Marine microbial cleaning and energy-saving agent and preparation method thereof

Abstract

The invention belongs to the technical field of fuel additives, and provides a marine microbial cleaning energy-saving agent and a preparation method thereof, and the marine microbial cleaning energy-saving agent comprises the following raw materials by mass: 4-6 parts of marine microbial active mother liquor, 85-105 parts of aviation kerosene, 7-9 parts of methyl tert-butyl ether, 4-6 parts of butylated hydroxytoluene, 15-25 parts of alkylated waste oil, and 2-4 parts of a compound enzyme system. 1-3 parts of a nano carrier material; the marine candida tropicalis bio-enzyme obtained through a high-throughput screening and gene modification technology shows efficient catalytic performance and wide substrate specificity, and the fuel cleaning effect and the combustion efficiency can be remarkably improved; meanwhile, carbon deposits and sediments in the engine can be effectively removed through the synergistic effect of the compound enzyme system, and the purposes of harmlessness and cleaning are achieved; in addition, in combination with reutilization of alkylated waste oil, environmental pollution is reduced, and the concept of sustainable development is met.

Description

Technical Field

[0001] The invention belongs to the technical field of fuel additives, in particular to a marine microbial cleaning and energy-saving agent and a preparation method thereof. Background Art

[0002] In modern automobiles and industrial equipment, internal combustion engines are the main power source, and their fuel efficiency and emission performance have attracted much attention. In order to improve the performance of internal combustion engines, a variety of fuel additives have been launched on the market. These additives are usually chemically synthesized to optimize the reaction efficiency and cleanliness of the fuel;

[0003] However, many existing fuel additives mainly rely on chemical synthesis, which has led to environmental pollution to a certain extent. The harmful byproducts produced during the chemical synthesis process not only have a negative impact on the ecological environment, but also may release toxic substances during use, affecting the health of users. Secondly, existing fuel additives often do not fully consider the complex environment in which the engine actually operates, resulting in limited cleaning and combustion efficiency improvement effects of some products. The catalytic performance of traditional additives is limited to specific substrates, which limits their ability to deal with carbon deposits and sediments inside the engine, and cannot effectively solve the problems of low fuel utilization and high pollutant emissions. In addition, although some additives can improve combustion performance to a certain extent, due to the lack of protection for the entire cycle of the engine, carbon deposits and sediments may still form inside the engine after long-term use, thereby increasing the wear and failure rate of the engine. This not only reduces the working efficiency of the engine, shortens its service life, and increases maintenance costs, but also leads to more frequent replacement of engine oil and other related parts, further increasing the economic burden on consumers.

[0004] To this end, technicians in this field have proposed a marine microbial cleaning and energy-saving agent and a preparation method thereof, aiming to have high efficiency, cleanliness and energy saving, reduce environmental pollution, comply with the concept of sustainable development, and improve fuel quality and engine performance. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a marine microbial cleaning and energy-saving agent and a preparation method thereof to solve the problems raised in the background technology.

[0006] According to the first aspect of the present disclosure, a marine microbial cleaning and energy-saving agent is proposed, which is composed of the following raw materials measured in parts by mass: 4-6 parts of marine microbial active mother liquor, 85-105 parts of aviation kerosene, 7-9 parts of methyl tert-butyl ether, 4-6 parts of di-tert-butyl cresol, 15-25 parts of alkylation waste oil, 2-4 parts of complex enzyme system, and 1-3 parts of nanocarrier material.

[0007] Preferably, the marine microbial active mother solution is composed of the following raw materials in parts by mass: 3 to 7 parts of marine tropical Candida bioenzyme, 12 to 18 parts of aviation kerosene, 2 to 4 parts of ethylene glycol monoethyl ether, 6 parts of tert-butanol, 6 to 9 parts of N,N-dimethylacetamide, and 1 to 4 parts of ferrous acetylacetonate.

