CN115152763A - Rod-like nanocrystallization method of epsilon-polylysine and application of epsilon-polylysine nanoparticles - Google Patents
Rod-like nanocrystallization method of epsilon-polylysine and application of epsilon-polylysine nanoparticles Download PDFInfo
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- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
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Abstract
The invention discloses a rod-like nanocrystallization method of epsilon-polylysine and application of epsilon-polylysine nanoparticles thereof, belonging to the technical field of epsilon-polylysine nanoparticles, comprising the following steps of (1) preparing attapulgite or bentonite into aqueous dispersion with the concentration of 1-5% w/v, and then dispersing for 15-30min by a homogenizer until stable suspension is achieved to obtain suspension water dispersion; (2) Dissolving polylysine in water, slowly adding the polylysine into the suspension aqueous dispersion under the condition of stirring, stirring for 2-4h, centrifuging, collecting and drying to obtain rod-shaped epsilon-polylysine nanoparticles, wherein the prepared epsilon-polylysine nanoparticles solve the problems of high cost and poor antibacterial effect of epsilon-polylysine in agricultural disease control at present.
Description
Technical Field
The invention relates to the technical field of epsilon-polylysine nanoparticles, in particular to a rodlike nanocrystallization method of epsilon-polylysine and application of epsilon-polylysine nanoparticles thereof.
Background
Epsilon-polylysine is a biological polypeptide formed by linking epsilon-amino and carboxyl of lysine based on amido bond, and is usually obtained by secretion of streptomycete, when the molecular weight of the Epsilon-polylysine is 3600-4300, the bacteriostatic activity of the Epsilon-polylysine is the best, and when the molecular weight is lower than 1300, the Epsilon-polylysine loses the bacteriostatic activity. Under the influence of lysine molecular structure, epsilon-polylysine contains a large number of amino groups in the molecular structure, and can actively adsorb with bacterial cell membranes to reduce the fluidity of the cell membranes so as to cause the rupture of bacterial thalli and finally cause the death of bacteria. Therefore, this polypeptide is commonly used as a food preservative and is widely used in japan, china, and the like, but its functions other than food preservatives are less studied.
At present, polylysine can also be prepared into polylysine-silicon nanoparticles (Zhushiguo et al, a novel non-viral DNA transfer vector: polylysine-silicon nanoparticles, scientific bulletin 2002) and polylysine starch nanoparticles (publication No. CN 1462763A) which are used as gene vectors to be applied, but when the polylysine nanoparticles are prepared, polylysine and anionic nanoparticles are synthesized by utilizing charge interaction between the polylysine and alpha-configuration polylysine, and the molecular weight of the alpha-configuration polylysine is more than 75000. At present, electrostatic interaction or compound synergism is adopted for preparing related medicaments from epsilon-polylysine, and the related medicaments are applied to agricultural related fields, but the application of epsilon-polylysine to agricultural production is not realized, and the reasons are mainly high cost and poor practicability. The market price of epsilon-polylysine is 2-6 yuan/g, the polylysine disease-resistant cost of spraying 1 time per 1 mu of land can reach 720 yuan at most according to the average field dosage of 40-60L/mu, and the cost is extremely high for agricultural production. In addition, the polylysine has poor foliage adhesion and poor adaptability due to easy rain wash loss, and the polylysine has poor antibacterial and anti-bacteriophagic properties under alkaline conditions, such as application of alkaline fertilizers such as nitro compound fertilizers and plant ash, or in the presence of anionic compounds such as metaphosphoric acid, and has poor practicability. Therefore, the key point of the application of the epsilon-polylysine in the agricultural disease control lies in reducing the dosage of the polylysine while maintaining the antibacterial effect of the polylysine.
Disclosure of Invention
In view of the above, the invention aims to provide a rod-like nanocrystallization method of epsilon-polylysine and application of epsilon-polylysine nanoparticles thereof, and solve the problems of high cost and poor antibacterial effect of epsilon-polylysine in agricultural disease control at present.
The invention solves the problems by the following technical scheme:
a rod-like nanocrystallization method of epsilon-polylysine, which comprises the following steps:
(1) Preparing attapulgite or bentonite into an aqueous dispersion with a concentration of 1-5% w/v, followed by dispersing for 15-30min by a homogenizer to stabilize the suspension to obtain an aqueous suspension dispersion;
(2) And dissolving polylysine in water, slowly adding the suspension aqueous dispersion under the stirring condition, stirring for 2-4h, centrifuging, collecting, and drying to obtain the rod-shaped epsilon-polylysine nanoparticles.
