Preparation method and application of antibacterial ovotransferrin fiber
By preparing antibacterial egg transferrin fibers, the problem of insufficient research on egg transferrin fibrosis was solved, and the significant antibacterial effect and intestinal microorganism regulation was achieved on different types of microorganisms. It was used to prevent diarrhea in infants and young children, providing a Safe, natural antibacterial products.
Patent Information
- Application Number
- CN202411269122.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-11
- Publication Date
- 2024-12-13
AI Technical Summary
There are few studies on egg transferrin fibrosis in the prior art, which limits its application in the field of antibacterial, especially the mechanism of action and influencing factors of different types of microorganisms.
By dissolving egg transferrin in deionized water, adjusting the pH to 1.5-2.0, and freeze-drying after heating and freeze-drying, antibacterial egg transferrin fibers were prepared, which significantly enhanced its antibacterial effect and was used to regulate intestinal microorganisms. To prevent diarrhea in infants and young children.
The prepared antibacterial egg transferrin fibers have significant antibacterial effects, which can regulate intestinal microorganisms by destroying bacterial cell membranes, reducing diarrhea and intestinal mucosal barrier damage, and provide a natural and safe antibacterial product.
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Figure CN119121433A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to a field, and in particular to a preparation method and application of antibacterial ovotransferrin fiber. Background Art
[0002] Ovotransferrin (OVT), as a key protein, accounts for about 12% of the protein content in chicken egg white and is functionally similar to transferrin in human serum, however, their sugar chain structures are significantly different. Ovotransferrin contains 0.8% mannose, no galactose and sialic acid, and has a molecular weight of 70,000-78,000 Da. Ovotransferrin is an iron-binding glycoprotein associated with many biological activities, with multiple biological benefits such as anticancer and antiviral properties, and exhibits a wide range of antibacterial properties, showing significant antibiotic effects against various types of bacteria, including various Gram-positive and negative strains such as Escherichia coli, Pseudomonas, Streptococcus, Staphylococcus aureus, and Salmonella enterica. Therefore, ovotransferrin has the potential for wide application in the design and engineering of functional foods.
[0003] Infant diarrhea is a gastrointestinal disorder in infancy, a syndrome characterized by diarrhea and vomiting. Some newborns and infants will be affected by intense and excessive colic. The pathogenic factors are divided into three aspects, namely constitution, infection and digestive dysfunction. The main clinical manifestations are increased bowel movements, loose stools and water and electrolyte disorders. If not treated in time, serious water and electrolyte disorders will occur, which may endanger the lives of infants and young children. According to statistics from the World Health Organization, in developing countries, infectious diarrhea is the infectious disease with the highest incidence in young children, with a mortality rate of about 20‰. In Asia, Africa and Latin America alone, it kills about 4.6 million infants and young children every year. Therefore, it is particularly important to provide immediate and targeted treatment for diarrhea in young children.
[0004] Intestinal microorganisms and their metabolites make important contributions to human health by promoting the development of the body's immune system, regulating adaptive immunity and activating B cells or T cells to produce immune responses. When the body is infected by foreign pathogens, the disorder of intestinal flora is usually accompanied by a decline in the body's immunity, which in turn leads to diarrhea and damage to the intestinal mucosal barrier.
[0005] A large number of studies have confirmed that the gastrointestinal mucosal system can not only obtain various nutrients during the digestion and absorption of food, but also phagocytize and present foreign substances (antigens) through dendritic cells and other antigen-presenting cells distributed in the intestinal mucosal system, and convey information to immune cells, thereby activating the body's innate immune system and regulating the body's immune function. Studies have found that ovotransferrin is an avian acute phase protein that regulates the function of macrophages and heterophils by stimulating the production of interleukin-6 (IL-6) and nitrite, and promotes the proliferation of spleen lymphocytes to produce immune responses. In addition, ovotransferrin has the function of promoting the maturation of intestinal dendritic cells, increasing the number of beneficial microorganisms such as Akkermansia in the intestine, and affecting the intestinal mucosal morphology by increasing the depth of the crypts, which is beneficial to the expression of tight junction proteins in the intestine. Therefore, as a natural immunomodulator, ovotransferrin can improve intestinal microorganisms and reduce diarrhea and damage to the intestinal mucosal barrier, thereby enhancing the body's immune function. However, the current chemical drugs for treating diarrhea, such as loperamide and metronidazole, while relieving diarrhea, have side effects such as dizziness, nausea, vomiting and constipation. Therefore, the development of green, non-toxic, safe and effective food-borne products for treating diarrhea has broad application prospects.
