CN108998289B

CN108998289B - Enzyme stabilizer, compound pesticide hydrolase liquid and preparation method thereof, and fruit and vegetable cleaning liquid and preparation method thereof

Info

Publication number: CN108998289B
Application number: CN201811121827.2A
Authority: CN
Inventors: 吴启涛; 詹志春; 周樱; 谌颉; 江安娜; 高智明
Original assignee: Wuhan Sunhy Biological Co ltd
Current assignee: Wuhan Sunhy Biological Co ltd
Priority date: 2018-09-26
Filing date: 2018-09-26
Publication date: 2020-09-08
Anticipated expiration: 2038-09-26
Also published as: CN108998289A

Abstract

The invention relates to the field of biological detergents, and provides an enzyme stabilizer which comprises the following components in parts by weight: 10-30 parts of polyhydric alcohol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine and 0.1-2 parts of sodium citrate. Also provides a compound pesticide hydrolase solution and a preparation method thereof, and the method comprises the following steps: and uniformly mixing the enzyme stabilizer and the pesticide hydrolase solution. A fruit and vegetable cleaning liquid and a preparation method thereof are disclosed, the method comprises: mixing the enzyme stabilizer, pesticide hydrolase solution, EDTA-2Na and surfactant. The enzyme stabilizer can improve the stability of the pesticide hydrolase liquid, the compound pesticide hydrolase liquid has good stability and no harm to human health, and the fruit and vegetable cleaning liquid has higher cleaning capability and good stability to pesticides and no harm to human bodies.

Description

Enzyme stabilizer, compound pesticide hydrolase liquid and preparation method thereof, and fruit and vegetable cleaning liquid and preparation method thereof

Technical Field

The invention relates to the field of biological detergents, in particular to an enzyme stabilizer, a compound pesticide hydrolase liquid and a preparation method thereof, and a fruit and vegetable cleaning liquid and a preparation method thereof.

Background

How to reduce the pesticide residue in agricultural products to the maximum extent is more and more concerned by people. The main folk methods are cleaning, peeling, cooking, storing, ozone methods and the like, but the methods have weak universality and large limitation. Zongronfen and the like are found by experiments of soaking green vegetables in a commercially available kitchen detergent, tap water, edible alkali, 1% edible salt, a kitchen detergent added with edible alkali and the like, that the removal rate of the commercially available kitchen detergent on methamidophos and dimethoate is 50% -68%, the removal rate of 1% edible salt is 69%, and the removal rate of the commercially available kitchen detergent after soaking in tap water is only 46%. Wangdongjia et al analyzed the degradation sequence of organophosphorus, carbamate and pyrethroid insecticides in ozone water-soaked pakchoi as organophosphorus, pyrethroid and carbamate.

So far, the pesticide hydrolase liquid is recognized as the most effective method for eliminating pesticide residues on fruits and vegetables because the pesticide hydrolase liquid overcomes the defects of high difficulty in physical and chemical treatment and restoration, high cost, secondary pollution and the like and has the characteristics of thorough pesticide residue removal, safety, rapid degradation and the like. An enzyme stabilizer system of a traditional enzyme cleaning agent mostly comprises borax, propylene glycol or glycerol and the like, the environment can be seriously polluted by using a large amount of borax, and the health hazard of the borax is great. More importantly, it is a significant technical problem to ensure that the high activity of the pesticide-degrading enzyme and its activity do not decrease over time.

In view of this, the present application is specifically made.

Disclosure of Invention

The invention provides an enzyme stabilizer, aiming at solving the problem that the stability of pesticide hydrolase liquid needs to be improved.

The invention also provides a preparation method of the compound pesticide hydrolase liquid, which can prepare the compound pesticide hydrolase liquid with good stability and no harm to health.

The invention also provides a compound pesticide hydrolase liquid which has good stability and no harm to human health.

The invention also provides a preparation method of the fruit and vegetable cleaning solution, which can prepare the fruit and vegetable cleaning solution which can be stored for a long time and still has higher pesticide cleaning capability, and the cleaning solution is harmless to human health.

The invention also provides a fruit and vegetable cleaning solution which can be stored for a long time and still has higher pesticide cleaning capability, and the cleaning solution is harmless to human health.

The invention is realized by the following steps:

an enzyme stabilizer comprises the following components in parts by weight:

10-30 parts of polyhydric alcohol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine and 0.1-2 parts of sodium citrate.

A preparation method of a compound pesticide hydrolase liquid comprises the following steps:

mixing 10-30 parts of polyhydric alcohol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine, 0.1-2 parts of sodium citrate and 10-20 parts of water with 30-70 parts of pesticide hydrolase liquid.

A compound pesticide hydrolase liquid is prepared by the preparation method of the compound pesticide hydrolase liquid.

