acid solution to obtain a chitosan solution, the Maillard reaction product essence prepared in step S6, lemon flavor, emulsifier, and gelatin are dissolved in water, emulsified, the pH value of the solution is adjusted, the chitosan solution is added, and the mixture is stirred to react, then β-cyclodextrin is added, and the mixture is continuously stirred, centrifuged, washed, and dried, thereby obtaining a gold orange lemon Maillard reaction product essence.

[0022] As a further improvement of the present application, the mass ratio of the egg white and water in step S1 is 1:5-10, the ultrafiltration membrane for the first time of ultrafiltration is a polyether sulfone ultrafiltration membrane with a molecular weight of 50-60 kDa, and the ultrafiltration membrane for the second time of ultrafiltration is a polyether sulfone ultrafiltration membrane with a molecular weight of 25-35 kDa.

[0023] As a further improvement of the present application, the mass ratio of the milk cream, egg white albumin, lysozyme, and complex protease in step S2 is 20-25:40-50:5-7:3-5, the complex protease is selected from at least two of papain, ficin, bromelain, trypsin, neutral protease, and alkaline protease, preferably, a mixture of papain and ficin with a mass ratio of 5-7:3, the enzymolysis temperature is 45-50°C, and the enzymolysis time is 1-3 h, and the temperature of the heating reaction is 65-70°C, and the heating reaction time is 0.5-1 h.

[0024] As a further improvement of the present application, the first fermentation bacteria in step S3 are Pichia pastoris and Lactobacillus plantarum, the inoculation amount of the bacteria seed liquid with a bacterial content of 10 8 -10 9 cfu / mL is 2-3 v / v% and 1-2 v / v% respectively, and the fermentation culture conditions are 32-35°C, 70-100 r / min, and 24-36 h of fermentation culture.

[0025] As a further improvement of the present application, the mass ratio of the gold orange and lemon slice in step S4 is 10-15:7-10, the heating and cooking temperature is 95-100°C, and the heating and cooking time is 2-4 h, the complex enzyme is selected from at least two of cellulase, pectinase, hemicellulase, and glucosidase, preferably, a mixture of cellulase and pectinase with a mass ratio of 7-10:3-5, the second fermentation bacteria are Bifidobacterium longum and Lactobacillus rhamnosus, and the inoculation amount of the bacteria seed liquid with a bacterial content of 10 8 -10 9The seed liquid of the bacteria is inoculated in an amount of 1-2v / v% and 2-4v / v% respectively, and the fermentation culture is carried out at 37-39 DEG C and 70-100r / min for 24-48h.

[0026] As a further improvement of the present application, the mass ratio of the complex amino acid, D-xylose and glucose in step S5 is 15-20:5-7:3-5, the complex amino acid comprises the following components in weight parts: glycine 10-20 parts, cysteine 20-30 parts, glutamic acid 10-25 parts, isoleucine 20-30 parts, leucine 10-15 parts; the mass ratio of the desensitization fermentation hydrolysate, gold orange lemon enzyme assisted fermentation product, additive and water in step S6 is 20-25:17-20:7-10:50-70, the temperature of the heater in the heating and stirring reaction is 100-110 DEG C, and the reaction time is 1-2h.

[0027] As a further improvement of the present application, the acid solution in step S7 is 1-2wt% acetic acid solution, the mass ratio of the chitosan, Maillard reaction product essence, lemon fragrance, emulsifier, gelatin, beta-cyclodextrin is 17-20:15-20:3-5:0.5-1:10-15:3-5, the pH value of the adjusting solution is 5.7-6.2, and the lemon fragrance comprises the following components in weight parts: white lemon oil 1-4 parts, sweet orange aldehyde 1-3 parts, bitter orange oil 1-4 parts, citronella aldehyde 0.1-1 part, myrcene 0.2-1.5 parts.

[0028] The present application further protects a gold orange lemon Maillard reaction product essence prepared by the above preparation method.

[0029] The present application further protects the application of the above gold orange lemon Maillard reaction product essence in the preparation of betel nut.

[0030] The present application has the following beneficial effects:

[0031] Maillard reaction is one of non-enzyme browning reactions, mainly refers to the complex reaction of amino compounds (amino acids, peptides and proteins) and carbonyl compounds (sugars) in food. Maillard reaction products are various, which have important contribution to the color and flavor of food, can improve the oxidative stability of food and prolong the shelf life.

[0032] Chicken ovalbumin is the highest content protein in egg white, and egg white contains a small amount of lysozyme, which is usually obtained by isoelectric point chemical precipitation method, but a large amount of salt needs to be added, and the effect is often poor. The present application adopts the method of ultrafiltration, removes the anti-biotin protein, ovomucin, ovotransferrin, serine protease inhibitor, G3 ovoglobulin, etc. by the first ultrafiltration, so as to obtain high-purity chicken ovalbumin and lysozyme.

[0033] Typically, milk cream contains certain allergenic proteins. Similarly, chicken ovalbumin and lysozyme are common allergens in eggs. This invention, by adding a complex protease to assist the enzymatic hydrolysis reaction of lysozyme, not only hydrolyzes large protein molecules (including milk protein, chicken ovalbumin, etc.) into small polypeptides and amino acids, but also alters the structure of allergen proteins, exposing or hiding allergen epitopes, thereby changing their allergenicity. Upon further heating, through the interaction of disulfide bonds on the protein, the digestibility and conformation of the antigenic epitopes are altered, exposing hydrophobic groups and burying or destroying the allergen epitopes, thus reducing the allergenicity of these proteins and obtaining small-molecule desensitized fermentation hydrolysates.

[0034] Furthermore, under the action of the primary fermentation bacteria (including Pichia pastoris and Lactobacillus plantarum), these protein peptides can be further fermented to produce small-molecule short peptides, amino acids, etc., thereby obtaining abundant amino acid and short peptide products, which can serve as amino acid and protein sources in the Maillard reaction.

