CN107549582B - Walnut polypeptide compound used as antibacterial agent - Google Patents
Walnut polypeptide compound used as antibacterial agent Download PDFInfo
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- CN107549582B CN107549582B CN201610515412.8A CN201610515412A CN107549582B CN 107549582 B CN107549582 B CN 107549582B CN 201610515412 A CN201610515412 A CN 201610515412A CN 107549582 B CN107549582 B CN 107549582B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
Abstract
The invention provides a method for compounding walnut polypeptide extract and bamboo leaf essential oil, which is characterized in that defatted walnut powder is taken as a raw material, the walnut polypeptide extract is prepared by crushing and extracting by adopting a neutral enzyme enzymolysis technology, and the walnut polypeptide extract and the bamboo leaf essential oil are mixed in different volume ratios to obtain a walnut polypeptide compound with better antibacterial performance, wherein the walnut polypeptide compound has an obvious antibacterial effect, and the antibacterial effect is improved by 2-4 times compared with that of the walnut polypeptide by single bacteriostasis. The walnut polypeptide compound is applied to a food system, so that the drug resistance of microorganisms is not easy to generate, and various microorganisms can be directly killed. The walnut polypeptide compounding technology is simple and convenient, the bacteriostasis effect of the obtained walnut polypeptide compound is obviously improved, and the walnut polypeptide compound can be used for food preservation and storage and provides better development for the application of plant-derived antibacterial peptide in the food industry.
Description
Technical Field
The invention relates to a preparation method of a walnut polypeptide compound serving as an antibacterial agent. The walnut antibacterial polypeptide is an antibacterial substance extracted and separated from walnut pulp of Juglans of Juglandaceae, has an obvious antibacterial effect on a specific strain, the bamboo leaf essential oil is extracted and separated from bamboo leaves of Bambusoideae of Gramineae, has a broad-spectrum antibacterial effect, and the compound has an obvious antibacterial effect by compounding the two components, so that the walnut antibacterial polypeptide can be used for food preservation and storage.
Background
Food spoilage problems persist in the food industry, and food losses due to spoilage account for around 20% of total losses worldwide each year, with enormous losses being reported. Of which the effects due to microorganisms are most pronounced. Chemical control methods are the leading methods used at present, but are questioned for their harm to the environment and to humans and animals. The natural green and natural returning are pursuits of modern life, and with the increasing importance of people on the health problems, the pursuit of safe, efficient and environment-friendly natural preservatives becomes a necessary trend. Researches find that the walnut polypeptide can be used as a plant-derived antibacterial peptide, but the antibacterial effect is low.
The invention provides a walnut polypeptide-bamboo leaf essential oil compound, the antibacterial efficacy of which is higher than that of a walnut polypeptide extract, and the compound has important practical significance for promoting the development and application of walnut polypeptide in food fresh-keeping storage.
Disclosure of Invention
One of the purposes of the invention is to provide a preparation method of a walnut polypeptide compound.
The invention also aims to provide a walnut polypeptide compound with broad-spectrum antibacterial effect for efficiently inhibiting the growth of gram-positive bacteria, gram-negative bacteria and fungi, and the compound is applied to food fresh-keeping and storage.
1. The invention provides a walnut polypeptide compound which comprises the following components in parts by mass: 60-70 parts of walnut polypeptide extract, 5.8-7.8 parts of hexadecanoic acid, 3.2-4.2 parts of plant alcohol, 3.0-4.0 parts of pentacosane and 1.4-1.9 parts of lauric acid.
2. The walnut polypeptide compound provided by the invention comprises a walnut polypeptide extract and bamboo leaf essential oil.
3. The mass ratio of the walnut polypeptide extract to the bamboo leaf essential oil is (60-70): (40-30).
4. The invention also provides a preparation method of the walnut polypeptide extract, which is characterized by adopting a neutral enzyme hydrolysis method for preparation.