[0006] The self-assembly fibrillation of ovotransferrin can not only enhance its biological activity, such as antibacterial activity, but also improve its physicochemical properties, such as solubility and thermal stability, thereby broadening its application in the food and biomedical fields. However, there are relatively few studies on the fibrillation of ovotransferrin, and there are basically no studies and inventions on the structure and functional properties of ovotransferrin fibers formed by thermal polymerization self-assembly. Secondly, the research on the mechanism of action of ovotransferrin fibers against different types of microorganisms and their influencing factors is still insufficient, which limits the potential of applying ovotransferrin fibers in the antibacterial field. Summary of the invention
[0007] The purpose of the present invention is to fill the gap in the current research field of ovotransferrin fibrillation, and provide a preparation method and application of antibacterial ovotransferrin fiber. Ovotransferrin fiber can be obtained by a simple and rapid method. It is a natural antibacterial product with good antibacterial effect, clarifies the mechanism of action of ovotransferrin in fibrosis, and provides a theoretical basis for the research and development of new antibacterial materials; at the same time, the antibacterial ovotransferrin fiber and its byproducts regulate intestinal microorganisms to achieve the purpose of preventing diarrhea in infants and young children.
[0008] To achieve the above purpose, the technical solution designed by the present invention is as follows:
[0009] The present invention provides a method for preparing antibacterial ovotransferrin fiber, comprising the following steps:
[0010] (1) dissolving ovotransferrin in deionized water to prepare an ovotransferrin solution;
[0011] (2) adjusting the pH of the ovotransferrin solution to 1.5-2.0;
[0012] (3) The ovotransferrin solution is heated, quickly placed in an ice water bath to terminate the reaction, and freeze-dried to obtain a powder, which is the antibacterial ovotransferrin fiber.
[0013] Furthermore, in the step (1), the mass fraction of the ovotransferrin solution is 1.5-2% (w / w);
[0014] In the step (2), the pH adjuster is 2M hydrochloric acid;
[0015] In the step (3), the heating treatment conditions are: 85°C < heating temperature ≤ 95°C, 0h < heating time ≤ 24h.
[0016] The present invention also provides an application of the antibacterial ovotransferrin fiber prepared by the above preparation method in antibacterial applications.
[0017] Furthermore, the antibacterial strain is Gram-spore bacteria.
[0018] Furthermore, the Gram-spore bacteria are Escherichia coli or Staphylococcus aureus.
[0019] The present invention also provides an application of the antibacterial ovotransferrin fiber prepared by the above preparation method in the preparation of an antibacterial agent for inhibiting Gram-bacteria.
[0020] The present invention also provides an antibacterial ovotransferrin fiber antibacterial agent, which includes antibacterial ovotransferrin fibers; wherein the antibacterial ovotransferrin fibers are prepared by the preparation method according to claim 1, and the concentration of the antibacterial ovotransferrin fibers in the antibacterial agent is 128-200 μg / mL.
[0021] The present invention also provides an application of the antibacterial ovotransferrin fiber prepared by the preparation method in preparing a medicine for treating diarrhea in infants and young children.
[0022] The present invention also provides a medicine for treating infantile diarrhea, which comprises an effective amount of the antibacterial ovotransferrin fiber prepared by the preparation method and auxiliary materials.
[0023] Furthermore, the drug comprises, by weight, 1 part of antibacterial ovotransferrin fiber and 0.9 to 2 parts of auxiliary materials.
[0024] Furthermore, in terms of weight, the auxiliary materials are 0.5 to 1 part of vitamin C, 0.2 to 0.5 part of sucrose and 0.2 to 0.5 part of maltose.
[0025] Furthermore, the drug includes, by weight, 1 part of antibacterial ovotransferrin fiber, 0.6 part of vitamin C, 0.2 part of sucrose and 0.2 part of maltose.
