Fusion membrane system for constructing artificial organelle and construction method of fusion membrane system

Abstract

The invention is applicable to the technical field of synthetic biology and membrane bioengineering, and provides a fusion membrane system for constructing an artificial organelle and a construction method thereof, the fusion membrane system comprises: a pigment body membrane for light-driven ATP synthesis; the erythrocyte membrane is used for consuming ATP to drive ion transport; the fusion promoting component comprises cholesterol-PEG (Polyethylene Glycol)-(EIAALEK) 4 and cholesterol-PEG-(KIAALKE) 4; the pigmented body membrane and the erythrocyte membrane form a fused vesicle structure under the action of the fusion promoting component. The steps of protein purification and artificial lipid assembly are avoided, the operation process is simplified, functional recombination of a heterologous natural membrane protein module is achieved, an energy-self-sufficiency artificial organelle is constructed, the ATP utilization efficiency is remarkably higher than that of a physical mixed membrane system and is improved by about 2-3 times, and the application prospect is wide. And a feasible framework is provided for energy management and artificial organelle design in a synthetic biological system.

Description

Technical Field

[0001] The present invention belongs to the technical field of synthetic biology and membrane bioengineering, and in particular relates to a fusion membrane system for constructing artificial organelles and a construction method thereof. Background Art

[0002] The cell membrane is a vital structural basis in the life process and is involved in multiple core functions such as material transport, signal transduction and energy conversion. These functions are highly dependent on the orderly distribution and dynamic collaboration of membrane proteins. The conformation and activity of membrane proteins are usually restricted by the natural lipid environment in which they are located. Therefore, maintaining the conformational integrity and functional synergy of membrane proteins is a prerequisite for realizing complex membrane functional networks. Especially in highly coupled physiological reactions involving energy cascades and transmembrane transport, the natural membrane environment provides the necessary microstructure and chemical background, which is a core element that is difficult to fully simulate by artificial systems.

[0003] One of the important goals of synthetic biology is to construct controllable artificial membrane systems to reproduce the functions of natural cell membranes in energy conversion, signal integration and metabolic regulation. Existing technologies mainly focus on reconstruction methods based on synthetic liposomes and purified membrane proteins, or on the encapsulation of functional modules through artificially synthesized multilayer membranes. Such systems are widely used in drug delivery, synthetic cells and artificial organelle design, and provide important technical support for the precise construction of biomimetic membranes. However, existing technologies have the following limitations: First, the extraction and purification of membrane proteins are usually cumbersome, time-consuming and costly, which not only limits the scalability of the system, but also increases the technical threshold and resource consumption of the construction process; second, during the purification and reconstruction process, membrane proteins are prone to lose their original spatial conformation and catalytic activity due to changes in the membrane protein environment, which is particularly significant in energy conversion systems whose functions are highly dependent on their conformation; in addition, synthetic lipid systems are usually single in composition and lack the complexity and distribution heterogeneity of natural membrane lipids, making it difficult to provide the microenvironment required to maintain membrane protein activity. In summary, due to the limitations of low protein yield and activity, simple membrane phospholipid composition, and poor functional integration efficiency, existing artificial membrane systems still find it difficult to replicate the efficiency and stability of natural membrane systems.

[0004] Therefore, there is an urgent need to develop a membrane system construction method that can retain the natural functions of membrane proteins, integrate multiple membrane source modules, simplify the reconstruction steps and improve the system integration efficiency to support the design and construction of multifunctional bionic membrane systems. Summary of the Invention

[0005] The purpose of the embodiments of the present invention is to provide a fusion membrane system for constructing artificial organelles, aiming to solve the problems raised in the above background technology.

[0006] The embodiment of the present invention is implemented as follows: a fusion membrane system for constructing an artificial organelle, comprising:

[0007] The first membrane component: the chromatin membrane, which contains photosystem I and FOF1-ATP synthase for light-driven ATP synthesis;

[0008] Second membrane component: erythrocyte membrane, which contains Na + / K + ATPase, Ca 2+ ATPase and GLUT1 membrane protein, which consume ATP to drive ion transport;

[0009] Fusion-promoting components: including cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4, used to induce specific fusion between two membranes;

[0010] The first membrane component and the second membrane component form a fusion vesicle structure under the action of the fusion promoting component. The fusion vesicle structure generates ATP by light induction and is used to drive ATP-dependent membrane proteins in the red blood cell membrane to achieve an energy cascade effect.

