CN115975747A

CN115975747A - Enzyme cleaning solution and use method thereof

Info

Publication number: CN115975747A
Application number: CN202211384842.2A
Authority: CN
Inventors: 裴娟
Original assignee: Hangzhou West Lake Sanitation And Disinfection Medical Device Co ltd
Current assignee: Hangzhou West Lake Sanitation And Disinfection Medical Device Co ltd
Priority date: 2022-11-07
Filing date: 2022-11-07
Publication date: 2023-04-18

Abstract

The application discloses an enzyme cleaning solution and a using method thereof, the enzyme cleaning solution is prepared from solution A and solution B with the volume ratio of 1:1, wherein the solution A comprises the following components: 1 part of modified lysozyme; 19-29 parts of a protective agent; 10-20 parts of protease; water; the concentration of the solution A is 30-50 g/L; the solution B adopts 20-30 g/L hydrogen peroxide solution; the modified lysozyme is obtained by performing Schiff base reaction modification on lysozyme and salicylaldehyde. The enzyme cleaning liquid prepared by the application has a strong cleaning effect and a lasting antibacterial effect, and can meet the use requirement of in-situ cleaning of the bed unit.

Description

Enzyme cleaning solution and use method thereof

Technical Field

The application relates to the field of disinfection solution, in particular to an enzyme cleaning solution and a using method thereof.

Background

The bed unit refers to basic articles (a sickbed, a mattress, a quilt, a mattress, a pillow, a quilt cover, a bed sheet, a pillow cover) and the like contained in a sickbed in a hospital. The bed unit is part of the patient's medical environment, the objects of the bed unit mostly being in high frequency contact with the patient. In the using process, the body fluid, blood, excrement and other pollutants of a patient are often polluted, and the pollutants often carry a large amount of pathogens, so that the pollutants are not timely cleaned and easily cause the infection of personnel or the secondary infection of the patient. Therefore, cleaning and disinfecting the bed unit is always the focus of medical hygiene.

However, at present, disinfectant solutions such as hypochlorous acid disinfectant solution, lysozyme cleaning solution and the like only have a killing effect on a small part of germs, and can not well clean stains brought by patients, so that the cleaning effect of the bed unit is affected.

Disclosure of Invention

In order to improve the problem of bed unit cleaning effect, the application provides an enzyme cleaning solution and a preparation method thereof.

An enzyme cleaning liquid is prepared from a liquid A and a liquid B with the volume ratio of 1:1,

the solution A comprises:

1 part of modified lysozyme;

19-29 parts of a protective agent;

10-20 parts of protease;

water;

the concentration of the solution A is 30-50 g/L;

the solution B adopts 20-30 g/L hydrogen peroxide solution;

the modified lysozyme is obtained by performing Schiff base reaction modification on lysozyme and salicylaldehyde. Through adopting above-mentioned technical scheme, the different A liquid of configuration component and B liquid reach better disinfection effect of disinfecting, and wherein the lysozyme that contains in the A liquid is an antibacterial enzyme that antibiotic effect is excellent, and the modified lysozyme that obtains through the lysozyme of salicylaldehyde modification can overcome the defect that the lysozyme can only kill gram-positive fungus, has improved the antibacterial activity of lysozyme simultaneously, has further improved the cleaning performance to medical bed unit. Meanwhile, the liquid A also comprises a protective agent, so that the service life of the lysozyme can be prolonged, and the bacteriostatic activity can be improved; the modified lysozyme and the protease in the solution A form a multienzyme system, so that the sterilization and disinfection effects are improved. The solution B adopts hydrogen peroxide which is a weak acid oxidation type sterilization disinfectant with a certain sterilization and disinfection effect, can provide a weak acid working environment for lysozyme, and when the hydrogen peroxide and the lysozyme are used together, the lysozyme is converted from a monomer to a dimer or a trimer, the catalytic activity of polymer lysozyme is stronger than that of lysozyme monomer, and the polymer lysozyme has strong bactericidal property optimization to gram-negative bacteria, and the modified lysozyme is prepared from lysozyme, salicylaldehyde and a reducing agent in a weight ratio of 1 (0.5-10) to 0.3-0.6.