[0026] Beneficial effects of the present invention:
[0027] 1. The preparation method of the present invention heats ovotransferrin to form self-assembled protein fibers, which have a significant antibacterial effect by destroying bacterial cell membranes.
[0028] 2. The antibacterial ovotransferrin fiber prepared by the present invention is a natural immunomodulator, which has the effects of promoting the maturation of intestinal dendritic cells, increasing the number of beneficial microorganisms such as Akkermansia in the intestine, reducing diarrhea and damage to the intestinal mucosal barrier, thereby regulating intestinal microorganisms and preventing diarrhea in infants and young children. Therefore, by developing antibacterial ovotransferrin fiber, a new type of biofunctional material with broad application prospects in medical treatment, food, bioengineering, etc. is prepared.
[0029] 3. The product of the present invention is administered at a therapeutically effective dose or a prophylactically effective dose. Regular ovotransferrin administration will allow for a more constant level of protein, resulting in a prolonged protective effect. BRIEF DESCRIPTION OF THE DRAWINGS
[0030] Figure 1 This is the maximum fluorescence intensity change diagram of OVTF at different heating times;
[0031] In the figure, Note: the reaction conditions were pH 2 and 90°C;
[0032] Figure 2 TEM images of OVTF at different heating times;
[0033] Figure 3 This is the change diagram of the surface hydrophobicity of OVTF at different heating times;
[0034] Figure 4 This is the Zeta potential change diagram of OVTF at different heating times;
[0035] Figure 5 The SDS-PAGE profiles of OVTF at different heating times;
[0036] In the figure, from left to right: protein marker and OVTF were heated for 0, 1, 2, 4, 8, 12, 16, 20, and 24 h, respectively;
[0037] Figure 6 The UV spectra of antibacterial ovotransferrin fiber at different heating times;
[0038] Figure 7 The infrared spectra of antibacterial ovotransferrin fiber at different heating times;
[0039] Figure 8 The antibacterial effects of OVT oligomers and fibers prepared by heating at pH 2.0, 37°C or 90°C for 16 h on E. coli (A) and S. aureus (B);
[0040] Fig. 9 The results of colony forming units (cfu) of E. coli (A) and S. aureus (B) under the treatment of antibacterial ovotransferrin fibers at concentrations of 2, 10, 25, 50, 100, and 200 μg / mL;
[0041] Fig.10 SEM images of E. coli and S. aureus after exposure to blank control (PBS buffer), OVT, oligomers and OVTF for 6 h;
[0042] Fig.11 This is a graph showing the effect of antibacterial ovotransferrin fiber on the frequency of diarrhea in infants and young children. DETAILED DESCRIPTION
[0043] The present invention is further described in detail below in conjunction with specific embodiments so that those skilled in the art can understand.
[0044] The method for preparing the antibacterial ovotransferrin fiber of the present invention comprises the following steps:
[0045] (1) dissolving ovotransferrin (hereinafter referred to as OVT) in deionized water to prepare an ovotransferrin solution (OVT solution) with a mass fraction of 1.5 to 2% (w / w);
[0046] (2) using 2 M hydrochloric acid to adjust the pH of the ovotransferrin solution to 1.5-2.0;
[0047] (3) The ovotransferrin solution is heated at a temperature greater than 85° C. and less than or equal to 95° C., and then quickly placed in an ice water bath to terminate the reaction. The powder obtained by freeze drying is the antibacterial ovotransferrin fiber (hereinafter referred to as OVTF).
[0048] Example 1
[0049] The protein structure characterization and determination of the antibacterial ovotransferrin fibers prepared based on the above method.
[0050] The antibacterial ovotransferrin fiber structure was characterized by ThT fluorescence spectroscopy analysis, transmission electron microscopy observation, surface hydrophobicity test, Zeta potential measurement, SDS-PAGE gel electrophoresis, and ultraviolet and infrared spectroscopy analysis.
[0051] 1. ThT fluorescence spectrum analysis
[0052] a. Prepare ThT concentrate by dissolving 8 mg ThT in 10 mL phosphate buffer (10 mM, pH 7.0) containing 150 mM NaCl. Remove undissolved particles with a 0.22 μm filter. The concentrate should be stored in a dark place at 4°C and covered with tin foil. Before each measurement, dilute the ThT concentrate 50 times with PBS to obtain the ThT working solution.