A preparation method of a fruit and vegetable cleaning solution comprises the following steps of: mixing 10-30 parts of polyhydric alcohol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine, 0.1-2 parts of sodium citrate, 20-40 parts of water, 3-10 parts of pesticide hydrolase liquid, 0.02-0.2 part of EDTA-2Na and 3-15 parts of surfactant uniformly.

A fruit and vegetable cleaning solution is prepared by adopting the preparation method of the fruit and vegetable cleaning solution.

The invention has the beneficial effects that: the components of the enzyme stabilizer are food-grade raw materials basically, so that the enzyme stabilizer is harmless to human health; the high viscosity of the xanthan gum in water enables the pesticide hydrolase liquid to be suspended in the solution, so that the contact between the pesticide hydrolase liquid and water molecules is reduced as much as possible, the pesticide hydrolase liquid is almost kept as a single entity in the solution, and the stability of the pesticide hydrolase liquid is facilitated; the trehalose has non-specific protection effect on biological enzyme; sufficient polyol also provides a better environment for the stability of the pesticide hydrolase solution; the appropriate amount of sodium citrate can perform chelation with various metal ions, so that the enzyme activity is prevented from being reduced or inactivated by the action of the metal ions on the enzyme; the pH value of the formula is synergistically adjusted by the sodium citrate and the triethanolamine, so that the protection effect on the enzyme is enhanced, and the enzyme stabilizer can nonspecifically protect the pesticide hydrolase solution by combining the reasonable proportion of the components, so that the stability is good.

The compound pesticide hydrolase liquid obtained by the design comprises the enzyme stabilizer and is prepared by a reasonable method, so that the compound pesticide hydrolase liquid has good stability and no harm to human health.

According to the preparation method of the compound pesticide hydrolase liquid, which is designed, the compound pesticide hydrolase liquid which is good in stability and harmless to human health can be prepared.

The fruit and vegetable cleaning solution obtained by the design comprises the pesticide hydrolase solution and the enzyme stabilizer, so that the fruit and vegetable cleaning solution can effectively clean the pesticide on the surfaces of the fruits and vegetables, has good stability and is harmless to human health.

By the preparation method of the fruit and vegetable cleaning solution, the pesticide for effectively cleaning the surfaces of the fruits and vegetables can be prepared, and the fruit and vegetable cleaning solution is good in stability and harmless to human health.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a graph of the relative rate of activity of pesticide hydrolase solution as a function of temperature;

FIG. 2 is a graph of the relative rate of activity of pesticide hydrolase solution as a function of pH;

FIG. 3 is a graph showing the effect of a compound enzyme solution on removing organophosphorus pesticides on the surface of lemon according to an embodiment of the present invention;

FIG. 4 is a schematic diagram of the compound enzyme solution provided by the embodiment of the invention after reacting with pyrethrin and contrast water after reacting with pyrethrin;

fig. 5 is a quick-test color card of pyrethroid pesticides.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The embodiments of the invention provide an enzyme stabilizer, a compound pesticide hydrolase liquid and a preparation method thereof, and a fruit and vegetable cleaning liquid and a preparation method thereof.

An enzyme stabilizer comprises the following components in parts by weight:

The xanthan gum in the components can be dissolved into a highly viscous solution in water, when the enzyme stabilizer provided by the invention is used in combination with the pesticide hydrolase liquid, the highly viscous property of the xanthan gum in water enables the pesticide hydrolase liquid to be suspended in the solution, the contact between the pesticide hydrolase liquid and water molecules is reduced as much as possible, the pesticide hydrolase liquid is almost kept as a single entity in the solution, and the stability of the pesticide hydrolase liquid is facilitated; (ii) a Trehalose has a non-specific protective effect on biological enzymes; sufficient polyol also provides a better environment for the stability of the pesticide hydrolase solution, and a proper amount of sodium citrate is chelated with various metals, so that the reduction or inactivation of the enzymatic activity caused by the action of metal ions on the enzymes is avoided; the sodium citrate and the triethanolamine are mutually cooperated to enhance the protection effect on the enzyme, and the enzyme stabilizer can nonspecifically protect the pesticide hydrolase liquid by combining the reasonable proportion of the components, so that the stability of the pesticide hydrolase liquid is good.

In the present invention, the pesticide hydrolase liquid refers to a beta-lactamase belonging to and having the property of an organophosphorus hydrolase, and the enzyme has a zinc (II) dimer, each subunit containing an active center consisting of two divalent zinc ions. The enzyme specifically comprises at least one of methyl parathion hydrolase MPH and phospholipase OPH.

When the enzyme stabilizer is a liquid preparation, 10-40 parts of water is further included for better stabilizing effect of the enzyme stabilizer on pesticide hydrolase liquid.