5. The invention provides a preparation method of a walnut polypeptide compound, which is characterized by mixing 60-70 parts by mass of a walnut polypeptide extract, 5.8-7.8 parts by mass of hexadecanoic acid, 3.2-4.2 parts by mass of plant alcohol, 3.0-4.0 parts by mass of pentacosane and 1.4-1.9 parts by mass of lauric acid to obtain the walnut polypeptide compound.
6. The application of the walnut polypeptide compound in the antibacterial agent is characterized in that the walnut polypeptide compound comprises the following components in parts by mass:
60-70 parts by mass of a walnut polypeptide extract,
5.8 to 7.8 parts by mass of palmitic acid,
3.2 to 4.2 parts by mass of plant alcohol,
3.0 to 4.0 parts by mass of pentacosane,
1.4-1.9 parts of lauric acid.
7. The walnut polypeptide compound provided by the invention can inhibit gram-positive bacteria, gram-negative bacteria and fungi.
8. The walnut polypeptide compound provided by the invention can be used for keeping fruits and vegetables fresh.
The walnut polypeptide compound can inhibit the growth of gram-positive bacteria, gram-negative bacteria and fungi, and has the antibacterial effect which is 3 to 4 times higher than that of a walnut polypeptide extract used alone on the data of the minimum antibacterial concentration of various strains, and the walnut polypeptide compound has the effects on bacillus subtilis, staphylococcus aureus and escherichia coli, the minimum inhibitory concentrations of the saccharomyces cerevisiae and the pichia pastoris are respectively 4.82-5.09mg/mL, 6.76-6.89mg/mL, 3.24-3.56mg/mL, 5.12-5.47mg/mL and 5.67-6.01mg/mL, and the minimum inhibitory concentrations of the walnut polypeptide extract are singly used (15.58 mg/mL of bacillus subtilis, 20.92mg/mL of staphylococcus aureus, 10.89mg/mL of escherichia coli, 10.56mg/mL of saccharomyces cerevisiae and 10.86mg/mL of pichia pastoris).
Detailed Description
The invention provides a preparation method and application of a walnut polypeptide compound, and a person skilled in the art can appropriately improve process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The invention provides a walnut polypeptide compound which comprises the following components in parts by mass:
60-70 parts by mass of a walnut polypeptide extract,
5.8 to 7.8 parts by mass of palmitic acid,
3.2 to 4.2 parts by mass of plant alcohol,
3.0 to 4.0 parts by mass of pentacosane,
1.4-1.9 parts of lauric acid.
Preferably, the walnut polypeptide compound comprises: 60-70 parts of walnut polypeptide extract, 5.8-7.8 parts of hexadecanoic acid, 3.2-4.2 parts of plant alcohol, 3.0-4.0 parts of pentacosane and 1.4-1.9 parts of lauric acid.
More preferably, the walnut polypeptide compound comprises: 62-68 parts of walnut polypeptide extract, 6.2-7.4 parts of hexadecanoic acid, 3.4-4.0 parts of plant alcohol, 3.2-3.8 parts of pentacosane and 1.5-1.8 parts of lauric acid.
Most preferably, the walnut polypeptide formulation comprises: 64-66 parts of walnut polypeptide extract, 6.6-7.0 parts of hexadecanoic acid, 3.6-3.8 parts of plant alcohol, 3.4-3.6 parts of pentacosane and 1.6-1.7 parts of lauric acid.
In some embodiments, the walnut polypeptide formulation comprises a walnut polypeptide extract and bamboo leaf essential oil.
In the invention, the walnut polypeptide compound is prepared by compounding a walnut polypeptide extract and bamboo leaf essential oil, wherein preferably, the mass ratio of the walnut polypeptide extract to the bamboo leaf essential oil is (60-70): (30-40), more preferably (62-68): (32-38), most preferably (64-66): (34-36).
The invention also provides a preparation method of the walnut polypeptide compound, which comprises the step of mixing 60-70 parts by mass of walnut polypeptide extract, 5.8-7.8 parts by mass of hexadecanoic acid, 3.2-4.2 parts by mass of plant alcohol, 3.0-4.0 parts by mass of pentacosane and 1.4-1.9 parts by mass of lauric acid to obtain the walnut polypeptide compound.