The invention relates to a rodlike nano epsilon-polylysine nanoparticle prepared by chelation of divalent ions and amido bonds on the surfaces of polylysine and attapulgite or bentonite. The active adsorption of attapulgite or bentonite rod-shaped nano materials on the surface of bacteria is utilized to increase the adsorption and destruction capacity of polylysine on the surface of bacteria, and meanwhile, the rod-shaped nano polylysine can also effectively improve the probability of the polylysine entering plants, so that the resistance inducing performance of the polylysine is improved, and finally, the disease control performance of the polylysine which is far superior to that of non-nanocrystallized polylysine is obtained.
Furthermore, the mass ratio of the polylysine to the attapulgite or the bentonite is (1-5): 40.
The rod-shaped epsilon-polylysine nano particles are prepared by combining the epsilon-polylysine with the attapulgite or the bentonite, so that the consumption of the epsilon-polylysine is obviously reduced, and the use cost is reduced.
Further, the rotating speed of the homogenizer is 13000-20000rpm.
Further, modifying the rod-shaped epsilon-polylysine nanoparticles, wherein the modifying treatment comprises the following raw materials: arginine, betaine, calcium chloride, protamine, gelatin and rod-shaped epsilon-polylysine nanoparticles.
Further, the raw materials comprise the following components in parts by mass: 0.8-1.2 parts of arginine, 0.5-1 part of betaine, 0.2-0.4 part of calcium chloride, 0.2-0.5 part of protamine, 0.1-0.2 part of gelatin and 10-12 parts of rod-shaped epsilon-polylysine nanoparticles.
Further, the modification treatment comprises the following steps:
(1) Mixing calcium chloride with water, stirring uniformly to obtain a 35wt% calcium chloride solution, heating to 30-35 ℃, adding rod-like epsilon-polylysine nanoparticles to obtain an epsilon-polylysine nanoparticle water dispersion, and keeping the temperature for later use;
(2) Mixing arginine, betaine and protamine, adding into the water dispersion of the epsilon-polylysine nano particles, carrying out ultrasonic oscillation for 10-15min, adding gelatin, heating to 50-55 ℃, stirring for 15-20min, carrying out freeze drying, and taking out to obtain the modified rod-shaped epsilon-polylysine nano particles.
The invention also discloses application of the rod-shaped epsilon-polylysine nano-particles or the modified epsilon-polylysine nano-particles in preventing and controlling bacterial leaf diseases of solanaceae plants.
The nano-sized epsilon-polylysine nanoparticles or the modified epsilon-polylysine nanoparticles have good antibacterial and resistance inducing performances, the antibacterial property of the nano-sized epsilon-polylysine nanoparticles is improved by more than 5 times compared with that of the non-nano-sized polylysine, the consumption and the cost of the polylysine for field disease control are greatly reduced, and the application of the polylysine in the field disease control is facilitated. Especially, the modified epsilon-polylysine nanoparticles can obviously increase the leaf surface adhesion of polylysine, expand the application range of polylysine, and can not reduce the pesticide effect when being applied with alkaline fertilizer, improve the control effect and increase the practicability of field application.
Further, the rod-shaped epsilon-polylysine nano-particles or the modified epsilon-polylysine nano-particles are applied to the prevention and the treatment of tomato leaf spot diseases.
Further, the rod-shaped epsilon-polylysine nano-particles or the modified epsilon-polylysine nano-particles are applied to pesticides.
Further, the rod-shaped epsilon-polylysine nano-particles or the modified epsilon-polylysine nano-particles are mixed with water and directly sprayed on the surface of a plant.
Has the advantages that:
1. the epsilon-polylysine nano-particles prepared by the method are constructed by bio-based polymers, are nontoxic and pollution-free to the environment, are constructed by utilizing the physical interaction among raw materials, can actively adsorb and destroy the cell membrane of pathogenic bacteria to cause the death of the pathogenic bacteria, can be used with alkaline fertilizers and rainy days, can still keep higher disease control performance, does not reduce the pesticide effect, and obviously increases the practicability.
