A method for preparing and applying a glutaraldehyde-crosslinked organosilicon cell coating

Abstract

The present invention discloses a method for preparing a glutaraldehyde cross-linked organosilicon cell coating. The method uses glutaraldehyde (GA) as a cross-linking agent, which is added to a bacterial suspension having an organosilicon coating. The organosilicon coating is an organosilicon layer containing silicon oxide particles. The bacteria having the organosilicon coating are cross-linked by stable chemical bonds. After the cross-linking reaction, a GA-cross-linked organosilicon cell coating is obtained, and finally a bacteria that is superior to the organosilicon coating in protecting the bacterial structure from being destroyed and improving the stability of the bacteria itself and intracellular proteins is obtained. During the preparation process, only a small amount of GA is added to the organosilicon cell coating, and the cross-linking degree of the GA cross-linked organosilicon coating can be regulated by changing the concentration of GA, the reaction time and the reaction temperature during the preparation process, thereby obtaining cells with different stabilities. The raw materials prepared by the present invention are cheap and easy to obtain, the preparation process is simple and easy, and it is suitable for large-scale production.

Description

Technical Field

[0001] The invention relates to the preparation and application of a cell coating, and in particular to a preparation method and application of a glutaraldehyde cross-linked organic silicon cell coating. Background Art

[0002] As an emerging field, synthetic biology introduces the concept of engineering into life sciences, designs biological systems and organisms to adjust existing biological systems or construct biological systems that do not exist in nature. The term synthetic biology appeared in the early 20th century, and synthetic biology is still in the early stages of rapid development. Cell modification is a very important link in the study of synthetic biology. In nature, all cell behaviors are directly or indirectly controlled or regulated by the interface between the cell surface and the external environment. Coatings on the cell surface can provide cells with some functions that cannot be achieved in the natural state, enhance the resistance of cells under adverse conditions, control cell activity and metabolism, and expand the application of cells in cell catalysis, biosensing, cell therapy, sustainable energy, etc.

[0003] The method for preparing cell coating based on organosilicon (CN 202110455041X) is to form a layer of organosilicon on the surface of bacteria by the reaction of 3-aminopropyltriethoxysilane (APTES) with the active groups on the surface of bacteria and its own condensation. The organosilicon coating can effectively protect the structure of the bacteria from being destroyed and improve the stability of the bacteria and intracellular proteins. Through optimization, it was found that the addition of tetraethyl orthosilicate (TEOS) before the addition of 3-aminopropyltriethoxysilane (APTES) combined with the modification of 3-aminopropyltriethoxysilane (APTES) on the silica particles generated by the hydrolysis of tetraethyl orthosilicate (TEOS) increased the silica particles in the organosilicon coating on the surface of the bacteria, making the ability of the organosilicon coating to maintain the stability of cells and their intracellular proteins more significant. However, experimental phenomena show that although silicone coatings can improve the stability of bacteria and intracellular proteins, the mechanical strength of silicone coatings is poor. After repeated use, the coatings tend to fall off and degrade, which deteriorates the stability of bacteria and intracellular proteins and limits their application in large-scale cell coatings. Summary of the invention

[0004] Compared with the organosilicon cell coating, the purpose of the present invention is to provide a method for preparing a glutaraldehyde cross-linked organosilicon cell coating. The method for preparing the glutaraldehyde cross-linked organosilicon cell coating provided by the present invention can significantly improve the stability of cells and intracellular proteins after adding a small amount of cheap glutaraldehyde. The raw materials for the preparation of the present invention are cheap and easy to obtain, the preparation process is simple and easy, and it is suitable for industrial production.

[0005] In order to solve the above technical problems, the present invention proposes a method for preparing a glutaraldehyde cross-linked silicone cell coating, which uses glutaraldehyde (GA) as a cross-linking agent and adds it to a bacterial suspension having an silicone coating, wherein the silicone coating is an silicone layer containing silicon oxide particles, and a glutaraldehyde cross-linked silicone cell coating is obtained after a cross-linking reaction.

[0006] Furthermore, in the preparation method of the glutaraldehyde cross-linked silicone cell coating described in the present invention, wherein the bacterial suspension is a gram-positive bacterial suspension or a gram-negative bacterial suspension, the application field and actual use of Bacillus subtilis and Escherichia coli in industrial production are higher than those of other bacterial species. The present invention preferably uses Bacillus subtilis as the representative species of gram-positive bacteria, and preferably uses Escherichia coli as the representative species of gram-negative bacteria.