In a most preferred scheme, walnut polypeptide is mixed with bamboo leaf essential oil to obtain a walnut polypeptide compound, wherein the mass ratio of the walnut polypeptide extract to the bamboo leaf essential oil is (60-70): (30-40), more preferably (62-68): (32-38), most preferably (64-66): (34-36).
The instruments adopted by the invention are all common commercial products and can be purchased in the market.
The walnut polypeptide compound is obtained by mixing the walnut polypeptide extract and the bamboo leaf essential oil according to a certain proportion. The specific operation method comprises the following steps:
1. and (3) crushing the degreased walnut pulp to a certain mesh number to obtain degreased walnut powder, wherein the particle size of the degreased walnut powder is in the range of 10 meshes to 100 meshes.
2. Preferably, the pulverized particle size is 10 to 20 mesh.
3. Weighing a certain mass of the defatted walnut powder in a round-bottom flask, and adding distilled water to ensure that the ratio of the volume of the water to the defatted walnut powder is 50: 1 to 30:1, wherein the volume unit is milliliter, and the mass unit is gram; adding neutral protease, and controlling the enzyme amount to be 5000-; adjusting the pH value to 5-8; controlling the enzymolysis temperature at 35-60 deg.C, and performing enzymolysis for 2-8h to obtain enzymolysis solution.
4. Preferably, the ratio of the volume of the water to the mass of the defatted walnut powder is 35-40: 1; adding enzyme at 6000-7000U/g defatted walnut powder; the pH value is between 6 and 8; the enzymolysis temperature is controlled at 40-50 deg.C, and the enzymolysis time is 3-5 h.
5. Inactivating enzyme at 85 deg.C for 10min, and centrifuging at 8000r/min for 20min to obtain supernatant.
6. Removing most of water from the supernatant with a rotary evaporator (controlling water bath temperature at 40-60 deg.C), transferring the concentrated solution to a culture dish, and drying in a freeze dryer to obtain semen Juglandis polypeptide extract.
7. Preferably, the water bath temperature of the rotary evaporator is controlled at 45-55 ℃.
8. Cleaning bamboo leaves, naturally drying, and crushing into a certain mesh number, wherein the particle size of the bamboo leaf powder is 10-100 meshes.
9. Preferably, the pulverized particle size is 10 to 20 mesh.
10. Weighing a certain mass of the bamboo leaf powder in a round-bottom flask, and adding distilled water to ensure that the ratio of the volume of the water to the mass of the bamboo leaf powder is 20: 1 to 40:1, wherein the volume unit is milliliter, and the mass unit is gram.
11. Preferably, the ratio of the volume of the water to the mass of the bamboo leaf powder is 20-30: 1.
12. Heating the water solution in the round-bottom flask by adopting a plant essential oil extraction device for 4-6 hours, cooling to room temperature, separating out an n-hexane layer, drying by using anhydrous sodium sulfate, and removing the n-hexane by using a rotary evaporator (the water bath temperature is 35 ℃) to obtain the bamboo leaf essential oil.
13. Preferably, the heating time is 4.5 hours.
14. Mixing the walnut polypeptide extract and the bamboo leaf essential oil according to different mass ratios (60-70): (40-30) respectively mixing to obtain the walnut polypeptide compound.
15. Preferably, three mass ratios of 64:36, 65:35 and 66:34 are respectively selected and mixed to obtain the walnut polypeptide compound.
The invention also provides an application of the walnut polypeptide compound in a bacteriostatic agent, wherein the walnut polypeptide compound comprises the following components in parts by mass:
60-70 parts by mass of a walnut polypeptide extract,
5.8 to 7.8 parts by mass of palmitic acid,
3.2 to 4.2 parts by mass of plant alcohol,
3.0 to 4.0 parts by mass of pentacosane,
1.4-1.9 parts of lauric acid.
