Bacteriostatic food additive and preparation method thereof

Abstract

The application discloses a bacteriostatic food additive and a preparation method thereof. The bacteriostatic food additive comprises G3, cinnamyl aldehyde and vinegar powder and the like. The food additive provided by the application has a synergistic bacteriostatic effect on staphylococcus aureus and escherichia coli, and has the advantages of low use concentration and strong bacteriostatic capacity.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microorganisms; in particular to a bacteriostatic food additive and a preparation method thereof. BACKGROUND

[0002] With the rapid development of modern economic society, pre-prepared food such as pre-prepared dishes are increasingly popular in the market; however, in hot weather, dishes are easily deteriorated due to the influence of microorganisms, and even harmful bacteria are bred, which endangers human health.

[0003] In order to preserve food, freezing or adding preservatives and bacteriostatic agents are generally used to extend the shelf life and sales period of food. Many existing bacteriostatic agents have not only the effect of preventing and inhibiting bacteria, but also cause harm to the human body when added in excess, such as nitrite.

[0004] There are many food additives on the market, which can not only provide rich color and flavor to food, but also have the effect of inhibiting bacteria, slowing down the growth rate of bacteria and extending the shelf life of fresh food without freezing preservation. However, the current additives are often used alone, and the concentration needs to be large to achieve the bacteriostatic effect, which easily affects the flavor and other characteristics of the food. SUMMARY

[0005] The purpose of the present application is to provide a bacteriostatic food additive to solve the technical problems in the prior art that adding preservatives in food is not conducive to health or a large amount of food additives need to be added for bacteriostasis.

[0006] To achieve the above purpose, the present application adopts the following technical solutions:

[0007] A bacteriostatic food additive, comprising: a composite seasoning G3 and cinnamaldehyde.

[0008] Preferably, the G3 is composed of the following components by weight parts:

[0009]

[0010] Preferably, the G3 is composed of the following components by weight parts:

[0011]

[0012] Preferably, the concentration of the G3 is greater than or equal to 2 mg / mL; and the concentration of the cinnamaldehyde is greater than or equal to 0.063 mg / mL.

[0013] Preferably, it further comprises vinegar powder.

[0014] Preferably, the G3 concentration is greater than 4 mg / mL; the cinnamaldehyde concentration is greater than 0.25 mg / mL; and the vinegar powder concentration is greater than 4 mg / mL.

[0015] Preferably, the bacteria inhibited by the bacteriostatic food additive are any of E. coli, S. aureus, Listeria, and Vibrio parahaemolyticus.

[0016] Preferably, the bacteriostatic agent inhibits S. aureus for a duration of 12 hours or less, and inhibits E. coli for a duration of 24 hours or less.

[0017] The present application also provides a method for preparing the bacteriostatic food additive, comprising:

[0018] (1) Each component is added to pure water and stirred to prepare a single-component mother liquor with a G3 concentration of 100 mg / mL, a vinegar powder concentration of 100 mg / mL, and a cinnamaldehyde concentration of 20 mg / mL, and then filtered and stored;

[0019] (2) The mother liquor is mixed and diluted according to the required concentration of the formula to obtain the required bacteriostatic food additive.

[0020] The present application has the following beneficial effects:

[0021] The bacteriostatic food additive provided by the present application has the beneficial effects of low use concentration, synergistic or additive effect between components; good bacteriostatic effect on E. coli within 24 hours and good synergistic bacteriostatic effect on S. aureus within 12 hours; and provides convenience for the daily preservation and sale of fresh food. At the same time, the bacteriostatic food additive provided by the present application and vinegar powder have additive bacteriostatic effect, which can further enhance the bacteriostatic effect of vinegar powder. BRIEF DESCRIPTION OF DRAWINGS

[0022] In order to clearly show the specific embodiments of the present application and certain detection techniques used in the experiments, the embodiments and the techniques used will be described below, mainly in the form of drawings.

[0023] Figure 1 Growth inhibition curves of vinegar powder on E. coli, S. aureus, Listeria, and Vibrio parahaemolyticus; wherein CF in the figure represents vinegar powder.

[0024] Figure 2 Growth inhibition curves of G3 on E. coli, S. aureus, Listeria, and Vibrio parahaemolyticus.

[0025] Figure 3 Inhibition of E. coli, S. aureus, Listeria, and Vibrio parahaemolyticus growth by the combination of vinegar powder and G3 for 12 hours.

[0026] Figure 4 G3 combined with cinnamaldehyde for 24 hours on the growth of E. coli and S. aureus.

[0027] Detailed mode of implementation

[0028] The specific embodiments of the present application are assisted by examples, except that the techniques used for detection do not make any form of limitation to the present application, part of the schemes in the described examples are part of the embodiments of the present application, the examples obtained by the ordinary technical operations of the person skilled in the art without creative achievements are all within the protection scope of the present application.

