Biological enzyme preparation which rapidly reduces pesticide residues and preparation method therefor

Abstract

Provided in the present invention are a biological enzyme preparation which rapidly reduces pesticide residues and a preparation method therefor, belonging to the technical field of pesticide residue degradation. The biological enzyme preparation which rapidly reduces pesticide residues provided in the present invention comprises the following components in parts by weight: 10-20 parts of a phytase, 10-50 parts of acetylcholinesterase, 5-10 parts of a protease, 10-20 parts of tyrosinase, 1-3 parts of a Platycodon grandiflorus extract, and 1-3 parts of an Aloe vera extract. Provided in the present invention is a biological enzyme preparation which rapidly and efficiently reduces pesticide residues; the biological enzyme preparation has a removal rate of 95% or higher with respect to organophosphorus pesticides on fruits and vegetables and a removal rate of 70% or higher with respect to pyrethroid pesticides on fruits and vegetables; moreover, the biological enzyme preparation provided in the present invention has a simple formulation, is easy to wash off, and does not form a secondary residue.

Description

A bio-enzyme preparation for rapidly reducing pesticide residues and its preparation method Technical Field

[0001] This invention relates to the field of pesticide residue degradation technology, and in particular to a bio-enzyme preparation for rapidly reducing pesticide residues and its preparation method. Background Technology

[0002] Fruits and vegetables are essential foods in daily life; however, to prevent pests and diseases during cultivation, most fruits and vegetables require repeated application of pesticides. This leads to the problem of pesticide residues in fruits and vegetables. Traditional solutions to pesticide residue problems include soaking and washing, alkaline water soaking and washing, boiling water rinsing and washing, rice water soaking and washing, salt water washing, water, baking soda and flour removal, peeling, storage, and sun drying. With the development of science and technology and a better understanding of pesticides, it has been discovered that enzymes secreted by soil microorganisms have a degrading effect on pesticides. This has opened up new avenues for using bio-enzyme preparations to degrade pesticide residues. As a result, many new microorganisms and their enzyme products designed to degrade pesticide residues have emerged in existing technologies. However, developing new and highly efficient microorganisms and enzymes is more difficult, and developing new bio-enzymes through genetic modification is too costly. Developing highly efficient bio-enzyme preparations based on existing bio-enzymes is relatively easier and cheaper. Summary of the Invention

[0003] The purpose of this invention is to provide a bio-enzyme preparation for rapidly reducing pesticide residues and its preparation method, which can achieve efficient removal of pesticide residues with a simple formula and will not cause secondary residues.

[0004] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0005] This invention provides a bio-enzyme preparation for rapidly reducing pesticide residues, comprising the following components in parts by weight:

[0006] Phytase 10-20 parts, acetylcholinesterase 10-50 parts, protease 5-10 parts, tyrosinase 10-20 parts, Platycodon grandiflorus extract 1-3 parts, Aloe vera extract 1-3 parts.

[0007] Preferably, the preparation method of the platycodon root extract includes the following steps:

[0008] (1) Take the leaves or roots of Platycodon grandiflorus, dry them, and then pulverize them to obtain Platycodon grandiflorus powder;

[0009] (2) Mix the Platycodon grandiflorus powder with an ethanol solution, extract, collect the extract and dry it to obtain Platycodon grandiflorus extract.

[0010] Preferably, the mass-to-volume ratio of the platycodon root powder to the ethanol solution is 1g:15-20mL, and the volume concentration of the ethanol solution is 40-50%.

[0011] Preferably, the extraction is microwave-assisted extraction, with a microwave power of 400-420W, a temperature of 40-50℃, a total extraction time of 40-60min, and microwave extraction every 8-10min for 20-40s each time.

[0012] Preferably, the drying process involves first rotary evaporating the extract to remove ethanol at a temperature of 60–70°C, and then drying it at 40–50°C until the moisture content is below 10%.

[0013] Preferably, the method for preparing the aloe vera extract includes the following steps:

[0014] (1) The aloe vera is dried, peeled, baked, and pulverized to obtain aloe vera powder;

[0015] (2) The aloe vera powder was mixed with an ethanol solution and extracted to obtain an extract;

[0016] (3) After the extract is dried, aloe vera extract is obtained.

[0017] Preferably, the mass-to-volume ratio of the aloe vera powder to the ethanol solution is 1g:20-30mL, and the concentration of the ethanol solution is 60-70%.

