Bacillus cereus-resistant antibacterial peptide, and preparation method and application thereof

Abstract

The application discloses an antibacterial peptide against Bacillus cereus and a preparation method and application thereof. The general formula of the amino acid sequence of the synthetic antibacterial peptide is [KR]GG[KR]XC[FWY]C[KR]X[KR][FWY]C[LIAV]CXG[KR][KR], wherein the antibacterial peptide XWC26 has strong antibacterial activity against the Bacillus cereus, can efficiently inhibit the spore germination of the Bacillus cereus, and can effectively control the pollution of the Bacillus cereus in food. The amino acid sequence of the antibacterial peptide is KGGKLCWCKNKWCICLGKR, the molecular weight is 2224.79 Da, the isoelectric point is 9.70, and the net charge number is 6. The antibacterial peptide has the advantages of good stability, difficulty in causing drug resistance, high synthesis efficiency and low production cost.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to characterization, preparation and application of an artificial synthetic antibacterial peptide against Bacillus cereus. BACKGROUND

[0002] Bacillus cereus is a gram-positive bacteria with facultative anaerobic spore-forming, which is a common foodborne pathogen. It can produce enterotoxins, leading to diarrhea and vomiting food poisoning, and even death in severe cases. It widely exists in soil, dust and water, and grows and reproduces in relatively loose conditions, and can produce defensive spores under harsh conditions. Therefore, Bacillus cereus contamination is found in various pre-packaged foods, such as rice products, dairy products and meat products. Studies have found that when the number of Bacillus cereus in the ingested food is >10 2 CFU / g(ml), there is a risk of causing foodborne illness, but it does not cause sensory changes, making it difficult for people to detect, so it is easy to be mis-eaten. China's eating habits mainly include rice, noodles and other starch products, and there is a habit of eating leftover food and overnight food, which makes food poisoning caused by Bacillus cereus occur frequently. According to reports, among bacterial food poisoning in China, food poisoning caused by Bacillus cereus ranks third in terms of the number of occurrences and the number of people affected.

[0003] Antimicrobial peptides (AMPs) are a class of polypeptides widely existing in nature, which have broad-spectrum antibacterial activity and can quickly kill various bacteria, fungi, viruses and parasites. The extensive use of antibiotics has led to the emergence of drug-resistant bacteria, which has prompted people to develop new antibacterial agents, and AMP is an excellent choice because of its unique biological activity and special action mechanism different from traditional antibiotics, and it has good bacteriostatic effect on antibiotic-resistant bacteria. Therefore, AMP has become one of the most potential alternatives to antibiotics, and its application in food, medicine, skin care and cosmetic preservatives is also becoming more and more widespread, and it has good development prospects.

[0004] Although antimicrobial peptides have universal advantages, there are also some obvious shortcomings, such as low antibacterial activity, poor stability, high toxicity, etc. of some antimicrobial peptides. The antimicrobial peptides obtained by modification or synthesis of existing antimicrobial peptides can greatly improve some of the above shortcomings to meet different application requirements and achieve the effect of reducing the amount and cost. The present application discloses a class of synthetic antibacterial peptides obtained by modification of host defense peptides, which have strong antibacterial effect and low inhibitory concentration for most bacteria. SUMMARY

[0005] In order to overcome the shortcomings and deficiencies of the prior art, the purpose of the present application is to provide a kind of synthetic antibacterial peptide, solve the problems in the above background art.

[0006] The purpose of the present application is achieved by the following technical solutions:

[0007] A kind of synthetic antibacterial peptide, its amino acid sequence general formula is [KR]GG[KR]XC[FWY]C[KR]X[KR][FWY]C[LIAV]CXG[KR][KR].

[0008] Expression [KR] in the bracket indicates that, in the first, fourth, ninth, eleventh, eighteenth and nineteenth positions in the syntax, lysine, arginine is also acceptable; [FWY] indicates that, in the seventh, twelfth position in the syntax, phenylalanine, tryptophan, tyrosine is also acceptable; [LIAV] indicates that, in the fourteenth position in the syntax, leucine, isoleucine, alanine, valine is also acceptable. Further, the amino acid sequence of the antibacillus cereus antibacterial peptide is as follows:

[0009]

[0010]

[0011]

[0012] Further, the molecular weight of the antibacillus cereus antibacterial peptide ranges from 1907.36 to 2583.08 Da, the isoelectric point ranges from 9.62 to 11.31, and the net charge number ranges from 6 to 8.