2. The nanocrystallized polylysine nanoparticles can more effectively induce the self-resistance of plants and improve the SOD content, thereby showing better comprehensive disease control performance than that of the non-nanocrystallized polylysine.
3. Compared with the traditional epsilon-polylysine, the consumption of the epsilon-polylysine nano-particles prepared by the invention is reduced by 4 times, but the epsilon-polylysine nano-particles have better control effect, can obviously reduce the agricultural production cost, and are beneficial to the popularization and expansion of novel pesticides.
Drawings
FIG. 1: the (left) attapulgite and the (right) epsilon-polylysine nanoparticles are enlarged;
FIG. 2: the control effect graphs of tomato leaf spot are respectively shown as a control graph of example 6, a control graph of comparative example 1, a control graph of comparative example 2, a control graph of comparative example 3, an epsilon-polylysine group and a control graph of CK clear water group from left to right.
Detailed Description
The invention will be described in detail below with reference to specific embodiments and the attached drawings:
example 1: preparation of rod-shaped epsilon-polylysine nanoparticles
Dissolving 1g of attapulgite in 100mL of deionized water, uniformly stirring, and shearing and dispersing for 15min at the rotating speed of 15000rpm by a homogenizer until the attapulgite is stably suspended to obtain a suspension water dispersion liquid;
dissolving 25mg of epsilon-polylysine in 20mL of deionized water, slowly adding the dissolved epsilon-polylysine into the attapulgite suspension dispersion at the stirring speed of 500rpm, continuously stirring the mixed solution for 2 hours, centrifuging at 5000rpm, pouring out the supernatant, and drying the precipitate at low temperature to obtain the rod-shaped epsilon-polylysine nanoparticles.
Example 2: preparation of rod-shaped epsilon-polylysine nanoparticles
Dissolving 1g of bentonite in 100mL of deionized water, uniformly stirring, and shearing and dispersing for 15min at the rotating speed of 15000rpm by using a homogenizer until the suspension is stably suspended to obtain a suspension water dispersion liquid;
dissolving 25mg of epsilon-polylysine in 20mL of deionized water, slowly adding the solution into the suspension dispersion of the bentonite at the stirring speed of 500rpm, continuously stirring the mixed solution for 2 hours, centrifuging the mixed solution at 5000rpm, pouring out the supernatant, and drying the precipitate at low temperature to obtain the rod-shaped epsilon-polylysine nanoparticles.
Example 3: preparation of rod-shaped epsilon-polylysine nanoparticles
Dissolving 20g of bentonite in 2000mL of deionized water, stirring uniformly, and shearing and dispersing for 30min at the rotating speed of 20000rpm by a homogenizer until the suspension is stably suspended to obtain a suspension water dispersion liquid;
dissolving 500mg of epsilon-polylysine in 400mL of deionized water, slowly adding the deionized water into the suspension dispersion liquid of the bentonite at the stirring speed of 500rpm, continuously stirring the mixed liquid for 2 hours, centrifuging at 5000rpm, pouring out the supernatant, and drying the precipitate at low temperature to obtain the rod-shaped epsilon-polylysine nanoparticles.
Example 4: preparation of rod-shaped epsilon-polylysine nanoparticles
Dissolving 20g of attapulgite in 2000mL of deionized water, uniformly stirring, and shearing and dispersing for 30min at the rotation speed of 18000rpm by a homogenizer until the attapulgite is stably suspended to obtain a suspended water dispersion liquid;
dissolving 2g of epsilon-polylysine in 1000mL of deionized water, slowly adding the deionized water into the attapulgite suspension dispersion liquid at the stirring speed of 500rpm, continuously stirring the mixed liquid for 2 hours, centrifuging at 5000rpm, pouring out the supernatant, and drying the precipitate at low temperature to obtain the rod-shaped epsilon-polylysine nanoparticles.