By adopting the technical scheme, under the action of a reducing agent, lysine residues on lysozyme can be combined with salicylaldehyde to form Schiff base, and the Schiff base has good bactericidal and antibacterial effects; in the reaction process, the spatial conformation of the lysozyme activity area is changed, the hydrophobic group of the lysozyme activity center is exposed, the exposure of the hydrophobic group is beneficial to the combination of the lysozyme and the bacterial cell wall, the killing property to the bacteria is stronger, and the catalytic activity of the lysozyme is improved; salicylaldehyde has unpleasant tar smell, is insoluble in water and has certain biotoxicity, so the salicylaldehyde has good bacteriostatic effect, but is not generally used as a sterilizing disinfectant for medical units, and the modified lysozyme formed by reacting the salicylaldehyde with the lysozyme has no unpleasant smell and can be dissolved in water, so that the bacteriostatic effect of the salicylaldehyde can be exerted. Compared with lysozyme or salicylaldehyde, the modified lysozyme has a wider antibacterial spectrum and stronger bactericidal activity, and the modified lysozyme is difficult to pass through the horny layer of a human body and cannot be absorbed by the human body through skin, so that the human health cannot be influenced.

Typically, but not by way of limitation, sodium borohydride is selected as the reducing agent.

Preferably, the modified lysozyme comprises lysozyme, salicylaldehyde and a reducing agent in a weight ratio of 1 (2-8) to (0.3-0.6).

By adopting the technical scheme, too much salicylaldehyde can cause too much lysine residues on lysozyme to react to block the active center of the lysozyme, so that the activity of the lysozyme is reduced, and too little salicylaldehyde can not obtain more Schiff base, so that the compatibility sterilization effect between the Schiff base and the lysozyme can be weakened, and the weight ratio of the modified lysozyme for preparing the raw materials is preferably selected.

Preferably, the preparation method of the modified lysozyme comprises the following steps:

A1. dissolving salicylaldehyde in tetrahydrofuran to obtain a first solution; dissolving lysozyme in a buffer solution to obtain a solution II;

A2. adding the solution I into the solution II, stirring for 30-40 min at room temperature, then cooling to 0-4 ℃, adding a reducing agent every 30min, mixing and stirring, and repeating for three times; then adding ammonium sulfate to 50-70% saturation, standing until the precipitate is not generated, and filtering to obtain the modified lysozyme.

By adopting the technical scheme, the solution I and the solution II are uniformly dispersed by stirring at room temperature, and then the reducing agent sodium borohydride is added in stages at low temperature, so that the reaction is continuously and stably carried out. Adding saturated ammonium sulfate to salt out to obtain modified lysozyme.

Preferably, the modified lysozyme obtained in the step A2 is subjected to high-pressure treatment for 30-40 min.

By adopting high-pressure treatment, the space structure of the modified lysozyme active center is changed, the hydrophobic group of the active center is exposed, the active group is easier to contact and combine with a substrate, and the enzyme activity of the lysozyme is improved.

Preferably, the protective agent comprises EDTA, D-potassium erythorbate and chitosan according to the weight ratio of 10 (2-3) to (1-3).

Through adopting above-mentioned technical scheme, lysozyme can destroy the cell wall of harmful bacterial strain for EDTA and chitosan are more easy to permeate to the cytoplasmic membrane on, EDTA and chitosan homoenergetic chelate with the lipopolysaccharide on cell membrane surface, make cell membrane become to be more destroyed, have improved the bactericidal effect of lysozyme, and under this effect, EDTA, chitosan or lysozyme can kill the bacterial strain that single component can't be killed, have strengthened the effect of disinfecting and have widened the antibacterial spectrum. Lysozyme can degrade chitosan into chitosan oligosaccharide with smaller molecular weight, and the chitosan oligosaccharide can enter the interior of cells to interfere the synthesis of DNA and RNA, thereby destroying the reproduction and survival of strains and improving the antibacterial effect and antibacterial spectrum. The potassium ions carried by the D-potassium ascorbate have a micro-activation effect on lysozyme, and can enhance the antibacterial activity of the lysozyme. The D-potassium ascorbate has stronger inoxidizability and can be used as a stabilizer of EDTA and lysozyme, and the lysozyme has better bacteriostatic activity and longer bacteriostatic duration through the synergy of the D-potassium ascorbate, the EDTA and the lysozyme.