[0013] Further, the synthetic antibacterial peptide XWC26 has an amino acid sequence general formula of KGGKLCWCKNKWCICLGKR, a molecular weight of 2224.79 Da, an isoelectric point of 9.70, and a net charge number of 6.

[0014] The present application provides a kind of synthetic antibacterial peptide in the application of antibacterial drug, feed additive or food preservative, the antibacterial drug, feed additive or food preservative is used to inhibit and / or kill bacillus cereus.

[0015] In the present application, the antibacillus cereus antibacterial peptide is used in the preparation of a microbial inhibitor. The microbial inhibitor is a food preservative, a cosmetic preservative or a drug for preventing and treating bacterial infection. The microorganism is bacillus cereus, as well as other gram-positive bacteria and gram-negative bacteria.

[0016] Another purpose of the present application is to provide a preparation method of the above-mentioned synthetic antibacterial peptide.

[0017] The application relates to a synthesis method of an antibacterial peptide against Bacillus cereus, which adopts CTC resin as a solid-phase carrier and adopts an Fmoc solid-phase synthesis method to synthesize a peptide with an amino acid sequence of a target product.

[0018] Compared with the prior art, the application has the following beneficial effects:

[0019] The application designs and screens a new polypeptide with an amino acid sequence through bioinformatics technology. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 is a cell morphology result diagram after antibacterial peptide XWC26 acts on Bacillus cereus.

[0021] Figure 2 is an inhibition effect result diagram of antibacterial peptide XWC26 on spore germination of Bacillus cereus.

[0022] Figure 3 is a detection result diagram of hemolytic activity of antibacterial peptide XWC1.

[0023] Figure 4 is a sterilization result diagram of antibacterial peptide XWC26 in food samples. DETAILED DESCRIPTION

[0024] The specific implementation of the application is further illustrated below in combination with examples, but the implementation and protection of the application are not limited to this. It should be pointed out that, if the following processes are not particularly detailed, they can be realized or understood by referring to the prior art by those skilled in the art. If the reagents or instruments used are not marked with manufacturers, they are regarded as conventional products that can be obtained by market purchase.

[0025] Example 1: Design and screening of antibacterial peptides

[0026] In view of the above-mentioned technical problems, the application designs and screens an antibacterial peptide library with strong antibacterial activity and capable of effectively inhibiting drug-resistant bacteria by modifying the sequence and spatial structure of host defense peptides in a certain conservation manner, and verifies the antibacterial activity.

[0027] The application is designed on the basis of the following general amino acid sequence:

[0028] [KR]GG[KR]XC[FWY]C[KR]X[KR][FWY]C[LIAV]CXG[KR][KR]

[0029] (The expression in the bracket [KR] means that, in the first position in the syntax, lysine, arginine is also acceptable)

[0030] Table 1 Sequence list of antibacterial peptides XCW1-XCW50

[0031]

[0032] Example 2: Synthesis of antibacterial peptides

[0033] The antibacterial peptide of the present application adopts conventional polypeptide solid-phase synthesis. Taking antibacterial peptide XWC2 as an example, its sequence is: KGGKLCWCKWKYCICLGKR.

[0034] Sequence characteristics: the sequence type is an amino acid sequence, contains 19 amino acid residues, the molecular weight size is 2273.89 Da, and the net charge number is 6.

