CN114134065B

CN114134065B - Biological cell membrane system and preparation method and application thereof

Info

Publication number: CN114134065B
Application number: CN202111164850.1A
Authority: CN
Inventors: 颜贵卉; 魏建良; 章鹏坤; 姚雨辰; 王旻子; 张明洲
Original assignee: Hangzhou Youmada Biological Co ltd
Current assignee: Hangzhou Youmada Biological Co ltd
Priority date: 2021-09-30
Filing date: 2021-09-30
Publication date: 2024-04-26
Anticipated expiration: 2041-09-30
Also published as: CN114134065A

Abstract

The invention discloses a biological cell membrane system and a preparation method and application thereof. The biological cell membrane system comprises a phospholipid bilayer, a polyamine complex and a polyisoprene compound. The polyamine complex includes a polyamine complex in a free or polymerized state. The polyamine complex is selected from any one or more of linear or branched alkylamine having two amino groups at both ends, linear or branched alkylamine having at least two amino groups at both ends and containing 1 to 4 secondary amines, linear or branched alkylamine having at least one amino group at one end and an imidazolyl group at the other end, and cyclic polyamine. The polyisoprene compounds are selected from any one or more of tetraterpene compounds and tetraterpene compound fatty acid ester derivatives. The biological cell membrane system provided by the invention can effectively reduce the content of glycolipid, and can obtain the polyamine complex, the polyisoprene compound and other beneficial components with higher content, thereby improving the biocompatibility and having wider application range.

Description

Biological cell membrane system and preparation method and application thereof

Technical Field

The invention relates to the technical field of biology and high polymer materials, in particular to a biological cell membrane system and a preparation method and application thereof.

Background

The biological membrane or biological membrane is a generic term for all membrane structures bordering cells, organelles and their environment, and serves to divide and isolate cells and organelles, and is also an important part in connection with intracellular communication. For early years, artificial membranes composed of single or several lipids have been often used for ease of research. At present, although artificial membranes have been widely used in practice, artificial membranes have the following disadvantages: is easy to oxidize, leak and has poor storability in vitro; the disadvantages of being easily degraded by some enzyme substances in vivo and being not capable of reaching target tissues due to phagocytosis by macrophages, and the like limit the application of the recombinant DNA as a carrier. In addition, the artificial membrane is implanted into human body as polymer material to avoid rejection and limit use.

Although the natural biological membrane can effectively avoid the defects of the artificial membrane, the natural biological membrane has complex components and high separation and purification difficulty, certain unfavorable components such as glycolipid and other substances can reduce the biocompatibility, and certain favorable components such as carotenoid, biological polyamine, SOD and the like are favorable for bioremediation. The positively charged polyamine structure can be mutually adsorbed with the surface of a negatively charged cell membrane to form a couple, so that the active ingredients wrapped by the biological membrane can be more effectively introduced into cells through endocytosis to play a role. However, the prior art is difficult to simultaneously reduce or even improve the content of the beneficial components on the premise of ensuring the lower content of the adverse components.

Disclosure of Invention

The invention aims to provide a biological cell membrane system which can effectively reduce the content of glycolipid and can obtain beneficial components such as polyamine complex, polyisoprene compound and the like with higher content, thereby improving the biocompatibility and having wider application range.

The technical scheme adopted by the invention is as follows: the invention provides a biological cell membrane system, which comprises a phospholipid bilayer, a polyamine complex grafted on the phospholipid bilayer and a polyisoprene compound inlaid on the phospholipid bilayer. Wherein the polyamine complex comprises any one or more of: linear or branched alkylamines having two amino groups at both ends, linear or branched alkylamines having at least two amino groups at both ends and comprising 1 to 4 secondary amines at both ends, linear or branched alkylamines having at least one amino group at one end and an imidazolyl group at the other end, cyclic polyamines. The polyisoprene compound is selected from any one or a combination of a plurality of tetraterpene compounds and tetraterpene compound fatty acid ester derivatives.