The walnut polypeptide compound can inhibit gram-positive bacteria, gram-negative bacteria and fungi. Preferably, the walnut polypeptide compound can inhibit fungi. The fungus is preferably Saccharomyces cerevisiae and/or Pichia pastoris. In certain embodiments of the invention, the walnut polypeptide formulation comprises (60-70) by mass: (30-40) the walnut polypeptide extract and the bamboo leaf essential oil.
The invention is further illustrated by the following examples:
example 1: preparation of walnut polypeptide extract
Crushing the degreased walnut pulp to 20 meshes to obtain degreased walnut powder, weighing the degreased walnut powder with a certain mass in a round-bottom flask, and adding distilled water to ensure that the ratio of the volume of the water to the mass of the degreased walnut powder is 35: 1, adding neutral protease, controlling the enzyme content at 6000U/g, adjusting the pH value to 7, controlling the enzymolysis temperature at 45 ℃ and the enzymolysis time for 4 hours; then inactivating enzyme at 85 deg.C for 10min, centrifuging at 8000r/min for 20min to obtain supernatant; removing most of water from the supernatant with a rotary evaporator (controlling the temperature of water bath at 45 deg.C), transferring the concentrated solution to a culture dish, and drying in a freeze dryer to obtain semen Juglandis polypeptide extract.
Example 2: preparation of bamboo leaf essential oil
The method comprises the following steps of taking moso bamboo leaves as a raw material, cleaning, drying and crushing the moso bamboo leaves, accurately weighing bamboo leaf powder with a certain mass average particle fineness of 10 meshes in a round-bottom flask according to an essential oil extraction device, and adding distilled water to ensure that the ratio of the volume of water to the mass of the bamboo leaf powder is 25: heating the aqueous solution in the round-bottom flask for 4.5 hours, cooling to room temperature, separating an n-hexane layer, drying with anhydrous sodium sulfate, and removing the n-hexane by using a rotary evaporator (the water bath temperature is 35 ℃) to obtain the bamboo leaf essential oil.
Example 3: walnut polypeptide extract and bamboo leaf essential oil compound
Mixing the walnut polypeptide extract prepared in the example 1 and the bamboo leaf essential oil prepared in the example 2 in 64 parts by mass: and mixing the raw materials in a proportion of 36 parts by mass fully to obtain the walnut polypeptide compound. Through the detection of a gas chromatography-mass spectrometry combined technology and an ultraviolet visible spectrophotometry, the walnut polypeptide compound comprises the following components: 64 parts of walnut polypeptide extract, 7.0 parts of hexadecanoic acid, 3.8 parts of plant alcohol, 3.6 parts of pentacosane and 1.7 parts of lauric acid.
Example 4: walnut polypeptide extract and bamboo leaf essential oil compound
Mixing the walnut polypeptide extract prepared in the example 1 and the bamboo leaf essential oil prepared in the example 2 in 65 parts by mass: and (3) fully mixing 35 parts by mass to obtain the walnut polypeptide compound. Through the detection of a gas chromatography-mass spectrometry combined technology and an ultraviolet visible spectrophotometry, the walnut polypeptide compound comprises the following components: 65 parts of walnut polypeptide extract, 6.8 parts of hexadecanoic acid, 3.7 parts of plant alcohol, 3.5 parts of pentacosane and 1.6 parts of lauric acid.
Example 5: walnut polypeptide extract and bamboo leaf essential oil compound
Mixing the walnut polypeptide extract prepared in the example 1 and the bamboo leaf essential oil prepared in the example 2 according to 66 parts by mass: and (3) fully mixing 34 parts by mass to obtain the walnut polypeptide compound. Through the detection of a gas chromatography-mass spectrometry combined technology and an ultraviolet visible spectrophotometry, the walnut polypeptide compound comprises the following components: 66 parts of walnut polypeptide extract, 6.6 parts of hexadecanoic acid, 3.6 parts of plant alcohol, 3.4 parts of pentacosane and 1.6 parts of lauric acid.