Preferably, the protease is papain.

Through the technical scheme, the papain can rapidly decompose protein or fat pollutants, and can form a multi-enzyme system with the modified lysozyme, so that a better sterilization and disinfection effect is achieved.

In a second aspect, a method of using an enzymatic rinse, comprising the steps of:

s1, subpackaging, namely taking the solution A and the solution B to be respectively stored in different independent cavities in a sprinkling can device through the sprinkling can device for enzyme cleaning solution;

s2, spraying: simultaneously spraying the solution A and the solution B on the bed unit to be cleaned through a spray pot device.

By adopting the technical scheme, as the hydrogen peroxide has certain oxidizability, the sterilizing effect of the hydrogen peroxide can be influenced when the hydrogen peroxide is stored together with the liquid A, and the protective effect of the protective agent in the liquid A is damaged. The liquid A and the liquid B are subpackaged, so that substances in the liquid A and the liquid B are separately stored and can be mixed in actual use, the effect reduction of enzyme cleaning liquid caused by the common storage of the liquid A and the liquid B can be reduced, and the liquid A and the liquid B can be mixed to play a better disinfection function in use.

Preferably, the watering can device comprises a can body and a nozzle, a partition board is fixedly connected in the can body and used for dividing an inner cavity of the can body into a first cavity and a second cavity which are independent, a first conduit for communicating the nozzle and the can body is arranged in the first cavity, and a second conduit for communicating the nozzle and the can body is arranged in the second cavity.

By adopting the technical scheme, the sprinkling can device capable of respectively storing the liquid A and the liquid B is provided, so that medical personnel can conveniently carry the separately-packaged liquid A and liquid B, and can simultaneously spray the liquid A and the liquid B with equal quantity, so that the cleaning effect of the enzyme cleaning liquid is better; the watering can adopts a detachable structure, so that the liquid A and the liquid B can be conveniently supplemented, and the watering can device can be conveniently maintained. The efficiency of the cleaning work is improved.

In summary, the present application has the following beneficial effects:

1. the application overcomes the defects that salicylaldehyde is used as a bactericide, is insoluble in water and has tar pungent smell and overcomes the defect that the lysozyme has a low antibacterial spectrum by modifying the lysozyme through the salicylaldehyde.

2. The protection liquid is prepared to protect the lysozyme from being inactivated too fast to lose the sterilization effect, and the EDTA, the chitosan, the D-potassium erythorbate and the lysozyme in the protection liquid have compatibility in the sterilization aspect, so that the sterilization effect of the disinfectant is improved.

3. Store A liquid and B liquid respectively through watering can device, then mix the blowout during use, can reduce because A liquid and B liquid mixing time overlength to the effect of A liquid or B liquid that leads to descends, can make A liquid and B liquid mix when using together again, play the effect of disinfecting in coordination.

Drawings

Fig. 1 is a schematic structural view of a watering can apparatus.

Fig. 2 is a structural sectional view of the watering can device.

Description of reference numerals:

100. a kettle body; 110. a partition plate; 101. a first cavity; 102. a second cavity; 210. a first conduit; 220. a second conduit; 300. and (4) a spray head.

Detailed Description

The present application will be described in further detail with reference to examples.

The raw materials used in the examples and preparation examples can be specified by commercial methods, and the hydrogen peroxide solution in the present application is obtained by diluting hydrogen peroxide solution commercially available.

Preparation of starting materials/intermediates

Preparation of the protective agent

Preparation examples 0 to 1, a protecting agent, comprising the following steps

10g of EDTA, 2.5g of D-potassium erythorbate and 2g of chitosan are taken to obtain the protective agent.

Preparation examples 0 to 2, a protecting agent, comprising the following steps

10g of EDTA, 3g of D-potassium erythorbate and 1g of chitosan are taken to obtain the protective agent.

Preparation examples 0 to 3, a protecting agent, comprising the following steps

10g of EDTA, 2g of D-potassium erythorbate and 3g of chitosan are taken to obtain the protective agent.