Among these, tetraterpene compounds include, but are not limited to carotenoids, mainly consisting of zeaxanthin, lutein fatty acid esters, β -cryptoxanthin eicosanoids.

The phospholipid bilayer has a structure similar to that of a cell membrane of skin, and can play a role in conditioning the skin. The polyamine complex has various functions of promoting radiation resistance, oxidation resistance, promoting cell growth and the like. The polyisoprene compound has various functions of radiation resistance, oxidation resistance and the like. In addition, the polyamine complex in the biological cell membrane system is positively charged, so that the affinity of cell membranes related to negative charge is improved, and the polyamine complex is more favorable for being used as a liposome for wrapping and can be used for cell transportation.

The phospholipid bilayer of the present invention includes, but is not limited to: bilayer phospholipid molecular structure, embedded membrane proteins, channels and membrane-associated organelles.

Long-chain polyamine-like proteins are inserted into the phospholipid bilayer, but the polyamine complex is mainly grafted onto the phosphatidic acid layer because the polyamine complex can be bound to the carboxyl groups of fatty acids or proteins, and also to the phosphate bonds of phospholipids. Carotenoids (polyisoprene) are predominantly inserted in mosaic form.

Optionally, the biological membrane system further comprises glycolipid, and the content of the glycolipid is less than or equal to 100ppm. The content of glycolipid is low.

Optionally, the biological membrane system further comprises a biologically active oxidoreductase having a metal ion in its active center; the metal ions are from any one or more of Mn, ca, mg, zn, cu, fe. Wherein Mn, ca, mg, zn, cu, fe ions are selected from common valence ions of the following metal elements: mn, ca, mg, zn, cu, fe. The oxidoreductase is selected from any one or more of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and Succinic Dehydrogenase (SDH). Specifically, the oxidoreductase is superoxide dismutase, and the active center of the oxidoreductase is provided with manganese ions. SOD has oxidation resistance. Wherein, the Mn-SOD has excellent heat stability.

Alternatively, the primary form of the polyamine complex is an acylated polymeric polyamine, which may specifically be one or more of histamine, agmatine, pentylamine, spermidine, putrescine, spermidine, norspermine, homospermine, spermine. The polyamine complex comprises a free state or a polymerized state. Wherein, the norspermine is N-aminopropyl norspermine, and the spermidine is N-aminopropyl-spermidine. The cyclic polyamine is cytochrome c containing porphyrin structure. Wherein the cytochromes are selected from bacterial pigments, and can be specifically mycolipin.

The invention also provides a preparation method of the biological cell membrane system, which comprises the following steps:

1) Performing high-metabolism culture of the strain in a sugar-free culture medium by any one or more of a temperature gradient method, an illumination intensity adjustment method and an osmotic pressure adjustment method;

2) Collecting thalli, re-suspending thalli by adopting a homogenate buffer solution, centrifuging, taking precipitated thalli for crushing, and obtaining a cell lysate;

3) Adding the mixture into a resuspension medium to obtain a resuspension, and purifying the biological cell membrane system in the layered resuspension by an ultracentrifugation method and a secondary density gradient centrifugation method to obtain the required biological cell membrane system.

The cell lysate was added to various concentrations of the resuspension medium, and then ultracentrifuged at 150,000-300,000 Xg, 60-90min, at 1-6deg.C, to collect the liquid C in the desired area. Diluting the liquid C with PBS buffer solution, ultracentrifugating at 1-6deg.C for 30-90min at 100,000-200,000 ×g, discarding supernatant, and collecting precipitate to obtain the desired biological membrane system.

When the resuspension medium is sucrose, sucrose solutions having an amount concentration of the substance of 0.3 to 0.9mol/L, 1 to 3mol/L and 0.01 to 0.3mol/L, respectively, may be selected. Wherein, because the densities of the sucrose solutions with different concentrations are different, the sucrose solutions with different concentrations are added step by step to form a density gradient during centrifugation.