Further, the enzyme stabilizer further comprises: 0.01-0.2 parts of preservative. The addition of the preservative is beneficial to preventing the enzyme stabilizer from going bad early and prolonging the storage time of the enzyme stabilizer. Preferably, the preservative comprises at least one of methylisothiazolinone, methylchloroisothiazolinone, paraben complex ester and paraben complex ester salt.

mixing 10-30 parts of polyhydric alcohol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine, 0.1-2 parts of sodium citrate and 10-20 parts of water with 30-70 parts of pesticide hydrolase liquid. The compound pesticide hydrolase liquid obtained by uniformly mixing the components has good stability, and the components of the compound pesticide hydrolase liquid are harmless to human health. Preferably, the pesticide hydrolase liquid comprises at least one of methyl parathion hydrolase MPH and phospholipase OPH.

Furthermore, the stability of the compound pesticide hydrolase liquid is better. 10-20 parts of polyol, 2-15 parts of trehalose, 0.05-0.2 part of xanthan gum, 0.1-1 part of triethanolamine, 0.1-1 part of sodium citrate, 10-20 parts of water and 30-70 parts of pesticide hydrolase liquid.

Further, the preparation method of the compound pesticide hydrolase liquid specifically comprises the following steps:

s1, mixing triethanolamine and water to obtain a first mixed solution.

Firstly, 10-30 parts of polyhydric alcohol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine, 0.1-2 parts of sodium citrate and 10-20 parts of water are weighed or measured for later use with 30-70 parts of pesticide hydrolase liquid, or 10-20 parts of polyhydric alcohol, 2-15 parts of trehalose, 0.05-0.2 part of xanthan gum, 0.1-1 part of triethanolamine, 0.1-1 part of sodium citrate and 10-20 parts of water are weighed or measured for later use with 30-70 parts of pesticide hydrolase liquid.

And mixing and stirring the triethanolamine and the water uniformly to obtain a first mixed solution. Further, when the enzyme stabilizer also comprises 0.01-0.2 part of preservative. The preservative is mixed with triethanolamine and water to obtain a first mixed solution. Furthermore, when the content of the preservative has minimal or no influence on the stability of the compound pesticide hydrolase liquid, the preservative is 0.03-0.1 part.

And S2, uniformly mixing xanthan gum, trehalose and sodium citrate with the first mixed solution to obtain a second mixed solution.

And pouring the xanthan gum, the trehalose and the sodium citrate in parts by weight into the first mixed solution, and uniformly stirring to obtain a second mixed solution.

S3, mixing the polyalcohol, the pesticide hydrolase solution and the second mixed solution uniformly;

pouring the polyol and the pesticide hydrolase liquid into the second mixed solution according to the parts by weight, and uniformly stirring. And obtaining the compound pesticide hydrolase liquid with the pH value of 7.0-9.0.

The compound pesticide hydrolase liquid is prepared by adopting the preparation method of the compound pesticide hydrolase liquid. It has the characteristics of good stability and no harm to human health.

A preparation method of a fruit and vegetable cleaning solution comprises the following steps of: mixing 10-30 parts of polyhydric alcohol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine, 0.1-2 parts of sodium citrate, 20-40 parts of water, 3-10 parts of pesticide hydrolase liquid, 0.02-0.2 part of EDTA-2Na and 3-15 parts of surfactant uniformly. The fruit and vegetable cleaning solution prepared by uniformly mixing the components has good stability, the effect of cleaning pesticides on the surfaces of fruits and vegetables after long-time storage is still good, and the components of the fruit and vegetable cleaning solution are harmless to human health. Because the prepared fruit and vegetable cleaning liquid contains the surfactant which is not beneficial to enzyme stability, and the water adding amount in the prepared fruit and vegetable cleaning liquid is slightly larger, the ratio of the enzyme stabilizer to the pesticide hydrolase liquid is relatively larger, namely, the stability of the pesticide hydrolase liquid is better, and the enzyme stabilizer is relatively more added in the fruit and vegetable cleaning liquid.

Furthermore, the stability of the fruit and vegetable cleaning solution is further improved. 15-30 parts of polyhydric alcohol, 8-15 parts of trehalose, 0.05-0.15 part of xanthan gum, 0.2-0.8 part of triethanolamine, 0.4-0.8 part of sodium citrate, 20-40 parts of water, 3-10 parts of pesticide hydrolase liquid, 0.02-0.2 part of EDTA-2Na and 3-15 parts of surfactant.

Further, in order to make the cleaning effect of the fruit and vegetable cleaning solution better, the surfactant comprises: 2-10 parts of nonionic surfactant and 1-5 parts of betaine surfactant. Still further, the nonionic surfactant includes at least one of polysorbate 80, an alkyl glycoside, and a fatty alcohol polyoxyethylene ether.

Furthermore, in order to make the fruit and vegetable cleaning solution have better smell, a proper amount of essence can be added.

Further, the preparation method of the fruit and vegetable cleaning solution specifically comprises the following steps:

s1, mixing triethanolamine and water uniformly to obtain a first intermediate solution.