[0029] Example 1: Experiment and method

[0030] 1. Determination of minimum inhibitory concentration (MIC)

[0031] The minimum antibacterial concentration (MIC) of cinnamaldehyde on E. coli, S. aureus, Listeria and Vibrio parahaemolyticus was determined according to the broth dilution method. The culture medium of S. aureus, E. coli and Vibrio parahaemolyticus was LB medium, and the culture medium of Listeria was 0.5% THYE medium. The specific steps are as follows:

[0032] (1) 1 × 10 6 CFU / mL of bacteria were mixed with different concentrations of cinnamaldehyde (from 0.0625 mg / mL to 2 mg / mL, two-fold dilution) and added to 96-well plates, and incubated in a 37℃ incubator for 24h.

[0033] (2) The absorbance value of each well at 600 nm was determined by UV spectrum, OD 600 <0.1, i.e. no visible bacterial growth, the minimum drug concentration corresponding to MIC was recorded.

[0034] In this embodiment, the specific ratio of the composite seasoning G3 is as follows: white granulated sugar 8-15 parts, preferably 12 parts; malt dextrin 6-12 parts, preferably 9 parts; glucose 0.6-1.8 parts, preferably 1.2 parts; yeast extract 0.1-1.2 parts, preferably 0.6 parts; sodium gluconate 0.1-2 parts, preferably 1 part; edible salt 0.1-1 part, preferably 0.5 part. The preferred weight ratio in the above embodiment is used in this embodiment. Specifically, the yeast extract is a product obtained by completely autolysis of yeast using its own hydrolytic enzyme system after plasmolysis, and the cell wall and insoluble molecules are removed; it meets the relevant index requirements of GB / T 20886.2-2021.

[0035] 2. Determination of growth curve

[0036] Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Vibrio parahaemolyticus were mixed with different concentrations of vinegar powder (0, 0.5, 1.0, 2.0, 4.0, 8.0 mg / mL) and G3 (0, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg / mL) and incubated at 37°C for 12 h. Their OD values ​​were measured at 4, 6, 8, 10, 12, and 24 hours. 600 The optical density values ​​were used to plot the antibacterial growth curves of acetic acid powder and G3 against Escherichia coli, Staphylococcus aureus, Listeria, and Vibrio parahaemolyticus.

[0037] 3. Chessboard dilution method

[0038] The checkerboard dilution method was used to detect the combined use of acetic acid powder with G3 or G3 with cinnamaldehyde. In the checkerboard dilution method, the drug concentrations in both the horizontal and vertical rows were 2×MIC, 1×MIC, 1 / 2×MIC, 1 / 4×MIC, 1 / 8×MIC, and 0. The interaction was determined by calculating the fractional inhibitory concentration (FIC). FIC = (MIC of drug A in combination / MIC of drug A alone) + (MIC of drug B in combination / MIC of drug B alone). FIC values ​​≤0.5, >0.5~1, >1~2, and >2 represent synergistic, additive, unrelated, and antagonistic effects, respectively.

[0039] Example 2: Results and Discussion

[0040] 1. Vinegar powder can inhibit the growth of Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Vibrio parahaemolyticus.

[0041] A vinegar powder concentration greater than 4 mg / mL can inhibit the growth of Escherichia coli within 12 hours. Figure 1 A); and when the concentration of vinegar powder is greater than 8 mg / mL, it can inhibit the growth of Staphylococcus aureus, Listeria monocytogenes and Vibrio parahaemolyticus within 10 hours. Figure 1 BD).

[0042] 2. G3 can inhibit the growth of Escherichia coli, Staphylococcus aureus, Listeria, and Vibrio parahaemolyticus.

[0043] G3 concentrations greater than 8 mg / mL can inhibit the growth of Escherichia coli within 10 hours. Figure 2 A) When the G3 concentration is greater than 16 mg / mL, it can inhibit the growth of Staphylococcus aureus, Listeria monocytogenes and Vibrio parahaemolyticus within 10 hours. Figure 2 BD).

[0044] 3. The inhibitory effect of combined use of vinegar powder and G3 on the growth of Escherichia coli, Staphylococcus aureus, Listeria, and Vibrio parahaemolyticus.

[0045] When 4 mg / mL acetic acid powder is used in combination with 4 mg / mL G3, it can inhibit the growth of Escherichia coli within 12 hours. Figure 3 A). When 8 mg / mL acetic acid powder is used in combination with 4 mg / mL G3, it can inhibit the growth of Staphylococcus aureus and Vibrio parahaemolyticus within 12 hours. Figure 3 B&D). When 8 mg / mL acetic acid powder is used in combination with 8 mg / mL G3, it can inhibit the growth of Listeria monocytogenes within 12 hours. Figure 3 C).

[0046] 4. Inhibition of the growth of Escherichia coli, Staphylococcus aureus, Listeria monocytogenes and Vibrio parahaemolyticus by cinnamaldehyde.

[0047] The MIC of cinnamaldehyde against Escherichia coli, Staphylococcus aureus, and Listeria was 0.25 mg / mL, and the MIC of cinnamaldehyde against Vibrio parahaemolyticus was 0.5 mg / mL.

[0048] 5. Inhibition of the growth of Escherichia coli and Staphylococcus aureus by the combined use of G3 and cinnamaldehyde.