Example 6: antibacterial effect of walnut polypeptide compound
The antibacterial effect of the walnut polypeptide compound obtained in example 3 is examined by adopting a double-layer plate punching method, and the walnut polypeptide compound is found to be capable of inhibiting the growth of gram-positive bacteria, gram-negative bacteria and fungi, and the minimum inhibitory concentrations to bacillus subtilis, staphylococcus aureus, escherichia coli, saccharomyces cerevisiae and pichia pastoris are 4.82mg/mL, 6.76mg/mL, 3.24mg/mL, 5.12mg/mL and 5.70mg/mL respectively. Compared with the lowest bacteriostatic concentration (15.58 mg/mL of bacillus subtilis, 20.92mg/mL of staphylococcus aureus, 10.89mg/mL of escherichia coli, 10.56mg/mL of saccharomyces cerevisiae and 10.86mg/mL of pichia pastoris) of the walnut polypeptide extract used alone, the bacteriostatic effect is improved by 2-4 times.
Example 7: antibacterial effect of walnut polypeptide compound
The antibacterial effect of the walnut polypeptide compound obtained in example 4 is examined by adopting a double-layer flat plate punching method, and the walnut polypeptide compound is found to be capable of inhibiting the growth of gram-positive bacteria, gram-negative bacteria and fungi, and the minimum inhibitory concentrations to bacillus subtilis, staphylococcus aureus, escherichia coli, saccharomyces cerevisiae and pichia pastoris are 4.95mg/mL, 6.83mg/mL, 3.39mg/mL, 5.30mg/mL and 5.89mg/mL respectively. Compared with the lowest bacteriostatic concentration (15.58 mg/mL of bacillus subtilis, 20.92mg/mL of staphylococcus aureus, 10.89mg/mL of escherichia coli, 10.56mg/mL of saccharomyces cerevisiae and 10.86mg/mL of pichia pastoris) of the walnut polypeptide extract used alone, the bacteriostatic effect is improved by 2-4 times.
Example 8: antibacterial effect of walnut polypeptide compound
The antibacterial effect of the walnut polypeptide compound obtained in example 5 is examined by adopting a double-layer flat plate perforation method, and the walnut polypeptide compound is found to be capable of inhibiting the growth of gram-positive bacteria, gram-negative bacteria and fungi, and the minimum inhibitory concentrations to bacillus subtilis, staphylococcus aureus, escherichia coli, saccharomyces cerevisiae and pichia pastoris are 5.09mg/mL, 6.89mg/mL, 3.56mg/mL, 5.47mg/mL and 6.02mg/mL respectively. Compared with the lowest bacteriostatic concentration (15.58 mg/mL of bacillus subtilis, 20.92mg/mL of staphylococcus aureus, 10.89mg/mL of escherichia coli, 10.56mg/mL of saccharomyces cerevisiae and 10.86mg/mL of pichia pastoris) of the walnut polypeptide extract used alone, the bacteriostatic effect is improved by 2-4 times.
Example 9: application of walnut polypeptide compound
Dissolving the walnut polypeptide compound obtained in the example 3 in 0.1% of tween-20, fully emulsifying, diluting to 10mg/mL with sterile water, selecting fresh cherry tomatoes without obvious defects such as tetanus, spraying a proper amount of walnut polypeptide compound to the surfaces of the cherry tomatoes at a fixed point every day (based on wetting of the surfaces of the fruits and vegetables and no adhesion of large water beads), and meanwhile, spraying distilled water daily as a control group, and storing and observing at room temperature (25 ℃). The cherry tomatoes in the control group are basically and completely rotten when being stored for 5 days, and the treatment group is still intact and is not rotten until 20 days, so that the preservative and fresh-keeping effects of the walnut polypeptide compound on the cherry tomatoes, which are obtained by the invention, are nearly 4 times of the preservative and fresh-keeping effects of marketers on the cherry tomatoes on weekdays.
