Preparation method and application of tomato enzyme

Abstract

The invention discloses a preparation method and application of a tomato enzyme, which optimizes and improves the preparation process and formula of the tomato enzyme by screening proper fermentation strains, optimizing the strain proportion and fermentation conditions and other processes in various processing links and taking SOD (superoxide dismutase) and LYC (LYC) as evaluation indexes so as to increase the nutritional value of the tomato enzyme. The stabilizing agent is added, so that the sensory quality is obviously improved, and the product is free of scattering and separation; the antioxidant activity of the tomato enzyme is tested by using H2O2 to induce oxidative stress of C2C12 cells, and it is found that cell viability is remarkably increased by 16.83% and 22.22% due to the concentration of 2 mg / mL of unfermented tomato juice and 4 mg / mL of tomato enzyme, it is indicated that compared with the unfermented tomato juice, the tomato enzyme has a more excellent effect in the aspect of protecting C2C12 cells from oxidative stress, and the antioxidant activity of the tomato enzyme is improved. The fermented tomato enzyme is delicious in taste, moderate in sour and sweet degrees and suitable for people of all ages to eat.

Description

Technical Field

[0001] The invention relates to the technical field of tomato enzyme process development, and in particular to the technical field of a preparation method of tomato enzyme and application thereof. Background Art

[0002] Microbial enzymes refer to functional microbial fermentation products that are rich in nutrients such as vitamins, minerals, enzymes, and secondary metabolites. They are generally made from one or more fresh fruits, vegetables, mushrooms, Chinese herbal medicines, etc., and are fermented by a variety of beneficial bacteria. They have antibacterial, antioxidant, immunity-enhancing, cardiovascular protection, and obesity-inhibiting effects. Since fermentation is a complex microbial metabolic process, through its own material metabolism, the fermentation raw materials are decomposed and structurally changed at the molecular level, resulting in complex intermediate metabolism or cross-metabolism products, realizing metabolic conversion between substances, thereby producing new bioactive ingredients, and metabolizing to produce a variety of biological enzymes. However, there are currently few tomato enzyme production companies, and it is difficult to produce tomato enzymes on a large scale. The product quality is unstable, resulting in poor taste and other problems. Therefore, the research on tomato enzymes is crucial to optimize its fermentation process, explore its biological activity, and study the dynamic changes of microbial communities during fermentation and metabolites. Summary of the invention

[0003] In view of the shortcomings of the existing tomato enzyme preparation process, which leads to the problems of unstable product quality, low production efficiency and poor taste, the present invention aims to provide a method for preparing tomato enzyme and its application. The present invention optimizes and improves the preparation process and formula of tomato enzyme with SOD and LYC as the main indicators, and solves the problem of long-term precipitation by screening suitable fermentation strains, optimizing strain ratios, fermentation time and fermentation temperature and other processing links, and then adding stabilizers. The nutritional value and sensory quality of the obtained tomato enzyme are improved; the juice is not separated, and the sour taste of the fermented product has a significant retention feeling in the mouth, the taste is delicious, and the acidity is moderate, which is suitable for people of all ages.

[0004] In order to achieve the above technical effects, the present invention is implemented through the following technical solutions.

[0005] The invention provides a method for preparing tomato enzyme, the method comprising: taking tomato enzyme according to a proportion for every 100g tomato paste, stirring and mixing evenly, adding white sugar, sterilizing at 90°C for 15min, and cooling to room temperature; inoculating bacterial liquid into tomato juice according to a proportion, fermenting; filtering, adding a stabilizer after filtering, and sterilizing at 100°C for 15min after homogenization;

[0006] The bacterial liquid is selected from any one or more of Lactobacillus fermentum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus casei, Pediococcus acidilactici, Lactobacillus bulgaricus and Pediococcus pentosaceus.

[0007] In the present invention, the formula of the tomato enzyme is that the mass ratio of tomato sauce to water is 1:2.6.

[0008] Preferably, the bacterial liquid is a combination of Pediococcus pentosaceus, Lactobacillus casei, Lactobacillus plantarum and Lactobacillus fermentum.

[0009] Preferably, the bacterial liquid comprises, by volume percentage, 53.79% Pediococcus pentosaceus, 13.17% Lactobacillus casei, 19.87% Lactobacillus plantarum, and 13.17% Lactobacillus fermentum.