[0035] The specific synthesis steps are as follows: 1) resin swelling: take the degree of substitution of 0.1 mmol 2-Chlorotrityl Chloride Resin resin, put the resin into the reaction tube, add DCM (15 ml / g) solvent, and shake for 40 min. 2) Connect the first amino acid: filter out the DCM solvent through the sand core, add 3 times the amount of Fmoc-Arg(Pbf)-OH amino acid, then add 2,4,6-trimethylpyridine, and finally add DCM for dissolution, shake for 12 h, and then wash with DMF for 3 times. 3) Deprotection: add 10 ml of 20% piperidine DMF solution (15 ml / g), shake for 6 min, then remove the piperidine DMF solution, add 10 ml of 20% piperidine DMF solution (15 ml / g), and shake for 20 min. 4) Detection: remove the piperidine DMF solution, take a dozen resins, wash with ethanol three times, add one drop of ninhydrin and phenol solution, heat at 105℃-110℃ for 3 min, and turn dark blue for positive reaction. 5) First washing: wash with DMF (10 ml / g) for 3 times. 6) Blocking: add methanol (10 ml / g) and 3 times the amount of DIEA, shake for 20 min, and then wash with DMF for 3 times. 7) Condensation (generate the second amino acid Lysine in the sequence): add 3 times the amount of Fmoc-Lys(boc)-OH amino acid, HCTU, and DIEA to the reaction tube, dissolve with as little DMF as possible, and react for 1 h. 8) Second washing: wash with DMF (10 ml / g) once, and then wash with DMF (10 ml / g) twice. 9) Repeat the above operation to connect the amino acids in the sequence from left to right. 10) After connecting the last amino acid, deprotection, and wash the resin according to the following method: DMF (10 ml / g) twice, DCM (10 ml / g) twice, and dry for 8 min. 11) Cut the polypeptide from the resin: the ratio of resin and cleavage solution is 10 ml / g, constant temperature shaking for 150 min. (The cleavage solution is prepared according to the volume percentage, which can be TFA 95%, water 2.5%, and TIS 2.5%). 12) Blow dry and wash: blow the cleavage solution as much as possible with nitrogen, chromatograph with ether, wash with ether six times, and then dry at room temperature to obtain the crude peptide. 13) Analyze and identify the polypeptide by HPLC-MS: (1) Prepare a solution of the crude peptide with a concentration of 1 mg / ml. (2) Filter the solution with a 0.45 μm filter membrane. (3) Analysis: take 10 μl for HPLC-MS analysis. The mobile phase is water and acetonitrile, and the time is 25 min, isocratic elution, first equilibrate the HPLC with isocratic gradient for 5 min, then inject the sample, gradient water 40%, acetonitrile 60%; get the purity and MS identification. 13) Freeze-dry the purified solution to obtain the finished product. 14) Seal the white powder-like polypeptide and store at -20 degrees.

[0036] Example 3: Determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)

[0037] The minimum inhibitory concentration (MIC) of 50 antimicrobial peptides that conform to the general formula predicted and synthesized in Examples 1 and 2 was determined, and the peptide with the lowest MIC value and the strongest antimicrobial activity was screened out.

[0038] In this experiment, the Bacillus cereus type strain ATCC14579 was selected as the test strain. Bacillus cereus was streaked onto antibiotic-free MH solid medium and incubated at 37°C for 16 hours. Three to five colonies were then inoculated into 3 mL of MH liquid medium and incubated overnight. The OD of the bacterial culture was measured the following morning using a spectrophotometer. 600 Value, and dilute it to 5×10 5 CFU / mL. 90 μL of diluted bacterial suspension was added to a 96-well plate. The antimicrobial peptide was diluted twofold, and 10 μL of each solution was added sequentially to wells containing bacterial suspension, resulting in final concentrations of 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2 μg / mL, 1 μg / mL, 0.5 μg / mL, and 0.25 μg / mL. Ampicillin was used as a control. The 96-well plates were incubated at 37°C for 16 h, and the OD values ​​of each well were measured. 600 The minimum inhibitory concentration (MIC) is the lowest concentration of antimicrobial peptide at which no bacterial growth is observed. To determine the minimum bactericidal concentration (MBC), 10 μL of suspension was drawn from each well and spotted onto an agar plate; the lowest peptide concentration that resulted in no bacterial colony growth was recorded as the MBC. This experiment was repeated three times, with three replicates each time.

[0039] The results are shown in Table 2. XWC26 had the most significant antibacterial activity against Bacillus cereus with a MIC value of 2 μg / mL. XWC1, XWC7, XWC35, and XWC46 all had MIC values ​​of 4 μg / mL against Bacillus cereus, and their antibacterial activity was superior to the other antimicrobial peptides.

[0040] The minimum bactericidal concentration (MBC) of antimicrobial peptides (XWC1, XWC7, XWC26, XWC35, and XWC46) with MIC ≤ 4 μg / mL was further determined. The results are shown in Table 3. Except for antimicrobial peptide XWC7, whose MBC value was twice the MIC value, the MBC values ​​of the other four peptides were consistent with their MIC values. The experiment shows that the above-mentioned antimicrobial peptides have strong antimicrobial activity against Bacillus cereus.