[0035] By enzymatic hydrolysis of dried kumquats and lemon slices using compound enzymes (including cellulase and pectinase) and fermentation by secondary fermentation bacteria (Bifidobacterium longum and Lactobacillus rhamnosus), the plant cell walls of kumquats and lemons can be broken down, and sugars such as polysaccharides and complex sugars can be dissolved and decomposed into small molecule monosaccharides and disaccharides, which can serve as the source of sugars in the Maillard reaction.

[0036] In addition, in the Maillard reaction of this invention, complex amino acids, D-xylose, and glucose are added. The complex amino acids include glycine, cysteine, glutamic acid, isoleucine, and leucine. Cysteine ​​helps to form a meaty aroma. The mechanism is that glutathione is cleaved at the positions of glutamyl and cysteine ​​to form 5-hydroxyproline or pyroglutamic acid, cysteine, and cysteine-glycine dipeptide. This process yields 2-methyl-3-furanthiophene, 2-methyl-thiophene, and 2-pentylthiophene derived from meaty aroma compounds such as thiamine. After the reaction of leucine and isoleucine, a cheese-like aroma is emitted. Meanwhile, after the fat in milk cream undergoes the Maillard reaction, the long-chain aldehydes produced by the oxidation and degradation of fat usually participate in the Maillard reaction to form long-chain thiophenes, which enrich the meaty aroma and improve the flavor profile of the Maillard reaction. Therefore, after the Maillard reaction is carried out, this invention obtains a compound flavor essence with lemon kumquat fruit aroma, cheese milk aroma and meaty aroma. The aroma is pure and rich. When added to the production process of betel nut, the resulting betel nut has a unique fruity aroma, milky aroma and meaty aroma. The aroma is attractive and the taste is unique, which is loved by consumers.

[0037] Finally, the present application embeds the Maillard reaction flavor essence and lemon flavor through complex coagulation method in the chitosan-gelatin-beta-cyclodextrin shell layer, which not only improves the appearance, color and other adverse factors of the Maillard reaction flavor essence, but also avoids the volatilization of volatile lemon flavor, reduces the lemon aroma, prolongs the shelf life of the essence, improves the storage stability, and widens the application range.

[0038] The prepared gold orange lemon Maillard reaction flavor essence has fruit aroma, milk aroma and meat aroma, rich, realistic and mellow fragrance, which attracts consumers, maximally retains the nutritional value, is green, healthy, nutritious and safe, has unique flavor, improves the appearance, color and other adverse factors of the Maillard reaction flavor essence, avoids the volatilization loss of volatile flavor substances, prolongs the shelf life of the essence, improves the storage stability, and has broad application prospects. Embodiment

[0039] The technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the present application.

[0040] The milk cream is Aolwei light cream, which is purchased from Shanghai Binglong Industry and Trade Co., Ltd.

[0041] Papain, 100,000 U / g, cellulase, 10,000 U / g, pectinase, 10,000 U / g, purchased from Nanning Dongheng Huadao Biological Technology Co., Ltd.; ficin, 10,000 U / g, purchased from Kezhi Xin (Nanjing) Biological Technology Co., Ltd.

[0042] Pichia pastoris, 100 billion cfu / g, purchased from Shanghai Jiachu Biological Engineering Co., Ltd., Lactobacillus plantarum, Bifidobacterium longum and Lactobacillus rhamnosus, 100 billion cfu / g, all purchased from Shandong Zhongke Jiayi Biological Engineering Co., Ltd.

[0043] The preparation method of the seed liquid of the strain of the present application is as follows: a single fermentation strain is inoculated into sterilized Gao's medium, activated and cultured at 35 DEG C, 100 r / min for 18-24 h to obtain a seed liquid of the strain containing 10 8 -10 9 cfu / mL of bacteria. Embodiment

[0044] The present embodiment provides a preparation method of a gold orange lemon Maillard reaction flavor essence, which comprises the following steps:

[0045] S1. Separation of chicken egg white albumin and lysozyme: 10 parts by weight of egg white was separated from egg yolk, mixed with 50 parts by weight of water, and stirred to be uniform, centrifuged, and the supernatant was collected. The supernatant was subjected to first ultrafiltration with a polyether sulfone ultrafiltration membrane with a molecular weight cut-off of 50 kDa, and the dialysate was subjected to second ultrafiltration with a polyether sulfone ultrafiltration membrane with a molecular weight cut-off of 25 kDa. The filtrate was collected and freeze-dried to obtain chicken egg white albumin. The dialysate was freeze-dried to obtain lysozyme.

[0046] S2. Enzymatic hydrolysis with enzyme and conformational change reaction of exogenous enzyme: 20 parts by weight of butterfat, 40 parts by weight of chicken egg white albumin prepared in step S1, and 5 parts by weight of lysozyme prepared in step S1 were mixed uniformly, added to 500 parts by weight of water, and 3 parts by weight of complex protease was added. Enzymatic hydrolysis was performed at 45°C for 1 h, and the temperature was increased to 65°C for 0.5 h to obtain a desensitization hydrolysate.

[0047] The complex protease is a mixture of papain and ficin with a mass ratio of 5:3.

[0048] S3. Fermentation: Pichia pastoris and Lactobacillus plantarum seed liquid were inoculated into the desensitization hydrolysate prepared in step S2, and the inoculation amount was 2 v / v% and 1 v / v%, respectively. The fermentation was carried out at 32°C and 70 r / min for 24 h, and then filtered and freeze-dried to obtain a desensitization fermentation hydrolysate.