[0010] In the present invention, the fermentation temperature is 27-47°C, the fermentation time is 14-30h, and the initial fermentation bacterial concentration is 1×10 6 ~5×10 6 CFU / mL.

[0011] Preferably, the amount of added white sugar is

[0012] In the present invention, the stabilizer is selected from a combination of soluble soybean polysaccharides, pullulan and chitosan.

[0013] Preferably, the added mass of the stabilizer is 0.15% soluble soybean polysaccharide+0.03% pullulan+0.15% chitosan.

[0014] Furthermore, the present application also provides the application of the above-mentioned tomato enzyme preparation method in the production of tomato enzyme.

[0015] Through the above technical scheme, the present invention obtains the following technical effects:

[0016] (1) The present invention provides a method for preparing tomato enzyme using SOD and LYC as the main evaluation indicators, wherein the fermentation time is 22 h, the fermentation temperature is 37° C., and the lactic acid bacteria inoculation amount is 3×10 6 CFU / mL, sugar addition amount Under the conditions of , the SOD activity of tomato enzyme was verified to be 496.67U / g, and the LYC content was 77.12μg / g, and the nutritional quality of the final product was significantly improved. The addition of stabilizers stabilized the body, without dispersion and separation, and reduced precipitation. The fermented product tasted delicious and had moderate acidity, suitable for people of all ages.

[0017] (2) The present invention provides a method for preparing tomato enzyme using SOD and LYC evaluation as main indicators. By measuring SOD and LYC under fermentation with Pediococcus pentosaceus, Lactobacillus casei, Lactobacillus plantarum, and Lactobacillus fermentum, the SOD and LYC of the product are improved without losing the original nutritional value of the tomato. Then, by adding a stabilizer, the product has better sensory quality. Through verification, 2 mg / mL unfermented tomato juice and 4 mg / mL tomato enzyme concentrations resulted in a significant increase in cell viability of 16.83% and 22.22%, indicating that compared with unfermented tomato juice, tomato enzyme showed better efficacy in protecting C2C12 cells from oxidative stress. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is a line graph of SOD and LYC content after fermentation of tomato enzyme under different conditions; Figure 1 A is a line graph of SOD and LYC contents measured after tomato enzyme fermentation at different fermentation temperatures; Figure 1 B is a line graph of SOD and LYC contents measured after tomato enzyme fermentation at different fermentation times; Figure 1 C is a line graph of SOD and LYC contents measured after tomato enzyme fermentation with different TSS; Figure 1 D is a line graph of SOD and LYC contents after tomato enzyme fermentation with different initial bacterial densities.

[0019] Figure 2 This is an example diagram of the effects of different stabilizers on tomato enzymes; Figure 2 A is an example of the effect of different stabilizers on the Zeta potential of tomato enzymes; Figure 2 B. An example diagram of the effect of different stabilizers on the precipitation rate of tomato enzymes.

[0020] Figure 3 This is an example diagram of the effect of different stabilizers on the shear force and apparent viscosity of tomato enzyme; Figure 3 A is an example of the effect of different stabilizers on the shear force of tomato enzyme; Figure 3B. An example diagram of the effect of different stabilizers on the apparent viscosity of tomato enzyme. In the figure, Group A is Blank; Group B is 0.15% SSPS; Group C is 0.2% SSPS; Group D is 0.25% SSPS; Group E is 0.02% PUL; Group F is 0.04% PUL; Group G is 0.06% PUL; Group H is 0.2% CS; Group I is 0.25% CS; Group J is 0.3% CS; Group K is 0.05% SSPS+0.01% PUL+0.05% CS; Group L is 0.1% SSPS+0.02% PUL+0.1% CS; Group M is 0.15% SSPS+0.03% PUL+0.15% CS (soluble soybean polysaccharide (SSPS), pullulan (PUL), chitosan (CS)).

[0021] Figure 4 This is an example of the effect of unfermented tomato juice and tomato enzymes on cell viability; Figure 4 A is an example of the effect of unfermented tomato juice and tomato enzyme on normal cell viability; Figure 4 B is an example of the effect of unfermented tomato juice and tomato enzyme on the survival rate of C2C12 cells stimulated by 500μmol / LH2O2 oxidation. DETAILED DESCRIPTION

[0022] The technical solutions in the embodiments of the present invention will be described clearly and completely below in combination with the contents in the embodiments of the present invention. However, the present invention is not limited to the following embodiments.