[0041] Table 2 shows the MIC (μg / mL) of XWC1-AMP50 against Bacillus cereus.

[0042]

[0043]

[0044] Table 3. MBC (μg / mL) of antimicrobial peptide XWC1 and other antimicrobial peptides against Bacillus cereus.

[0045]

[0046] Example 4: Antibacterial spectrum determination

[0047] The antimicrobial activity of the antimicrobial peptides (XWC1, XWC7, XWC26, XWC35, XWC46) with MIC ≤ 4 μg / mL in Example 3 was determined, and their antimicrobial activity against Gram-positive bacteria (such as Staphylococcus aureus) and Gram-negative bacteria (such as Escherichia coli) was evaluated.

[0048] Each test strain was streaked onto antibiotic-free MH solid medium and incubated at 37°C for 16 hours. Three to five colonies were then inoculated into 3 mL of MH liquid medium and incubated overnight. The OD of the bacterial culture was measured the following morning using a spectrophotometer. 600 Value, and dilute it to 5×10 5 CFU / mL. 90 μL of diluted bacterial suspension was added to a 96-well plate. The antimicrobial peptide was diluted twofold, and 10 μL of each solution was added sequentially to wells containing the bacterial suspension, resulting in final concentrations of 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2 μg / mL, 1 μg / mL, 0.5 μg / mL, and 0.25 μg / mL. Ampicillin was used as a control. The 96-well plates were incubated at 37°C for 16 h, and the OD values ​​of each well were measured. 600 The minimum inhibitory concentration (MIC) is the lowest concentration of antimicrobial peptide at which bacterial growth is not visible. This experiment was repeated three times, with three replicates each time.

[0049] The results, shown in Table 4, indicate that the antimicrobial peptides exhibited stronger antimicrobial activity against tested Gram-positive bacteria (such as Staphylococcus aureus and Bacillus subtilis) than against Gram-negative bacteria (such as Escherichia coli and Pseudomonas aeruginosa). The experiments demonstrate that the aforementioned antimicrobial peptides possess broad-spectrum antimicrobial activity and are particularly effective against Gram-positive bacteria.

[0050] Table 4. Determination of the antimicrobial spectrum of antimicrobial peptides such as XWC1.

[0051]

[0052] Example 5: Observation of Bacillus cereus cell morphology before and after treatment with antimicrobial peptide XWC26

[0053] Bacillus cereus ATCC14579 was inoculated into antibiotic-free LB liquid medium and cultured overnight. The OD of the bacterial culture was measured using a spectrophotometer the following morning. 600 Value, and dilute it to 1×10 8 CFU / mL of antimicrobial peptide XWC26 was added to a final concentration of 2×MIC. Using PBS as a negative control, the mixture was incubated at 37°C for 1 h. Then, the cells were centrifuged at 8000 rpm for 3 min, the supernatant was discarded, and the cells were washed three times with PBS buffer. Next, 3% (v / v) glutaraldehyde was added, and the mixture was fixed overnight at 4°C. After thorough fixation, the cells were centrifuged at 8000 rpm for 5 min, washed three times with PBS buffer, and then dehydrated sequentially with 30%, 50%, 70%, 80%, and 90% ethanol solutions for 15 min each time, followed by two cycles of dehydration with 100% ethanol. Finally, the cells were critically dried, sputter-coated with gold, and observed under a scanning electron microscope.

[0054] Scanning electron microscopy results as follows Figure 1 As shown, untreated Bacillus cereus cells have normal morphology, intact cell membranes, and smooth surfaces. However, after treatment with the antimicrobial peptide XWC26, the bacterial membrane surface undergoes significant changes, becoming wrinkled and rough, with obvious membrane rupture and leakage of cytoplasmic components. The experiment demonstrates that the antimicrobial peptide XWC26 can disrupt the Bacillus cereus cell membrane, alter cell membrane permeability, and ultimately lead to bacterial death.