Preparation examples 0 to 4, a protectant, were different from preparation examples 0 to 1 in that EDTA was replaced with the same amount of chitosan.

Preparation examples 0 to 5, a protectant, were different from preparation examples 0 to 1 in that potassium D-erythorbate was replaced with an equal amount of chitosan.

Preparation examples 0 to 6, a protectant, were different from preparation examples 0 to 1 in that chitosan was replaced with the same amount of EDTA.

Preparation of modified lysozyme

The phosphate buffer in preparation example had a pH of 7.4 and a concentration of 0.001mol/L.

Preparation example 1-1, a modified lysozyme, comprising the following preparation steps:

A1. dissolving 5g of salicylaldehyde in 10ml of tetrahydrofuran to obtain a first solution; dissolving 1g of lysozyme in 1L of phosphate buffer solution to obtain a solution II;

A2. adding the solution I in the step A1 into the solution II in the step A1, stirring at room temperature for 40min, then cooling to 0 ℃, adding 0.15g of sodium borohydride once every 30min, stirring and dissolving, and repeating for three times; then adding ammonium sulfate to 60% saturation, standing until no precipitate is generated, filtering, and treating under 100MPa for 35min to obtain modified lysozyme.

Preparation examples 1-2, a modified lysozyme, comprising the following preparation steps:

A1. dissolving 8g of salicylaldehyde in 10ml of tetrahydrofuran to obtain a first solution; dissolving 1g of lysozyme in 1L of phosphate buffer solution to obtain a solution II;

A2. adding the solution I in the step A1 into the solution II in the step A1, stirring at room temperature for 35min, then cooling to 4 ℃, adding 0.2g of sodium borohydride once every 30min, stirring and dissolving, and repeating for three times; then adding ammonium sulfate to 50% saturation, standing until the precipitate is almost not generated, filtering, and treating under 100MPa for 40min to obtain modified lysozyme.

Preparation examples 1 to 3, a modified lysozyme, comprising the following preparation steps:

A1. dissolving 2g of salicylaldehyde in 10ml of tetrahydrofuran to obtain a first solution; dissolving 1g of lysozyme in 1L of phosphate buffer solution to obtain a solution II;

A2. adding the solution I in the step A1 into the solution II in the step A1, stirring at room temperature for 40min, then cooling to 2 ℃, adding 0.1g of sodium borohydride once every 30min, stirring and dissolving, and repeating for three times; then adding ammonium sulfate to 70% saturation degree, standing until the precipitate is hardly generated, filtering, and treating under 100MPa for 30min to obtain the modified lysozyme.

Preparation examples 1-4, a modified lysozyme, were different from the preparation example 1-1 in that salicylaldehyde was used in an amount of 10g.

Preparation examples 1-5, a modified lysozyme, were different from preparation examples 1-1 in that salicylaldehyde was used in an amount of 0.5g preparation examples 1-6, and a modified lysozyme was different from preparation example 1-1 in that sodium borohydride was added directly at room temperature (23. + -. 2 ℃ C.).

Preparation examples 1-7, a modified lysozyme, were different from preparation examples 1-1 in that 0.45g of sodium borohydride was added in one portion.

Preparation examples 1 to 8, a modified lysozyme, were different from preparation examples 1 to 1 in that no high pressure treatment was conducted.

Preparation of the liquids

Preparation example 2-1, a solution A, was prepared by the following steps:

and (2) blending 1g of modified lysozyme, 24g of protective agent and 15g of papain, adding a small amount of deionized water for dilution and dissolution, adjusting the pH to 6.0 by using 1mol/L hydrochloric acid solution, then adding deionized water for dilution to 1L, and preserving heat at 35 ℃ for storage to obtain solution A.

Modified lysozyme was prepared in preparation example 1-1, and a protective solution was prepared in preparation example 0-1.

Preparation example 2-2, a solution A, comprising the following preparation steps:

and (2) blending 1g of modified lysozyme, 29g of protective agent and 20g of papain, adding a small amount of deionized water for dilution and dissolution, adjusting the pH to 6.2 by using 1mol/L hydrochloric acid solution, adding deionized water for dilution to 1L, and preserving heat at 37 ℃ for storage to obtain solution A.