[0007] The specific steps of the preparation method of the glutaraldehyde cross-linked silicone cell coating are:

[0008] Step 1: Add glutaraldehyde to the bacterial suspension with silicone coating, the final concentration of glutaraldehyde is 0.05%-0.3%; place the bacterial suspension with silicone coating containing glutaraldehyde on a shaker at a temperature of 4°C-37°C and shake for 20min-180min to obtain a mixed solution of glutaraldehyde cross-linked silicone cell coating.

[0009] Step 2: centrifuge the mixed solution of the glutaraldehyde cross-linked silicone cell coating, remove the supernatant, and wash with deionized water to finally obtain the glutaraldehyde cross-linked silicone cell coating on the surface of the bacteria.

[0010] The advantages of the preparation method of the glutaraldehyde cross-linked silicone cell coating proposed in the present invention are: the preparation raw materials are cheap and easy to obtain, the preparation conditions are mild, the preparation process is simple and easy, only a small amount of glutaraldehyde (GA) is added to the silicone cell coating, and the cross-linking degree of the glutaraldehyde cross-linked silicone coating can be adjusted by changing the concentration of glutaraldehyde (GA), reaction time and reaction temperature during the preparation process, thereby obtaining cells with different stabilities.

[0011] The stability of the cell@organic silicon@glutaraldehyde prepared by the present invention is significantly improved. Compared with the cells with organosilicon coatings that have not been cross-linked with glutaraldehyde and bacteria in the prior art, the glutaraldehyde cross-linked organosilicon cell coating prepared by the preparation method of the present invention is more effective in maintaining the activity of cells and their intracellular proteins in an adverse external environment (such as high temperature). Under the condition of 50-75°C, the half-life is significantly extended compared with the organosilicon cell coating that has not been cross-linked with glutaraldehyde and bacteria. The half-life at 50°C is extended by 95.82-256.24 times, at 60°C by 105.74-180.48 times, and at 75°C by 15.97-46.04 times. In the cell catalysis process, the glutaraldehyde cross-linked organosilicon cell coating improves the stability of the cells and the reusability of the cells more significantly than the organosilicon cell coating. The activity of the enzyme of Bacillus subtilis after 55 cycles is 41.18% of the initial activity, and the activity of the enzyme of Escherichia coli after 45 cycles is 40.82% of the initial activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is the residual activity diagram after the comparative example 1 and the embodiment 1 are recycled.

[0013] Figure 2 This is the residual activity diagram after the comparative example 2 and the embodiment 7 are recycled. DETAILED DESCRIPTION

[0014] The present invention proposes a method for preparing a glutaraldehyde cross-linked silicone cell coating. The design idea is: adding glutaraldehyde (GA), a cross-linking agent with high chemical activity, into cells with silicone coating to cross-link the silicone, thereby obtaining a coating with a denser coating structure and higher mechanical stability, so that the GA cross-linked silicone coating has a more significant ability to maintain the stability of cells and their intracellular proteins.

[0015] The basic scheme of the preparation method of the organosilicon cell coating of the present invention is: first, GA is added to the bacterial suspension with the organosilicon cell coating and shaken for reaction; then, the obtained bacterial suspension is centrifuged, the supernatant is removed, and the suspension is washed by centrifugation with deionized water to obtain cell@organosilicon@glutaraldehyde.

[0016] The technical solution of the present invention is further described in detail below in conjunction with the accompanying drawings, comparative examples and specific embodiments. The described specific embodiments are only used to explain the present invention and are not intended to limit the present invention.

[0017] Comparative Example 1

[0018] Preparation of Bacillus subtilis with catalytic activity and surface coated with silicone (enzyme@Bacillus subtilis@silicon)

[0019] Step 1: Prepare a Bacillus subtilis suspension with an organosilicon coating (Bacillus subtilis@organosilicon):

[0020] To a concentration of 100 OD 600 671.7 μl of tetraethyl orthosilicate (TEOS) was added to the suspension of Bacillus subtilis to make the final concentration of TEOS 150 mM. -1 Incubate in a 30°C water bath shaker for 20 min;

[0021] Then, 164.7 μl of 3-aminopropyltriethoxysilane (APTES) was added to the bacterial suspension to make the final concentration of 3-aminopropyltriethoxysilane (APTES) 35.15 mM. The Bacillus subtilis suspension containing 3-aminopropyltriethoxysilane (APTES) was placed at 120 r min. -1 The mixture was reacted in a 30°C water bath shaker for 160 min. The mixture was centrifuged to remove the supernatant and washed with deionized water to finally obtain Bacillus subtilis@organic silicon.