[0049] S4. Enzyme-assisted fermentation of gold orange and lemon: 10 parts by weight of dried gold orange and 7 parts by weight of lemon slices were washed and dried, added to 100 parts by weight of water, heated to 95°C, and cooked for 2 h. The mixture was cooled to room temperature to obtain a gold orange and lemon mixture, and a complex enzyme was added in an amount of 2 wt% of the total system. Enzymatic hydrolysis was performed at 50°C for 1 h, and Bifidobacterium longum and Lactobacillus rhamnosus seed liquid were inoculated in an amount of 1 v / v% and 2 v / v%, respectively. The fermentation was carried out at 37°C and 70 r / min for 24 h, and then filtered and dried to obtain an enzyme-assisted fermentation product of gold orange and lemon.

[0050] The complex enzyme is a mixture of cellulase and pectinase with a mass ratio of 7:3.

[0051] S5. Preparation of additives: 15 parts by weight of complex amino acids, 5 parts by weight of D-xylose, and 3 parts by weight of glucose were mixed uniformly to obtain additives.

[0052] The complex amino acids include the following components: 10 parts of glycine, 20 parts of cysteine, 10 parts of glutamic acid, 20 parts of isoleucine, and 10 parts of leucine.

[0053] S6. Maillard reaction: 20 parts by weight of the desensitization fermentation hydrolysate prepared in step S3, 17 parts by weight of the Jincheng lemon enzyme-assisted fermentation product prepared in step S4, and 7 parts by weight of the additive prepared in step S5 are mixed, 50 parts by weight of water is added, and the heater is heated to a temperature of 100°C, and stirred for 1 h to obtain a Maillard reaction product essence;

[0054] S7. Embedding: 17 parts by weight of chitosan is dissolved in 200 parts by weight of a 1 wt% acetic acid solution to obtain a chitosan solution, 15 parts by weight of the Maillard reaction product essence prepared in step S6, 3 parts by weight of a lemon flavor, 0.5 parts by weight of lecithin, and 10 parts by weight of gelatin are dissolved in water, emulsified at 10,000 r / min for 10 min, the pH value of the solution is adjusted to 5.7, the chitosan solution is added, stirred for 30 min, and then 3 parts by weight of β-cyclodextrin is added, and the stirring is continued for 2 h, centrifuged, washed, and dried to obtain a Jincheng lemon Maillard reaction product essence;

[0055] The lemon flavor includes the following components in parts by weight: 1 part of white lemon oil, 1 part of sweet orange aldehyde, 1 part of bitter orange oil, 0.1 part of citronellal, and 0.2 part of myrcene. Embodiment

[0056] The embodiment provides a preparation method of a Jincheng lemon Maillard reaction product essence, which comprises the following steps:

[0057] S1. Separation of chicken egg white protein and lysozyme: 10 parts by weight of egg white is separated from egg yolk, mixed uniformly in 100 parts by weight of water, centrifuged, and the supernatant is collected. The supernatant is subjected to first ultrafiltration by using a 60 kDa polyether sulfone ultrafiltration membrane, and the dialysate is subjected to second ultrafiltration by using a 35 kDa polyether sulfone ultrafiltration membrane. The filtrate is frozen and dried to obtain chicken egg white protein, and the dialysate is frozen and dried to obtain lysozyme;

[0058] S2. Enzyme-assisted hydrolysis and conformational change reaction of exogenous enzymes: 25 parts by weight of butter, 50 parts by weight of chicken egg white protein prepared in step S1, and 7 parts by weight of lysozyme prepared in step S1 are uniformly mixed, 500 parts by weight of water is added, 5 parts by weight of complex protease is added, and enzymolysis is performed at 50°C for 3 h. The temperature is increased to 70°C, and reaction is performed for 1 h to obtain a desensitization hydrolysate;

[0059] The complex protease is a mixture of papain and ficin, and the mass ratio is 7:3;

[0060] S3. Fermentation: seed liquids of Pichia pastoris and Lactobacillus plantarum are inoculated into the desensitization hydrolysate prepared in step S2, and the inoculation amounts are 3 v / v% and 2 v / v% respectively. Fermentation culture is performed at 35°C and 100 r / min for 36 h, filtration is performed, and freeze-drying is performed to obtain a desensitization fermentation hydrolysate;

[0061] S4. Enzyme-assisted fermentation of kumquat lemon: 15 parts by weight of dried kumquat and 10 parts by weight of lemon slices were washed and dried, then added to 100 parts by weight of water, heated to 100℃, and cooked for 4 hours. The mixture was cooled to room temperature to obtain a kumquat lemon mixture, then a composite enzyme was added in an amount of 2wt% of the total mass of the system, and enzyme hydrolysis was performed at 50℃ for 1 hour. Seed liquids of Lactobacillus rhamnosus and Bifidobacterium longum were inoculated in an amount of 2v / v% and 4v / v% respectively, and fermentation was carried out at 39℃ and 100r / min for 48 hours. After filtration, drying was performed to obtain the enzyme-assisted fermentation product of kumquat lemon;

[0062] The composite enzyme is a mixture of cellulase and pectinase in a mass ratio of 10:5.

[0063] S5. Preparation of additives: 20 parts by weight of composite amino acids, 7 parts by weight of D-xylose, and 5 parts by weight of glucose were uniformly mixed to obtain the additives.

[0064] The composite amino acids include the following components in parts by weight: glycine 20 parts, cysteine 30 parts, glutamic acid 25 parts, isoleucine 30 parts, and leucine 15 parts.

[0065] S6. Maillard reaction: 25 parts by weight of the desensitization fermentation hydrolysate prepared in step S3, 20 parts by weight of the enzyme-assisted fermentation product of kumquat lemon prepared in step S4, and 10 parts by weight of the additives prepared in step S5 were mixed and added to 70 parts by weight of water. The heater was heated to a temperature of 110℃, and stirring was performed for 2 hours to obtain a Maillard reaction product essence.