[0055] Example 6: Antimicrobial peptides inhibit the germination of Bacillus cereus spores

[0056] Bacillus cereus ATCC14579 was inoculated into antibiotic-free LB liquid medium and cultured for 48 h. After dilution, 100 μL of the bacterial suspension was spread onto spore medium and cultured for 5 days. Cells were then scraped from the culture plate and washed three times with PBS buffer, centrifuged at 4000 rpm for 20 min, and resuspended in PBS buffer. The spore suspension was incubated at 80°C for 10 min to kill vegetative cells, followed by washing three times with PBS buffer to remove vegetative cells. The spore suspension was diluted with an appropriate amount of LB medium. 200 μL of the spore suspension and 200 μL of peptide solutions of different concentrations were added to 96-well plates and incubated overnight. A 10 μL sample was then diluted and spread. After 16 h of incubation, plate counting was performed. Plates without antimicrobial peptides served as controls. This experiment was repeated three times, with three replicates per well.

[0057] like Figure 2As shown, the spore germination rate was 75% when the B. cereus spores were treated with the antibacterial peptide XWC26 at a concentration of 2 μg / mL, and the spore germination rate decreased to 15% when the concentration increased to 8 μg / mL. When the concentration increased to 16 μg / mL, the spore germination rate was less than 10%. The experiment showed that the antibacterial peptide XWC26 at a high concentration could effectively inhibit the germination of B. cereus spores.

[0058] Example 7: Stability determination of antibacterial peptides

[0059] Thermal stability determination: The antibacterial peptides (XWC1, XWC7, XWC26, XWC35, XWC46) were respectively placed at 25, 50, 75 and 100°C for 1 h, and then restored to room temperature. The MIC of the antibacterial peptides against B. cereus was determined, and the untreated antibacterial peptides (4°C) were used as a control. The experiment was repeated 3 times, with 3 parallel samples each time.

[0060] Acid-base stability determination: Buffer solutions with pH values of 2 (50 mM, glycine-HCl buffer), 6 (50 mM, sodium phosphate buffer) or 10 (50 mM, glycine-NaOH buffer) were prepared. The above antibacterial peptides were dissolved in the buffer solutions and incubated at 37°C for 1 h. The MIC of the antibacterial peptides against B. cereus was determined, and the antibacterial peptides dissolved in PBS buffer were used as a control. The experiment was repeated 3 times, with 3 parallel samples each time.

[0061] Protease stability determination: Pepsin, trypsin and protease-K were respectively dissolved in PBS buffer to a final concentration of 20 μg / mL. The above antibacterial peptides were mixed with the above proteases at a volume ratio of 1:1 at 37°C for 1 h. Then, the peptide-protease mixture was heated at 60°C for 15 min to inactivate the protease activity. The MIC of the antibacterial peptides against B. cereus was determined, and the antibacterial peptides mixed in PBS buffer were used as a control. The experiment was repeated 3 times, with 3 parallel samples each time.

[0062] Table 5 is the temperature tolerance result. As can be seen from the table, the activities of the above 5 antibacterial peptides remain unchanged after being treated at a temperature of up to 100°C, indicating that the above antibacterial peptides have good thermal stability and can tolerate heat treatment during food processing.

[0063] Table 6 is the acid-base tolerance result. The antibacterial activity of XWC7 is not affected by pH changes; XWC1, XWC26, XWC35 and XWC46 maintain antibacterial activity at pH 2-6, and the activity slightly decreases at pH 10, indicating that the antibacterial peptides have good stability under acidic and neutral conditions. In alkaline conditions, precipitation occurs due to reaching the isoelectric point of the protein, which affects the antibacterial activity to a certain extent.

[0064] Table 7 is the protease tolerance results, XWC1, XWC26, XWC35, XWC46 remain unchanged in the presence of pepsin, XWC7 activity is slightly reduced, while in the presence of trypsin or proteinase-K, the activity of the above antibacterial peptides is reduced by 8-16 times, indicating that the above antibacterial peptides remain good stability in the presence of pepsin, and poor stability in the presence of trypsin and / or proteinase-K.