[0018] Preferably, the extraction is performed 2 to 3 times, with each extraction lasting 40 to 60 minutes and at a temperature of 80 to 90°C.

[0019] Preferably, the drying process involves first rotary evaporating the extract to remove ethanol at a temperature of 60–70°C, and then drying it at 40–50°C until the moisture content is below 10%.

[0020] The present invention also provides a method for preparing the bio-enzyme preparation for rapidly reducing pesticide residues, wherein phytase, acetylcholinesterase, protease and tyrosinase are mixed with Platycodon grandiflorus extract and Aloe vera extract to obtain the bio-enzyme preparation for rapidly reducing pesticide residues.

[0021] This invention provides a rapid and efficient bio-enzyme preparation for reducing pesticide residues. It achieves a clearance rate of over 95% for organophosphorus pesticides on fruits and vegetables and over 70% for pyrethroid pesticides on fruits and vegetables. Furthermore, the bio-enzyme preparation provided by this invention has a simple formula, with selected components being bio-enzymes and plant extracts. It is easy to wash off and will not leave secondary residues. Detailed Implementation

[0022] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0023] The phytase used in the following examples was purchased from Shanxi Zhongnuo Biotechnology Co., Ltd., with an enzyme activity of 50,000 U / g; acetylcholinesterase was purchased from Shanghai Kanglang Biotechnology Co., Ltd., with an enzyme activity of 200 U / g; protease was purchased from Hebei Saiyi Biotechnology Co., Ltd., with an enzyme activity of 50,000 U / g; tyrosinase was purchased from Hefei Bomei Biotechnology Co., Ltd., with an enzyme activity of 25,000 U / g; and amylase was purchased from Hubei Haijia Biotechnology Co., Ltd., with an enzyme activity of 100,000 U / g.

[0024] The Platycodon grandiflorus extract, Aloe vera extract, and Sapindus mukorossi extract used in the following examples were prepared according to the following methods.

[0025] Platycodon grandiflorus extract: Platycodon grandiflorus leaves were dried at 50℃ until the moisture content was less than 15%, pulverized through an 80-mesh sieve, and the sieve residue was collected to obtain Platycodon grandiflorus powder. The Platycodon grandiflorus powder was weighed and mixed with 50% ethanol solution at a ratio of 1g:20mL. Microwave-assisted extraction was performed at a power of 408W and an extraction temperature of 45℃. Microwaves were applied every 10 minutes for 30 seconds each time, for a total extraction time of 60 minutes. The mixture was filtered, and the extract was collected and rotary evaporated at 65℃ until the ethanol was completely evaporated. Then, it was transferred to a dryer and dried at 45℃ until the moisture content was less than 10% to obtain Platycodon grandiflorus extract.

[0026] Aloe vera extract: After drying aloe vera and removing the skin, dry it in a 45℃ oven until the moisture content is below 5%. Then, pulverize it through a 100-mesh sieve and collect the sieve residue to obtain aloe vera powder. Weigh the aloe vera powder and mix it with a 65% ethanol solution at a ratio of 1g:30mL. Extract at 85℃ for 1 hour. Collect the extract and then extract the residue again under the same material-liquid ratio and extraction conditions. Collect the extracts and combine the two extracts. Perform rotary evaporation at 65℃ until the ethanol is completely evaporated. Then, transfer the extract to a dryer and dry it at 45℃ until the moisture content is below 10% to obtain the aloe vera extract.

[0027] Sapindus mukorossi extract: The dried fruit of Sapindus mukorossi was pulverized and passed through a 40-mesh sieve. The sieve residue was collected to obtain Sapindus mukorossi powder. The Sapindus mukorossi powder was weighed and mixed with 50% ethanol solution at a mass-to-volume ratio of 1:10 mL. Ultrasonic extraction was performed at a power of 200 W, a temperature of 80℃, and a time of 1 h. The mixture was centrifuged (8000 rpm, 10 min), and the supernatant was collected. The supernatant was then freeze-dried under vacuum (-80℃, 24 h) to obtain Sapindus mukorossi extract.

[0028] Example 1

[0029] Phytase 15 parts by weight, acetylcholinesterase 30 parts by weight, protease 8 parts by weight, tyrosinase 15 parts by weight, platycodon root extract 2 parts by weight, aloe vera extract 2 parts by weight.

[0030] Mix the components according to the above-mentioned weight proportions to obtain the biological enzyme preparation.