Example 5: preparation of modified rod-like epsilon-polylysine nanoparticles
(1) Mixing 0.04g of calcium chloride with water, stirring uniformly to obtain a 35wt% calcium chloride solution, heating to 35 ℃, adding 1g of the rod-shaped epsilon-polylysine nanoparticles prepared in example 3 to obtain an epsilon-polylysine nanoparticle water dispersion, and keeping the temperature at 35 ℃ for later use;
(2) 0.08g of arginine, 0.05g of betaine and 0.02g of protamine are mixed and added into the aqueous dispersion of the epsilon-polylysine nano particles, 0.01g of gelatin is added after ultrasonic oscillation is carried out for 10min under the ultrasonic frequency of 20KHz, the mixture is heated to 55 ℃, stirred for 20min until the gelatin is dissolved, and then freeze drying is carried out under the conditions of-10 ℃ and 10pa, and the modified rod-shaped epsilon-polylysine nano particles are obtained after being taken out.
Example 6: preparation of modified rod-like epsilon-polylysine nanoparticles
(1) Mixing 0.02g of calcium chloride with water, stirring uniformly to obtain a 35wt% calcium chloride solution, heating to 35 ℃, adding 1.2g of the rod-shaped epsilon-polylysine nanoparticles prepared in example 4 to obtain an epsilon-polylysine nanoparticle aqueous dispersion, and keeping the temperature at 35 ℃ for later use;
(2) 0.12g of arginine, 0.1g of betaine and 0.05g of protamine are mixed and added into the aqueous dispersion of the epsilon-polylysine nano particles, 0.02g of gelatin is added after ultrasonic oscillation is carried out for 15min under the ultrasonic frequency of 20KHz, the mixture is heated to 55 ℃, stirred for 20min until the gelatin is dissolved, and then freeze drying is carried out under the conditions of-10 ℃ and 10pa, and the modified rod-shaped epsilon-polylysine nano particles are obtained after being taken out.
Comparative example 1:
the invention is compared with example 6, the main difference being that no calcium chloride is used in the modification step (1), and the step (2) is the same as example 6, wherein the step (1) is specifically operated as follows:
(1) Mixing 1.2g of the rod-shaped epsilon-polylysine nano particles prepared in the example 4 with 6g of water, heating to 35 ℃ to obtain an epsilon-polylysine nano particle water dispersion, and keeping the temperature for later use;
step (2) was the same as in example 6.
Comparative example 2:
the present invention is compared with example 6, the main difference is that arginine and protamine are not used in the modification step (2), and the step (1) is the same as example 6, wherein the step (2) is specifically operated as follows:
step (1) was the same as in example 6;
(2) And (2) mixing 0.1g of betaine, adding the mixture into the aqueous dispersion of the epsilon-polylysine nanoparticles, carrying out ultrasonic oscillation for 15min at the ultrasonic frequency of 20KHz, adding 0.02g of gelatin, heating to 55 ℃, stirring for 20min until the gelatin is dissolved, carrying out freeze drying at the temperature of-10 ℃ and the pressure of 10pa, and taking out to obtain the modified rod-shaped epsilon-polylysine nanoparticles.
Comparative example 3:
the present invention is compared with example 6, and the main difference is that no betaine is used in the modification step (2), and the step (1) is the same as example 6, wherein the step (2) is specifically operated as follows:
step (1) was the same as in example 6;
(2) 0.12g of arginine and 0.05g of protamine are mixed and added into the aqueous dispersion of the epsilon-polylysine nano particles, 0.02g of gelatin is added after ultrasonic oscillation is carried out for 15min under the ultrasonic frequency of 20KHz, the mixture is heated to 55 ℃, stirred for 20min until the gelatin is dissolved, and freeze drying is carried out at the temperature of-10 ℃ and the temperature of 10pa, and the modified rod-shaped epsilon-polylysine nano particles are obtained after being taken out.
Comparative example 4:
the present invention is compared with example 6, the main difference is that gelatin is not used in the modification step (2), and the step (1) is the same as example 6, wherein the step (2) is specifically operated as follows:
step (1) was the same as in example 6;
(2) 0.12g of arginine, 0.1g of betaine and 0.05g of protamine are mixed and added into the aqueous dispersion of the epsilon-polylysine nano particles, the mixture is subjected to ultrasonic oscillation for 15min at the ultrasonic frequency of 20KHz, and then is subjected to freeze drying at the temperature of-10 ℃ and the temperature of 10pa, and the modified rod-shaped epsilon-polylysine nano particles are obtained after being taken out.