[0011] Another object of the present invention is to provide a method for constructing a fusion membrane system for constructing an artificial organelle, comprising the following steps:

[0012] The plastid membranes were isolated from the photosynthetic bacterium Rhodobacter sphaeroides, washed, resuspended in Tris-HCl buffer, and purified to obtain purified plastid membranes retaining photosystem I and F0F1-ATP synthase.

[0013] Separate red blood cell membranes from anticoagulated whole blood and process them to obtain purified red blood cell membranes, retaining their Na + / K + ATPase, Ca 2 + ATPase and GLUT1 membrane protein;

[0014] Cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4 were added to the chromatin membrane and erythrocyte membrane suspensions, respectively, to integrate them into the membrane surface in a cholesterol-anchored manner, forming two pre-fusion membranes, chromatin-E and erythrocyte membrane-K.

[0015] Chromatosome-E and erythrocyte membrane-K were mixed and incubated to promote specific recognition of E / K peptides, thereby inducing the formation of fusion vesicles;

[0016] The fusion vesicles are placed under light conditions to construct an artificial organelle - the pigment metabolosome, so that the ATP synthesized by the pigment membrane components is utilized by the membrane proteins in the red blood cell membrane components on the fusion vesicle membrane, forming a complete energy cascade system.

[0017] Preferably, in the step of purifying to obtain purified chromatin membranes, the purification operations are specifically high-pressure crushing and ultracentrifugation.

[0018] Preferably, in the step of obtaining purified red blood cell membranes through treatment, the treatment operations specifically include hypotonic treatment and washing centrifugation.

[0019] Preferably, in the step of adding cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4 to the chromatin membrane and erythrocyte membrane suspensions respectively, the concentrations of cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4 are 100 μM respectively.

[0020] Preferably, in the step of mixing the chromatin-E and the erythrocyte membrane-K, the chromatin-E and the erythrocyte membrane-K are mixed at a phospholipid molar ratio of 1:1.

[0021] Preferably, the incubation temperature is 30° C. and the incubation time is 30 min.

[0022] The present invention provides a fusion membrane system for constructing artificial organelles. By directional fusion of the chloroplast membrane of photosynthetic bacteria and the erythrocyte membrane of mammals, the functions of light-driven ATP synthesis and ATP-driven ion transport are efficiently coupled in a single closed membrane structure. The artificial organelle-pigment metabolite is successfully constructed, wherein the chloroplast membrane provides photosystem I and ATP synthase, and the erythrocyte membrane retains Na + / K + ATPase, Ca 2+ The fusion of proteins such as ATPase and GLUT1 is induced by cholesterol-modified coiled peptide E / K pairing, retaining the natural microenvironment and function of membrane proteins without relying on protein purification and artificial lipid reconstruction; the artificial organelles prepared in the embodiments of the present invention exhibit good energy cascade efficiency and system sustainability; the method provided in the embodiments of the present invention does not require protein purification or complex membrane engineering, is applicable to a variety of natural membrane sources, and has wide adaptability and application potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Characterization of the morphology and particle size of chromatin membranes and erythrocyte membranes provided in the embodiments of the present invention;

[0024] Figure 2 The results of the membrane functionality verification experiment provided by the embodiment of the present invention;

[0025] Figure 3 Schematic diagram of the lipidated peptide used in the membrane fusion process provided in an embodiment of the present invention;

[0026] Figure 4 Schematic diagram of the molecular structures of two fluorescent peptides used to verify whether CPE and CPK are inserted into phospholipid membranes, provided in an embodiment of the present invention;

[0027] Figure 5 Schematic diagram of using fluorescent peptides to verify the integration of lipopeptides CPE or CPK into chromatin or erythrocyte membranes according to an embodiment of the present invention;

[0028] Figure 6 Fluorescent images characterizing the formation of chromatin-E and erythrocyte membrane-K provided in an embodiment of the present invention;

[0029] Figure 7 A schematic diagram of the fusion process and the formation of an energy cascade provided by an embodiment of the present invention;

[0030] Figure 8 Characterization of the morphology and particle size of the pigment metabolites formed by fusion provided in the embodiments of the present invention;

[0031] Figure 9 A structured illumination microscopy (SIM) image of a pigment metabolite provided in an embodiment of the present invention;

[0032] Figure 10 The present invention provides a method for analyzing membrane fusion dynamics based on fluorescence resonance energy transfer (FRET).