[0049] In the checkerboard dilution method, interactions are determined by calculating the fractional inhibitory concentration (FIC). FIC = (MIC of drug A in combination / MIC of drug A alone) + (MIC of drug B in combination / MIC of drug B alone). FIC values ​​≤0.5, >0.5–1, >1–2, and >2 represent synergistic, additive, unrelated, and antagonistic effects, respectively. Figure 4 As shown in Figure A, 2 mg / mL (1 / 4 × MIC) G3 and 0.063 mg / mL (1 / 4 × MIC) cinnamaldehyde inhibited the growth of *E. coli* within 24 hours. The FIC of the combined use of G3 and cinnamaldehyde was 0.50, indicating a synergistic effect between the two drugs on *E. coli* growth. Furthermore, the FIC of the combined use of 2 mg / mL (1 / 8 × MIC) G3 and 0.063 mg / mL (1 / 4 × MIC) cinnamaldehyde on *Staphylococcus aureus* growth after 12 hours was 0.375, indicating a synergistic inhibitory effect between the two drugs on *Staphylococcus aureus* growth after 12 hours. Figure 4 B); and the FIC of the combined administration of 2 mg / mL (1 / 8×MIC) G3 and 0.125 mg / mL (1 / 2×MIC) cinnamaldehyde for 24 hours on the growth inhibition of Staphylococcus aureus was 0.625, indicating that the combined administration of G3 and cinnamaldehyde had an additive inhibitory effect on the growth of Staphylococcus aureus. Figure 4 C).

[0050] The measured E. coli cinnamaldehyde MIC: 0.25 mg / ml; E. coli G3 MIC: 8 mg / ml; the combination drug experiment was designed, and the data in the following table were the specific results of the influence of different time periods and different concentrations on bacterial growth, wherein M represents MIC; h represents the detection time after inoculation, the measured value is the OD value of the bacteria, 95LB+5 bacteria is the blank control group without adding drugs; see Table 1 for details. Similarly, see Table 2 for the detection of Staphylococcus aureus; the detected Staphylococcus aureus cinnamaldehyde MIC: 0.5 mg / ml; Staphylococcus aureus G3 MIC: 16 mg / ml.

[0051] Table 1, inhibition effect table of G3-cinnamaldehyde combination drug on E. coli.

[0052]

[0053]

[0054]

[0055]

[0056]

[0057] Table 2, inhibition effect table of G3-cinnamaldehyde combination drug on Staphylococcus aureus.

[0058]

[0059]

[0060]

[0061]

[0062] All the documents mentioned in the present application are cited as references in the present application, just as each document is cited as a reference individually. In addition, it should be understood that various modifications or changes can be made to the present application by those skilled in the art after reading the above teaching of the present application, and these equivalent forms also fall within the scope defined by the claims attached to the present application.

Claims

1. A bacteriostatic food additive, characterized by Comprising: a complex seasoning G3 having a concentration of equal to or greater than 2 mg / mL and cinnamaldehyde having a concentration of equal to or greater than 0.063 mg / mL; the complex seasoning G3 is composed of the following components by weight parts: white granulated sugar 8-15 parts malt dextrin 6-12 parts glucose 0.6-1.8 parts yeast extract 0.1-1.2 parts sodium gluconate 0.1-2 parts edible salt 0.1-1 part.

2. The bacteriostatic food additive of claim 1, wherein: The complex seasoning G3 is composed of the following components by weight parts: white granulated sugar 12 parts malt dextrin 9 parts glucose 1.2 parts yeast extract 0.6 parts sodium gluconate 1 part edible salt 0.5 parts.

3. The bacteriostatic food additive of claim 1, wherein: Also comprising vinegar powder.

4. The bacteriostatic food additive of claim 3, wherein: The complex seasoning G3 has a concentration of equal to or greater than 4 mg / mL; cinnamaldehyde has a concentration of equal to or greater than 0.25 mg / mL; vinegar powder has a concentration of equal to or greater than 4 mg / mL.

5. The bacteriostatic food additive according to any one of claims 1 to 4, characterized in that: The bacteria inhibited by the bacteriostatic food additive are any one of Escherichia coli, Staphylococcus aureus, Listeria and Vibrio parahaemolyticus.

6. The bacteriostatic food additive according to any one of claims 1 to 4, characterized in that: The bacteriostatic food additive inhibits Staphylococcus aureus for a duration of equal to or less than 12 hours.

7. The bacteriostatic food additive according to any one of claims 1 to 4, characterized in that: The bacteriostatic food additive inhibits Escherichia coli for a duration of equal to or less than 24 hours.

8. A process for the preparation of the bacteriostatic food additive as claimed in claim 4, characterized in that Comprising: (1) Each component is added to pure water and stirred to make a single component stock solution of complex seasoning G3 with a concentration of 100 mg / mL, vinegar powder with a concentration of 100 mg / mL, and cinnamaldehyde with a concentration of 20 mg / mL, and then filtered and stored; (2) The stock solution is mixed and diluted according to the required concentration of the formula to obtain the required bacteriostatic food additive.

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