1. Control experiment 1:
(1) Inoculating pseudomonas syringae on the surface of tomato leaves by a puncture inoculation method to obtain tomato plants infected with tomato leaf rashes, and respectively performing control experiments on the infected plants by adopting the rod-shaped epsilon-polylysine nanoparticles prepared in examples 3-4, the modified epsilon-polylysine nanoparticles prepared in examples 5-6, the raw material epsilon-polylysine and clear water control for three times;
(2) After 1 day of inoculation, the prepared product is prepared into 1mg/mL dispersion liquid to be sprayed on inoculated leaves for 3 days continuously, 1 time of spraying is carried out every day, 30mL is sprayed every time, and the clear water group is sprayed according to the same method, and 30mL of clear water is sprayed every time. After 3 days, the spraying is stopped, inoculated plants are cultured for 3 days under the conditions that the temperature is 25-28 ℃ and the humidity is 75-80%, and the diameter of the lesion of the inoculated leaves is measured after 3 days, and the obtained results are shown in the table 1:
TABLE 1
2. Control experiment 2
Tomato plants infected with tomato leaf spot disease were prepared using the same method as in experiment 1. If the mass ratio of the raw material attapulgite to epsilon-polylysine in example 1 is 1g.
The rod-shaped polylysine nanoparticles are prepared into 1g/L dispersion liquid to be used for spraying inoculated leaves, the dispersion liquid is continuously sprayed for 3 days for 1 time every day, 30mL of the dispersion liquid is sprayed every time, and a control group is sprayed according to the same method by using the concentration of 0.1g/L of epsilon-polylysine, and the three steps are repeated. After 3 days, spraying is stopped, inoculated leaves are cultured for 3 days, the lesion diameter of the inoculated leaves is measured, and the content of SOD (superoxide dismutase) in the leaves is detected by using an SOD test kit, and the obtained data are shown in Table 2:
TABLE 2
| Spraying concentration | Diameter of lesion | SOD content | |
| Example 1 | 1g/L | 0.25±0.05cm | 1550±50U/g |
| Group of epsilon-polylysines | 0.1g/L | 1.20±0.10cm | 1200±100U/g |
3. Experiment of antibacterial stability
Tomato plants infected with tomato leaf spot disease were prepared using the same method as in experiment 1. The modified epsilon-polylysine nanoparticles prepared in examples 5-6 and comparative examples 1-4, the raw material epsilon-polylysine and clear water are respectively used for carrying out an antibacterial stability experiment on infected plants, wherein the clear water group is a blank control and is repeated for three times.
After tomato plants are inoculated for 1 day, the prepared product is prepared into 1mg/mL dispersion liquid to be sprayed on inoculated leaves, 20mL of plant ash leachate with the concentration of 10mg/mL is sprayed immediately after 30mL is sprayed each time, and 20mL of plant ash leachate with the concentration of 10mg/mL is sprayed after 30mL of clear water is sprayed each day by the clear water group, and the plant ash leachate is sprayed for 3 days continuously according to the method and is sprayed for 1 time each day. The spraying was stopped after 3 days, and the inoculated plants were cultured for 3 days under the conditions of a temperature of 25 to 28 ℃ and a humidity of 75 to 80%, and the lesion diameter of the inoculated leaves was measured after 3 days, and the obtained results are shown in table 3, wherein the control patterns of example 6, comparative example 1, comparative example 2, comparative example 3, epsilon-polylysine group, and clear water group are shown in fig. 2:
TABLE 3
| Experimental group | Spraying concentrateDegree of rotation | Diameter of lesion |
| Example 5 | 1mg/mL | 0.23±0.05cm |
| Example 6 | 1mg/mL | 0.21±0.04cm |
| Comparative example 1 | 1mg/mL | 0.82±0.03cm |
| Comparative example 2 | 1mg/mL | 1.49±0.05cm |
| Comparative example 3 | 1mg/mL | 0.76±0.07cm |
| Comparative example 4 | 1mg/mL | 0.98±0.01cm |
| Group of epsilon-polylysines | 1mg/mL | 1.81±0.07cm |
| Clean water group | 30mL | 2.10±0.13cm |
And (3) data analysis:
1. from the results shown in Table 1, it can be seen that the rod-shaped epsilon-polylysine nanoparticles prepared in examples 3-4 or the modified rod-shaped epsilon-polylysine nanoparticles prepared in examples 5-6 of the present invention have good tomato leaf spot control ability, and the diameter of the inoculated leaf spot is significantly smaller than that of the clear water group. And the control capacity of the embodiments 3-4 is improved by more than 4 times compared with that of the epsilon-polylysine group which is not nano-sized, which shows that the rod-shaped epsilon-polylysine nanoparticles prepared by mixing the epsilon-polylysine with the attapulgite or the bentonite have obvious synergistic effect, the modified epsilon-polylysine nanoparticles prepared by the embodiments 5-6 have stronger synergistic effect, the diameter of a lesion is reduced by nearly 6 times compared with that of the epsilon-polylysine group, and the control effect of the modified epsilon-polylysine nanoparticle method is better.