[0023] The white sugar, bacterial liquid, stabilizer, etc. used in the present invention can all be purchased through public channels, and the equipment and instruments used in the process are all common equipment in the field.

[0024] All materials, practical and instrumental determination methods selected in the present invention are well known in the art but do not limit the implementation of the present invention. Other reagents and equipment well known in the art can be applicable to the implementation of the following embodiments of the present invention.

[0025] Example 1: Preparation method of tomato enzyme

[0026] A method for preparing a tomato enzyme comprises raw materials and water, wherein the raw material formula is: tomato juice (ketchup: water=1:2.6), bacterial liquid comprises 53.79% of Pediococcus pentosaceus, 13.17% of Lactobacillus casei, 19.87% of Lactobacillus plantarum, 13.17% of Lactobacillus fermentum, 0.02% of pullulan, and white sugar is added to make the soluble solid content of

[0027] Taking 100g of tomato sauce as the standard, take tomato enzyme according to the proportion and stir to mix evenly, add white sugar and sterilize at 90℃ for 15min, and cool to room temperature; inoculate the four bacterial liquids into tomato juice according to the proportion, ferment at 37℃ for 22h; filter, add stabilizer after filtration, homogenize, and sterilize at 100℃ for 15min.

[0028] Example 2: Preparation method of tomato enzyme

[0029] Based on Example 1, this embodiment provides a method for preparing a tomato enzyme with SOD activity and LYC content as main evaluation indicators, wherein the tomato enzyme comprises raw materials and water, wherein the raw material formula is: tomato juice (ketchup: water = 1:2.6), the bacterial liquid comprises 53.79% Pediococcus pentosaceus, 13.17% Lactobacillus casei, 19.87% Lactobacillus plantarum, and 13.17% Lactobacillus fermentum. The product fermentation temperature is 37°C, the fermentation time is 22h, and the initial fermentation bacterial concentration is 1×10 6 CFU / mL, add pullulan 0.04%, add white sugar to make the soluble solid content

[0030] Example 3: Preparation method of tomato enzyme

[0031] Based on Example 1, this embodiment provides a method for preparing tomato enzyme with SOD activity and LYC content as main evaluation indicators. The tomato sour soup includes raw materials and water, wherein the raw material formula is: tomato juice (ketchup: water = 1:2.6), the bacterial liquid includes 53.79% of Pediococcus pentosaceus, 13.17% of Lactobacillus casei, 19.87% of Lactobacillus plantarum, and 13.17% of Lactobacillus fermentum. The product fermentation temperature is 37°C, the fermentation time is 22h, and the initial fermentation bacterial concentration is 2×10 6 CFU / mL, add 0.05% soluble soybean polysaccharide, 0.01% pullulan, 0.05% chitosan, and add white sugar to make the soluble solid content

[0032] Example 4: Preparation method of tomato enzyme

[0033] Based on Example 1, this embodiment provides a method for preparing a tomato enzyme with SOD and LYC as the main evaluation indicators. The tomato enzyme includes raw materials and water, wherein the raw material formula is: tomato juice (ketchup: water = 1:2.6), tomato juice (ketchup: water = 1:2.6), and the bacterial liquid includes 53.79% of Pediococcus pentosaceus, 13.17% of Lactobacillus casei, 19.87% of Lactobacillus plantarum, and 13.17% of Lactobacillus fermentum. The product fermentation temperature is 37°C, the fermentation time is 22h, and the initial fermentation bacterial concentration is 3×10 6CFU / mL, add 0.1% soluble soybean polysaccharide, 0.02% pullulan, 0.1% chitosan, and add white sugar to make the soluble solid content

[0034] Example 5: Preparation method of tomato enzyme

[0035] Based on Example 1, this embodiment provides a method for preparing a tomato enzyme with SOD and LYC as main evaluation indicators, wherein the tomato enzyme includes raw materials and water, wherein the raw material formula is: tomato juice (ketchup: water = 1: 2.6), tomato juice (ketchup: water = 1: 2.6), tomato juice (ketchup: water = 1: 2.6), and the bacterial liquid includes 53.79% of Pediococcus pentosaceus, 13.17% of Lactobacillus casei, 19.87% of Lactobacillus plantarum, and 13.17% of Lactobacillus fermentum. The product fermentation temperature is 37°C, the fermentation time is 22h, and the initial fermentation bacterial concentration is 4×10 6 CFU / mL. Add 0.15% soluble soybean polysaccharide, 0.03% pullulan, 0.15% chitosan, and add white sugar to make the soluble solid content

[0036] Example 6: Optimization of the preparation method of tomato enzyme

[0037] Design-Expert.13 software was used for experimental design and variance analysis, Origin2021 software was used for drawing, and SPSS26 was used for variance analysis, with a significant difference level of 0.05. Each group of experiments was repeated 3 times, and the results were expressed as mean ± standard deviation.