In addition to the above preparation method, the biological cell membrane system of the present invention may further comprise the following preparation method, which specifically comprises the following steps:

3) Adding an organic solvent into the cell lysate to extract lipid, drying the extracted lipid into a film, adding water into the film, mixing uniformly, centrifuging, discarding the precipitate, and collecting supernatant A;

4) And (3) performing ultracentrifugation on the supernatant A, discarding the supernatant, and collecting the precipitate B to obtain the required biological cell membrane system.

Wherein the organic solvent is chloroform-methanol solution, and can extract fat-soluble components, thereby reducing the content of water-soluble components such as glycolipid. In the extraction of the biological cell membrane system, the extracted lipid is firstly dried into a film, and then the film is added with water and is centrifuged after being uniformly mixed, and the drying purpose is that the organic solvent can be removed by drying. The resuspension medium is prepared from at least one of sucrose, glycerol, polysucrose, cesium chloride, sodium chloride, tri-iodo benzoyl glucosamine, silica sol and fluorocarbon.

Wherein the strain is one or more of Thermus thermophilus, streptococcus thermophilus, thermus aquaticus, thermus acidophilus, sulfolobus, thermobifida, archaea, thermococcus equi, yeast, and Bacillus subtilis.

The strain in the application adopts severe culture conditions such as sugar-free, high osmotic pressure, illumination and the like, so that the glycolipid metabolism of the strain is accelerated in the culture process. Stringent culture conditions refer to improving the metabolism of the strain on the premise that the strain can survive.

The carbon source in the sugar-free culture medium is sugar components such as polyhydroxy aldehyde or polyhydroxy ketone, polycondensates thereof, certain derivatives and the like, namely various sugar, starch, dextrin and sugar alcohol are not contained, and the carbon source can be dairy products or proteins or a small amount of cellulose or yeast extract and the like.

Alternatively, step 1) of the two preparation methods above regulates the osmotic pressure of the fermentation broth to 350-3100mOsmol/kg by adding an osmotic pressure regulating solute, which is a salt or glycerol. The salt may be NaCl or CaCl ₂. Specifically, the osmotic pressure regulating solute is 1% -10% (w/v) NaCl solution or 1% -15% (w/v) CaCl ₂ solution or 5-30% (w/v) glycerol solution (prepared by glycerol and deionized water). Under the condition of high osmotic pressure of the culture medium, the metabolism of glycolipid can be accelerated.

Alternatively, the temperature gradient method in step 1) of the two preparation methods adopts a gradual heating mode for culturing for 24 hours within the temperature range of 60-90 ℃.

Optionally, the adjustment range of the illumination intensity in the step 1) in the two preparation methods is 500-1500lx, and the illumination time is 24-72h. Thermophilic thermus is a growth bacterium which does not need illumination, and adopts a gradient enhanced illumination method or a constant illumination method with lower illumination to promote bacteria to adjust growth metabolism so as to adapt to new environment in order to improve the illumination growth adaptation process of the bacterium.

The contents of the components of each substance in the biological cell membrane system prepared by the two methods are shown in the following table 1.

TABLE 1 composition of biological cell membrane system

Substance (B)	Content of
		Film relative Density	1.160-1.290
Protein (%)	5-15
		Phospholipid (mg/g)	40-200
Total polyamine (mg/g)	20-95 (Mass percent)
		Carotenoids (ppm)	50-500
RNA(％)	0.03-1.0
		SOD(U/g)	20-400
Glycolipid (ppm)	0.5-95

The biological cell membrane system provided by the invention combines active substances mainly through one or more combination modes of intra-membrane encapsulation, surface adsorption, surface cross-linking, inter-membrane embedding and targeting.

The phospholipid bilayer of the biological cell membrane system of the application forms spherical or vesicular liposome, has wrapping function and targeting function, can be used as a carrier such as a carrier, a cell organelle vesicle and the like, and can also be used as a natural liposome; the biological enzyme functional enzyme can partially carry biological components (active running and passive movement).

In particular, the active substance may be a vaccine, an immunomodulator, a cosmetic ingredient, a pharmaceutical ingredient, a genetic material, a cell or a cell tissue.