2. Analysis of the data in table 2 shows that when the compounding ratio of attapulgite or bentonite to epsilon-polylysine reaches 40:1, the content of the epsilon-polylysine in 1g/L of aqueous dispersion prepared from the epsilon-polylysine nanoparticles is 0.025g/L (25 mg/mL), compared with the content of the epsilon-polylysine of an epsilon-polylysine group of 0.1g/L, the dosage of the epsilon-polylysine is reduced by 4 times, but the control capability of the epsilon-polylysine is outstanding and is superior to that of the epsilon-polylysine group with large dose, and the control cost is obviously reduced when the epsilon-polylysine nanoparticle is used in the field. In addition, SOD has a significant positive correlation with the stress resistance of the plants, and has been widely used for indicating the resistance capability of the plants, and the SOD content of example 1 is significantly higher than that of epsilon-polylysine group, which shows that the tomato plants infected by example 1 have higher vitality and better tomato resistance inducing performance, and can reduce the infection probability of the tomato plants.
3. Analysis of the data in tables 1 and 3 shows that the lesion diameter of the embodiments 5 and 6 is significantly reduced compared with the lesion diameter of the epsilon-polylysine group, and the lesion diameter of the epsilon-polylysine group is also significantly increased compared with the data in table 1, which indicates that the antibacterial performance and the prevention and treatment effect are reduced when the epsilon-polylysine and the alkaline plant ash leachate are simultaneously applied, while the difference of the embodiments 5 to 6 is not large compared with the lesion diameter in table 1, which indicates that the modified epsilon-polylysine nanoparticles can be simultaneously used with the alkaline fertilizer without reducing the pesticide effect.
4. Example 6 is compared with comparative examples 1 to 4, and compared with comparative example 1, in comparative example 1, calcium chloride is not used, arginine and protamine are not used, betaine is not used in comparative example 3, and gelatin is not used in comparative example 4, the antibacterial effect is obviously reduced, the lesion diameter is obviously increased, particularly, the lesion diameter of comparative example 2 reaches 1.49cm, and the result is shown in fig. 2. The antibacterial action is achieved because the antibacterial principle of epsilon-polylysine is that microorganisms are adsorbed to the surface of cells through electrostatic absorption, then cell membranes are physically cracked, and inner substances leak to kill bacteria, but epsilon-polylysine is a polycation compound, the content of anions is too much under alkaline conditions, the cationic charges of epsilon-polylysine are reduced, the anions adsorbed to the cell membranes of the microorganisms are correspondingly reduced, and the adsorption and damage capability of epsilon-polylysine to the surfaces of the bacteria are weakened, so that the antibacterial activity is reduced. The modification of the epsilon-polylysine nanoparticles is that arginine and protamine are combined with the epsilon-polylysine nanoparticles under the action of betaine to form anion complexing points to further fix and bind anions, the influence of the anions on the epsilon-polylysine nanoparticles is reduced, a proper amount of calcium ions are added, partial phosphate radicals in most fertilizers can also be combined, the application range of the epsilon-polylysine nanoparticles is further expanded, the added gelatin can further bind arginine, protamine, betaine and calcium ions on the surfaces of the epsilon-polylysine nanoparticles, a certain viscosity can be increased, the leaf surface adhesion of the epsilon-polylysine nanoparticles is remarkably increased, the rain or fertilizer erosion is further prevented, the prevention and treatment effect is improved, and the pesticide effect is increased.
Although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the spirit and scope of the invention as defined in the appended claims. The techniques, shapes, and configurations not described in detail in the present invention are all known techniques.