Example 10: application of walnut polypeptide compound
Dissolving the walnut polypeptide compound obtained in the example 4 in 0.1% of tween-20, fully emulsifying, diluting to 10mg/mL with sterile water, selecting fresh cucumbers without obvious defects such as tetanus, spraying a proper amount of walnut polypeptide compound to the surfaces of the cucumbers at fixed points every day (based on wetting the surfaces of the fruits and the vegetables and no large water beads attached), and meanwhile, taking daily spraying of distilled water as a control group, and storing and observing at room temperature (25 ℃). The cherry tomatoes in the control group are basically and completely rotten when stored for 7 days, and the treatment group is still intact and is not rotten until 25 days, so that the preservative and fresh-keeping effects of the walnut polypeptide compound on cucumbers, which is obtained by the invention, are nearly 3-4 times of the preservative and fresh-keeping effects of marketers on the cherry tomatoes on weekdays.
The walnut polypeptide compound has a far greater antibacterial effect than that of the walnut polypeptide alone, and compared with the walnut polypeptide alone, the antibacterial performance of the walnut polypeptide compound is improved by 3-4 times.
The walnut polypeptide compound designed by the invention can effectively inhibit the growth and reproduction of gram-positive bacteria, gram-negative bacteria and fungi, can be widely applied to food fresh-keeping storage, and has wide practicability.
The walnut polypeptide compound obtained by the invention can be used for corrosion prevention and fresh keeping of more fruits and vegetables, such as grapes, oranges, apples, pears, kiwifruits, cucumbers, white gourds, loofah, pumpkins and other fruits and vegetables, and has wide practicability.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (3)
1. The walnut polypeptide compound is characterized by being prepared by mixing a walnut polypeptide extract and bamboo leaf essential oil; the walnut polypeptide extract is prepared by the following method: crushing the degreased walnut pulp to 20 meshes to obtain degreased walnut powder, weighing the degreased walnut powder with a certain mass in a round-bottom flask, and adding distilled water to ensure that the ratio of the volume of the water to the mass of the degreased walnut powder is 35: 1, adding neutral protease, controlling the enzyme content at 6000U/g, adjusting the pH value to 7, controlling the enzymolysis temperature at 45 ℃ and the enzymolysis time for 4 hours; then inactivating enzyme at 85 deg.C for 10min, centrifuging at 8000r/min for 20min, concentrating the obtained supernatant, and freeze drying to obtain semen Juglandis polypeptide extract; the preparation method of the bamboo leaf essential oil comprises the steps of taking moso bamboo leaves as raw materials, cleaning, drying and crushing the moso bamboo leaves, accurately weighing moso bamboo leaf powder with a certain mass average particle fineness of 10 meshes in a round-bottom flask according to an essential oil extraction device, adding distilled water to ensure that the ratio of the volume of the water to the mass of the bamboo leaf powder is 25:1, wherein the volume unit is milliliter and the mass unit is gram, heating an aqueous solution in the round-bottom flask for 4.5 hours, cooling to room temperature, separating out an n-hexane layer, drying with anhydrous sodium sulfate, and removing the n-hexane by using a rotary evaporator to obtain the bamboo leaf essential oil; the mass ratio of the walnut polypeptide extract to the bamboo leaf essential oil is (60-70): (40-30), the walnut polypeptide compound comprises the following components in parts by mass: 60-70 parts of walnut polypeptide extract, 5.8-7.8 parts of hexadecanoic acid, 3.2-4.2 parts of plant alcohol, 3.0-4.0 parts of pentacosane and 1.4-1.9 parts of lauric acid.