[0008] Preferably, the marine tropical Candida albicans bioenzyme comprises the following preparation steps:

[0009] S101, by isolating a variety of marine microbial strains from the marine environment, conducting high-throughput screening, and sequencing the genomes of the screened high-activity strains;

[0010] S102, using CR I SPR gene editing technology to genetically modify the target strain, construct an expression vector, clone the enzyme gene into a selective vector, and transform it into Candida tropicalis;

[0011] S103, adding 6-12 parts of agar, 4-6 parts of sodium alginate, 3-6 parts of glyceryl stearate, 2-4 parts of ethylene glycolamine, and 1-4 parts of ferrous acetylacetonate to the culture medium, and preparing a transformation culture solution at a culture temperature of 30-35° C., a pH value of 5.5-7.0, and a dissolved oxygen saturation of 60%-80%;

[0012] S104, adding trehalose and cyclohexane acid to the culture medium to induce the expression of the target enzyme, and the fermentation culture time can be set to 24-48 hours;

[0013] S105. Collect cells in the culture medium by centrifugation, lyse the cells with phosphate buffer, extract intracellular enzymes, and purify the extracted enzyme solution by ultrafiltration, precipitation or chromatography.

[0014] Preferably, the complex enzyme system is composed of the following raw materials in parts by weight: 5-15 parts of lipase, 5-10 parts of esterase, 7-12 parts of cellulase, 5-15 parts of protease, and 8-10 parts of amylase;

[0015] The preparation step of the composite enzyme system comprises: S201, mixing lipase, esterase, cellulase, protease, amylase with 2-5 parts of antioxidant and 3-6 parts of stabilizer in a ratio of 10:7:10:10:9:4:5, stirring to make the mixture uniform, the stirring time is 30 minutes to 1 hour, and the rotation speed is 300-500 rpm;

[0016] S202, culturing and fermenting at 30-35°C and pH 5.5-7.0, inoculating the mixture into a liquid culture medium, setting the culture time to 24-48 hours, and regularly monitoring the growth of the bacteria and the accumulation of metabolites;

[0017] S203, separating the microbial cells and metabolites by centrifugation at 8000 rpm for 10 minutes, removing the cell residues by filtration and precipitation, and performing chromatography purification.

[0018] Preferably, the antioxidant is BHT, and the stabilizer is polyvinyl alcohol.

[0019] Preferably, the nanocarrier material is modified natural polysaccharide nanoparticles, and the complex enzyme system is loaded onto the nanocarrier.

[0020] According to the second aspect of the present disclosure, a method for preparing a marine microbial cleaning and energy-saving agent is proposed, comprising the following preparation steps:

[0021] S1. First prepare the marine tropical Candida bioenzyme, then mix the marine tropical Candida bioenzyme with aviation kerosene, ethylene glycol monoethyl ether, tert-butyl alcohol, N,N-dimethylacetamide, and ferrous acetylacetonate, and stir under magnetic shear for 24 hours at 37° C. to obtain a marine microbial active mother solution;

[0022] S2, constructing a composite enzyme system, and loading the composite enzyme system onto a nanocarrier of modified natural polysaccharide nanoparticles;

[0023] S3, adding the prepared marine microbial active mother liquid, aviation kerosene, methyl tert-butyl ether and di-tert-butyl cresol into a reaction container, and mixing the raw materials thoroughly and evenly under magnetic shear stirring at 35-40°C;

[0024] S4, slowly adding the alkylation waste oil to the mixed raw materials, while continuing to stir to make it evenly dispersed, and then adding the nano-carrier material loaded with the complex enzyme system;

[0025] S5, stirring and reacting the mixed materials at 35-45°C for 24-48 hours to allow all materials to fully react to form a stable mixture. After the reaction is completed, the mixture is aged;

[0026] S6. Filter the aged mixture to remove impurities and unreacted raw materials. The clear liquid after filtration is the prepared marine microbial cleaning and energy-saving agent.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] 1. The marine tropical Candida bioenzyme obtained by high-throughput screening and genetic modification technology in the present invention has efficient catalytic performance, can significantly enhance the fuel purification effect, effectively remove carbon deposits and sediments inside the engine, and improve the combustion efficiency of the fuel. The composite enzyme system composed of multiple enzymes has a wide range of substrate specificity and can work synergistically to further promote the fuel purification and energy-saving effects. At the same time, the addition of antioxidants and stabilizers improves the stability and service life of the composite enzyme system.

[0029] 2. The present invention not only realizes the reuse of waste resources but also reduces environmental pollution by adding alkylation waste oil, which is in line with the concept of sustainable development. The bio-enzyme preparation technology is adopted to avoid the environmental pollution problems that may be caused by traditional chemical methods, and has higher environmental protection.

