Bio-enzyme preparation capable of rapidly reducing pesticide residues and preparation method of bio-enzyme preparation

Abstract

The invention provides a biological enzyme preparation for rapidly reducing pesticide residues and a preparation method thereof, and belongs to the technical field of pesticide residue degradation. The invention provides a biological enzyme preparation capable of rapidly reducing pesticide residues. The biological enzyme preparation is prepared from the following components in parts by weight: 10 to 20 parts of phytase, 10 to 50 parts of acetylcholin esterase, 5 to 10 parts of protease, 10 to 20 parts of tyrosinase, 1 to 3 parts of radix platycodonis extract and 1 to 3 parts of aloe extract. According to the biological enzyme preparation capable of rapidly and efficiently reducing the pesticide residues, the removal rate of organophosphorus pesticides on fruits and vegetables is 95% or above, the removal rate of pyrethroid pesticides on the fruits and vegetables is 70% or above, and the biological enzyme preparation is simple in formula, easy to clean and free of secondary residues.

Description

Technical Field

[0001] The invention relates to the technical field of pesticide residue degradation, and in particular to a biological enzyme preparation for rapidly reducing pesticide residues and a preparation method thereof. Background Art

[0002] Fruits and vegetables are indispensable foods in daily life. However, in order to avoid pests and diseases during the planting process, most fruits and vegetables need to be continuously and repeatedly applied with pesticides. Therefore, the problem of pesticide residues in fruits and vegetables is caused. For the problem of pesticide residues, the traditional solutions are mainly soaking and washing, alkaline water soaking and washing, boiling water blanching and washing, rice washing water soaking and washing, salt water washing, water baking soda and flour removal, peeling, storage, airing, etc. With the development of science and technology and the improvement of the understanding of pesticides, people have found that the enzymes secreted by soil microorganisms have a degrading effect on pesticides, which has opened up a new idea for people to use biological enzyme preparations to degrade agricultural residues. For this reason, many new microorganisms and their enzyme products for the purpose of degrading agricultural residues have emerged in the prior art. However, it is more difficult to continue to develop new and efficient microorganisms and enzymes, and the cost of developing new biological enzymes by genetic modification is too high. On the basis of existing biological enzymes, it is relatively easier and less costly to develop efficient biological enzyme preparations. Summary of the invention

[0003] The purpose of the present invention is to provide a biological enzyme preparation for rapidly reducing pesticide residues and a preparation method thereof, which can achieve the purpose of efficiently removing pesticide residues with a simple formula and will not form secondary residues.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0005] The present invention provides a biological enzyme preparation for rapidly reducing pesticide residues, comprising the following components in parts by weight:

[0006] 10-20 parts of phytase, 10-50 parts of acetylcholinesterase, 5-10 parts of protease, 10-20 parts of tyrosinase, 1-3 parts of platycodon grandiflorum extract, and 1-3 parts of aloe extract.

[0007] Preferably, the preparation method of the Platycodon grandiflorum extract comprises the following steps:

[0008] (1) drying and crushing the leaves or roots of Platycodon grandiflorum to obtain Platycodon grandiflorum powder;

[0009] (2) Mixing the platycodon powder with an ethanol solution, extracting, collecting the extract and drying it to obtain a platycodon extract.

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

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

[0012] Preferably, the drying is to first subject the extract to rotary evaporation to remove ethanol, the rotary evaporation temperature is 60-70°C, and then dry at 40-50°C until the water content is less than 10%.

[0013] Preferably, the preparation method of the aloe extract comprises the following steps:

[0014] (1) drying, peeling, drying and crushing aloe vera to obtain aloe vera powder;

[0015] (2) mixing the aloe vera powder with an ethanol solution, extracting to obtain an extract;

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

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

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

[0019] Preferably, the drying is to first subject the extract to rotary evaporation to remove ethanol, the rotary evaporation temperature is 60-70°C, and then dry at 40-50°C until the water content is less than 10%.