[0066] S7. Embedding: 20 parts by weight of chitosan was dissolved in 200 parts by weight of a 2wt% acetic acid solution to obtain a chitosan solution. 20 parts by weight of the Maillard reaction product essence prepared in step S6, 5 parts by weight of lemon base, 1 part by weight of lecithin, and 15 parts by weight of gelatin were dissolved in water, emulsified at 10000r / min for 10 minutes, the pH value of the solution was adjusted to 6.2, the chitosan solution was added, stirring was performed for 30 minutes, then 5 parts by weight of β-cyclodextrin was added, and stirring was continued for 2 hours. After centrifugation, washing, and drying, the kumquat lemon Maillard reaction product essence was obtained.

[0067] The lemon base includes the following components in parts by weight: white lemon oil 4 parts, sweet orange aldehyde 3 parts, bitter orange oil 4 parts, citronellal 1 part, and myrcene 1.5 parts. Embodiment

[0068] The embodiment provides a preparation method of a kumquat lemon Maillard reaction product essence, which comprises the following steps:

[0069] S1. Separation of chicken egg white albumin and lysozyme: 10 parts by weight of egg white was separated from egg yolk, mixed with 70 parts by weight of water, stirred and mixed uniformly, centrifuged, and the supernatant was collected. The supernatant was subjected to first ultrafiltration with a polyether sulfone ultrafiltration membrane with a molecular weight cut-off of 55 kDa, and the dialysate was subjected to second ultrafiltration with a polyether sulfone ultrafiltration membrane with a molecular weight cut-off of 30 kDa. The filtrate was frozen and dried to obtain chicken egg white albumin, and the dialysate was frozen and dried to obtain lysozyme;

[0070] S2. Enzymatic hydrolysis with enzyme and conformational change reaction of exogenous enzyme: 22 parts by weight of butterfat, 45 parts by weight of chicken egg white albumin prepared in step S1, and 6 parts by weight of lysozyme prepared in step S1 were mixed uniformly, added to 500 parts by weight of water, and 4 parts by weight of complex protease was added. Enzymatic hydrolysis was carried out at 47°C for 2h, and the temperature was raised to 67°C for 1h to obtain a desensitization hydrolysate;

[0071] The complex protease is a mixture of papain and ficin with a mass ratio of 6:3;

[0072] S3. Fermentation: Pichia pastoris and Lactobacillus plantarum seed liquid were inoculated into the desensitization hydrolysate prepared in step S2, and the inoculation amount was 2.5% and 1.5% respectively. The fermentation was carried out at 33°C and 85r / min for 30h, and then filtered and freeze-dried to obtain a desensitization fermentation hydrolysate;

[0073] S4. Enzyme-assisted fermentation of gold orange and lemon: 12 parts by weight of dried gold orange and 8.5 parts by weight of lemon slices were washed and dried, added to 100 parts by weight of water, heated to 97°C, and cooked for 3h. The mixture was cooled to room temperature to obtain a gold orange and lemon mixture, and a complex enzyme was added in an amount of 2wt% of the total system. Enzymatic hydrolysis was carried out at 50°C for 1h, and Bifidobacterium longum and Lactobacillus rhamnosus seed liquid were inoculated in an amount of 1.5% and 3% respectively. The fermentation was carried out at 38°C and 85r / min for 36h, and then filtered and dried to obtain an enzyme-assisted fermentation product of gold orange and lemon;

[0074] The complex enzyme is a mixture of cellulase and pectinase with a mass ratio of 8.5:4;

[0075] S5. Preparation of additives: 17 parts by weight of complex amino acids, 6 parts by weight of D-xylose, and 4 parts by weight of glucose were mixed uniformly to obtain additives;

[0076] The complex amino acids include the following components: glycine 15 parts, cysteine 25 parts, glutamic acid 20 parts, isoleucine 25 parts, and leucine 12 parts;

[0077] S6. Melanosis reaction: 22 parts by weight of the desensitization fermentation hydrolysate prepared in step S3, 18.5 parts by weight of the Jinjie lemon enzyme-assisted fermentation product prepared in step S4, and 8.5 parts by weight of the additive prepared in step S5 are mixed, 60 parts by weight of water is added, and the heater is heated to a temperature of 105°C, and the mixture is stirred for 1.5 h to obtain a melanosis reaction essence;

[0078] S7. Embedding: 18.5 parts by weight of chitosan is dissolved in 200 parts by weight of a 1.5 wt% acetic acid solution to obtain a chitosan solution, 17 parts by weight of the melanosis reaction essence prepared in step S6, 4 parts by weight of a lemon base, 0.7 parts by weight of lecithin, and 12 parts by weight of gelatin are dissolved in water, emulsified at 10,000 r / min for 10 min, the pH value of the solution is adjusted to 6, the chitosan solution is added, and the mixture is stirred for 30 min, then 4 parts by weight of β-cyclodextrin is added, and the stirring is continued for 2 h, followed by centrifugation, washing, and drying to obtain a Jinjie lemon melanosis reaction essence;

[0079] The lemon base comprises the following components in parts by weight: white lemon oil 2 parts, sweet orange aldehyde 2 parts, bitter orange oil 3 parts, citronellal 0.5 part, and myrcene 0.8 part. Example

[0080] Compared with Example 3, the difference lies in that the complex protease is a single papain. Example

[0081] Compared with Example 3, the difference lies in that the complex protease is a single papain. Example

[0082] Compared with Example 3, the difference lies in that the complex protease is a single papain. Example

[0083] Compared with Example 3, the difference lies in that the complex protease is a single papain. Example

[0084] Compared with Example 3, the difference lies in that the complex protease is a single papain.

[0085] The complex amino acid comprises the following components in parts by weight: glycine 15 parts, glutamic acid 20 parts, isoleucine 25 parts, and leucine 12 parts. Example

[0086] Compared with Example 3, the difference lies in that the complex protease is a single papain.

[0087] The complex amino acid comprises the following components in parts by weight: glycine 15 parts, glutamic acid 20 parts, isoleucine 25 parts, and leucine 12 parts. Example

[0088] The difference compared with Example 3 is that no complex amino acid is added in the additive.