[0022] Step 2: Place the Bacillus subtilis@organic silicon obtained in step 1 at 120 r min -1 The reaction was carried out in a water bath shaker at 70°C for 30 minutes to obtain enzyme@Bacillus subtilis@organic silicon with catalytic activity.

[0023] The catalytically active enzyme @ Bacillus subtilis @ organosilicon obtained in Comparative Example 1 was incubated at 50°C, 60°C, and 75°C, respectively, and the incubated enzyme @ Bacillus subtilis @ organosilicon was taken out, and substrate starch was added to carry out catalytic reaction, and the content of tagatose in the product was detected by high performance liquid chromatography. Calculate the half-life of the enzyme, where E t is the residual activity of the enzyme after incubation at a specific temperature, E0 is the initial activity of the enzyme before incubation at a specific temperature, and K d is the thermal inactivation constant at a specific temperature, and t is the incubation time. The activity of the enzyme is calculated based on the activity of the intracellular protein in catalyzing the production of tagatose from the substrate starch. The half-life of the intracellular enzyme is the incubation time when the intracellular enzyme activity remains 50%.

[0024] After calculation, the half-life of the enzyme @ Bacillus subtilis @ silicone coating prepared in Comparative Example 1 at 50°C is 225.42 min; the half-life at 60°C is 149.34 min; the half-life at 75°C is 37.32 min; Figure 1 As shown, the activity of the enzyme after 20 cycles was 51.39% of the initial activity.

[0025] Example 1

[0026] Preparation of Bacillus subtilis with catalytic activity and surface coated with glutaraldehyde cross-linked silicone coating (enzyme @ Bacillus subtilis @ silicone @ glutaraldehyde)

[0027] Step 1: preparing a Bacillus subtilis suspension having an organosilicon coating and containing glutaraldehyde (GA) (Bacillus subtilis@organosilicon@glutaraldehyde), comprising:

[0028] Step 1-1) Add 100 OD 600 671.7 μl of tetraethyl orthosilicate (TEOS) was added to the suspension of Bacillus subtilis to make the final concentration of TEOS 150 mM. -1 Incubate in a 30°C water bath shaker for 20 min;

[0029] Step 1-2) 164.7 μl of 3-aminopropyltriethoxysilane (APTES) was added to the bacterial suspension to make the final concentration of 3-aminopropyltriethoxysilane (APTES) 35.15 mM. The Bacillus subtilis suspension containing 3-aminopropyltriethoxysilane (APTES) was placed at 120 r min. -1 The mixture was reacted in a water bath shaker at 30°C for 160 min to obtain a bacterial suspension of Bacillus subtilis@organic silicon;

[0030] Step 1-3) Add 100 μl of glutaraldehyde (GA) to the bacterial suspension obtained in step 1-2) to make the final concentration of GA 0.25%. Place the Bacillus subtilis suspension containing GA at 120 r min. -1 The mixture was reacted in a water bath shaker at 37°C for 60 min. The mixture was centrifuged to remove the supernatant and washed with deionized water to finally obtain Bacillus subtilis@organic silicon@glutaraldehyde.

[0031] Step 2: Place the above-mentioned Bacillus subtilis@organic silicon@glutaraldehyde at 120 r min -1 The reaction was carried out in a water bath shaker at 70°C for 30 minutes to obtain enzyme@Bacillus subtilis@organic silicon@glutaraldehyde with catalytic activity.

[0032] The catalytically active enzyme @ Bacillus subtilis @ organosilicon @ glutaraldehyde obtained in Example 1 was incubated at 50°C, 60°C, and 75°C, the incubated enzyme @ Bacillus subtilis @ organosilicon was taken out, substrate starch was added for catalytic reaction, and the content of the product tagatose was detected by high performance liquid chromatography. Calculate the half-life of the enzyme, where E t is the residual activity of the enzyme after incubation at a specific temperature, E0 is the initial activity of the enzyme before incubation at a specific temperature, and K dis the thermal inactivation constant at a specific temperature, and t is the incubation time. The activity of the enzyme is calculated based on the activity of the intracellular protein in catalyzing the production of tagatose from the substrate starch. The half-life of the intracellular enzyme is the incubation time when the intracellular enzyme activity remains 50%.