[0020] The present invention also provides a method for preparing the biological enzyme preparation for rapidly reducing pesticide residues, wherein phytase, acetylcholinesterase, protease and tyrosinase are mixed with platycodon extract and aloe extract to obtain the biological enzyme preparation for rapidly reducing pesticide residues.

[0021] The present invention provides a biological enzyme preparation for rapidly and efficiently reducing pesticide residues, with a removal rate of more than 95% for organophosphorus pesticides on fruits and vegetables, and a removal rate of more than 70% for pyrethroid pesticides on fruits and vegetables. The biological enzyme preparation provided by the present invention has a simple formula, and the selected components are biological enzymes and plant extracts, which are easy to clean and will not form secondary residues. DETAILED DESCRIPTION

[0022] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope 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 grandiflorum extract, aloe vera extract and soapberry extract used in the following examples were prepared according to the following method.

[0025] Platycodon grandiflorum extract: Take Platycodon grandiflorum leaves, dry them at 50°C until the water content is less than 15%, crush them through an 80-mesh sieve, take the sieve to obtain Platycodon grandiflorum powder. Weigh Platycodon grandiflorum powder and mix it with 50% ethanol solution in a ratio of 1g:20mL, perform microwave-assisted extraction, the microwave power is 408W, the extraction temperature is controlled at 45°C, microwave once every 10 minutes, each microwave is 30s, the total extraction time is 60 minutes, filter, collect the extract, rotary evaporate at 65°C until the ethanol evaporates completely, and then transfer to a dryer and dry at 45°C until the water content is less than 10%, to obtain Platycodon grandiflorum extract.

[0026] Aloe vera extract: Take aloe vera, dry it, peel it, put it in a 45℃ oven to dry it until the water content is less than 5%, then crush it through a 100-mesh sieve, take the sieve to obtain aloe vera powder. Weigh the aloe vera powder and mix it with 65% ethanol solution in a ratio of 1g:30mL, extract it at 85℃ for 1h, collect the extract, extract the residue again according to the same material-liquid ratio and extraction conditions, collect the extract, combine the two extracts, perform rotary evaporation, evaporate at 65℃ until the ethanol evaporates completely, then transfer it to a dryer and dry it at 45℃ until the water content is less than 10%, and obtain aloe vera extract.

[0027] Soapberry extract: The dried fruit of Soapberry is crushed, passed through a 40-mesh sieve, and the sieved material is taken to obtain Soapberry powder. The Soapberry powder is weighed and mixed with a 50% ethanol solution at a mass volume ratio of 1:10 mL, and ultrasonic extraction is performed at a power of 200 W. The temperature during the extraction is 80°C for 1 h, and the extraction is centrifuged (8000 rpm, 10 min), and the supernatant is collected and vacuum freeze-dried (-80°C, 24 h) to obtain the Soapberry extract.

[0028] Example 1

[0029] 15 parts by weight of phytase, 30 parts by weight of acetylcholinesterase, 8 parts by weight of protease, 15 parts by weight of tyrosinase, 2 parts by weight of Platycodon grandiflorum extract, and 2 parts by weight of aloe extract.

[0030] The components are mixed according to the above weight proportions to obtain a bioenzyme preparation.

[0031] Example 2

[0032] 10 parts by weight of phytase, 10 parts by weight of acetylcholinesterase, 5 parts by weight of protease, 20 parts by weight of tyrosinase, 1 part by weight of Platycodon grandiflorum extract, and 1 part by weight of aloe extract.

[0033] The components are mixed according to the above weight proportions to obtain a bioenzyme preparation.

[0034] Example 3

[0035] 20 parts by weight of phytase, 50 parts by weight of acetylcholinesterase, 10 parts by weight of protease, 10 parts by weight of tyrosinase, 3 parts by weight of Platycodon grandiflorum extract, and 3 parts by weight of aloe extract.