2. The preparation method of the walnut polypeptide compound is characterized in that a walnut polypeptide extract is mixed with bamboo leaf essential oil, and the walnut polypeptide extract is prepared according to the following method: crushing the degreased walnut pulp to 20 meshes to obtain degreased walnut powder, weighing the degreased walnut powder with a certain mass in a round-bottom flask, and adding distilled water to ensure that the ratio of the volume of the water to the mass of the degreased walnut powder is 35: 1, adding neutral protease, controlling the enzyme content at 6000U/g, adjusting the pH value to 7, controlling the enzymolysis temperature at 45 ℃ and the enzymolysis time for 4 hours; then inactivating enzyme at 85 deg.C for 10min, centrifuging at 8000r/min for 20min, concentrating the obtained supernatant, and freeze drying to obtain semen Juglandis polypeptide extract; the preparation method of the bamboo leaf essential oil comprises the steps of taking moso bamboo leaves as raw materials, cleaning, drying and crushing the moso bamboo leaves, accurately weighing moso bamboo leaf powder with a certain mass average particle fineness of 10 meshes in a round-bottom flask according to an essential oil extraction device, adding distilled water to ensure that the ratio of the volume of the water to the mass of the bamboo leaf powder is 25:1, wherein the volume unit is milliliter and the mass unit is gram, heating an aqueous solution in the round-bottom flask for 4.5 hours, cooling to room temperature, separating out an n-hexane layer, drying with anhydrous sodium sulfate, and removing the n-hexane by using a rotary evaporator to obtain the bamboo leaf essential oil; the mass ratio of the walnut polypeptide extract to the bamboo leaf essential oil is (60-70): (40-30), the walnut polypeptide compound comprises the following components in parts by mass: 60-70 parts of walnut polypeptide extract, 5.8-7.8 parts of hexadecanoic acid, 3.2-4.2 parts of plant alcohol, 3.0-4.0 parts of pentacosane and 1.4-1.9 parts of lauric acid.
3. The application of the walnut polypeptide compound in the antibacterial agent is characterized in that the walnut polypeptide compound is prepared by mixing a walnut polypeptide extract and bamboo leaf essential oil; the walnut polypeptide extract is prepared by the following method: crushing the degreased walnut pulp to 20 meshes to obtain degreased walnut powder, weighing the degreased walnut powder with a certain mass in a round-bottom flask, and adding distilled water to ensure that the ratio of the volume of the water to the mass of the degreased walnut powder is 35: 1, adding neutral protease, controlling the enzyme content at 6000U/g, adjusting the pH value to 7, controlling the enzymolysis temperature at 45 ℃ and the enzymolysis time for 4 hours; then inactivating enzyme at 85 deg.C for 10min, centrifuging at 8000r/min for 20min, concentrating the obtained supernatant, and freeze drying to obtain semen Juglandis polypeptide extract; the preparation method of the bamboo leaf essential oil comprises the steps of taking moso bamboo leaves as raw materials, cleaning, drying and crushing the moso bamboo leaves, accurately weighing moso bamboo leaf powder with a certain mass average particle fineness of 10 meshes in a round-bottom flask according to an essential oil extraction device, adding distilled water to ensure that the ratio of the volume of the water to the mass of the bamboo leaf powder is 25:1, wherein the volume unit is milliliter and the mass unit is gram, heating an aqueous solution in the round-bottom flask for 4.5 hours, cooling to room temperature, separating out an n-hexane layer, drying with anhydrous sodium sulfate, and removing the n-hexane by using a rotary evaporator to obtain the bamboo leaf essential oil; the mass ratio of the walnut polypeptide extract to the bamboo leaf essential oil is (60-70): (40-30); the walnut polypeptide compound comprises the following components in parts by mass: 60-70 parts of walnut polypeptide extract, 5.8-7.8 parts of hexadecanoic acid, 3.2-4.2 parts of plant alcohol, 3.0-4.0 parts of pentacosane and 1.4-1.9 parts of lauric acid; the walnut polypeptide compound can inhibit bacillus subtilis, staphylococcus aureus, escherichia coli, saccharomyces cerevisiae and pichia pastoris.
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| 核桃蛋白酶水解物的抑菌性能探讨;刘昭明 等;《四川食品与发酵》;20081231;第44卷(第5期);20-23 * |
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