[0053] b. Mix 40 μL of sample (20 mg / mL) with 4 mL of ThT working solution. After reacting for 2 min, measure ThT fluorescence using IF-5401 fluorescence spectrophotometer (Shimadzu Corporation, Japan). Set the excitation wavelength to 460 nm, the emission wavelength range to 450-600 nm, and the slit width to 5 nm. Record the maximum absorption peak height at around 489 nm.
[0054] The results are as follows Figure 1 As shown: When the heating time was 16 h, the antibacterial ovotransferrin fibers showed the highest fluorescence intensity and formed the largest number of fibers.
[0055] 2. Transmission electron microscopy observation
[0056] The prepared protein fiber solution was diluted to a protein concentration of 0.1% (w / v), and 1 to 2 drops of the sample to be tested were adsorbed onto a copper mesh plate and dried for about 10 minutes. The microscopic morphology of the sample was observed using an H-7650 transmission electron microscope (Hitachi, Tokyo, Japan) using an operating voltage of 100 kV.
[0057] The results are as follows Figure 2 As shown: After heating for 16 hours, the largest number of long and short fibers were observed.
[0058] 3. Surface hydrophobicity test
[0059] 8-phenylamino-1-naphthalenesulfonic acid (ANS) is used as a fluorescent probe for detection; the specific process is as follows:
[0060] First, various protein samples were diluted to a concentration of 0.01 to 0.05 mg / mL with 50 mM phosphate buffer (pH 7.2). Then, 20 μL of ANS solution (concentration of 8 mM) was added to 4 mL of the protein dilution solution. When recording the fluorescence emission spectrum, 390 nm was selected as the excitation wavelength and 470 nm as the emission wavelength, and the slit width was set to 5 nm. By analyzing the initial slope of the trend line of fluorescence intensity changing with protein concentration, the change in hydrophobicity of the protein surface was observed.
[0061] The results are as follows Figure 3 As shown in Figure 2, the surface hydrophobicity increased significantly during the first 8 h of heat treatment at pH 2. After 8 h of heating, the surface hydrophobicity gradually decreased.
[0062] 4. Zeta potential measurement
[0063] The protein fiber samples were diluted with buffer solution (5 mM PBS, pH 2.0) at a 20-fold dilution concentration to avoid multiple scattering effects. The samples were equilibrated at 25°C for 60 seconds in a dynamic light scattering particle size analyzer (Malvern Instruments Ltd., UK). Data were collected for at least 12 consecutive readings.
[0064] The results are as follows Figure 4 As shown: After 16 hours of heating treatment, the highest charge was observed, indicating that there was a significant electrostatic repulsion between the protein fibers, forming a self-assembled protein fiber structure, which led to the enhanced stability of the solution.
[0065] 5. SDS-PAGE gel electrophoresis
[0066] First, the sample was diluted to a concentration of 2 mg / mL with deionized water and used together with the initial protein solution for gel electrophoresis using a DYCZ-24DN gel electrophoresis instrument (Six One Factory, Beijing). 80 μL of the sample was placed in a 200 μL centrifuge tube and an appropriate amount of loading buffer was added. Before spotting, the sample was heated in boiling water for 5 min to achieve protein binding and denaturation with SDS, and the spotting volume was set to 10 μL. During the gel electrophoresis, a constant voltage mode was used. In the initial stage, a fixed voltage of 80 V was used for electrophoresis separation of the concentrated gel. When the sample entered the separation gel, the voltage was adjusted to 120 V. After the electrophoresis was completed, the film needed to be immersed in a solution containing ethanol and acetic acid for 30 min. Subsequently, the film was transferred to a staining solution (containing 0.25% Coomassie Brilliant Blue R250, 50% methanol, and 10% acetic acid) for 1 h of staining. A decolorizing solution prepared by mixing methanol, acetic acid and water in a volume ratio of 2:2:9 was used for decolorizing until the stripes were clearly visible and the background color completely disappeared.
[0067] The results are as follows Figure 5 As shown: As the heating treatment time increases, the stripes in the molecular weight range gradually weaken and become thinner, and more low molecular weight stripes appear, indicating that the protein is hydrolyzed into smaller fragments and develops into mature self-assembled protein fibers.