The beneficial effects of the invention are as follows: the invention provides a preparation method of a biological cell membrane system, which is characterized in that a sugar-free culture medium is adopted for culturing a strain under the severe conditions of illumination and high osmotic pressure, so that the strain consumes glycolipid as much as possible in the propagation process and advantageous components such as polyamine complex, carotenoid, SOD and the like are produced. Meanwhile, in the biological cell membrane extraction process, because the glycolipid is a water-soluble substance, and the polyamine complex, the carotenoid, the SOD and the like are fat-soluble substances, the content of the glycolipid in an extracted biological cell membrane system can be effectively reduced by an organic solvent extraction or secondary density gradient centrifugation method, and the contents of beneficial components such as the polyamine complex, the carotenoid, the SOD and the like are higher.

Drawings

FIG. 1 is a graph showing the results of protein electrophoresis in different biological membrane systems obtained in examples 3, 5 and 6 of the present invention;

FIG. 2 is a graph showing comparative photo-damage repair of HaCaT cells by different biological cell membrane systems according to example 10 of the present invention;

FIG. 3 is a graph showing the comparison of anti-inflammatory capacity of the biological membrane system of example 11 of the present invention in LPS model.

Detailed Description

The present invention will now be described in more detail and fully hereinafter with reference to the accompanying drawings, in which embodiments of the invention are shown.

In the application, the enzyme activity of SOD is measured according to the method of GB5009.171, and the enzyme quantity corresponding to the inhibition of the pyrogallol autoxidation rate reaching 50% is calculated as one SOD activity unit (U).

The total polyamine content was determined by the acid chloride reaction method (reaction of Polyamine (PAS) with benzoyl chloride under alkaline conditions, attaching benzoyl groups to primary and secondary amines) at a wavelength of 254 nm. The total polyamine content includes the sum of the contents of all polyamine complexes of histamine, agmatine, pentylamine, spermidine, putrescine, spermidine, norspermine, homospermine, spermine, and mycolipin.

The carotenoid content is measured by using a GB/T5009.83 method.

The glycolipid content is calculated as 3-deoxy-D-manno-2-octanonic acid (Kdo) and determined by thiobarbituric acid method.

The content detection of metal ions was performed by Atomic Absorption Spectrometry (AAS) for elemental analysis.

The strains adopted in the embodiment of the application are thermophilic thermus, and are separated and purified from a yellow stone national park hot spring pool with the preservation number of ATCC27634. Thermophilic thermus species are able to survive in severe environments with high osmotic pressure. Thermus thermophilus is inoculated into the sugarless culture medium according to the ratio of 1:100.

The strain used in example 1 was Thermus thermophilus.

1) Strain culture: the thermophilic thermus strain is inoculated into a culture medium according to the proportion of 1:100, and the yield of the polyamine complex, carotenoid and SOD is improved by utilizing a method of combining a gradient high-temperature fermentation method with high pressure and high permeation. Adding NaCl to a concentration of 1.35%, regulating osmotic pressure of the fermentation broth to 460mOsmol/kg, gradually heating at 60-70deg.C, 70-75deg.C and 75-90deg.C every 8H, and culturing for 24 hr.

2) Breaking the wall: after fermentation, centrifugally collecting thalli at 4,000rpm for 30min and 4 ℃; the cells were resuspended in homogenization buffer (20 mmol/L Tris-Cl pH8.0, 100mmol/L NaCl,2mmol/L MgCl ₂, 1mmol/L DTT) and the supernatant was centrifuged at 6,000rpm,10min at 4 ℃; adding homogenizing buffer solution to resuspend precipitate (about 1g of 10ml buffer solution), adding PMSF with final concentration of 1mmol/L, and performing ice bath ultrasonic disruption for 30min (amplitude of 55%, ultrasonic disruption for 5s and stopping for 8 s) to obtain cell lysate.