[0038] Example 7: Effects of fermentation with different strains on the quality of tomato enzyme SOD and LYC

[0039] Different strains were added to the sterilized and cooled tomato juice in a certain proportion, stirred evenly, kept at a constant temperature, and fermented under closed conditions. The fermentation was carried out at 37°C for 28 hours, and SOD and LYC were measured every 4 hours during the fermentation.

[0040] By fermenting ten kinds of bacteria, including Lactobacillus fermentum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus casei, Pediococcus acidilactici, Lactobacillus bulgaricus and Pediococcus pentosaceus, the SOD, LYC and sensory properties of the fermented tomato enzymes were measured to analyze the effects of fermentation with different bacterial species on the SOD, LYC and other properties of tomato enzymes, and the most suitable fermentation bacteria were selected. The specific data are shown in Table 1.

[0041] Table 1: Indicators of tomato enzymes fermented by ten bacteria

[0042]

[0043]

[0044] From the data in Table 1, we can see that fermentation of different strains has a certain effect on the SOD activity and LYC content of tomato enzymes. Among them, the SOD activity of tomato enzymes fermented by Lactobacillus casei, Lactobacillus plantarum, Lactobacillus fermentum and Lactobacillus paracasei is higher, and the lycopene content of Pediococcus pentosaceus, Lactobacillus fermentum, Pediococcus acidilactici, Lactobacillus plantarum and four kinds of lycopene is higher. Combined with sensory, pH and TSS, the four most suitable fermented lactic acid bacteria were finally selected: Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus casei and Pediococcus pentosaceus.

[0045] Example 8: Effects of different fermentation strain ratios on tomato enzyme SOD and LYC

[0046] According to the four strains obtained by the initial screening in Example 7, the inoculation ratios of the four bacterial solutions were studied through a uniform design experiment, the effects of different strain ratios on the SOD and LYC of tomato enzymes were analyzed, and the optimal strain ratio was selected. For specific data, see Table 2.

[0047] Table 2: Test results of uniform ratio design of tomato enzyme fermentation strains

[0048]

[0049]

[0050] From the data in Table 2, we can see that different bacterial species ratios have an effect on the SOD of tomato enzyme and lycopene LYC. 10 )Uniform design table and uniform design table usage table, design a uniform experiment with four factors and ten levels. Use SPSS26 to fit the SOD in Table 2, and get the regression equation:

[0051] Y=267.485+1284.208X1-288.073X22-2436.117X32-8973.864X1X4-6613.659X2X3+5530.703X2X4+12747.912X3X4.

[0052] Through optimization, the optimal inoculation ratio of tomato enzyme fermented by four lactic acid bacteria is 53.79% of Pediococcus pentosaceus, 13.17% of Lactobacillus casei, 19.87% of Lactobacillus plantarum, and 13.17% of Lactobacillus fermentum.

[0053] Example 9: Effects of different fermentation temperatures on tomato enzymes SOD and LYC

[0054] Under the premise of keeping the basic formula unchanged, the fermentation temperature was set to five gradients: 27℃, 32℃, 37℃, 42℃, and 47℃. The effects of different fermentation temperatures on tomato enzymes SOD and LYC were analyzed, and the optimal fermentation temperature was selected. For specific data, see Figure 1 A.