[0089] The details are as follows:

[0090] S5. Preparation of the additive: 6 parts by weight of D-xylose and 4 parts by weight of glucose are mixed uniformly to prepare the additive.

[0091] Comparative Example 1

[0092] The difference compared with Example 3 is that no ultrafiltration in step S1 is performed.

[0093] The details are as follows:

[0094] S1. Separation of egg white: 10 parts by weight of egg white is separated from egg yolk to obtain egg white.

[0095] Comparative Example 2

[0096] The difference compared with Example 3 is that no complex protease is added in step S2.

[0097] The details are as follows:

[0098] S2. Enzymatic hydrolysis assisted by exogenous enzymes and conformational change reaction: 22 parts by weight of butterfat, 45 parts by weight of chicken egg albumin prepared in step S1, and 6 parts by weight of lysozyme prepared in step S1 are mixed uniformly, 500 parts by weight of water is added, and the mixture is reacted at 47°C for 2 h, and then the temperature is increased to 67°C and reacted for 1 h to obtain a desensitized hydrolysate.

[0099] Comparative Example 3

[0100] The difference compared with Example 3 is that no temperature increase reaction in step S2.

[0101] The details are as follows:

[0102] S2. Enzymatic hydrolysis assisted by exogenous enzymes and conformational change reaction: 22 parts by weight of butterfat, 45 parts by weight of chicken egg albumin prepared in step S1, and 6 parts by weight of lysozyme prepared in step S1 are mixed uniformly, 500 parts by weight of water is added, and the mixture is reacted at 47°C for 2 h, and then the temperature is increased to 67°C and reacted for 1 h to obtain a desensitized hydrolysate.

[0103] Comparative Example 4

[0104] The difference compared with Example 3 is that no Pichia pastoris seed liquid is inoculated in step S3.

[0105] The details are as follows:

[0106] S3. Fermentation: inoculate the desensitization hydrolysate prepared in step S2 with Pichia pastoris seed liquid, inoculation amount is 4 v / v%, 33℃, 85 r / min, fermentation culture for 30h, filter, freeze-drying, to obtain desensitization fermentation hydrolysate.

[0107] Comparative Example 5

[0108] Compared with Example 3, the difference is that no Pichia pastoris seed liquid is inoculated in step S3.

[0109] Specifically as follows:

[0110] S3. Fermentation: inoculate the desensitization hydrolysate prepared in step S2 with Pichia pastoris seed liquid, inoculation amount is 4 v / v%, 33℃, 85 r / min, fermentation culture for 30h, filter, freeze-drying, to obtain desensitization fermentation hydrolysate.

[0111] Comparative Example 6

[0112] Compared with Example 3, the difference is that step S3 is not performed.

[0113] Specifically as follows:

[0114] S1. Isolation of chicken egg white protein and lysozyme: 10 parts by weight of egg white is separated from egg yolk, added to 70 parts by weight of water and stirred to mix uniformly, centrifuged, and the supernatant is collected. The supernatant is subjected to first ultrafiltration using a polyether sulfone ultrafiltration membrane with a molecular weight cut-off of 55 kDa, and the dialysate is subjected to second ultrafiltration using a polyether sulfone ultrafiltration membrane with a molecular weight cut-off of 30 kDa. The filtrate is collected and freeze-dried to obtain chicken egg white protein. The dialysate is freeze-dried to obtain lysozyme.

[0115] S2. Enzymatic hydrolysis with exogenous enzymes and conformational modification reaction: 22 parts by weight of butter, 45 parts by weight of chicken egg white protein prepared in step S1, and 6 parts by weight of lysozyme prepared in step S1 are mixed uniformly, added to 500 parts by weight of water, and 4 parts by weight of complex protease is added. Enzymatic hydrolysis is performed at 47℃ for 2h, and then the temperature is raised to 67℃ and reaction is performed for 1h to obtain a desensitization hydrolysate.

[0116] The complex protease is a mixture of papain and ficin with a mass ratio of 6:3.

[0117] S3. Enzymatic fermentation of gold orange lemon: 12 parts by weight of dried gold orange and 8.5 parts by weight of lemon slices are washed and dried, added to 100 parts by weight of water, heated to 97℃, and cooked for 3h. The mixture is cooled to room temperature to obtain a gold orange lemon mixture, and then complex enzymes are added. Enzymatic hydrolysis is performed at 50℃ for 1h, and then P. acidipiscicola and L. rhamnosus seed liquid are inoculated with inoculation amounts of 1.5 v / v% and 3 v / v% respectively. Fermentation culture is performed at 38℃ and 85 r / min for 36h. The product is filtered and dried to obtain a gold orange lemon enzymatic fermentation product.

[0118] The complex enzyme is a mixture of cellulase and pectinase, with a mass ratio of 8.5:4;

[0119] S4. Preparation of the additive: 17 parts by weight of the complex amino acid, 6 parts by weight of D-xylose, 4 parts by weight of glucose are uniformly mixed to prepare the additive;

[0120] The complex amino acid comprises the following components in parts by weight: glycine 15 parts, cysteine 25 parts, glutamic acid 20 parts, isoleucine 25 parts, leucine 12 parts;

[0121] S5. Maillard reaction: 22 parts by weight of the desensitization hydrolysate prepared in step S2, 18.5 parts by weight of the gold orange lemon enzyme-assisted fermentation product prepared in step S3, and 8.5 parts by weight of the additive prepared in step S4 are mixed, 60 parts by weight of water is added, and the heater is heated to a temperature of 105°C. Stirring reaction for 1.5h, the Maillard reaction product essence is obtained;

[0122] S6. Embedding: 18.5 parts by weight of chitosan is dissolved in 200 parts by weight of 1.5wt% acetic acid solution to obtain a chitosan solution, 17 parts by weight of the Maillard reaction product essence prepared in step S5, 4 parts by weight of lemon base, 0.7 parts by weight of lecithin, and 12 parts by weight of gelatin are dissolved in water, emulsified at 10000r / min for 10min, the pH value of the solution is adjusted to 6, the chitosan solution is added, and stirring reaction for 30min, then 4 parts by weight of β-cyclodextrin is added, and stirring for 2h, centrifugation, washing, and drying to obtain the gold orange lemon Maillard reaction product essence;

[0123] The lemon base comprises the following components in parts by weight: white lemon oil 2 parts, sweet orange aldehyde 2 parts, bitter orange oil 3 parts, citronellal 0.5 parts, myrcene 0.8 parts.