3) Biological cell membrane system extraction: 10ml of cell lysate was extracted from cells with chloroform-methanol solution and the non-lipid extract was separated with distilled water. The extracted lipids were dried to a thin film in a 10ml round flask and vigorously shaken with water by vortex mixer (0.5% w/v solution). Then, the obtained lipid was sonicated for 10 minutes at a temperature of 4℃under centrifugation of 3000 Xg, the supernatant was taken, dissolved in 5ml of PBS buffer solution, centrifuged at a high speed at 15,000-30,000 Xg for 10-30 minutes at a temperature of 1-6℃and the supernatant was discarded.

4) Taking the precipitate obtained by centrifugation under the condition of high rotation speed for the first time; then, the precipitate is dissolved by PBS buffer solution, and ultracentrifuged at the temperature of between 1 and 6 ℃ with 100,000-200,000 Xg and 30 to 90min, the supernatant is discarded, and the precipitate B is collected, thus obtaining the biological cell membrane system required to be extracted.

Example 2

1) Strain culture: thermus thermophilus is inoculated into a culture medium according to the proportion of 1:100, and the yield of the polyamine complex and the carotenoid is improved by utilizing a method of combining a gradient high-temperature fermentation method with high pressure and high permeation. Adding NaCl to a concentration of 1.35%, regulating osmotic pressure of the fermentation broth to 460mOsmol/kg, gradually heating at 60-70deg.C, 70-75deg.C and 75-90deg.C every 8H, and culturing for 24 hr.

3) The extraction operation is performed by a secondary density gradient centrifugation method and an ultracentrifugation method.

Specifically, the density gradient centrifugation method includes the steps of: adding the obtained cell lysate into sucrose solutions with mass percent concentration of 20%, 30%, 40%, 50%, 60% and 70% respectively, re-suspending to obtain heavy suspension, ultracentrifugating at 150,000-300,000 ×g and 60-90min at 1-6deg.C, and collecting liquid C with mass percent concentration of 40% -50% in the sucrose solution. Diluting the liquid C with PBS buffer solution, ultracentrifugating at 1-6deg.C for 30-90min at 100,000-200,000 ×g, discarding supernatant, and collecting precipitate to obtain the desired biological membrane system.

TABLE 2 comparison of the vesicles of the resulting biological systems by density gradient centrifugation (secondary) at different sucrose concentrations

As is clear from Table 2, in the sucrose solution having a mass percentage concentration of 40% -50%, the activity of biological enzymes such as superoxide dismutase (SOD) was the strongest.

Example 3

The difference from example 1 is that step 1) strain culture: inoculating thermophilic Thermus into culture medium at ratio of 1:100, irradiating with 500lx fluorescent lamp under the condition that DO dissolved oxygen value is less than 15% (controlling oxygen passage), starting from 65deg.C, raising 2 deg.C per hour, gradient fermenting, and continuously culturing under final condition (75deg.C) for 24 hr, and collecting thallus.

Example 4

The difference from example 3 is that in step 1) the strain is cultivated by a fluorescent lamp gradient irradiation method: and adjusting the total time to 1500lx after adjusting the fermentation time to 1000lx for 0-2h with 500lx and 2-8h until the fermentation is finished for 24 h.

TABLE 3 comparison of the levels of polyamines in class Hu Bosu of the illuminated versus non-illuminated biological systems

Index (I)	Non-illuminated group	Example 3 (500 lx)	Example 4 (gradient irradiation method)
				Total polyamine (mg/g)	6.19±0.68	39.18±1.76	52.43±3.12
Carotenoids (ppm)	198.0±22.0	305.5±18.4	299.2±10.1
				Mn²⁺(ppm)	0.54±0.08	3.82±0.13	5.14±0.18
SOD(U/g)	39.2±2.5	125.2±4.9	133.62±7.8
				Fe³⁺(ppm)	1.58±0.19	13.34±0.37	14.58±0.21

As can be seen from Table 3, the illumination groups using 500lx and gradient irradiation method can effectively increase the total polyamine, carotenoid, SOD and Mn ²⁺ and Fe ³⁺ metal ion contents, wherein the total polyamine content in the biological cell membrane system under 500lx condition is increased by 5 times, and the carotenoid content is increased by 52.29%.