[0055] Depend on Figure 1 A It can be seen that within 27℃-37℃, as the fermentation temperature of tomato enzyme increases, the SOD of tomato enzyme fermented by mixed bacteria gradually increases, and when the fermentation temperature reaches 37℃, the SOD content of tomato enzyme reaches the maximum value; but when the fermentation temperature exceeds 37℃, the SOD content of tomato enzyme gradually decreases. When fermented at 27℃ and 32℃, since the fermentation temperature does not reach the optimal fermentation temperature, the lactic acid bacteria in the fermentation liquid grow slowly and the fermentation speed is also relatively slow; when fermented at 42℃ and 47℃, since the fermentation temperature is higher than the optimal fermentation temperature, some lactic acid bacteria in the fermentation liquid die under high temperature environment, resulting in a slowdown in fermentation speed; both temperature conditions lead to a low content of tomato enzyme SOD when it is finally measured. However, when the fermentation temperature is 37℃, the lactic acid bacteria in the fermentation liquid are at the optimal fermentation temperature, the lactic acid bacteria multiply rapidly, and the fermentation liquid ferments rapidly. In the final determination, SOD reaches 488.78U / g, which is the tomato enzyme with the highest SOD activity at five temperatures.

[0056] At the same time by Figure 1 A It can be seen that fermentation at different temperatures also has a certain effect on the LYC of tomato enzyme. When the temperature reaches 37°C, the LYC content reaches the maximum. Therefore, combined with the SOD activity of tomato enzyme, the optimal fermentation temperature of tomato enzyme is: 37°C.

[0057] Example 10: Effects of different fermentation times on tomato enzymes SOD and LYC

[0058] Under the premise of keeping the basic formula unchanged, the fermentation time was set to five gradients: 14h, 18h, 22h, 26h, and 30h. The effects of different fermentation times on tomato enzymes SOD and LYC were analyzed, and the optimal fermentation time was selected. For specific data, see Figure 1 B.

[0059] Depend on Figure 1B It can be seen that the fermentation time will affect the SOD activity and LYC content. When the fermentation time is too short, the growth and reproduction of lactic acid bacteria are insufficient, the accumulation of metabolites is insufficient, and the SOD activity and LYC content are low. If the time is too long, the nutrients required for the growth and reproduction of lactic acid bacteria are insufficient, the number of dead bacteria increases, and metabolic waste accumulates in the fermentation liquid, which will also lead to a decrease in SOD activity and LYC content. SOD activity increases rapidly between 14h and 22h, reaches its maximum value of 493.06U / g at 22h, and then decreases. LYC reaches a maximum value of 72.20mg / g at 26h, and then decreases with time.

[0060] Therefore, the optimal fermentation time of tomato enzyme is 22h.

[0061] Example 11: Effect of TSS on tomato enzymes SOD and LYC

[0062] Under the premise of keeping the basic formula unchanged, set TSS to These five gradients were used to analyze the effects of different TSS concentrations on tomato enzymes SOD and LYC, and to select the optimal TSS concentration. For specific data, see Figure 1 C.

[0063] like Figure 1 C, when the TSS content is adjusted to When SOD activity reaches its maximum value of 486.95U / g, it is adjusted to LYC reached 71.14 mg / g.

[0064] Example 12: Effects of different initial fermentation bacterial densities on tomato enzyme SOD and LYC

[0065] Under the premise of keeping the basic formula unchanged, the initial fermentation bacteria density is set to: 1×10 6 , 2×10 6 , 3×10 6 , 4×10 6 , 5×10 6 CFU / mL, analyze the effects of different initial fermentation bacterial densities on tomato enzyme SOD and LYC, and select the optimal initial fermentation bacterial density. For specific data, see Figure 1 D.

[0066] like Figure 1 D, when the inoculum concentration is 10 6 CFU / ml, the bacterial count was insufficient and the bacterial metabolism rate was slow, resulting in SOD activity. However, as the inoculum size increased, the value of the indicator also increased. When the inoculum size was 3×10 6CFU / mL, the maximum SOD activity is 486.92U / g. 6 CFU / mL may lead to over-fermentation, resulting in instability of terpenoids and loss of enzyme activity. Therefore, the optimal inoculation range is 3×10 6 CFU / mL.

[0067] Depend on Figure 1 D shows that when the inoculum size is 2×10 6 CFU / mL, LYC reached 72.43μg / g. 6 CFU / mL may lead to over-fermentation, resulting in instability of terpenoids and loss of enzyme activity. Therefore, the optimal inoculation range is 3×10 6 CFU / mL.

[0068] Therefore, the optimal initial bacterial density of tomato enzyme fermentation is: 3×10 6 CFU / mL.