[0124] Comparative Example 7

[0125] Compared with Example 3, the difference is that no complex enzyme is added in step S4.

[0126] Specifically as follows:

[0127] S4. Enzyme-assisted fermentation of gold orange lemon: 12 parts by weight of dried gold orange and 8.5 parts by weight of lemon slices are washed and dried, then added to 100 parts by weight of water, heated to 97°C, and cooked for 3h, then cooled to room temperature to obtain a mixed pulp of gold orange and lemon, inoculated with seed liquid of Bifidobacterium longum and Lactobacillus rhamnosus, with inoculation amounts of 1.5v / v% and 3v / v% respectively, fermented at 38°C and 85r / min for 36h, then filtered and dried to obtain the enzyme-assisted fermentation product of gold orange lemon.

[0128] Comparative Example 8

[0129] The difference compared with Example 3 is that no fermentation is performed in step S4.

[0130] The details are as follows:

[0131] S4. Enzymatic hydrolysis of kumquat lemon: 12 parts by weight of kumquat dry and 8.5 parts by weight of lemon slices are washed and dried, added to 100 parts by weight of water, heated to 97℃, and cooked for 3h, and then cooled to room temperature to obtain a kumquat lemon mixed pulp, and then a composite enzyme is added, and enzymatic hydrolysis is performed at 50℃ for 37h, and then filtered, dried, and the kumquat lemon enzymatic hydrolysis product is prepared;

[0132] The composite enzyme is a mixture of cellulase and pectinase, and the mass ratio is 8.5:4.

[0133] Comparative Example 9

[0134] The difference compared with Example 3 is that no lemon fragrance is added in step S7.

[0135] The details are as follows:

[0136] S7. Embedding: 18.5 parts by weight of chitosan is dissolved in 200 parts by weight of a 1.5wt% acetic acid solution to obtain a chitosan solution, 17 parts by weight of the Maillard reaction product essence prepared in step S6, 0.7 parts by weight of lecithin, and 12 parts by weight of gelatin are dissolved in water, emulsified at 10000r / min for 10min, the pH value of the solution is adjusted to 6, the chitosan solution is added, and stirring is performed for 30min, and then 4 parts by weight of β-cyclodextrin is added, and stirring is continued for 2h, and then centrifugation, washing, and drying are performed to prepare the kumquat lemon Maillard reaction product essence.

[0137] Comparative Example 10

[0138] The difference compared with Example 3 is that no embedding is performed in step S7, and the components are directly mixed.

[0139] The details are as follows:

[0140] S7. Embedding: 17 parts by weight of the Maillard reaction product essence prepared in step S6 and 4 parts by weight of lemon fragrance are uniformly mixed to prepare the kumquat lemon Maillard reaction product essence.

[0141] The lemon fragrance includes the following components in parts by weight: 2 parts of white lemon oil, 2 parts of sweet orange aldehyde, 3 parts of bitter orange oil, 0.5 part of citronellal, and 0.8 part of myrcene.

[0142] Test Example 1: Antioxidant performance

[0143] The kumquat lemon Maillard reaction product essence prepared in the examples and comparative examples of the present application is prepared into a sample solution of 5mg / mL, and antioxidant performance testing is performed.

[0144] Determination of DPPH radical scavenging capacity:

[0145] Mix 3 mL of 100 μmol / L DPPH ethanol solution with 1 mL of sample solution, and place in dark at room temperature for 25 min. Measure the absorbance (A1) at 517 nm with ethanol as blank. Measure the absorbance of 1 mL of ethanol and 3 mL of DPPH ethanol solution mixture at 517 nm, and record as A0. In addition, measure the absorbance of 1 mL of sample solution and 3 mL of ethanol mixture at 517 nm, and record as A2.

[0146] The calculation formula of DPPH radical scavenging capacity is as follows:

[0147] DPPH radical scavenging rate (%) = [1 - (A1-A2) / A0] x 100%

[0148] Determination of superoxide anion radical scavenging capacity:

[0149] Mix 9 mL of 50 mmol / L Tris-HCl buffer solution with pH of 8.2 with 0.5 mL of sample solution, and heat in water bath at 25 °C for 20 min. Immediately mix with 0.04 mL of 45 mmol / L pyrogallol solution (0.45 mmol of pyrogallol dissolved in 10 mL of 10 mmol / L hydrochloric acid) preheated at 25 °C, and measure the absorbance (A1) at 327 nm after standing for 3 min. Replace 0.5 mL of sample solution mixture with equal amount of water, and measure the absorbance (A0) at 327 nm.

[0150] The calculation formula of superoxide anion radical scavenging capacity is as follows:

[0151] Superoxide anion radical scavenging rate (%) = (A0-A1) / A0 x 100%

[0152] The results are shown in Table 1.

[0153] Table 1

[0154]

[0155] From the above table, it can be seen that the gold orange lemon Maillard reaction product essence prepared in Examples 1-3 has good antioxidant performance.

[0156] Test Example 2 Sensory evaluation

[0157] The gold orange lemon Maillard reaction product essence prepared in the examples and the comparative examples is added into the betel gum base at an amount of 5 wt%, and then stirred and mixed uniformly, and the flavor characteristics are compared. The average value is recorded by 20 professional personnel internal evaluation, and the gold orange lemon Maillard reaction product essence prepared is stored at 55±5℃ and humidity of 70±10% for 3 months, and then the same sensory evaluation is carried out. The results are shown in Table 2.