Modified lysozyme was prepared in accordance with preparation examples 1-2, and a protective solution was prepared in accordance with preparation examples 0-2.

Preparation examples 2 to 3, a solution A, comprising the following preparation steps:

and (2) blending 1g of modified lysozyme, 19g of protective agent and 10g of papain, adding a small amount of deionized water for dilution and dissolution, adjusting the pH to 6.3 by using 1mol/L hydrochloric acid solution, then adding deionized water for dilution to 1L, and preserving heat at 30 ℃ for storage to obtain solution A.

Modified lysozyme was prepared according to preparation examples 1 to 3, and the protective solution was prepared according to preparation examples 0 to 3.

Preparation examples 2-4, a solution A, were different from preparation example 2-1 in that the protective solution was prepared using 0-4.

Preparation examples 2-5, a solution A, were different from preparation example 2-1 in that the protective solution was prepared using 0-5.

Preparation examples 2-6, a solution A, were different from preparation example 2-1 in that the protective solution was prepared using 0-6.

Preparation examples 2-7, a solution A, were different from preparation example 2-1 in that modified lysozyme was prepared using 1-4.

Preparation examples 2-8, a solution A, were different from preparation example 2-1 in that modified lysozyme was prepared using 1-5.

Preparation examples 2 to 9, a solution A, were different from preparation example 2 to 1 in that modified lysozyme was prepared using 1 to 6.

Preparation examples 2 to 10, a solution A, were different from preparation example 2 to 1 in that modified lysozyme was prepared using 1 to 7.

Preparation examples 2 to 11, A liquid A, were different from preparation example 2 to 1 in that modified lysozyme was prepared using 1 to 8.

Preparation examples 2-12, a solution A, were different from preparation example 2-1 in that papain was replaced with the same amount of subtilisin or pepsin.

Examples

Example 1, a method of using an enzyme wash, was used as follows:

s1, subpackaging, namely respectively storing 1L A liquid and 1L B liquid in different independent cavities in a spraying pot device through the spraying pot device for enzyme cleaning liquid;

s2, spraying: the spraying pot device sprays the same amount of the liquid A and the liquid B on the bed unit to be cleaned.

S3, cleaning: cleaning of the bed unit is performed.

The solution A was taken from preparation example 2-1,B as an aqueous solution of hydrogen peroxide having a concentration of 25 g/L.

In addition, it should be noted that the specific structure of the watering can device is as follows:

referring to fig. 1 and 2, the watering can assembly includes a can body 100, a spray head 300, a first conduit 210 and a second conduit 220.

Referring to fig. 1 and 2, the kettle body 100 includes a kettle body, a kettle lid, a partition board 110 and a baffle, the kettle body is in threaded connection with the kettle lid, an air inlet is formed in the side wall of the kettle body, the partition board 110 is vertically fixedly connected in the kettle body and separates the cavity inside the kettle body into a first cavity 101 and a second cavity 102, the baffle is located inside the kettle body and horizontally supports against the inner wall of the kettle body through a rubber ring, and the baffle is provided with a baffle hole.

Referring to fig. 2, a spray head 300 is threadedly coupled to an end of the lid remote from the ground.

Referring to fig. 2, one end of the first guide pipe 210 is communicated with the spray head 300, the other end of the first guide pipe 210 is communicated with the first cavity 101, the first guide pipe 210 passes through the baffle hole and is located in the first cavity 101, one end of the first guide pipe 210 far away from the spray head 300 is fixedly connected with an anti-dropping piece, and the diameter of the anti-dropping piece is greater than that of the baffle hole; one end of second conduit 220 is communicated with spray head 300, the other end of second conduit 220 is communicated with second cavity 102, second conduit 220 passes through the baffle hole and is located in the second cavity, and one end of second conduit 220 far away from spray head 300 is fixedly connected with an anti-drop piece. When liquid needs to be filled, the first guide pipe 210 and the second guide pipe 220 are pulled, so that the anti-falling part moves towards the baffle plate, the baffle plate is driven to be separated from the kettle body, and the enzyme cleaning liquid can be poured into the first cavity 101 and the second cavity 102 respectively. When disinfectant is required to be sprayed, the opening of the kettle body is blocked by the mounting baffle plate to prevent liquid leakage.