Firstly, 10-30 parts of polyhydric alcohol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine, 0.1-2 parts of sodium citrate, 20-40 parts of water and 3-10 parts of pesticide hydrolase liquid are weighed or measured for later use. Or weighing or measuring 15-30 parts of polyhydric alcohol, 8-15 parts of trehalose, 0.05-0.15 part of xanthan gum, 0.2-0.8 part of triethanolamine, 0.4-0.8 part of sodium citrate, 20-40 parts of water and 3-10 parts of pesticide hydrolase liquid for later use.

And mixing and stirring the triethanolamine and the water uniformly to obtain a first intermediate solution. Further, when the enzyme stabilizer also comprises 0.01-0.2 part of preservative. The preservative is mixed with triethanolamine and water to obtain a first intermediate solution. Furthermore, in order to minimize or not influence the content of the preservative on the stability of the fruit and vegetable cleaning solution, 0.03-0.1 part of the preservative is added, and if the prepared fruit and vegetable cleaning solution has fragrance, essence is required to be added, and a proper amount of the essence is uniformly mixed with triethanolamine, water and the preservative.

S2, mixing xanthan gum, trehalose, sodium citrate, EDTA-2Na and the first intermediate solution uniformly to obtain a second intermediate solution.

Adding the xanthan gum, the trehalose, the sodium citrate and the EDTA-2Na in parts by weight into the first intermediate solution, and uniformly mixing and stirring to obtain a second intermediate solution.

S3, mixing the polyalcohol, the pesticide hydrolase solution and the second intermediate solution uniformly to obtain a third intermediate solution.

And adding the polyol and the pesticide hydrolase liquid in parts by weight into the second intermediate liquid, and mixing and stirring uniformly to obtain a third intermediate liquid.

And S4, mixing the surfactant and the third intermediate solution uniformly.

And adding the surfactant in parts by weight into the third intermediate solution, and uniformly stirring and mixing to obtain the fruit and vegetable cleaning solution.

A fruit and vegetable cleaning solution is prepared by adopting the preparation method of the fruit and vegetable cleaning solution. The cleaning effect of the pesticide on the surfaces of the fruits and vegetables is good after the fruits and vegetables are stored for a long time, and the components of the cleaning agent are harmless to human health.

The following describes an enzyme stabilizer provided by the present invention with reference to specific examples.

Example 1

The embodiment of the invention provides an enzyme stabilizer, a compound pesticide hydrolase liquid, a preparation method of the compound pesticide hydrolase liquid, a fruit and vegetable cleaning liquid and a preparation method of the fruit and vegetable cleaning liquid.

An enzyme stabilizer comprises 10g glycerol, 15g trehalose, 0.01g xanthan gum, 2g triethanolamine and 0.1g sodium citrate.

The preparation method of the compound pesticide hydrolase liquid comprises the step of mixing and uniformly stirring the enzyme stabilizer, 10g of water and 30g of phospholipase to obtain the compound pesticide hydrolase liquid provided by the embodiment.

The preparation method of the fruit and vegetable cleaning solution comprises the step of mixing and uniformly stirring the enzyme stabilizer, 20g of water, 3g of phospholipase, 0.02g of EDTA-2Na, 2g of polysorbate 80 and 1g of betaine surfactant to obtain the fruit and vegetable cleaning solution provided by the embodiment.

Example 2

An enzyme stabilizer comprises 30g ethylene glycol, 1g trehalose, 0.3g xanthan gum, 0.1g triethanolamine, 2g sodium citrate and 0.01g paraben complex ester.

The preparation method of the compound pesticide hydrolase liquid comprises the step of mixing and uniformly stirring the enzyme stabilizer, 20g of water and 70g of methyl parathion hydrolase to obtain the compound pesticide hydrolase liquid provided by the embodiment.

The preparation method of the fruit and vegetable cleaning solution comprises the step of uniformly mixing and stirring the enzyme stabilizer, 40g of water, 10g of methyl parathion hydrolase, 0.2g of EDTA-2Na, 10g of alkyl glycoside and 5g of betaine surfactant to obtain the fruit and vegetable cleaning solution provided by the embodiment.

Example 3

The embodiment of the invention provides an enzyme stabilizer, a compound pesticide hydrolase liquid and a preparation method thereof.

An enzyme stabilizer comprises 20g of glycerol, 5g of trehalose, 0.05g of xanthan gum, 1g of triethanolamine, 1g of sodium citrate and 0.05g of methylchloroisothiazolinone.

the above components of the enzyme stabilizer were weighed out in the above weight, and 15g of water and 40g of methylparathion hydrolase were weighed out for use.

Mixing and stirring the triethanolamine, the water and the methylchloroisothiazolinone with the above weights uniformly to obtain a first mixed solution; pouring the xanthan gum, the trehalose and the sodium citrate into the first mixed solution by weight, and uniformly stirring to obtain a second mixed solution; the polyhydric alcohol and the methyl parathion hydrolase in the weight are poured into the second mixed solution to obtain the compound pesticide hydrolase liquid provided by the embodiment.