[0033] After calculation, the half-life of the enzyme @ Bacillus subtilis @ silicone @ glutaraldehyde coating prepared in Example 1 at 50°C is 57761.62 min, which is 256.24 times longer than that in Comparative Example 1; the half-life at 75°C is 1718.96 min, which is 46.04 times longer than that in Comparative Example 1; Figure 1 As shown, the activity of the enzyme after 55 cycles was 41.18% of the initial activity.

[0034] Example 2

[0035] The steps of Example 2 are basically the same as those of Example 1, except that in the process operation of steps 1-3), after adding GA, the final concentration of GA in the solution is changed from 0.25% to 0.3%, and the Bacillus subtilis suspension containing GA is placed at 120 rmin. -1 The reaction was continued in a 37°C water bath shaker for 60 min. The suspension of Bacillus subtilis containing GA was placed in a 120 r min -1 Incubate the mixture in a 4°C water bath shaker for 180 min.

[0036] After calculation, the half-life of the enzyme@Bacillus subtilis@organic silicon@glutaraldehyde coating prepared in Example 2 at 60°C is 26952.88 min, which is 180.48 times longer than that in Comparative Example 1; the half-life at 75°C is 1702.54 min, which is 45.62 times longer than that in Comparative Example 1.

[0037] Example 3

[0038] The steps of Example 3 are basically the same as those of Example 1, except that in the process operation of steps 1-3), after adding GA, the final concentration of GA in the solution is changed from 0.25% to 0.05%, and the Bacillus subtilis suspension containing GA is placed at 120 rmin. -1 The reaction was continued in a 37°C water bath shaker for 60 min. The suspension of Bacillus subtilis containing GA was placed in a 120 r min -1 Incubate in a 20°C water bath shaker for 120 min.

[0039] After calculation, the half-life of the enzyme@Bacillus subtilis@organic silicon@glutaraldehyde coating prepared in Example 3 at 75°C is 28269.92 min, which is 125.41 times longer than that in Comparative Example 1; the half-life at 75°C is 785.96 min, which is 21.06 times longer than that in Comparative Example 1.

[0040] Example 4

[0041] The steps of Example 4 are basically the same as those of Example 1, except that in the process operation of steps 1-3), after adding GA, the final concentration of GA in the solution is changed from 0.25% to 0.1%, and the Bacillus subtilis suspension containing GA is placed at 120 rmin. -1 The reaction was continued in a 37°C water bath shaker for 60 min. The suspension of Bacillus subtilis containing GA was placed in a 120 r min -1 Incubate in a 37°C water bath shaker for 20 min.

[0042] After calculation, the half-life of the enzyme@Bacillus subtilis@organic silicon@glutaraldehyde coating prepared in Example 4 at 60°C is 19597.89 min, which is 131.23 times longer than that in Comparative Example 1; the half-life at 75°C is 1209.91 min, which is 32.42 times longer than that in Comparative Example 1.

[0043] Example 5

[0044] The steps of Example 5 are basically the same as those of Example 1, except that in the process operation of step 1-3), after adding GA, the final concentration of GA in the solution is changed from 0.25% to 0.15%, and the Bacillus subtilis suspension containing GA is placed at 120 rmin -1 The reaction was continued in a 37°C water bath shaker for 60 min. The suspension of Bacillus subtilis containing GA was placed in a 120 r min -1 Incubate in a 30°C water bath shaker for 60 min.

[0045] After calculation, the half-life of the enzyme@Bacillus subtilis@organic silicon@glutaraldehyde coating prepared in Example 5 at 50°C is 49829.09 min, which is 221.05 times longer than that in Comparative Example 1; the half-life at 75°C is 1362.55 min, which is 36.51 times longer than that in Comparative Example 1.

[0046] Table 1 shows the thermal stability of enzyme@Bacillus subtilis@organic silicon@glutaraldehyde coatings prepared under different conditions.

[0047] Table 1

[0048]

[0049] Comparative Example 2: Preparation of Escherichia coli with catalytic activity and coated with organic silicon coating (enzyme @ Escherichia coli @ organic silicon). The preparation method is basically the same as the steps in Comparative Example 1, except that the concentration of 100 OD 600 The concentration of Bacillus subtilis suspension was changed to 100 OD 600 of E. coli, and finally obtained enzyme@E. coli@organic silicon.