[0069] Example 13: Response Surface Optimization of Tomato Enzyme Fermentation Process

[0070] Based on the single factor experiment, taking SOD and LYC as the response values, four factors that significantly affected the fermentation of tomatoes with mixed bacteria of Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus casei and Pediococcus pentosaceus were fermentation temperature (32, 37 and 42℃), inoculum size (2×10 6 CFU / mL, 3×10 6 CFU / mL and 4×10 6 CFU / mL), fermentation time (18, 22 and 26 h) and TSS The response surface results of tomato enzyme fermentation process were analyzed as experimental factors to find the optimal fermentation process of tomato enzyme.

[0071] The experimental design was performed using the Design-expert11 analysis software Box-Behnken. The SOD and LYC in Table 3 were fitted using the Design-expert11 software to obtain the regression equation:

[0072] Y1(SOD)=503.10-0.1538A+10.48B-4.47C-9.97D-8.49AB+6.19AC-0.0775AD-6.53BC+7.90BD+5.19CD-29.59A2-12.11B2-31.52C2-32.09D2

[0073] Y2(LYC)=77.25+1.03A+0.7420B+1.02C+0.4004D-0.5100AB-1.02AC-0.2075AD-1.17BC+0.5587BD-0.5475CD-3.18A 2 -0.17B 2 -1.15C 2 -0.9473D 2

[0074] Table 3: Box-Behnken test results for tomato enzyme fermentation process optimization

[0075]

[0076]

[0077] The significance test of the model regression equation coefficient of Y1, see Table 4 for specific data, main terms B, D, interaction terms AB, AD, BD, BC, quadratic term A 2 , B 2 , C 2 , D 2 The Y2 model regression equation coefficients are highly significant, and the other terms are not significant. The significance test of the Y2 model regression equation coefficients is shown in Table 4. The main terms A, B, C, the interaction terms AC, BD, and the quadratic term A 2 , B 2 , C 2 , D 2 The correlation coefficient of SOD activity was r 2 The value is 0.9877, and the r of LYC 2 The value is 0.9605, which shows that the measured value and the predicted value have a good fit, and the process parameters of tomato enzyme can be analyzed and predicted.

[0078] Table 4: Analysis of variance of response surface test results

[0079]

[0080]

[0081] Note: * indicates significant, ** indicates extremely significant.

[0082] The optimal treatment method was obtained through the prediction of the regression model: the fermentation temperature was 37.09℃, the fermentation time was 22.20h, the lactic acid bacteria inoculation amount was 2.95×10 6 CFU / mL, sugar addition amount At this time, the predicted value of tomato enzyme SOD activity was 505.79U / g, and the predicted value of LYC was 77.33μg / g. Considering the actual production, the optimal conditions were modified to fermentation time of 22h, fermentation temperature of 37℃, lactic acid bacteria inoculation amount of 3×10 6 CFU / mL, sugar addition amount Under these conditions, the SOD activity of tomato enzyme was verified to be 496.67U / g and the LYC content was 77.12μg / g.

[0083] (1) Effect of stabilizers on tomato enzymes

[0084] Based on the tomato enzyme fermented by the method adopted in the above embodiment, experiments were conducted on the tomato enzyme by adding different proportions of soluble soybean polysaccharides, pullulan and chitosan, and the optimal ratio was determined using rheological properties, Zeta potential, etc. as indicators.

[0085] In this experiment, the addition amount of each stabilizer was set as follows: soluble soybean polysaccharides were added at 0.15%, 0.2%, and 0.3%, chitosan was added at 0.15%, 0.2%, and 0.3%, pullulan was added at 0.02%, 0.04%, and 0.06, and the compound ratio was soluble soybean polysaccharides: pullulan: chitosan = 1:0.2:1, which were added at 0.05%:0.01%:0.0.5%, 0.1%:0.02%:0.1%, and 0.15%:0.03%:0.15%, respectively.

[0086] like Figure 2 A It can be seen that the absolute values ​​of potential of all experimental groups are improved compared with the blank group. When the added amount is 0.15% soluble soybean polysaccharide + 0.03% pullulan + 0.15% chitosan, the larger the absolute value of the potential, the more it indicates that the system is more stable. Figure 2 It can also be seen in B that when the addition amount is 0.15% SSP + 0.03% PLL + 0.15% KGM, the sedimentation rate is the smallest. The size of the sedimentation rate can also indicate the stability of the system. The smaller the sedimentation rate, the more stable the system. Figure 3 The rheological properties of different thickeners can be clearly observed. The shear stress also increases with the increase of shear rate, but the ratio of shear stress to shear rate decreases with the increase of shear rate.