Example 4

An enzyme stabilizer comprises 15g ethylene glycol, 10g trehalose, 0.1g xanthan gum, 0.5g triethanolamine, 0.5g sodium citrate and 0.2g sodium paraben complex.

the above components of the enzyme stabilizer were weighed in the above weight, and 12g of water and 50g of methylparathion hydrolase were weighed for use.

Mixing and stirring the triethanolamine, the water and the paraben composite ester uniformly to obtain a first mixed solution; pouring the xanthan gum, the trehalose and the sodium citrate into the first mixed solution by weight, and uniformly stirring to obtain a second mixed solution; the compounded pesticide hydrolase liquid provided by the embodiment is obtained by pouring the ethylene glycol and the methyl parathion hydrolase with the weight into the second mixed solution.

Example 5

An enzyme stabilizer comprises 18g of 2-propylene glycol, 12g of trehalose, 0.15g of xanthan gum, 0.8g of triethanolamine, 0.8g of sodium citrate and 0.15g of methylisothiazolinone.

the above components of the enzyme stabilizer were weighed out by the above weight, and 18g of water and 60g of phospholipase were weighed out for use.

Mixing and stirring the triethanolamine, the water and the methylisothiazolinone uniformly to obtain a first mixed solution; pouring the xanthan gum, the trehalose and the sodium citrate into the first mixed solution by weight, and uniformly stirring to obtain a second mixed solution; the 2-propylene glycol and the phospholipase with the weight are poured into the second mixed solution to obtain the compound pesticide hydrolase solution provided by the embodiment.

Example 6

An enzyme stabilizer comprises 15g of glycerol, 2g of trehalose, 0.1g of xanthan gum, 0.3g of triethanolamine, 0.5g of sodium citrate and 0.1g of methylisothiazolinone.

the above components of the enzyme stabilizer were weighed by the above weight, and 22g of water and 60g of methyl parathion hydrolase MPH were weighed for use.

Mixing and stirring the triethanolamine, the water and the preservative uniformly to obtain a first mixed solution; pouring the xanthan gum, the trehalose and the sodium citrate into the first mixed solution by weight, and uniformly stirring to obtain a second mixed solution; the glycerin and the methyl parathion hydrolase MPH with the weight are poured into the second mixed solution to obtain the compound pesticide hydrolase liquid provided by the embodiment.

Example 7

The embodiment of the invention provides an enzyme stabilizer, a fruit and vegetable cleaning solution and a preparation method thereof.

An enzyme stabilizer comprises 15g ethylene glycol, 8g trehalose, 0.05g xanthan gum, 0.2g triethanolamine, 0.4g sodium citrate and 0.03 g paraben complex ester.

A preparation method of a fruit and vegetable cleaning solution comprises the following steps:

the ingredients of the enzyme stabilizer were weighed out in the above amounts, and 25g of water, 6g of phospholipase OPH, 0.1g of EDTA-2Na, 3g of alkyl glycoside and 2g of dodecylhydroxysultaine were weighed out for use.

Mixing and stirring the triethanolamine, the water and the methylisothiazolinone uniformly to obtain a first intermediate solution; adding xanthan gum, trehalose, sodium citrate and EDTA-2Na into the first intermediate solution by weight, and uniformly stirring to obtain a second intermediate solution; pouring the ethylene glycol and the phospholipase into the second intermediate solution to obtain a third intermediate solution; and pouring alkyl glycoside and dodecyl hydroxyl sulfobetaine into the third intermediate solution to obtain the fruit and vegetable cleaning solution provided by the embodiment.

Example 8

An enzyme stabilizer comprises 20g of pentaerythritol, 10g of trehalose, 0.15g of xanthan gum, 0.8g of triethanolamine, 0.8g of sodium citrate and 0.15g of paraben complex ester.

the ingredients of the enzyme stabilizer are weighed according to the weight, and 35g of water, 7g of methyl parathion hydrolase, 0.08g of EDTA-2Na, 6g of fatty alcohol polyoxyethylene ether and 3g of dodecyl dimethyl betaine are weighed for later use.

Mixing and stirring the triethanolamine, the water and the methylisothiazolinone uniformly to obtain a first intermediate solution; pouring the xanthan gum, the trehalose, the sodium citrate and the EDTA-2Na into the first intermediate solution by weight, and uniformly stirring to obtain a second intermediate solution; pouring the pentaerythritol and the methyl parathion hydrolase into the second intermediate liquid to obtain a third intermediate liquid; and pouring alkyl glycoside and dodecyl dimethyl betaine into the third intermediate solution to obtain the fruit and vegetable cleaning solution provided by the embodiment.

Example 9

An enzyme stabilizer comprises 25g glycerol, 12g trehalose, 0.12g xanthan gum, 0.6g triethanolamine, 0.7g sodium citrate and 0.17g paraben complex ester.

the ingredients of the enzyme stabilizer are weighed according to the weight, and 35g of water, 7g of methyl parathion hydrolase, 0.12g of EDTA-2Na, 9g of fatty alcohol polyoxyethylene ether and 4g of dodecyl dimethyl betaine are weighed for later use.