Claims (10)
1. A rod-like nanocrystallization method of epsilon-polylysine, which is characterized by comprising the following steps:
(1) Preparing attapulgite or bentonite into an aqueous dispersion with a concentration of 1-5% w/v, followed by dispersing for 15-30min by a homogenizer to stabilize the suspension to obtain an aqueous suspension dispersion;
(2) And dissolving polylysine in water, slowly adding the polylysine into the suspension aqueous dispersion under the condition of stirring, stirring for 2-4h, centrifuging, collecting, and drying to obtain the rod-shaped epsilon-polylysine nanoparticles.
2. The rod-like nanocrystallization method of epsilon-polylysine according to claim 1, wherein the mass ratio of polylysine to attapulgite or bentonite is (1-5): 40.
3. The rod-like nanocrystallization method of epsilon-polylysine as recited in claim 1, wherein the rotation speed of said homogenizer is 13000-20000rpm.
4. The rod-like nanocrystallization method of epsilon-polylysine as set forth in claim 1, wherein rod-like epsilon-polylysine nanoparticles are subjected to a modification treatment comprising the following raw materials: arginine, betaine, calcium chloride, protamine, gelatin and rod-shaped epsilon-polylysine nanoparticles.
5. The rod-like nanocrystallization method of epsilon-polylysine according to claim 4, wherein the raw materials comprise the following components in parts by mass: 0.8-1.2 parts of arginine, 0.5-1 part of betaine, 0.2-0.4 part of calcium chloride, 0.2-0.5 part of protamine, 0.1-0.2 part of gelatin and 10-12 parts of rod-shaped epsilon-polylysine nanoparticles.
6. The rod-like nanocrystallization method of epsilon-polylysine according to claim 5, wherein the modification treatment comprises the following steps:
(1) Mixing calcium chloride with water, stirring uniformly to obtain a 35wt% calcium chloride solution, heating to 30-35 ℃, adding rod-like epsilon-polylysine nanoparticles to obtain an epsilon-polylysine nanoparticle water dispersion, and keeping the temperature for later use;
(2) Mixing arginine, betaine and protamine, adding into the water dispersion of the epsilon-polylysine nano particles, carrying out ultrasonic oscillation for 10-15min, adding gelatin, heating to 50-55 ℃, stirring for 15-20min, carrying out freeze drying, and taking out to obtain the modified rod-shaped epsilon-polylysine nano particles.
7. Use of epsilon-polylysine nanoparticles prepared according to any of claims 1 or 4-6, wherein the rod-shaped epsilon-polylysine nanoparticles or modified epsilon-polylysine nanoparticles are used for controlling bacterial leaf diseases of solanaceae plants.
8. The use of epsilon-polylysine nanoparticles of claim 7 wherein the rod-shaped epsilon-polylysine nanoparticles or modified epsilon-polylysine nanoparticles are used in the control of tomato leaf spot disease.
9. Use of epsilon-polylysine nanoparticles prepared according to any of claims 1 or 4, characterized in that the rod-shaped epsilon-polylysine nanoparticles or modified epsilon-polylysine nanoparticles are used in pesticides.
10. The use of epsilon-polylysine nanoparticles according to claim 9, wherein the rod-like epsilon-polylysine nanoparticles or modified epsilon-polylysine nanoparticles are mixed with water and sprayed directly onto the surface of plants.
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| CN108184873A (en) * | 2018-01-29 | 2018-06-22 | 吴元华 | A kind of medicament composition of the prevention phytobacterial disease containing epsilon-polylysine |
| CN113397081A (en) * | 2021-06-21 | 2021-09-17 | 扬州大学 | Preparation method of epsilon-polylysine-Arabic gum nano-particles |
| CN113881154A (en) * | 2021-09-29 | 2022-01-04 | 山东小麦歌环保科技有限公司 | Bacteriostatic food packaging box and production method thereof |
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| CN108184873A (en) * | 2018-01-29 | 2018-06-22 | 吴元华 | A kind of medicament composition of the prevention phytobacterial disease containing epsilon-polylysine |
| CN113397081A (en) * | 2021-06-21 | 2021-09-17 | 扬州大学 | Preparation method of epsilon-polylysine-Arabic gum nano-particles |
| CN113881154A (en) * | 2021-09-29 | 2022-01-04 | 山东小麦歌环保科技有限公司 | Bacteriostatic food packaging box and production method thereof |
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