The implementation principle of the watering can device is as follows: and operating the spray head to spray the liquid in the first cavity and the liquid in the second cavity to the position to be cleaned in the same amount from the spray head.

Example 2, a method of using an enzyme wash, according to the following:

s1, subpackaging, namely storing 1L A liquid and 1L B liquid in different independent cavities in a spraying pot device respectively through the spraying pot device for enzyme cleaning liquid;

s2, spraying: spraying the same amount of solution A and solution B onto the bed unit to be cleaned.

S3, cleaning: cleaning of the bed unit is performed.

The solution A was taken from preparation example 2-2,B as a 20g/L aqueous hydrogen peroxide solution.

The watering can device in this embodiment is the watering can device of embodiment 1.

Example 3, a method of using an enzyme wash, according to the following:

s1, subpackaging, namely respectively storing 500mL of solution A and 500mL of solution B in different independent cavities in a spraying pot device for enzyme cleaning solution;

s2, spraying: spraying the same amount of solution A and solution B on the bed unit to be cleaned through a spray pot device.

S3, cleaning: cleaning of the bed unit is performed.

The solution A was taken from preparation example 2-3,B as a 30g/L aqueous hydrogen peroxide solution.

Example 4, a method of using an enzyme wash, differs from example 1 in that solution A was taken from preparation examples 2-4.

Example 5, a method of using an enzyme wash, differs from example 2 in that solution A was taken from preparation examples 2-5.

Example 6, a method of using an enzyme wash, differs from example 3 in that solution A was taken from preparation examples 2-6.

Example 7, a method of using an enzyme wash, differs from example 3 in that solution A was taken from preparation examples 2-7.

Example 8, a method of using an enzyme wash, differs from example 1 in that solution A was taken from preparation examples 2-8.

Example 9, a method of using an enzyme wash, differs from example 1 in that solution A was taken from preparation examples 2-9.

Example 10, a method of using an enzyme wash, differs from example 1 in that solution A was taken from preparation examples 2-10.

Example 11, an enzyme wash, was used in a manner different from that of example 1 in that solution A was taken from preparation examples 2-11.

Example 12, a method of using an enzyme wash, differs from example 1 in that solution A was taken from preparation examples 2-12.

Example 13, a use method of enzyme cleaning solution, with the difference of example 1, does not use the watering can device, but mix 1L A liquid and 1L B liquid, spray on the bed unit to be cleaned within 10-15 min, carry on the cleaning of the bed unit.

Comparative example

Comparative example 1, a method of using an enzyme cleaning solution, differs from example 1 in that solution A is replaced with an equal amount of solution B.

Comparative example 2, a method of using an enzyme cleaning solution, differs from example 1 in that solution B is replaced with an equal amount of solution A.

Comparative example 3, a method of using an enzyme wash, differs from example 1 in that the modified lysozyme was replaced with an equal amount of unmodified lysozyme.

Comparative example 4, a method of using an enzyme wash, comprising the steps of:

the method comprises the following steps: and mixing the solution A and the solution B and storing for 2 hours to obtain an enzyme cleaning solution.

Step two: and (4) spraying the enzyme cleaning solution in the step one on the bed unit to be cleaned by using a spray can device.

Step three: the bed unit is cleaned.

The watering can apparatus used was the watering can apparatus of example 1.

Comparative example 5, a method of using an enzyme wash, differs from example 1 in the preparation of an enzyme wash comprising the steps of: the enzyme cleaning solution comprises 1.5% of polyethylene glycol, 5% of sodium sulfite, 3% of lysozyme, 80.5% of deoxidized water and 10% of glutaraldehyde; the enzyme cleaning solution can be obtained by weighing the components according to the formula and then uniformly mixing the components.

The enzyme activity of the lysozyme is 50000U/mmg.

Performance test

Test subjects: the bed units after treatment of examples 1-13 and comparative examples 1-5, for a total of 18 experimental groups.

Test 1: artificially simulating a pollutant test: the method refers to the appendix F of the industrial standard of the medical cleaning agent,

test 2: and (3) bacteriostatic test:

refer to GB15979-2002 appendix C4 of sanitary Standard for Disposable sanitary articles and Disinfection technical Specification (2002 edition, edited by Ministry of health of the people's republic of China).