[0068] 6. Ultraviolet spectrum analysis
[0069] The samples were diluted to a concentration of 0.5 mg / mL with deionized water and scanned using a Nanodrop 2000C UV spectrophotometer (Thermo Fisher Scientific, USA) in the wavelength range of 230 to 350 nm, with 295 nm set as the excitation wavelength, and deionized water was used as the buffer solution blank control. Each sample was scanned three times, and the average of the three scans was used as the final result, and deionized water was used as the reference background for analysis.
[0070] The results are as follows Figure 6 As shown: With the increase of heating time and temperature, the UV absorption intensity of OVT shows an upward trend, confirming that heating causes the protein structure to unfold and form fibers.
[0071] 7. Infrared spectroscopy analysis
[0072] The ovotransferrin fiber sample was freeze-dried to turn it into a powder state, and then 1 to 3 mg of dry sample powder was taken out. Next, the extracted sample was mixed with dry potassium bromide (KBr) powder at a ratio of 1:100 and ground into powder for tablet preparation. Then, the prepared tablets were scanned and analyzed using a NEXUS670 Fourier transform infrared spectrometer (Thermo Fisher Scientific, USA), and the measurement parameters were set as follows: the scanning range was 4000 to 400 cm at room temperature 25°C. -1 , the instrument resolution is set to 4cm -1 , 32 scans were performed, using air as the scanning background. Background subtraction was performed before scanning each sample.
[0073] The results are as follows Figure 7 As shown: within the initial 8 hours of heating, the secondary structure of ovotransferrin has changed, the content of α-helix structure has decreased, and the content of β-fold has increased.
[0074] Table 1 shows that compared with the unheated samples, the heating treatment significantly changed the secondary structure of the OVT samples, thereby promoting the formation of protein fibers. It is particularly noteworthy that the sample heated for 16 hours has the highest β-fold content, reaching 27.42%. In addition, too high a temperature or too long a heating time may also lead to a reduction in the proportion of β-fold structure.
[0075] Table 1 Secondary structure distribution of antibacterial ovotransferrin fibers at different heating times
[0076]
[0077] Example 2
[0078] The method for preparing antibacterial ovotransferrin fiber comprises the following steps:
[0079] (1) Dissolve 1 g of ovotransferrin in 50 mL of deionized water to obtain an OVT solution with a mass fraction of 2% (w / w).
[0080] (2) Add 2 M hydrochloric acid to the OVT solution until the pH value is 2.0.
[0081] (3) The OVT solution was transferred to a water bath at 90°C and heated with stirring for 16 h, then quickly placed in an ice water bath to terminate the reaction, and freeze-dried to obtain a powder, which is the antibacterial ovotransferrin fiber (OVTF).
[0082] Comparative Example 1
[0083] Preparation method of ovotransferrin oligomer
[0084] (1) Dissolve 1 g of OVT in 50 mL of deionized water to obtain a 2% (w / w) OVT solution.
[0085] (2) Add 2 M hydrochloric acid to the OVT solution and adjust its pH to 2.0 using a pH meter.
[0086] (3) The OVT solution was transferred to a water bath, heated and stirred at 37°C, and immediately placed in an ice water bath after 16 h to terminate the reaction. The resulting powder was freeze-dried to obtain ovotransferrin oligomer (hereinafter referred to as Oligomer).
[0087] Example 3
[0088] Detection of antibacterial activity of ovotransferrin fibers
[0089] 1. The antibacterial effect of antibacterial ovotransferrin fiber was evaluated by the disc diffusion method, which includes the following steps:
[0090] (1) Cultivate bacteria E. coli and S. aureus and adjust the concentration to 1×10 6 CFU / mL, and spread the bacterial suspension evenly on the surface of LB agar medium.
[0091] (2) A paper sheet with a diameter of 6 mm was placed on the LB agar medium, and a 50 μg / mL ovotransferrin oligomer solution and a 50 μg / mL antibacterial ovotransferrin fiber solution were dripped onto the paper sheet, respectively, with a dosage of 10 μL for each.
[0092] (3) The culture dish was placed in a constant temperature incubator at 37°C for 12 h, and the antibacterial ability was determined by measuring the size of the inhibition zone around the paper.