[0031] Example 2

[0032] Phytase 10 parts by weight, acetylcholinesterase 10 parts by weight, protease 5 parts by weight, tyrosinase 20 parts by weight, Platycodon grandiflorus extract 1 part by weight, and aloe vera extract 1 part by weight.

[0033] Mix the components according to the above-mentioned weight proportions to obtain the biological enzyme preparation.

[0034] Example 3

[0035] Phytase 20 parts by weight, acetylcholinesterase 50 parts by weight, protease 10 parts by weight, tyrosinase 10 parts by weight, Platycodon grandiflorus extract 3 parts by weight, and Aloe vera extract 3 parts by weight.

[0036] Mix the components according to the above-mentioned weight proportions to obtain the biological enzyme preparation.

[0037] Comparative Example 1

[0038] Phytase 15 parts by weight, acetylcholinesterase 30 parts by weight, protease 8 parts by weight, tyrosinase 15 parts by weight.

[0039] Mix the components according to the above-mentioned weight proportions to obtain the biological enzyme preparation.

[0040] Comparative Example 2

[0041] Phytase 15 parts by weight, acetylcholinesterase 30 parts by weight, amylase 8 parts by weight, tyrosinase 15 parts by weight, platycodon root extract 2 parts by weight, aloe vera extract 2 parts by weight.

[0042] Mix the components according to the above-mentioned weight proportions to obtain the biological enzyme preparation.

[0043] Experimental Example 1

[0044] Experiments were conducted on the removal of pesticide residues from fruits and vegetables using the bio-enzyme preparations prepared in Examples 1-3 and Comparative Examples 1-2.

[0045] All fruits and vegetables used in this experiment were organic and had not been treated with pesticides during their growth to ensure the accuracy of the test results.

[0046] The fruit used in the experiment was strawberry, and the vegetable was lettuce.

[0047] Preparation of pesticide solution: Phoxim concentration is 0.2 mg / L.

[0048] Pesticide treatment: The experimental fruits and vegetables were immersed in the prepared pesticide solution for 15 minutes, then removed and air-dried. The air-dried fruits and vegetables were divided into six groups, with three replicates in each group. The pesticide residue at this point was measured and recorded as the initial pesticide residue (mg / kg) in Table 1.

[0049] Bioenzyme preparation treatment: The bioenzyme preparations prepared in Examples 1-3 and Comparative Examples 1-2 were dissolved in water at a ratio of 1g:1.5L and stirred evenly. Then, the corresponding experimental fruits and vegetables were immersed in the corresponding bioenzyme preparation solutions for 30 minutes. The control group was replaced with water. After being taken out, they were rinsed with running water for 1 minute, air-dried naturally, and the pesticide residue (mg / kg) was measured and recorded in Table 1.

[0050] The pesticide residue removal rate (%) was calculated as follows: [(initial pesticide residue - treated pesticide residue) / initial pesticide residue] * 100%. The results are shown in Table 1.

[0051] The method for determining pesticide residues is in accordance with NY / T 761-2008.

[0052] Table 1. Pesticide residue removal rate after treatment with bio-enzyme preparations

[0053] As shown in Table 1, the bio-enzyme preparations provided in Examples 1-3 have a high clearance rate for organophosphorus pesticides. In comparison, the clearance rate of organophosphorus pesticides in Comparative Example 1, which uses enzymes alone, is greatly reduced. In Comparative Example 2, the clearance rate of organophosphorus pesticides is also significantly affected after replacing the protease with amylase. It can be seen that the bio-enzyme preparation formulations provided in this application are relatively reasonable.

[0054] Experimental Example 2

[0055] To investigate the effect of plant extracts on the clearance rate of biological enzyme preparations, the following biological enzyme preparations were prepared in this experiment. For the pesticide residue experiment, please refer to Experiment Example 1. The experiment was conducted only on strawberries, and the final calculated pesticide residue clearance rate is shown in Table 2.

[0056] Table 2. Effects of plant extracts on pesticide residue removal rate

[0057] As shown in Table 2, the scavenging effect of Platycodon grandiflorus extract and Aloe vera extract is not as good as that of the present application when they are combined with enzyme components alone or with Sapindus mukorossi extract.

[0058] Experimental Example 3

[0059] This experiment investigated the effects of different combinations of enzymes and plant extracts on the pesticide residue removal rate of biological enzyme preparations. The pesticide residue experiment is described in Experiment Example 1, and the experiment was conducted only on strawberries. The final calculated pesticide residue removal rate is shown in Table 3.