Test strains: escherichia coli, candida albicans, staphylococcus aureus and Pseudomonas aeruginosa provided by China center for strain preservation are selected to prepare bacterial suspension for later use.

Culture medium: common nutrient agar culture medium, sabouraud's agar culture medium

The average inhibitory rates (%) of the effects on Escherichia coli, candida albicans, staphylococcus aureus and helicobacter pylori over different periods of time (minutes) measured according to the above method are shown in Table 2 below

Table 1: test 1 test results

	Presence or absence of residual material visible to the naked eye	Rate of removal of contaminants
			Example 1	Is composed of	＞994％
Example 2	Is free of	＞99.4％
			Example 3	Is free of	＞99.2％
Example 4	Is free of	＞99.2％
			Example 5	Is free of	＞99.1％
Example 6	Is free of	＞99.0％
			Example 7	Is composed of	＞97.6％
Example 8	Is free of	＞98.8％
			Example 9	Is composed of	＞98.4％
Example 10	Is free of	＞985％
			Example 11	Is free of	＞99.0％
Example 12	Is composed of	＞96.6％
			Example 13	Is free of	＞99.2％
Comparative example 1	Partial residue is left	＞70.6％
			Comparative example 2	Partial residue is left	＞615％
Comparative example 3	Is composed of	＞96.6％
			Comparative example 4	Is free of	＞98.9％
Comparative example 5	Is composed of	＞917

Table 2: test 2 test results

It can be seen from the combination of examples 1-6 and table 2 that when EDTA, D-potassium erythorbate and chitosan are present in the protective solution at the same time, the prepared enzyme cleaning solution has better bacteriostatic activity and longer time effect, because of the synergistic effect among EDTA, D-potassium erythorbate and chitosan.

Combining example 1, example 7 and example 8 and table 2, it can be seen that when the weight ratio of lysozyme to salicylaldehyde is 1:2-8, the prepared enzyme cleaning solution has better antibacterial activity and wider antibacterial spectrum, because when the amount of salicylaldehyde is too large, the activity area of lysozyme is blocked due to too much combination of lysozyme and salicylaldehyde, so that the activity of lysozyme is reduced too much, and the antibacterial activity is reduced, and when the amount of salicylaldehyde is too small, the formed schiff base is reduced, so that the antibacterial effect of the finally prepared enzyme cleaning solution is reduced.

It can be seen from the combination of example 1, example 9 and example 10 and the combination of table 2 that when the reducing agent is added at room temperature or added all at once, the bacteriostatic activity of the prepared enzyme cleaning solution is reduced because when sodium borohydride is added at room temperature or added all at once, the reaction of lysozyme and salicylaldehyde is too fast, resulting in that part of lysozyme does not participate in the reaction, and part of lysozyme binds too much salicylaldehyde, resulting in the reduction of the whole bacteriostatic activity.

Combining with the embodiment 1, the embodiment 11 and the table 2, it can be seen that the modified lysozyme is not subjected to high pressure treatment, and the bacteriostatic activity of the prepared enzyme cleaning solution is reduced, because the spatial structure of the active center of the modified lysozyme is changed by the high pressure treatment, the hydrophobic group of the active center is exposed, the active group is more easily contacted and combined with the substrate, and the enzyme activity of the lysozyme is improved.

In combination with example 1, example 12 and table 2, it can be seen that, when the solution B uses subtilisin or pepsin which can only decompose protein, but not papain, the removal rate of the pollutants is obviously reduced and the residual substances are visible to the naked eye because papain has the effect of decomposing fat and protein, and subtilisin or pepsin can only decompose protein pollutants; the bacteriostatic effect of the enzyme disinfectant is reduced because bacteria carried on the unremoved contaminants are difficult to be eliminated by lysozyme.

When the enzyme cleaning solutions prepared by using only the solutions A and B are used in combination with example 1, comparative example 1 and comparative example 2, and table 2, the effects are not as good as those of the enzyme cleaning solutions prepared by using only the solutions A and B. The reason is that the solution A and the solution B have compatible effect.