[0065] Table 5 Temperature tolerance determination of antibacterial peptides XWC1 and the like

[0066]

[0067] Table 6 Acid-base tolerance determination of antibacterial peptides XWC1 and the like

[0068]

[0069] Table 7 Protease tolerance determination of antibacterial peptides XWC1 and the like

[0070]

[0071] Example 8: Hemolytic activity determination of antibacterial peptides

[0072] Take 100 μL of 6% (v / v) mouse red blood cell suspension (Shanghai Yuan Ye) and add it to a 96-well plate, then add 100 μL of different concentrations of antibacterial peptides (XWC1, XWC7, XWC26, XWC35, XWC46) to a final concentration of 1-128 μg / mL, and take the same amount of PBS and 1% (v / v) Triton X-100 as negative and positive control groups, respectively. Incubate the 96-well plate at 37°C for 1 hour, centrifuge at 1000 r / min for 3 min, take the supernatant to a new 96-well plate, measure the OD 540 nm value of each well, and calculate the hemolysis rate, the hemolysis rate calculation formula is as follows: Hemolysis rate (%) = (sample OD 540 nm-negative control OD 540 nm) / (positive control OD 540 nm-negative control OD 540 nm) x 100

[0073] Figure 3 The hemolytic activity of the above antibacterial peptides changes with the concentration. As can be seen from the figure, the antibacterial peptide concentration is below 2 μg / mL, almost no hemolytic activity, when the concentration is increased to 4 μg / mL, it begins to show hemolytic activity, and when the concentration is increased to 8 μg / mL, it shows obvious hemolytic activity, and the hemolysis rate is higher than 10%. It is shown that the above antibacterial peptides have good biological safety at their minimum inhibitory concentration (MIC≤4 μg / mL), but still need to be improved.

[0074] Example 9: Application of the embodiment in food

[0075] The cooked rice was chosen as the experimental object to study the preservative effect of the antibacterial peptide XWC26 in food. The rice sample was taken from a student canteen and heated in a microwave oven at high heat for 3 min to kill microorganisms. 10 g of cooked rice was diluted with 40 mL of distilled water and shaken for 10 min. Then, 5 x 10 5 The rice sample was artificially contaminated with 5 x 105CFU / mL of Bacillus cereus and left at 25 °C for 1 h. The antibacterial peptide was added to the sample to a final concentration of 8 μg / mL and incubated at 25 °C for 3 h, and the sample without the antibacterial peptide was used as a control. Sampling was performed every 1 h, and the sample was diluted and plated. The plates were incubated at 37 °C for 16 h, and the average number of colonies was counted. The experiment was performed 3 times.

[0076] The results, as shown in Figure 4 Figure 2, show that the sample treated with the antibacterial peptide XWC26 decreased by about 2 orders of magnitude within 2 h, indicating that it can inhibit and kill the bacteria in the rice sample in a short time and can be used for food preservation.

[0077] The above embodiments are only the preferred embodiments of the present application, which are used to explain the present application, but not to limit the present application. The changes, replacements, modifications, etc. made by those skilled in the art without departing from the spirit and essence of the present application should belong to the protection scope of the present application.

Claims

1. An antimicrobial peptide against Bacillus cereus, characterized in that: The amino acid sequence of the antimicrobial peptide against Bacillus cereus is as follows: 。 2. The antimicrobial peptide against Bacillus cereus according to claim 1, characterized in that: The molecular weight of the antibacterial peptide against Bacillus cereus ranges from 1907.36 to 2583.08 Da, the isoelectric point ranges from 9.62 to 11.31, and the net charge number ranges from 6 to 8.

3. The antimicrobial peptide against Bacillus cereus according to claim 1, characterized in that, One of the synthetic antimicrobial peptides, XWC26, has a molecular weight of 2224.79 Da, an isoelectric point of 9.70, and a net charge of 6.

4. The method for synthesizing the antimicrobial peptide against Bacillus cereus according to any one of claims 1 to 3, wherein the peptide having the amino acid sequence of the target product is synthesized by using CTC resin as a solid phase carrier and Fmoc solid phase synthesis method.

5. The application of the antibacterial peptide against Bacillus cereus according to any one of claims 1 to 3 in inhibiting the activity of Bacillus cereus and inhibiting the germination of Bacillus cereus spores.

6. The application of the antimicrobial peptide against Bacillus cereus according to claim 1 in the preparation of microbial inhibitors; wherein the microorganism is Bacillus cereus, and other Gram-positive and Gram-negative bacteria.

7. The application according to claim 6, characterized in that: The aforementioned microbial inhibitors are food preservatives, cosmetic preservatives, or drugs for preventing and treating bacterial infections.

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