[0030] 3. The present invention can improve the combustion performance of fuel, reduce the emission of harmful substances, increase the utilization rate of fuel and the working efficiency of the engine by adding the active mother liquid of marine microorganisms and the complex enzyme system. In addition, the active ingredients in the cleaning and energy-saving agent can remove carbon deposits and sediments inside the engine, reduce the wear and failure rate of the engine, and extend the service life of the engine. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a flow chart of the method for preparing the marine microbial cleaning and energy-saving agent of the present invention;

[0032] Figure 2 This is a flow chart of the method for preparing the marine tropical Candida bioenzyme of the present invention;

[0033] Figure 3 The present invention is a flow chart of the method for preparing the composite enzyme system. DETAILED DESCRIPTION

[0034] The following embodiments of the present invention are described in further detail in conjunction with the accompanying drawings and examples. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0035] As attached Figure 1 To Attachment Figure 3 As shown:

[0036] Embodiment 1: The present invention provides a marine microbial cleaning and energy-saving agent, which is composed of the following raw materials measured by mass: 4-6 parts of marine microbial active mother liquor, 85-105 parts of aviation kerosene, 7-9 parts of methyl tert-butyl ether, 4-6 parts of di-tert-butyl cresol, 15-25 parts of alkylation waste oil, 2-4 parts of composite enzyme system, and 1-3 parts of nano-carrier material. Alkylation waste oil contains certain organic components, which can produce synergistic effects when interacting with other raw materials such as marine microbial active mother liquor and composite enzyme system, further enhancing the cleaning effect of the cleaning and energy-saving agent. It can effectively remove carbon deposits and sediments inside the engine, keep the fuel system clean, thereby improving the combustion efficiency of the fuel and the working efficiency of the engine. The addition of alkylation waste oil can also improve the performance of the fuel. It can react chemically with other components in the fuel to form a more stable and uniform fuel mixture.

[0037] The marine microbial active mother solution is composed of the following raw materials in parts by mass: 3-7 parts of marine tropical Candida bioenzyme, 12-18 parts of aviation kerosene, 2-4 parts of ethylene glycol monoethyl ether, 6 parts of tert-butanol, 6-9 parts of N,N-dimethylacetamide, and 1-4 parts of ferrous acetylacetonate.

[0038] The preparation process of the marine tropical Candida albicans enzyme comprises the following steps:

[0039] S101, by isolating a variety of marine microbial strains from the marine environment, conducting high-throughput screening, and sequencing the genomes of the screened high-activity strains;

[0040] S102, using CR I SPR gene editing technology to genetically modify the target strain, construct an expression vector, clone the enzyme gene into a selective vector, and transform it into Candida tropicalis;

[0041] S103, adding 6-12 parts of agar, 4-6 parts of sodium alginate, 3-6 parts of glyceryl stearate, 2-4 parts of ethylene glycolamine, and 1-4 parts of ferrous acetylacetonate to the culture medium, and preparing a transformation culture solution at a culture temperature of 30-35° C., a pH value of 5.5-7.0, and a dissolved oxygen saturation of 60%-80%;

[0042] S104, adding trehalose and cyclohexane acid to the culture medium to induce the expression of the target enzyme, and the fermentation culture time can be set to 24-48 hours;

[0043] S105. Collect cells in the culture medium by centrifugation, lyse the cells with phosphate buffer, extract intracellular enzymes, and purify the extracted enzyme solution by ultrafiltration, precipitation or chromatography.

[0044] The complex enzyme system is composed of the following raw materials in parts by weight: 5-15 parts of lipase, 5-10 parts of esterase, 7-12 parts of cellulase, 5-15 parts of protease, and 8-10 parts of amylase;

[0045] The steps of preparing the composite enzyme system include: S201, mixing lipase, esterase, cellulase, protease, amylase with 2-5 parts of antioxidant and 3-6 parts of stabilizer in a ratio of 10:7:10:10:9:4:5, stirring to make the mixture uniform, the stirring time is 30 minutes to 1 hour, and the rotation speed is 300-500rpm; the antioxidant is BHT, and the stabilizer is polyvinyl alcohol.

[0046] S202, culturing and fermenting at 30-35°C and pH 5.5-7.0, inoculating the mixture into a liquid culture medium, setting the culture time to 24-48 hours, and regularly monitoring the growth of the bacteria and the accumulation of metabolites;

[0047] S203, separating the microbial cells and metabolites by centrifugation at 8000 rpm for 10 minutes, removing the cell residues by filtration and precipitation, and performing chromatography purification.