Mixing and stirring the triethanolamine, the water and the methylisothiazolinone uniformly to obtain a first intermediate solution; adding xanthan gum, trehalose, sodium citrate and EDTA-2Na into the first intermediate solution by weight, and uniformly stirring to obtain a second intermediate solution; pouring the pentaerythritol and the methyl parathion hydrolase into the second intermediate liquid to obtain a third intermediate liquid; and pouring alkyl glycoside and dodecyl dimethyl betaine into the third intermediate solution to obtain the fruit and vegetable cleaning solution provided by the embodiment.

Example 10

An enzyme stabilizer comprises 19.35g glycerol, 13.68g trehalose, 0.144g xanthan gum, 0.477g triethanolamine, 0.315g sodium citrate and 0.189g paraben complex ester.

the enzyme stabilizer is prepared by weighing 39.717g of water, 5.4g of methyl parathion hydrolase, 0.072g of EDTA-2Na, 7.2g of fatty alcohol polyoxyethylene ether, 3.6g of amidohydroxysulfopropyl betaine and 0.072g of essence according to the weight for later use.

Mixing and stirring the triethanolamine, the water, the methylisothiazolinone and the essence uniformly to obtain a first intermediate solution; adding xanthan gum, trehalose, sodium citrate and EDTA-2Na into the first intermediate solution by weight, and uniformly stirring to obtain a second intermediate solution; pouring the glycerol and the methyl parathion hydrolase into the second intermediate liquid to obtain a third intermediate liquid; and pouring the alkyl glycoside and the amide hydroxysulfopropyl betaine into the third intermediate solution to obtain the fruit and vegetable cleaning solution provided by the embodiment.

Experimental example 1

The method comprises the following steps of verifying the activity of methyl parathion hydrolase at different temperatures, verifying the activity of the methyl parathion hydrolase at different pH values and verifying the influence of different metal ions and chemical reagents on the activity of the methyl parathion hydrolase.

The enzyme activity was evaluated by comparing the degree of enzymatic reactions carried out at 15 ℃, 20 ℃, 25 ℃, 30 ℃, 45 ℃, 55 ℃, 65 ℃, 75 ℃ and the like, and the optimum temperature was determined, and the results are shown in FIG. 1. Meanwhile, the degree of the enzyme participating in the reaction after heat preservation at 45 ℃ and 50 ℃ for 4 hours is tracked, and the thermal stability of the enzyme is further evaluated. Therefore, the enzyme has the optimum temperature of 45 ℃ and good heat-resistant stability at the temperature of 25-55 ℃.

The enzyme activity was evaluated by comparing the degree of enzymatic reactions carried out at different pH's of 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, determining the optimum pH and at the same time tracking the pH stability of the enzyme, see Table 2. As can be seen, the optimum pH value of the enzyme is 9, and the activity can still be maintained above 80% within 24h between 6 and 11.

Comparative Ca²⁺、Mg²⁺、Na⁺、K⁺And the influence of triethanolamine, borax, urea and trehalose on the enzyme activity is that 0.5mmol/L of metal ions and 5mmol/L of chemical reagents are added according to the weight of a terminal test sample. As a result, it was found that Na⁺The enzyme activity can be activated by glycerol, triethanolamine, trehalose, citrate ions and the like, wherein the trehalose and the glycerol are most obvious; ca²⁺、Mg²⁺Borax and urea are not good for enzyme performance, and others have no influence on enzyme protein.

The enzyme activity was determined according to the following method: method for measuring pesticide hydrolase liquid enzyme activity

Definition of enzyme activity: the enzyme amount required to produce 1. mu. mol of p-nitrophenol per minute in a buffer system at 37 ℃ and pH9.5 is one enzyme activity unit, and is expressed by 1U.

Absorbing 3160 μ L of 0.1M Tris-HCl buffer (pH 9.5) and 40 μ L of methyl parathion mother liquor (10mmol/L), mixing to obtain reaction solution, preheating the reaction solution and enzyme diluent in a water bath kettle at 37 deg.C for 5min and 2min, adding 80 μ L of enzyme diluent into the reaction solution, timing, mixing rapidly, and determining initial absorbance A at 405nm₁Continuously reacting for 2min, and measuring the absorbance value at 405nm as A₂。

The activity of the pesticide degrading enzyme in the sample is calculated according to the following formula:

in the formula: x < - > the activity of the pesticide degrading enzyme in the liquid sample; vt- -total volume of reaction (2050. mu.L);

vs — sample volume (50 μ Ι _); 17.7 millimolar extinction coefficient (cm) of p-nitrophenol at 405nm²/mmol)；

1.0- -optical path (cm); delta A- - - -A₂-A₁(ii) a N- - -dilution factor; t- - -2 min;

1000-p-nitrophenol converted in molar extinction coefficient units at 405nm, 1 mmol-1000. mu. mol.