Example 5

The difference from example 1 is that in step 1) a 3.4% CaCl ₂ hypertonic solution was used to adjust the osmotic pressure of the fermentation broth to 600mOsmol/kg.

Example 6

The difference from example 1 is that the fermentation broth of step 1) is subjected to an isotonic pressure.

FIG. 1 is a graph of the results of electrophoresis of total proteins from different biological systems, M-1 is a biological system obtained by CaCl ₂ method (example 5), M-2 is a biological system obtained by isotonic method (example 6), M-3 is a biological system obtained by 500lx light fermentation (example 3), and M-4 is lecithin.

Example 7

The difference from example 1 is that in step 1) a 9% CaCl ₂ hypertonic solution is used to adjust the osmotic pressure of the fermentation broth to 1500mOsmol/kg.

Example 8

The difference from example 1 is that in step 1) a 10% glycerol hypertonic solution is used and the osmotic pressure of the fermentation broth is adjusted to 2100mOsmol/kg.

TABLE 4 osmotic pressure and the effects of osmotic pressure on regulating solutes

As is clear from Table 4, the higher the osmotic pressure, the higher the strain resistance of the strain against high osmotic pressure, the more polyamine complex and carotenoid are produced, and the lower the glycolipid content.

Example 9

The difference from example 2 is that: when the resuspension medium is silica sol (Percoll): centrifuging at 4000rpm at low temperature for 1 hr, collecting supernatant, adding into Percoll density gradient layering liquid with density gradient specific gravity according to equal volume, centrifuging at room temperature, and centrifuging at 1200r/min for 25min; and respectively sucking out each layer of cells, putting the cells into 1 test tube, adding PBS heavy suspension, washing for 3 times at 4 ℃, centrifuging for 10min at 4000r/min each time, and discarding the supernatant to obtain the biological cell membrane system required to be extracted.

The total polyamine in the obtained biological cell membrane system is 31.43+/-0.86 mg/g, the carotenoid is 248.0+/-25.1 ppm, and the glycolipid is 67.3+/-10.9 ppm. As can be seen from a comparison of the total polyamine and carotenoid contents in example 2 in Table 4, the effects of resuspension with sucrose solutions were also obtained with Percoll resuspension.

Example 10

The biological membrane system prepared according to example 4 (light process) was compared (no light and isotonic) and the biological membrane system obtained under the same other process conditions was tested in a photodamage repair model. And (3) irradiating the HaCaT cells through a simulated UVB light source to establish a photodamage repair model, adding a test object, and detecting the repair capability of the test object to the photodamaged cells. Log phase cells were collected, suspension of HaCaT cells was prepared with RPMI-1640 medium containing 10% fbs, 1% sk, 100 μl/well was inoculated at 5×10 ⁴/mL in 96 well plates, incubated at 37 ℃, 5% (v/v) CO ₂, and wells around the plates were filled with sterile PBS to seal the edges. After cell culture for 24h, the culture was aspirated, washed once with PBS, and then 20uL of PBS buffer was added for UVB light irradiation (UVB irradiation time: UVB lamp, 10 min after the lamp was turned on, with ultraviolet irradiation intensity of 1.05mW/cm ². Irradiation dose was controlled to 30mJ/cm ², irradiation time (30 s), samples of different volume fractions (0.5%, 1%, 3%, 5%) were added, 100ppm dexamethasone was used as positive control group, and culture without drug sample was used as control group, and culture was continued for 24 h. MTT assay was performed as a method for detecting cell viability and growth based on the principle that succinic dehydrogenase in live cell mitochondria could reduce exogenous MTT to water insoluble blue-violet crystal formazan (Formazan) and deposited in cells, whereas dead cells had no such function.

The ability of the samples to repair photodamage was evaluated by cell viability was recorded, and the biological cell membrane system prepared in example 3 had better ability to repair photodamage, as shown in FIG. 2.