[0048] The nanocarrier material adopts modified natural polysaccharide nanoparticles, and the complex enzyme system is loaded onto the nanocarrier, which improves the dispersibility and stability of the enzyme, and is also conducive to the recovery and reuse of the enzyme.

[0049] Embodiment 2: The present invention also provides a method for preparing a marine microbial cleaning and energy-saving agent, which is applied in Embodiment 1 and includes the following preparation steps:

[0050] S1. First prepare the marine tropical Candida bioenzyme, then mix the marine tropical Candida bioenzyme with aviation kerosene, ethylene glycol monoethyl ether, tert-butyl alcohol, N,N-dimethylacetamide, and ferrous acetylacetonate, and stir under magnetic shear for 24 hours at 37° C. to obtain a marine microbial active mother solution;

[0051] S2, constructing a composite enzyme system, and loading the composite enzyme system onto a nanocarrier of modified natural polysaccharide nanoparticles;

[0052] S3, adding the prepared marine microbial active mother liquid, aviation kerosene, methyl tert-butyl ether and di-tert-butyl cresol into a reaction container, and mixing the raw materials thoroughly and evenly under magnetic shear stirring at 35-40°C;

[0053] S4. Slowly add alkylation waste oil to the mixed raw materials, continue stirring to make it evenly dispersed, and then add the nano-carrier material loaded with the complex enzyme system; the nano-carrier material can help the complex enzyme system to be better dispersed in the system and improve its catalytic efficiency; the addition of alkylation waste oil not only realizes the reuse of waste resources, but also reduces environmental pollution, which is in line with the concept of sustainable development.

[0054] S5, stirring and reacting the mixed materials at 35-45°C for 24-48 hours to allow all materials to fully react to form a stable mixture. After the reaction is completed, the mixture is aged to further improve its stability and performance;

[0055] S6. Filter the aged mixture to remove impurities and unreacted raw materials. The clear liquid after filtration is the prepared marine microbial cleaning and energy-saving agent.

[0056] Importantly, it should be noted that the construction and arrangement of the present application shown in a plurality of different exemplary embodiments are only exemplary. Although only a few embodiments are described in detail in this disclosure, it should be readily understood by those who refer to this disclosure that many modifications are possible (e.g., the size, scale, structure, shape and proportion of various elements, as well as parameter values ​​(e.g., temperature, pressure, etc.), mounting arrangements, use of materials, color, directional changes, etc.) without substantially departing from the novel teachings and advantages of the subject matter described in the application. For example, the element shown as integrally formed can be composed of multiple parts or elements, the position of the element can be inverted or otherwise changed, and the nature or number or position of the discrete element can be changed or changed. Therefore, all such modifications are intended to be included in the scope of the present invention. The order or sequence of any process or method steps can be changed or reordered according to alternative embodiments. In the claims, any "device plus function" clause is intended to cover the structure described herein that performs the function, and is not only structurally equivalent but also equivalent structure. Without departing from the scope of the present invention, other replacements, improvements, changes and omissions can be made in the design, operating conditions and arrangement of the exemplary embodiments. Therefore, the invention is not limited to a specific embodiment, but extends to numerous modifications still falling within the scope of the appended claims.

[0057] Additionally, in order to provide a concise description of exemplary embodiments, all features of an actual embodiment (ie, those features that are not relevant to the best mode presently contemplated for carrying out the invention or those that are not relevant to implementing the invention) may not be described.

[0058] It should be understood that in the development of any actual implementation, as in any engineering or design project, numerous implementation-specific decisions may be made. Such a development effort may be complex and time-consuming, but for those of ordinary skill having the benefit of this disclosure, without undue experimentation, the development effort will be a routine task of design, fabrication, and production.

[0059] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A marine microbial cleaning and energy-saving agent, characterized in that: The invention is composed of the following raw materials measured by weight: 4-6 parts of marine microbial active mother liquor, 85-105 parts of aviation kerosene, 7-9 parts of methyl tert-butyl ether, 4-6 parts of di-tert-butyl cresol, 15-25 parts of alkylation waste oil, 2-4 parts of complex enzyme system and 1-3 parts of nano-carrier material.