[0036] The components are mixed according to the above weight proportions to obtain a bioenzyme preparation.

[0037] Comparative Example 1

[0038] 15 parts by weight of phytase, 30 parts by weight of acetylcholinesterase, 8 parts by weight of protease, and 15 parts by weight of tyrosinase.

[0039] The components are mixed according to the above weight proportions to obtain a bioenzyme preparation.

[0040] Comparative Example 2

[0041] 15 parts by weight of phytase, 30 parts by weight of acetylcholinesterase, 8 parts by weight of amylase, 15 parts by weight of tyrosinase, 2 parts by weight of Platycodon grandiflorum extract, and 2 parts by weight of aloe extract.

[0042] The components are mixed according to the above weight proportions to obtain a bioenzyme preparation.

[0043] Experimental Example 1

[0044] The bioenzyme preparations prepared in Examples 1 to 3 and Comparative Examples 1 to 2 were subjected to an experiment for removing pesticide residues from fruits and vegetables.

[0045] The fruits and vegetables used in this experiment are all organic fruits and vegetables, and no pesticides have been applied during the growth process to ensure the accuracy of the measurement results.

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

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

[0048] Pesticide treatment: Immerse the experimental fruits and vegetables in the prepared pesticide solution for 15 minutes, then take them out and let them dry naturally. Divide the naturally dried fruits and vegetables into six groups, with three replicates in each group, and measure the pesticide residue at this time as the initial pesticide residue (mg / kg), which is recorded in Table 1.

[0049] Bioenzyme preparation treatment: The bioenzyme preparations prepared in Examples 1 to 3 and Comparative Examples 1 to 2 were dissolved in water at a ratio of 1 g: 1.5 L, stirred evenly, and then the corresponding experimental fruits and vegetables were immersed in the corresponding bioenzyme preparation solution for 30 min. The control group was replaced with clean water. After taking out, they were rinsed with running clean water for 1 min. After naturally drying, the pesticide residue (mg / kg) was determined and recorded in Table 1.

[0050] Calculate the pesticide residue removal rate (%) = [(initial pesticide residue amount - pesticide residue amount after treatment) / initial pesticide residue amount] * 100%. The results are shown in Table 1.

[0051] The determination method of pesticide residues shall refer to NY / T 761-2008.

[0052] Table 1 Pesticide residue removal rate treated with bio-enzyme preparations

[0053]

[0054] As can be seen from Table 1, the bioenzyme preparations provided in Examples 1 to 3 have a high clearance rate for organophosphorus pesticides. In comparison, the clearance rate of organophosphorus in Comparative Example 1 using only enzymes as a formula is greatly reduced, and the organophosphorus clearance rate in Comparative Example 2 is also significantly affected after protease is replaced by amylase. It can be seen that the formula of the bioenzyme preparation provided in this application is relatively reasonable.

[0055] Experimental Example 2

[0056] In order to explore the effect of plant extracts on the removal rate of bio-enzyme preparations, this experiment configured the following bio-enzyme preparations. The pesticide residue experiment refers to Experimental Example 1. Only strawberries were experimented. The final calculated pesticide residue removal rate is shown in Table 2.

[0057] Table 2 Effect of plant extracts on pesticide residue removal rate

[0058]

[0059]

[0060] As can be seen from Table 2, Platycodon grandiflorum extract and Aloe vera extract are fixed combinations, and the clearance effect of the biological enzyme preparation prepared by combining the enzyme components of the Platycodon grandiflorum extract and Aloe vera extract alone or in combination with Sapindus mukorossi is not as good as that of the present application.

[0061] Experimental Example 3

[0062] This experiment explored the effects of different enzyme combinations with plant extracts on the pesticide residue removal rate of bioenzyme preparations. The pesticide residue experiment refers to Experimental Example 1, which only experiments on strawberries. The final calculated pesticide residue removal rate is shown in Table 3.