[0158] The score standard (full score 100 points) is as follows:

[0159] Faint aroma, peculiar smell, unnatural - 0-29 points;

[0160] Slightly faint aroma, unnatural, slightly milky, meaty or fruity - 30-69;

[0161] Pure and thick aroma, unique milky, meaty and fruity, fresh, natural and soft - 70-100 points.

[0162] Table 2

[0163]

[0164] As shown in the above table, the gold orange lemon Maillard reaction product essence prepared in examples 1-3 has good sensory evaluation, and has fruity, milky and meaty aroma, rich, realistic and mellow, and has good storage stability. The addition amount of the essence can be different according to specific conditions and consumer demand, and the essence can be applied not only to betel products, but also to beverages or other flavored candies.

[0165] Compared with example 3, the complex protease in example 4 and 5 is single papain or ficin. Compared with example 3, no complex protease is added in step S2 of comparative example 2, and the antioxidant property is reduced. The milk butter contains certain allergenic proteins, and similarly, chicken egg albumin and lysozyme are also common allergens in chicken eggs. By adding the complex protease, the enzymatic reaction of lysozyme is assisted, the large molecular protein (including milk protein, chicken egg albumin, etc.) is hydrolyzed into small molecular polypeptide, amino acid, etc., at the same time, the structure of the allergen protein is changed, the allergen epitope is exposed or hidden, so that the allergenicity is changed and the allergenicity is reduced.

[0166] Example 6, 7 and example 3, the complex enzyme is a single cellulase or pectinase. Comparative example 7 and example 3, no complex enzyme is added in step S4. Comparative example 8 and example 3, no fermentation is carried out in step S4, the sensory evaluation is reduced, and the antioxidant property is reduced. Under the action of the enzymolysis of the complex enzyme (including cellulase and pectinase) and the second fermentation bacteria (Bifidobacterium longum and Lactobacillus rhamnosus), the plant cell wall of the kumquat and lemon is broken, the polysaccharide and complex sugar are dissolved, and are decomposed into small molecule monosaccharide and disaccharide, etc., as the sugar source in the Maillard reaction.

[0167] Example 8 and example 3, no cysteine is added in the complex amino acid. Example 9 and example 3, no isoleucine and leucine are added in the complex amino acid. Example 10 and example 3, no complex amino acid is added in the additive, and the sensory evaluation is reduced. In the Maillard reaction of the application, the complex amino acid, D-xylose and glucose are also added, wherein the complex amino acid includes glycine, cysteine, glutamic acid, isoleucine and leucine, wherein the cysteine helps to form meaty aroma, and the mechanism is that the glutathione is cleaved at the glutamyl and cysteine positions to form 5-hydroxyproline or pyroglutamic acid, cysteine and cysteine-glycine dipeptide, 2-methyl-3-furan thiophene, 2-methyl-thiophene and 2-pentyl thiophene are obtained, which are derived from sulfur-containing meaty aroma flavor substances such as thiamine, and the leucine and isoleucine will emit a roasted cheese smell after reaction. At the same time, the long-chain aldehyde substances produced by the oxidation and degradation of the fat in the milk cream after the Maillard reaction will usually participate in the Maillard reaction to form long-chain thiophene substances, enrich the meaty aroma and improve the flavor profile of the Maillard reaction, so that the complex flavor essence with lemon kumquat fruit aroma, cheese milk aroma and meaty aroma is obtained after the Maillard reaction, the flavor is pure and mellow, and the betel nut prepared by adding the betel nut in the preparation process has unique fruit aroma, milk aroma and meaty aroma, attractive aroma and unique taste, which is deeply loved by consumers.

[0168] Comparative example 1 and example 3, no ultrafiltration is carried out in step S1, the sensory evaluation is reduced, and the antioxidant property is reduced. In the application, the ultrafiltration method is used, and the deantibiotin, ovomucoid, ovotransferrin, serine protease inhibitor and G3 ovoglobulin are removed by the first ultrafiltration, so that the chicken egg white protein and lysozyme with high purity are obtained.

[0169] Comparative example 3 and example 3, the temperature is not increased in step S2, and the antioxidant property is reduced. In the application, after heating, the digestion and the conformation of the antigen epitope of the protein are changed by the interaction of the disulfide bond, the hydrophobic group is exposed, the allergen epitope is buried or destroyed, so that the sensitization of these proteins is reduced, and the small molecule desensitization fermentation hydrolysate is obtained.

[0170] Compared with Example 3, the inoculation of Pichia pastoris or Lactobacillus plantarum seed liquid is not carried out in step S3 of Comparative Example 4 and 5. Compared with Example 3, step S3 is not carried out in Comparative Example 6, and the sensory evaluation and antioxidant property decrease. Under the action of the first fermentation bacteria (including Pichia pastoris and Lactobacillus plantarum), the protein peptides can be further fermented to produce small molecules such as short peptides and amino acids, so as to obtain rich amino acid and short peptide products as the amino acid and protein source in the Maillard reaction.

[0171] Compared with Example 3, the addition of lemon fragrance is not carried out in step S7 of Comparative Example 9, and the sensory evaluation and antioxidant property decrease. The addition of lemon fragrance makes the aroma abundance of the prepared Jincheng lemon Maillard reactant essence more abundant, and improves the antioxidant property, so that the prepared essence has more full flavor.

[0172] Compared with Example 3, the embedding is not carried out in step S7 of Comparative Example 10, and the components are directly mixed, and the sensory evaluation obviously decreases after 3 months of storage. In the present application, the Maillard reactant essence and lemon fragrance are embedded in the chitosan-gelatin-β-cyclodextrin shell layer by complex coacervation, which not only improves the appearance and color of the Maillard reactant essence and other adverse factors, but also avoids the volatilization of the volatile lemon fragrance, reduces the lemon aroma, prolongs the shelf life of the essence, improves the storage stability of the essence, and widens the application range of the essence.