Example 11

Using the biological membrane system obtained in example 1, and selecting different volume fractions (1%, 3%, 5%, 10%, 15%, 20%, 25%) as samples, 50ppm hydrocortisone was used as positive control, LPS was used as control, and the evaluation was performed in an anti-inflammatory capacity model of RAW cells (NO) to produce NO by LPS induction. As can be seen from FIG. 3, the anti-inflammatory capacity of the biological cell membrane system prepared in example 1 was increased with increasing concentration, and gradually decreased when the volume fraction of the biological cell membrane system was 20%, and the anti-inflammatory capacity close to 50ppm hydrocortisone was achieved when the volume fraction of the biological cell membrane system was 25%. Thus, the biological cell membrane system prepared in the embodiment 1 of the invention has obvious anti-inflammatory capability.

The foregoing description is only of the preferred embodiments of the present invention, and is not intended to limit the scope of the invention, but rather is intended to cover all equivalent structures modifications made by the teachings of the present invention, either directly or indirectly, within the scope of the invention.

Claims

1. The preparation method of the biological cell membrane system is characterized by comprising the following steps of:

3) Adding chloroform-methanol solution into cell lysate to extract lipid, drying the extracted lipid into film, adding water into the film, mixing well, centrifuging, discarding precipitate, and collecting supernatant A;

4) Ultracentrifugation is carried out on the supernatant A, the supernatant is discarded, and the sediment B is collected, so that a needed biological cell membrane system is obtained;

wherein the strain is Thermus thermophilus, and the preservation number is ATCC27634;

Wherein, the temperature gradient method adopts a gradual heating mode to culture for 24 hours in the temperature range of 60-90 ℃; the illumination intensity is adjusted by adopting single illumination intensity or gradient illumination intensity, the adjustment range of the illumination intensity is 500-1500lx, and the illumination time is 24-72h; the osmotic pressure is regulated to 350-3100mOsmol/kg by adding osmotic pressure regulating solute, and the osmotic pressure regulating solute is salt or glycerol.

2. The preparation method of the biological cell membrane system is characterized by comprising the following steps of:

3) Adding the cell lysate into a resuspension medium to obtain a resuspension, and purifying the biological cell membrane system in the layered resuspension by ultracentrifugation and density gradient centrifugation to obtain a required biological cell membrane system;

3. The method of preparing a biological membrane system according to claim 1 or 2, wherein the biological membrane system comprises a phospholipid bilayer and a polyamine complex grafted to the phospholipid bilayer and a polyisoprene-based compound embedded on the phospholipid bilayer, wherein the polyamine complex comprises any one or more of:

Linear or branched alkylamines having two amino groups at both ends,

Linear or branched alkylamines having at least two amino groups at each end and comprising 1 to 4 secondary amines,

Linear or branched alkylamines having at least one amino group at one end and an imidazolyl group at the other end,

A cyclic polyamine;

The polyisoprene compound is selected from any one or a combination of a plurality of tetraterpene compounds and tetraterpene compound fatty acid ester derivatives;

the content of glycolipid in the biological cell membrane system is less than or equal to 100ppm.

4. The method of preparing a biological membrane system according to claim 3, wherein the biological membrane system further comprises a biologically active oxidoreductase having a metal ion in the active center; the metal ions are from any one or more of Mn, ca, mg, zn, cu, fe.

5. The method for preparing a biological cell membrane system according to claim 4, wherein the oxidoreductase is superoxide dismutase and has manganese ions in the active center.

6. The method of claim 3, wherein the polyamine complex is one or more of histamine, agmatine, pentylamine, spermidine, putrescine, spermidine, norspermine, homospermine, spermine, and cytochrome c.

7. The method of claim 2, wherein the resuspension medium is formulated from at least one of sucrose, glycerol, polysucrose, cesium chloride, sodium chloride, tri-iodo-benzoyl-glucosamine, silica sol, and fluorocarbon.