[0063] Table 3 Effect of enzyme components on pesticide residue removal rate

[0064]

[0065] As shown in Table 3, phytase and acetylcholinesterase are the main enzymes that affect the removal effect of biological enzyme preparations. Although protease and tyrosinase have low removal rates for pesticide residues, their combined use with phytase and acetylcholinesterase can further improve the removal rate of biological enzyme preparations for organophosphorus pesticide residues.

[0066] Example 4

[0067] Based on the above conclusions, this experimental example measured the removal rate of pyrethroid pesticides by the biological enzyme preparation prepared in Example 1. The experimental method was the same as that in Experimental Example 1, wherein the pyrethroid pesticide was selected as deltamethrin, with a concentration of 0.5 mg / L, and the selected fruits and vegetables were still strawberries and lettuce, and the pesticide treatment and biological enzyme preparation treatment were the same as in Experimental Example 1. The results are shown in Table 4, and the removal rate of deltamethrin by the biological enzyme preparation prepared in Example 1 was above 70%.

[0068] Table 4 Removal rate of pyrethroid pesticide residues by bioenzyme preparations

[0069] deal with Clearance rate (%) strawberry 75.98％ lettuce 70.25％

[0070] It can be seen from the above embodiments that the present invention provides a biological enzyme preparation for quickly and efficiently reducing pesticide residues, with a removal rate of more than 95% for organophosphorus on fruits and vegetables and a removal rate of more than 70% for pyrethroid pesticides on fruits and vegetables. In addition, the biological enzyme preparation provided by the present invention has a simple formula, is easy to clean, and will not form secondary residues.

[0071] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A biological enzyme preparation for rapidly reducing pesticide residues, characterized in that: The composition comprises the following components in parts by weight: 10-20 parts of phytase, 10-50 parts of acetylcholinesterase, 5-10 parts of protease, 10-20 parts of tyrosinase, 1-3 parts of platycodon grandiflorum extract, and 1-3 parts of aloe extract.

2. The bioenzyme preparation according to claim 1, characterized in that The preparation method of the Platycodon grandiflorum extract comprises the following steps: (1) drying and crushing the leaves or roots of Platycodon grandiflorum to obtain Platycodon grandiflorum powder; (2) Mixing the platycodon powder with an ethanol solution, extracting, collecting the extract and drying it to obtain a platycodon extract.

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

4. The bioenzyme preparation according to claim 3, characterized in that The extraction is microwave-assisted extraction, the microwave power is 400-420W, the temperature is 40-50°C, the total extraction time is 40-60min, the microwave is applied every 8-10min, and the time of each microwave is 20-40s.

5. The bioenzyme preparation according to claim 4, characterized in that The drying step is to firstly subject the extract to rotary evaporation to remove ethanol, the rotary evaporation temperature is 60-70° C., and then dry the extract at 40-50° C. until the water content is less than 10%.

6. The bioenzyme preparation according to claim 1, characterized in that The preparation method of the aloe extract comprises the following steps: (1) drying, peeling, drying and crushing aloe vera to obtain aloe vera powder; (2) mixing the aloe vera powder with an ethanol solution, extracting to obtain an extract; (3) After the extract is dried, an aloe extract is obtained.

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

8. The bioenzyme preparation according to claim 7, characterized in that The extraction is performed 2 to 3 times, each extraction time is 40 to 60 minutes, and each extraction temperature is 80 to 90°C.

9. The bioenzyme preparation according to claim 8, characterized in that The drying step is to firstly subject the extract to rotary evaporation to remove ethanol, the rotary evaporation temperature is 60-70° C., and then dry the extract at 40-50° C. until the water content is less than 10%.

10. A method for preparing a biological enzyme preparation for rapidly reducing pesticide residues according to any one of claims 1 to 9, characterized in that: Phytase, acetylcholinesterase, protease and tyrosinase are mixed with platycodon grandiflorum extract and aloe vera extract to obtain the biological enzyme preparation for rapidly reducing pesticide residues.

Images