[0093] The results are as follows Figure 8 As described above, an inhibition zone appeared around the paper sheet to which ovotransferrin oligomer and antibacterial ovotransferrin fiber were dropped, and compared with ovotransferrin oligomer, antibacterial ovotransferrin fiber had a larger and more obvious inhibition zone, indicating that antibacterial ovotransferrin fiber had higher antibacterial activity.
[0094] 2. The colony counting method was used to determine the colony forming units to evaluate the inhibitory effect of the antibacterial ovotransferrin fiber on E. coli and S. aureus, including the following steps:
[0095] (1) E. coli and S. aureus were inoculated into LB medium and cultured overnight under an aerobic environment for proliferation. The concentrations of E. coli and S. aureus were adjusted to approximately 10% using Tris-HCl buffer at pH 7.2. 6 The density of CFU / mL.
[0096] (2) Prepare ovotransferrin oligomer solutions and antibacterial ovotransferrin fiber solutions with concentrations of 2, 10, 25, 50, 100, and 200 μg / mL, respectively.
[0097] (3) Incubate the cells with the same volume of ovotransferrin oligomer solution and the antibacterial ovotransferrin fiber solution at 30°C for about 10 min. 6 CFU / mL of the above bacteria for 6h.
[0098] (4) After the incubation, the same volume of the sample was diluted tenfold, and 100 μL of the diluted sample was spread on an LB agar plate containing 1.5% (w / v) agar. After culturing at 37° C. for 24 h, the cell survival rate was quantitatively analyzed by CFU counting method.
[0099] The results are as follows Fig. 9 As shown, it was revealed that the antibacterial ovotransferrin fibers exhibited significant antibacterial advantages against E. coli and S. aureus in the concentration range of 10 to 200 μg / mL, which was superior to ovotransferrin and ovotransferrin oligomers.
[0100] Example 4
[0101] Determination of minimum inhibitory concentration of different substances
[0102] First, each sample (OVTF, OVT and Oligomer, respectively) was diluted to 1 mg / mL with phosphate buffer, and then 2-fold serial dilutions were performed using LB broth on a 96-well microplate. Next, 10 μL of bacterial suspension was inoculated in each well. After incubation at 37°C for 24 h, the lowest concentration at which no growth was observed was recorded as the MIC value by visual observation and detection with an M 200Pro NanoQuant microplate reader (Tecan, Switzerland). MIC is defined as the minimum sample concentration that prevents bacterial growth.
[0103] The results are shown in Table 2. The MIC value of OVTF against E. coli was 128 μg / mL, while the MIC values of OVT and Oligomer were both over 256 μg / mL. In the test against S. aureus, OVTF showed the best inhibitory effect, with a MIC of only 64 μg / mL. Overall, OVTF has the highest antibacterial activity against these two bacteria.
[0104] Table 2 MIC (μg / mL) of OVT, Oligomer and OVTF against E. coli and S. aureus
[0105]
[0106] Example 5
[0107] The antibacterial ovotransferrin fiber antibacterial agent (OVTF antibacterial agent) includes the antibacterial ovotransferrin fiber prepared in Example 2, wherein the concentration of the antibacterial ovotransferrin fiber in the antibacterial agent is 150 μg / mL.
[0108] Comparative Example 2
[0109] OVT antimicrobial agent: Dissolve OVT in deionized water to prepare a 150 μg / mL solution to obtain the OVT antimicrobial agent.
[0110] Comparative Example 3
[0111] Oligomer antimicrobial agent: Dissolve the oligomer in deionized water to prepare a 150 μg / mL solution to obtain the oligomer antimicrobial agent.
[0112] 2. Scanning electron microscopy analysis
[0113] Quanta 250FEG scanning electron microscope (Thermo Fisher Scientific, USA) was used to analyze and observe the effects of sample solutions on bacterial cell morphology and surface structure. Bacterial cells were treated with PBS buffer, OVTF antimicrobial agent, OVT antimicrobial agent and Oligomer antimicrobial agent, respectively, and incubated at 37°C for 2 h.
[0114] After collection and washing with PBS, the bacterial cells were fixed with 2.5% (w / v) glutaraldehyde at 4°C overnight, followed by washing the cells with 0.1M PBS, and then dehydrated through a series of increasing concentrations of ethanol (30%, 50%, 70%, 90%, 95% and 100%), each dehydration step lasting 15 min. After dehydration, the bacterial cells were coated with gold-palladium alloy and observed at a voltage of 15 kV.