Combining example 1, comparative example 3 and table 2, it can be seen that the enzyme cleaning solution prepared by replacing modified lysozyme with the same amount of unmodified lysozyme has no antibacterial activity or antibacterial spectrum as the enzyme cleaning solution prepared by modified lysozyme. The reason is that the modified lysozyme obtained by modifying the lysozyme through the salicylaldehyde can overcome the defect that the lysozyme can only resist gram-positive bacteria, simultaneously improves the bacteriostatic activity of the lysozyme, and further improves the cleaning effect on a bed unit.

In combination with example 1, comparative example 4 and example 13 and in combination with tables 1 and 2, it can be seen that if the enzyme cleaning solution prepared by mixing solution A and solution B has reduced bacteriostatic activity, because too long mixing time of solution A and solution B will result in reduced effect of the components in solution A and solution B, thereby resulting in reduced bacteriostatic activity of the enzyme cleaning solution and reduced time for maintaining the disinfecting effect. But because the washing of the inside bed unit of hospital needs to consume longer time, prefabricate enzyme washing liquid in advance and can lead to actual disinfection effect to descend, and because the enzyme washing liquid is that liquid is inconvenient to carry and transport, consequently choose for use watering can device both can reduce the effect of the enzyme antiseptic solution that prefabricates in advance and descend, also can improve the efficiency of bed unit disinfection.

By combining the example 1, the comparative example 5 and the tables 1 and 2, the enzyme cleaning solution prepared by the method has higher bacteriostatic activity and antibacterial spectrum than the conventional enzyme cleaning solution, and has better pollutant removal treatment effect.

The specific embodiments are only for explaining the present application and are not limiting to the present application, and those skilled in the art can make modifications to the embodiments without inventive contribution as required after reading the present specification, but all the embodiments are protected by patent law within the scope of the claims of the present application.

Claims

1. An enzyme cleaning solution, which is characterized in that the enzyme cleaning solution is prepared from solution A and solution B with the volume ratio of 1:1,

the liquid A comprises:

1 part of modified lysozyme;

19-29 parts of a protective agent;

10-20 parts of protease;

water;

the concentration of the solution A is 30-50 g/L;

the solution B adopts 20-30 g/L hydrogen peroxide solution;

the modified lysozyme is obtained by performing Schiff base reaction modification on lysozyme and salicylaldehyde.

2. The enzyme cleaning solution according to claim 1, wherein: the modified lysozyme is prepared from lysozyme (0.5-10 wt.%) and salicylaldehyde (0.3-0.6 wt.%) and reducer.

3. An enzyme cleaning solution according to claim 2, wherein: the modified lysozyme comprises lysozyme, salicylaldehyde and a reducing agent in a weight ratio of 1 (2-8) to 0.3-0.6.

4. The enzyme cleaning solution according to any one of claims 1 to 3, wherein the preparation method of the modified lysozyme comprises the following steps:

5. The enzyme cleaning solution according to claim 4, wherein: and D, carrying out high-pressure treatment on the modified lysozyme obtained in the step A2 for 30-40 min.

6. The enzymatic cleaning solution of claim 1, wherein: the protective agent comprises EDTA, D-potassium erythorbate and chitosan in a weight ratio of 10 (2-3) to 1-3.

7. The enzymatic cleaning solution of claim 1, wherein: the protease is papain.

8. The method of using an enzymatic cleaning solution according to any of claims 1 to 7, comprising the steps of:

s1, subpackaging, namely respectively storing the solution A and the solution B in mutually independent cavities in a watering can device through the watering can device for enzyme cleaning solution;

s2, spraying: spraying equal amounts of the solution A and the solution B on the bed unit to be cleaned through a spray pot device.

9. The method of using an enzymatic cleaning solution of claim 8, wherein: the watering can device comprises a can body (100) and a spray head (300), a partition plate (110) is fixedly connected in the can body (100), the partition plate (110) is used for dividing an inner cavity of the can body (100) into a first cavity (101) and a second cavity (102) which are mutually independent, a first guide pipe (210) used for communicating the spray head (300) with the can body (100) is arranged in the first cavity (101), and a second guide pipe (220) used for communicating the spray head (300) with the can body (100) is arranged in the second cavity (102).