Experimental example 2

Storing the pure methyl parathion hydrolase, the compound pesticide hydrolase liquid provided in the embodiment 6 and the fruit and vegetable cleaning liquid provided in the embodiment 10 at 40 ℃ for 30 days, measuring the activity of the enzyme and recording the change condition of the activity of the enzyme. As shown in table 1.

TABLE 1 stability comparison of pesticide hydrolase liquid enzyme formulations (accelerated aging test)

According to the table 1, the compound pesticide hydrolase liquid and the fruit and vegetable cleaning liquid provided by the embodiment of the invention have low enzyme activity reduction degree at 40 ℃, have relatively high stability, and meet the requirement that the enzyme activity is at least kept at 50% after 30 days at 40 ℃.

The compound pesticide hydrolase liquid and the fruit and vegetable cleaning liquid provided in the embodiments 1 to 10 are stored for 30 days at 40 ℃, and then the activity of the enzyme is measured and the change condition of the enzyme activity is recorded. As shown in table 2.

Table 2 comparison of stability of the compounded pesticide hydrolase solution and the fruit and vegetable cleaning solution provided in the examples

As can be seen from Table 2, the activity of the compound pesticide hydrolase solution provided by the embodiment of the invention is not greatly reduced after being stored for 30 days at 40 ℃, and the activity is still maintained above 70%. In particular, the activity of the compound pesticide hydrolase liquid provided in embodiments 3 to 6 is maintained to be more than 75% and close to 80%, which indicates that the compound pesticide hydrolase liquid provided by the invention has better stability when the components are in the range of 10 to 20 parts of polyol, 5 to 15 parts of trehalose, 0.05 to 0.2 part of xanthan gum, 0.1 to 1 part of triethanolamine, 0.1 to 1 part of sodium citrate, 10 to 20 parts of water and 30 to 70 parts of pesticide hydrolase liquid.

The activity of the fruit and vegetable cleaning solution provided by the embodiment is not greatly reduced after being stored for 30 days at 40 ℃, and the activity is still kept above 75%. Particularly, the vitality of the fruit and vegetable cleaning liquid provided in embodiments 7 to 10 is maintained at 80% or more, which indicates that the fruit and vegetable cleaning liquid provided by the present invention has better stability when the polyol is 15 to 30 parts, the trehalose is 8 to 15 parts, the xanthan gum is 0.05 to 0.15 part, the triethanolamine is 0.2 to 0.8 part, and the sodium citrate is 0.4 to 0.8 part.

Experimental example 3

The lemon with the organophosphorus pesticide attached to the surface is washed and soaked by the compound pesticide hydrolase liquid provided by the embodiment 6 of the invention. And (3) dripping the pesticide hydrolase liquid compounded before and after the washing and soaking into a pesticide quick test card for color development. The color development is shown in FIG. 3.

As can be seen from FIG. 3, the change of the compound pesticide hydrolase solution before and after washing is large, which indicates that the compound pesticide hydrolase solution can clean a large amount of organophosphorus pesticide on the surface of the lemon.

Experimental example 4

The compound pesticide hydrolase liquid provided by the embodiment 6 is mixed with the pyrethroid 1:1 for reaction, and water is reacted with the pyrethroid 1:1 as a control. After the reaction is completed, the pyrethroid pesticide is subjected to color comparison by using a rapid test kit. As shown in fig. 4 and 5. The left side of the figure 4 is the liquid after the mixed pesticide hydrolase liquid and the pyrethrin 1:1 are mixed and reacted, which are provided by the embodiment 6; on the right of fig. 4 is the liquid after the reaction of water with pyrethrin 1: 1.

As can be seen by comparing fig. 4 with fig. 5, the content of the pyrethroid after the mixed pesticide hydrolase liquid and the pyrethroid are mixed and reacted at a ratio of 1:1 is lower than 0.2 mg/L.

Experimental example 5

The purpose is as follows: verifying whether the compound pesticide hydrolase solution provided by the embodiment 6 of the invention can effectively degrade specific pesticides (such as dichlorvos, chlorpyrifos, omethoate, cypermethrin, deltamethrin and the like)

The method comprises the following steps: GC-MS (gas chromatography-Mass spectrometer) method for agricultural marking institute in Hubei province

1) Dilution of 10mg/L pesticide standard the 5 pesticides were diluted with methanol or acetone (acetone dilution GC group);

2) preparing three gradient pesticide diluent substrates of 0.05mg/L, 0.2mg/L and 0.5mg/L, respectively placing 20mL of distilled water in four 100mLPE tubes, adding 100uL, 400uL and 1000uL of 10mg/L pesticide standard samples one by one, taking 1000uL of addition as a blank control sample, and uniformly mixing for later use;

3) respectively adding 200ul of 48u/mL compound pesticide hydrolase solution into the substrate, covering and uniformly mixing, reacting in water bath at 30 ℃ for 20min, and not adding the compound pesticide hydrolase solution into a blank sample;

4) taking out the sample 3), adding 20mL of acetonitrile and excessive salt, standing for 5min for layering, and taking 1mL of supernatant of each pesticide gradient sample to prepare for GC-MS.