[0060] Table 3. Effects of enzyme components on pesticide residue removal rate

[0061] As shown in Table 3, phytase and acetylcholinesterase are the main enzymes affecting the removal effect of biological enzyme preparations. Although protease and tyrosinase have relatively low removal rates of pesticide residues on their own, their combined use with phytase and acetylcholinesterase can further improve the removal rate of organophosphorus pesticide residues by biological enzyme preparations.

[0062] Example 4

[0063] Based on the above conclusions, this experiment determined the scavenging rate of the bio-enzyme preparation prepared in Example 1 against pyrethroid pesticides. The experimental method was the same as in Example 1, except that deltamethrin was selected as the pyrethroid pesticide at a concentration of 0.5 mg / L, and the selected fruits and vegetables were still strawberries and lettuce. The pesticide treatment and bio-enzyme preparation treatment were the same as in Example 1. The results are shown in Table 4. The scavenging rate of the bio-enzyme preparation prepared in Example 1 against deltamethrin was over 70%.

[0064] Table 4. Scavenging rate of pyrethroid pesticide residues by bio-enzyme preparations

[0065] As can be seen from the above embodiments, the present invention provides a rapid and efficient bio-enzyme preparation for reducing pesticide residues. The removal rate of organophosphates on fruits and vegetables is over 95%, and the removal rate of pyrethroid pesticides on fruits and vegetables is over 70%. Moreover, the bio-enzyme preparation provided by the present invention has a simple formula, is easy to clean, and will not form secondary residues.

[0066] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A bio-enzyme preparation for rapidly reducing pesticide residues, characterized in that, It includes the following components in parts by weight: Phytase 10-20 parts, acetylcholinesterase 10-50 parts, protease 5-10 parts, tyrosinase 10-20 parts, Platycodon grandiflorus extract 1-3 parts, Aloe vera extract 1-3 parts.

2. The biological enzyme preparation according to claim 1, characterized in that, The preparation method of the Platycodon grandiflorus extract includes the following steps: (1) Take the leaves or roots of Platycodon grandiflorus, dry them, and then pulverize them to obtain Platycodon grandiflorus powder; (2) Mix the Platycodon grandiflorus powder with an ethanol solution, extract, collect the extract and dry it to obtain Platycodon grandiflorus extract.

3. The biological enzyme preparation according to claim 2, characterized in that, The mass-to-volume ratio of the platycodon root powder to the ethanol solution is 1g:15-20mL, and the volume concentration of the ethanol solution is 40-50%.

4. The biological enzyme preparation according to claim 3, characterized in that, The extraction was microwave-assisted extraction, with a microwave power of 400-420W, a temperature of 40-50℃, a total extraction time of 40-60 minutes, and microwaves were applied every 8-10 minutes for 20-40 seconds each time.

5. The biological enzyme preparation as described in claim 4, characterized in that, The drying process involves first rotary evaporating the extract to remove ethanol at a temperature of 60–70°C, and then drying it at 40–50°C until the moisture content is below 10%.

6. The biological enzyme preparation according to claim 1, characterized in that, The preparation method of the aloe vera extract includes the following steps: (1) The aloe vera is dried, peeled, baked, and pulverized to obtain aloe vera powder; (2) The aloe vera powder was mixed with an ethanol solution and extracted to obtain an extract; (3) After the extract is dried, aloe vera extract is obtained.

7. The biological enzyme preparation according to claim 6, characterized in that, The mass-to-volume ratio of aloe vera powder to ethanol solution is 1g:20-30mL, and the concentration of ethanol solution is 60-70%.

8. The biological enzyme preparation according to claim 7, characterized in that, The extraction is performed 2 to 3 times, with each extraction lasting 40 to 60 minutes and at a temperature of 80 to 90°C.

9. The biological enzyme preparation according to claim 8, characterized in that, The drying process involves first rotary evaporating the extract to remove ethanol at a temperature of 60–70°C, and then drying it at 40–50°C until the moisture content is below 10%.

10. A method for preparing a bio-enzyme preparation for rapidly reducing pesticide residues as described in any one of claims 1 to 9, characterized in that, Phytase, acetylcholinesterase, protease, and tyrosinase are mixed with Platycodon grandiflorus extract and Aloe vera extract to obtain the aforementioned bio-enzyme preparation for rapidly reducing pesticide residues.

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