[0050] The catalytically active enzyme @ E. coli @ organosilicon obtained in Comparative Example 2 was incubated at 50°C, 60°C, and 75°C, respectively, and the incubated enzyme @ E. coli @ organosilicon was taken out, and substrate starch was added for catalytic reaction, and the content of the product tagatose was detected by high performance liquid chromatography. Calculate the half-life of the enzyme, where E t is the residual activity of the enzyme after incubation at a specific temperature, E0 is the initial activity of the enzyme before incubation at a specific temperature, and K d is the thermal inactivation constant at a specific temperature, and t is the incubation time. The activity of the enzyme is calculated based on the activity of the intracellular protein in catalyzing the production of tagatose from the substrate starch. The half-life of the intracellular enzyme is the incubation time when the intracellular enzyme activity remains 50%.

[0051] After calculation, the half-life of the enzyme @ Escherichia coli @ silicone coating prepared in this comparative example 2 at 50°C is 110.98min; the half-life at 60°C is 71.25min; and the half-life at 75°C is 17.64min. Figure 2 As shown, the activity of the enzyme after 12 cycles was 50.72% of the initial activity.

[0052] Example 6

[0053] Preparation of Escherichia coli with catalytic activity and coated with glutaraldehyde cross-linked silicone coating (enzyme @ Escherichia coli @ silicone @ glutaraldehyde coating), Example 6 is basically the same as Example 1, except that: the concentration in step 1-1) is 100 OD 600 The concentration of Bacillus subtilis suspension was changed to 100 OD 600 coli, and finally produced enzyme@Escherichia coli@organic silicon@glutaraldehyde coating.

[0054] After calculation, the half-life of the enzyme@Escherichia coli@organic silicon@glutaraldehyde coating prepared in Example 6 at 50°C is 22309.20 min, which is 201.02 times longer than that in Comparative Example 2; the half-life at 75°C is 593.94 min, which is 33.67 times longer than that in Comparative Example 2.

[0055] Example 7

[0056] The steps of Example 7 are basically the same as those of Example 6, except that in the process operation of steps 1-3), after adding GA, the final concentration of GA in the solution was changed from 0.25% to 0.3%, and the Escherichia coli suspension containing GA was placed at 120 r min - 1 The reaction was changed from 60 min in a 37°C water bath shaker to a 120 r min shaker. -1 Incubate the mixture in a shaking water bath at 18°C ​​for 100 min.

[0057] After calculation, the half-life of the enzyme @ Escherichia coli @ silicone @ glutaraldehyde coating prepared in Example 7 at 60°C is 10106.10 min, which is 141.84 times longer than that in Comparative Example 2; the half-life at 75°C is 621.10 min, which is 35.21 times longer than that in Comparative Example 2. Figure 2 As shown, the activity of the enzyme after 45 cycles was 40.82% of the initial activity.

[0058] Example 8

[0059] The steps of Example 8 are basically the same as those of Example 6, except that in the process operation of steps 1-3), after adding GA, the final concentration of GA in the solution is changed from 0.25% to 0.05%, and the Escherichia coli suspension containing GA is placed at 120 r min -1 The reaction was changed from 60 min in a 37°C water bath shaker to a 120 r min shaker. -1 Incubate in a 37°C water bath shaker for 20 min.

[0060] After calculation, the half-life of the enzyme@Escherichia coli@organic silicon@glutaraldehyde coating prepared in Example 8 at 50°C is 10634.10 min, which is 95.82 times longer than that in Comparative Example 2; the half-life at 75°C is 281.71 min, which is 15.97 times longer than that in Comparative Example 2.

[0061] Example 9

[0062] The steps of Example 9 are basically the same as those of Example 6, except that in the process operation of steps 1-3), after adding GA, the final concentration of GA in the solution was changed from 0.25% to 0.1%, and the Escherichia coli suspension containing GA was placed at 120 r min - 1 The reaction was changed from 60 min in a 37°C water bath shaker to a 120 r min shaker. -1 Incubate the mixture in a 4°C water bath shaker for 180 min.