[0173] The above only describes the preferred embodiments of the present application, and is not used to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing a kumquat-lemon Maillard reaction flavoring, characterized in that, Includes the following steps: S1. Separation of chicken egg white and lysozyme: Separate the egg white from the yolk, add water and stir to mix evenly, centrifuge, take the supernatant, perform the first ultrafiltration on the supernatant, collect the dialysate for the second ultrafiltration, freeze-dry the filtrate to obtain chicken egg white, freeze-dry the dialysate to obtain lysozyme; S2. Enzymatic hydrolysis and conformational change reaction of exogenous enzyme: Mix milk cream, chicken egg albumin obtained in step S1, and lysozyme obtained in step S1 evenly, add water, add complex protease, enzymatically hydrolyze, and heat to obtain desensitized hydrolysate; wherein, the complex protease is a mixture of papain and fig protease, with a mass ratio of 5-7:

3. S3. Fermentation: The first fermentation strain is inoculated into the desensitized hydrolysate obtained in step S2, fermented, filtered, and freeze-dried to obtain the desensitized fermented hydrolysate; wherein, the first fermentation strain is Pichia pastoris and Lactobacillus plantarum; S4. Enzymatic fermentation of kumquat and lemon: Dried kumquats and lemon slices are washed and dried, then added to water, heated and steamed, and cooled to room temperature to obtain a mixed kumquat and lemon pulp. A compound enzyme is added, and the mixture is enzymatically hydrolyzed. A second fermentation bacteria are inoculated, and the mixture is fermented, filtered, and dried to obtain the enzymatic fermentation product of kumquat and lemon. The compound enzyme is a mixture of cellulase and pectinase in a mass ratio of 7-10:3-5. The second fermentation bacteria are Bifidobacterium longum and Lactobacillus rhamnosus. S5. Preparation of additives: Mix the compound amino acids, D-xylose and glucose evenly to prepare the additives; wherein the compound amino acids are composed of the following components in parts by weight: glycine 10-20 parts, cysteine ​​20-30 parts, glutamic acid 10-25 parts, isoleucine 20-30 parts, and leucine 10-15 parts. S6. Maillard reaction: The desensitized fermentation hydrolysate obtained in step S3, the kumquat lemon enzyme-assisted fermentation product obtained in step S4, and the additive obtained in step S5 are mixed, added to water, heated and stirred to react, and the Maillard reaction product flavoring is obtained. S7. Encapsulation: Chitosan is dissolved in acid to obtain a chitosan solution. The Maillard reaction product flavoring, lemon flavor base, emulsifier, and gelatin obtained in step S6 are dissolved in water, emulsified, and the pH value of the solution is adjusted. The chitosan solution is added, and the reaction is stirred. Then β-cyclodextrin is added, and stirring is continued. The mixture is centrifuged, washed, and dried to obtain the kumquat lemon Maillard reaction product flavoring.

2. The preparation method according to claim 1, characterized in that, In step S1, the mass ratio of egg white to water is 1:5-10, the ultrafiltration membrane for the first ultrafiltration is a 50-60 kDa polyethersulfone ultrafiltration membrane, and the ultrafiltration membrane for the second ultrafiltration is a 25-35 kDa polyethersulfone ultrafiltration membrane.

3. The preparation method according to claim 1, characterized in that, In step S2, the mass ratio of milk cream, chicken egg albumin, lysozyme, and complex protease is 20-25:40-50:5-7:3-5. The enzymatic hydrolysis temperature is 45-50℃ and the time is 1-3 hours. The heating reaction temperature is 65-70℃ and the time is 0.5-1 hours.

4. The preparation method according to claim 1, characterized in that, In step S3, the inoculum content of the first fermentation bacteria is 10. 8 -10 9 The inoculum was prepared at cfu / mL, with inoculum amounts of 2-3 v / v% and 1-2 v / v%, respectively. The fermentation conditions were 32-35℃, 70-100 r / min, and fermentation for 24-36 h.

5. The preparation method according to claim 1, characterized in that, In step S4, the mass ratio of dried kumquats to lemon slices is 10-15:7-10; the heating and steaming temperature is 95-100℃; the time is 2-4 hours; and the inoculation concentration of the second fermentation bacteria is 10. 8 -10 9 The inoculum was prepared at cfu / mL, with inoculum amounts of 1-2 v / v% and 2-4 v / v%, respectively. The fermentation conditions were 37-39℃, 70-100 r / min, and fermentation for 24-48 h.

6. The preparation method according to claim 1, characterized in that, In step S5, the mass ratio of the compound amino acids, D-xylose, and glucose is 15-20:5-7:3-5; in step S6, the mass ratio of the desensitized fermentation hydrolysate, the kumquat lemon enzyme-assisted fermentation product, the additive, and water is 20-25:17-20:7-10:50-70; and in the heating and stirring reaction, the heater temperature is 100-110℃, and the reaction time is 1-2 hours.

7. The preparation method according to claim 1, characterized in that, The acid solution mentioned in step S7 is a 1-2 wt% acetic acid solution. The mass ratio of chitosan, Maillard reaction flavoring, lemon flavor base, emulsifier, gelatin, and β-cyclodextrin is 17-20:15-20:3-5:0.5-1:10-15:3-5. The pH value of the solution is adjusted to 5.7-6.

2. The lemon flavor base is composed of the following components in parts by weight: 1-4 parts white lemon oil, 1-3 parts sweet orange aldehyde, 1-4 parts bitter orange oil, 0.1-1 parts citronellol, and 0.2-1.5 parts myrcene.

8. A kumquat lemon Maillard reaction flavoring prepared by the method according to any one of claims 1-7.

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