2. A marine microbial cleaning and energy-saving agent as claimed in claim 1, characterized in that: The marine microorganism active mother solution is composed of the following raw materials in parts by mass: 3-7 parts of marine tropical Candida bioenzyme, 12-18 parts of aviation kerosene, 2-4 parts of ethylene glycol monoethyl ether, 6 parts of tert-butyl alcohol, 6-9 parts of N,N-dimethylacetamide, and 1-4 parts of ferrous acetylacetonate.

3. A marine microbial cleaning and energy-saving agent as claimed in claim 2, characterized in that: The marine tropical Candida albicans bioenzyme comprises the following preparation steps: S101, by isolating a variety of marine microbial strains from the marine environment, conducting high-throughput screening, and sequencing the genomes of the screened high-activity strains; S102, using CRISPR gene editing technology to genetically modify the target strain, construct an expression vector, clone the enzyme gene into a selective vector, and transform it into Candida tropicalis; S103, adding 6-12 parts of agar, 4-6 parts of sodium alginate, 3-6 parts of glyceryl stearate, 2-4 parts of ethylene glycolamine, and 1-4 parts of ferrous acetylacetonate to the culture medium, and preparing a transformation culture solution at a culture temperature of 30-35° C., a pH value of 5.5-7.0, and a dissolved oxygen saturation of 60%-80%; S104, adding trehalose and cyclohexane acid to the culture medium to induce the expression of the target enzyme, and the fermentation culture time can be set to 24-48 hours; S105. Collect cells in the culture medium by centrifugation, lyse the cells with phosphate buffer, extract intracellular enzymes, and purify the extracted enzyme solution by ultrafiltration, precipitation or chromatography.

4. A marine microbial cleaning and energy-saving agent as claimed in claim 1, characterized in that: The complex enzyme system is composed of the following raw materials in parts by weight: 5-15 parts of lipase, 5-10 parts of esterase, 7-12 parts of cellulase, 5-15 parts of protease, and 8-10 parts of amylase; The preparation step of the composite enzyme system comprises: S201, mixing lipase, esterase, cellulase, protease, amylase with 2-5 parts of antioxidant and 3-6 parts of stabilizer in a ratio of 10:7:10:10:9:4:5, stirring to make the mixture uniform, the stirring time is 30 minutes to 1 hour, and the rotation speed is 300-500 rpm; S202, culturing and fermenting at 30-35°C and pH 5.5-7.0, inoculating the mixture into a liquid culture medium, setting the culture time to 24-48 hours, and regularly monitoring the growth of the bacteria and the accumulation of metabolites; S203, separating the microbial cells and metabolites by centrifugation at 8000 rpm for 10 minutes, removing the cell residues by filtration and precipitation, and performing chromatography purification.

5. A marine microbial cleaning and energy-saving agent as claimed in claim 4, characterized in that: The antioxidant is BHT, and the stabilizer is polyvinyl alcohol.

6. A marine microbial cleaning and energy-saving agent as claimed in claim 1, characterized in that: The nano-carrier material adopts modified natural polysaccharide nano-particles, and the complex enzyme system is loaded on the nano-carrier.

7. A method for preparing a marine microbial cleaning and energy-saving agent, applied to a marine microbial cleaning and energy-saving agent as claimed in any one of claims 1 to 6, characterized in that: The method comprises the following preparation steps: S1. First prepare the marine tropical Candida bioenzyme, then mix the marine tropical Candida bioenzyme with aviation kerosene, ethylene glycol monoethyl ether, tert-butyl alcohol, N,N-dimethylacetamide, and ferrous acetylacetonate, and stir under magnetic shear for 24 hours at 37° C. to obtain a marine microbial active mother solution; S2, constructing a composite enzyme system, and loading the composite enzyme system onto a nanocarrier of modified natural polysaccharide nanoparticles; S3, adding the prepared marine microbial active mother liquid, aviation kerosene, methyl tert-butyl ether and di-tert-butyl cresol into a reaction container, and mixing the raw materials thoroughly and evenly under magnetic shear stirring at 35-40°C; S4, slowly adding the alkylation waste oil to the mixed raw materials, while continuing to stir to make it evenly dispersed, and then adding the nano-carrier material loaded with the complex enzyme system; S5, stirring and reacting the mixed materials at 35-45°C for 24-48 hours to allow all materials to fully react to form a stable mixture. After the reaction is completed, the mixture is aged; S6. Filter the aged mixture to remove impurities and unreacted raw materials. The clear liquid after filtration is the prepared marine microbial cleaning and energy-saving agent.

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