[0115] The results show that Fig.10 It was revealed that for Gram-negative E. coli, the integrity of its cell wall was relatively maintained after treatment with OVT and Oligomer antimicrobial agents. However, when exposed to OVTF antimicrobial agents, E. coli showed larger pore formation and obvious cell collapse. In Gram-positive S. aureus, only OVT antimicrobial treatment maintained cell structural integrity similar to that of the control group. In contrast, S. aureus treated with Oligomer and OVTF antimicrobial agents showed volume reduction and collapse, and OVTF antimicrobial treatment also led to more obvious cell aggregation.
[0116] Example 6
[0117] Drugs for antibacterial ovotransferrin fiber diarrhea
[0118] This drug is an oral compound preparation, and its preparation method is as follows:
[0119] 10 g of antibacterial ovotransferrin fiber, 6 g of vitamin C, 2 g of sucrose and 2 g of maltose were weighed and mixed to obtain a medicine.
[0120] According to actual conditions, add the above ratio into the solvent to prepare oral solution.
[0121] Example 7
[0122] Testing the effect of drugs on antibacterial ovotransferrin fiber diarrhea
[0123] The dosage of the drug for antibacterial ovotransferrin fiber diarrhea varies according to the age of the infant: 250 mg for infants aged 3 months, 300 mg for infants aged 6 months, and 400 mg for infants aged 9 months; the frequency of use is 3 times / day.
[0124] Expected results: Fig.11 As shown, the antibacterial ovotransferrin fiber diarrhea drug can significantly reduce the incidence of diarrhea in infants and young children.
[0125] Although the above embodiments have been described in detail, they are only a part of the embodiments of the present invention, not all of them. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all belong to the protection scope of the present invention.
Claims
1. A method for preparing antibacterial ovotransferrin fiber, characterized in that: The following steps are involved: (1) dissolving ovotransferrin in deionized water to prepare an ovotransferrin solution; (2) adjusting the pH of the ovotransferrin solution to 1.5-2.0; (3) The ovotransferrin solution is heated, quickly placed in an ice water bath to terminate the reaction, and freeze-dried to obtain a powder, which is the antibacterial ovotransferrin fiber.
2. The preparation method according to claim 1, characterized in that: In the step (1), the mass fraction of the ovotransferrin solution is 1.5-2%; In the step (2), the pH adjuster is 2M hydrochloric acid; In the step (3), the heating treatment conditions are: 85°C < heating temperature ≤ 95°C, 0h < heating time ≤ 24h.
3. Use of the antibacterial ovotransferrin fiber prepared by the preparation method as claimed in claim 1 in antibacterial applications.
4. The application according to claim 3, characterized in that: The antibacterial strain is Gram-like bacteria.
5. The application according to claim 4, characterized in that: The Gram bacteria are Escherichia coli or Staphylococcus aureus.
6. Use of the antibacterial ovotransferrin fiber prepared by the preparation method as claimed in claim 1 in the preparation of an antibacterial agent for inhibiting Gram-like bacteria.
7. An antibacterial ovotransferrin fiber antibacterial agent, characterized in that: The antibacterial agent comprises antibacterial ovotransferrin fibers; wherein the antibacterial ovotransferrin fibers are prepared by the preparation method according to claim 1, and the concentration of the antibacterial ovotransferrin fibers in the antibacterial agent is 128-200 μg / mL.
8. Use of the antibacterial ovotransferrin fiber prepared by the preparation method as claimed in claim 1 in the preparation of a drug for treating diarrhea in infants and young children.
9. A medicine for treating diarrhea in infants and young children, characterized in that: The medicine comprises an effective amount of the antibacterial ovotransferrin fiber prepared by the preparation method as claimed in claim 1 and auxiliary materials.
10. The drug according to claim 9, characterized in that: In terms of weight, the drug comprises 1 part of antibacterial ovotransferrin fiber and 0.9-2 parts of auxiliary materials; the auxiliary materials are 0.5-1 part of vitamin C, 0.2-0.5 part of sucrose and 0.2-0.5 part of maltose.