[0063] After calculation, the half-life of the enzyme@Escherichia coli@organic silicon@glutaraldehyde coating prepared in Example 9 at 50°C is 7533.98 min, which is 105.74 times longer than that in Comparative Example 2; the half-life at 75°C is 465.17 min, which is 26.37 times longer than that in Comparative Example 2.

[0064] Table 2 shows the thermal stability of enzyme@Escherichia coli@organic silicon@glutaraldehyde coatings prepared under different conditions.

[0065] Table 2

[0066]

[0067] In summary, the preparation method described in the present invention utilizes cells coated with an organosilicon coating, and by adding glutaraldehyde (GA), a cross-linking agent with high chemical activity and containing symmetrical aldehyde groups, the bacteria with the organosilicon coating are cross-linked using stable chemical bonds, thereby obtaining bacteria that are superior to the organosilicon coating in protecting the bacterial structure from destruction and improving the stability of the bacteria themselves and intracellular proteins. The present invention only adds a small amount of GA to the organosilicon cell coating to make the organosilicon coating more capable of maintaining the stability of cells and their intracellular proteins. The present invention can regulate the degree of cross-linking of the glutaraldehyde-cross-linked organosilicon coating by changing the concentration of GA, the reaction time and the reaction temperature during the preparation process, thereby ultimately improving the stability of intracellular proteins.

[0068] Although the glutaraldehyde cross-linked silicone cell coating of the present invention and the ideas and methods for its application in improving the stability of intracellular proteins are described above in conjunction with the accompanying drawings, the present invention is not limited to the above-mentioned specific embodiments. The above-mentioned specific embodiments are merely illustrative and not restrictive. Under the guidance of the present invention, ordinary technicians in this field can make many variations without departing from the purpose of the present invention, which are all within the protection of the present invention.

Claims

1. A method for preparing a glutaraldehyde cross-linked organosilicon cell coating, characterized in that: Glutaraldehyde is used as a crosslinking agent and added to a bacterial suspension having an organosilicon coating, wherein the organosilicon coating is an organosilicon layer containing silicon oxide particles, to obtain a glutaraldehyde crosslinked organosilicon cell coating; the preparation steps of the glutaraldehyde crosslinked organosilicon cell coating are as follows: Step 1, adding glutaraldehyde to the bacterial suspension with the organosilicon coating, the final concentration of glutaraldehyde being 0.05%-0.25%; placing the bacterial suspension with the organosilicon coating containing glutaraldehyde on a shaker at a temperature of 37° C. for a shaking time of 20 min-60 min to obtain a mixed solution of glutaraldehyde cross-linked organosilicon cell coating; Step 2: centrifuge the mixed solution of the glutaraldehyde cross-linked silicone cell coating, remove the supernatant, and wash with deionized water to finally obtain the glutaraldehyde cross-linked silicone cell coating on the surface of the bacteria.

2. The method for preparing the glutaraldehyde cross-linked silicone cell coating according to claim 1, characterized in that: The bacterial suspension is a gram-positive bacterial suspension or a gram-negative bacterial suspension.

3. The method for preparing the glutaraldehyde cross-linked organosilicon cell coating according to claim 2, characterized in that: The Gram-positive bacterial suspension is Bacillus subtilis.

4. The method for preparing the glutaraldehyde cross-linked organosilicon cell coating according to claim 2, characterized in that: The Gram-negative bacterial suspension is Escherichia coli.

5. The glutaraldehyde cross-linked organosilicon cell coating obtained according to the preparation method according to any one of claims 1 to 4.

6. The glutaraldehyde cross-linked organosilicon cell coating according to claim 5, characterized in that: The glutaraldehyde cross-linked silicone cell coating, under the condition of 50-75°C, has a half-life extended by 95.82-256.24 times at 50°C, by 105.74-180.48 times at 60°C, and by 15.97-46.04 times at 75°C, compared with the silicone cell coating that is not cross-linked with bacteria by glutaraldehyde. The enzyme activity of the glutaraldehyde cross-linked silicone cell coating is 41.18% of the initial activity after 55 cycles of use, and is 40.82% of the initial activity after 45 cycles of use.

7. An application of glutaraldehyde cross-linked organosilicon cell coating, characterized in that: The glutaraldehyde cross-linked silicone cell coating according to claim 5 or 6 is used to maintain the activity of cells and their intracellular proteins under adverse external environments; during the cell catalysis process, the stability of the cells is improved, thereby improving the reusability of the cells.

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