[0087] Example 14: Study on the efficacy of tomato enzyme

[0088] Based on the tomato enzyme fermented by the method of the above embodiment, the efficacy experiment of the tomato enzyme was carried out.

[0089] (1) Effect of tomato enzyme on H2O2-induced cytotoxicity of C2C12 cells

[0090] In order to more comprehensively study the antioxidant effect of tomato enzymes in cells, we chose to use H2O2 to induce oxidative stress in C2C12 cells and established unfermented tomato juice as a control to test the antioxidant activity of tomato enzymes. Figure 4 As shown in A, in the normal cell group, the tomato enzyme concentration below 4 mg / mL did not show obvious toxic effects on the cells, and even had a certain degree of growth-promoting effect. When the tomato enzyme concentration was 1 mg / mL, the cell viability increased by 15.45%. After the model group was treated with 500 μmol / L H2O2 for 24 hours, the cell viability decreased significantly by 5.15%. In the drug group, the researchers observed that the cell viability of the drug treatment group increased in a dose-dependent manner, such as Figure 4 B. Comparing the drug group with the model group, we observed that 2 mg / mL unfermented tomato juice and 4 mg / mL tomato enzyme concentrations resulted in a significant increase in cell viability by 16.83% and 22.22%, respectively. These findings suggest that tomato enzyme exhibits superior efficacy in protecting C2C12 cells from oxidative stress compared with unfermented tomato juice.

[0091] Based on the above data, the tomato enzyme preparation method provided by the present invention optimizes and improves the preparation process and formula of tomato enzyme by optimizing the fermentation process of tomato enzyme and the proportion of adding stabilizer after fermentation, etc., and takes LYC and SOD as evaluation indicators. The nutritional quality and sensory quality of the tomato enzyme finally obtained are significantly improved, and it is suitable for consumption by people of all ages.

[0092] The above embodiments are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the present invention.

Claims

1. A method for preparing tomato enzyme, characterized in that: The method comprises the following steps: taking tomato enzyme according to a proportion for every 100g of tomato paste, stirring and mixing evenly, adding white sugar, sterilizing at 90°C for 15min, and cooling to room temperature; inoculating bacterial liquid into tomato juice according to a proportion, fermenting; filtering, adding a stabilizer after filtration, and sterilizing at 100°C for 15min after homogenization; the bacterial liquid is selected from any one or more of fermented lactobacillus, lactobacillus acidophilus, lactobacillus rhamnosus, lactobacillus reuteri, lactobacillus plantarum, lactobacillus paracasei, lactobacillus casei, pediococcus acidilactici, lactobacillus bulgaricus, and pediococcus pentosaceus.

2. The method for preparing a tomato enzyme according to claim 1, characterized in that: The formula of the tomato enzyme is that the mass ratio of tomato sauce to water is 1:2.

6.

3. The method for preparing a tomato enzyme according to claim 1, characterized in that: The bacterial liquid is a combination of Pediococcus pentosaceus, Lactobacillus casei, Lactobacillus plantarum and Lactobacillus fermentum.

4. The method for preparing a tomato enzyme according to claim 1, characterized in that: The bacterial liquid comprises, by volume percentage, 53.79% of Pediococcus pentosaceus, 13.17% of Lactobacillus casei, 19.87% of Lactobacillus plantarum and 13.17% of Lactobacillus fermentum.

5. The method for preparing a tomato enzyme according to claim 1, characterized in that: The fermentation temperature is 27-47°C, the fermentation time is 14-30h, and the initial fermentation bacterial concentration is 1×10 6 ~5×10 6 CFU / mL.

6. The method for preparing a tomato enzyme according to claim 1, characterized in that: The amount of added white sugar is 7. The method for preparing tomato enzyme according to claim 1, characterized in that: The stabilizer is selected from a combination of soluble soybean polysaccharides, pullulan and chitosan.

8. The method for preparing a tomato enzyme according to claim 7, characterized in that: The added mass of the stabilizer is 0.15% soluble soybean polysaccharide+0.03% pullulan+0.15% chitosan.

9. Use of the tomato enzyme preparation method according to any one of claims 1 to 8 in the production of tomato enzyme.

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