The results of the experiment are reported in table 3.

Table 3 GC-MS method for detecting removal effect of pesticide hydrolase MPH on common pesticides

Note: the exact values in table 3 refer to: under the condition of the same pesticide addition amount as the control group, the pesticide hydrolysis enzyme liquid MPH has higher numerical reliability on the removal rate of the pesticide.

As can be seen from Table 3, the compound pesticide hydrolase solution provided by the invention has obvious degradation effect on dichlorvos, chlorpyrifos, omethoate and the like (organophosphorus pesticides), and the removal rate can reach more than 98.6%; the degradation effect on the pyrethroid pesticide is general, but the removal rate can also reach more than 50 percent.

In conclusion, when the enzyme stabilizer provided by the invention is used in combination with the pesticide hydrolase solution, the pesticide hydrolase solution can be kept as a single entity in the solution; trehalose has a non-specific protective effect on biological enzymes; sufficient polyalcohol also provides a better environment for the stability of the pesticide hydrolase liquid, a proper amount of sodium citrate and triethanolamine can perform chelation with various metals, so that the reduction or inactivation of enzyme activity caused by the action of metal ions on the enzyme is avoided, the protection effect on the enzyme is enhanced by the mutual coordination of the sodium citrate and triethanolamine ammonia, and the enzyme stabilizer can nonspecifically protect the pesticide hydrolase liquid by combining the reasonable mixture ratio of the components, so that the stability of the pesticide hydrolase liquid is good.

The compound pesticide hydrolase liquid provided by the invention comprises the enzyme stabilizer and is prepared by adopting a reasonable method, so that the compound pesticide hydrolase liquid has good stability and no harm to human health.

The preparation method of the compound pesticide hydrolase liquid provided by the invention can prepare the compound pesticide hydrolase liquid which has small stability and is harmless to human bodies.

The fruit and vegetable cleaning solution provided by the invention comprises the pesticide hydrolase solution and the enzyme stabilizer provided by the invention, so that the fruit and vegetable cleaning solution can effectively clean the pesticide on the surfaces of the fruits and vegetables, and has good stability and no harm to human health.

The preparation method of the fruit and vegetable cleaning solution provided by the invention can be used for preparing the pesticide for effectively cleaning the surfaces of the fruits and vegetables, and the fruit and vegetable cleaning solution is good in stability and harmless to human health.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. The preparation method of the fruit and vegetable cleaning solution is characterized by comprising the following steps of: uniformly mixing 10-30 parts of polyol, 1-15 parts of trehalose, 0.01-0.3 part of xanthan gum, 0.1-2 parts of triethanolamine, 0.1-2 parts of sodium citrate, 20-40 parts of water, 3-10 parts of pesticide hydrolase liquid, 0.02-0.2 part of EDTA-2Na and 3-15 parts of surfactant, wherein the pesticide hydrolase liquid is selected from at least one of methyl parathion hydrolase and phospholipase, and the surfactant comprises: 2-10 parts of a nonionic surfactant and 1-5 parts of a betaine surfactant, wherein the nonionic surfactant is selected from at least one of polysorbate 80, alkyl glycoside and fatty alcohol-polyoxyethylene ether.

2. The method for preparing fruit and vegetable cleaning solution according to claim 1, characterized in that,

15-30 parts of polyol, 8-15 parts of trehalose, 0.05-0.15 part of xanthan gum, 0.2-0.8 part of triethanolamine, 0.4-0.8 part of sodium citrate, 20-40 parts of water, 3-10 parts of pesticide hydrolase liquid, 0.02-0.2 part of EDTA-2Na and 3-15 parts of surfactant.

3. The preparation method of the fruit and vegetable cleaning solution as claimed in claim 1, which is characterized by comprising the following steps:

uniformly mixing the triethanolamine and the water to obtain a first intermediate solution;

uniformly mixing the xanthan gum, the trehalose, the sodium citrate, the EDTA-2Na and the first intermediate solution to obtain a second intermediate solution;

uniformly mixing the polyalcohol, the pesticide hydrolase solution and the second intermediate solution to obtain a third intermediate solution;

and uniformly mixing the surfactant and the third intermediate solution.

4. The method for preparing fruit and vegetable cleaning solution according to claim 3, characterized in that,

and mixing the triethanolamine with the water uniformly, and simultaneously mixing 0.01-0.2 part of preservative with the triethanolamine and the water uniformly to obtain the first intermediate solution.

5. The preparation method of the fruit and vegetable cleaning solution as claimed in claim 4, wherein the preservative is 0.03-0.1 part.

6. A fruit and vegetable cleaning liquid, which is characterized by being prepared by the preparation method of the fruit and vegetable cleaning liquid as claimed in any one of claims 1 to 5.