[0033] Figure 11 Fluorescence co-localization analysis of pigment metabolites provided in the embodiments of the present invention;

[0034] Figure 12 Functional characterization of the pigment metabolites provided in the embodiments of the present invention;

[0035] Figure 13 The ion pump activity characterization results provided by the embodiments of the present invention;

[0036] Figure 14 The ATP decay rate experimental results provided by the embodiment of the present invention;

[0037] Figure 15 This is a sustainability verification of the pigment metabolite system provided in the embodiments of the present invention. DETAILED DESCRIPTION

[0038] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0039] A fusion membrane system for constructing an artificial organelle, the construction method of which comprises the following steps:

[0040] (1) The chromatin membrane of the photosynthetic bacterium Rhodobacter sphaeroides was selected as a light-driven module to provide photosystem I and F0F1-ATP synthase. Functional chromatin was extracted and purified, and its physicochemical characterization was performed to ensure that the ATP synthesis capacity was intact.

[0041] (2) Extract the red blood cell membrane and retain its Na + / K + ATPase, Ca 2+ Membrane proteins such as ATPase and GLUT1, which serve as energy utilization modules, are separated from erythrocyte membranes by hypotonic swelling and centrifugation techniques, while maintaining the integrity of their membrane proteins.

[0042] (3) Using a simplified SNARE system, namely, peptide E (its amino acid sequence is shown in SEQ ID NO. 1) and peptide K (its amino acid sequence is shown in SEQ ID NO. 2), which are modified with PEG-cholesterol (cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4, respectively named CPE and CPK), attached to the membrane surface to mimic the natural SNARE complex structure to induce membrane fusion;

[0043] (4) The plastid membrane modified with E peptide and K peptide respectively is mixed and incubated with the red blood cell membrane, and the specific recognition pairing between E / K peptide is used to promote the docking of the two membranes and gradually tighten the membrane structure, thereby achieving efficient fusion;

[0044] (5) The composite membrane system formed by fusion realizes the spatial coupling of energy generation and utilization: the chromatin membrane components synthesize ATP under light and transfer it to the red blood cell membrane components as a local energy source, driving its membrane proteins such as Na + / K + ATPase, Ca 2+ The ion transport function of ATPase. As an artificial organelle, this structure effectively reconstructs and extends the energy cascade network of natural organelles, showing excellent ATP conversion and utilization efficiency.

[0045] The specific implementation of the present invention is described in detail below with reference to specific embodiments.

[0046] Example 1: A fusion membrane system for constructing an artificial organelle, the construction method of which comprises the following steps:

[0047] (1) The photoheterotrophic culture of Rhodobacter sphaeroides was centrifuged at 15,300 × g for 15 minutes at 4°C to collect the bacterial pellet, which was then washed twice with 10 mM Tris-HCl buffer (pH 8.0) to remove the culture medium residue; the cells were then resuspended in 50 mL of 10 mM Tris-HCl buffer (pH 8.0) containing 5 mM MgCl2, 50 mM KCl, 5 mM L-ascorbic acid (vitamin C) and 10% sucrose, and 250 μL of DNase I and 500 μL of protease inhibitors were added, and then incubated on ice for 15 minutes to degrade nucleic acids and inhibit protein hydrolysis; the cells were then disrupted using a French high-pressure cell disruptor at 1280 psi and 4°C, and the disrupted liquid was centrifuged at 15,300 × g for 15 minutes at 4°C to remove unbroken bacteria and large molecular fragments. The supernatant was collected and centrifuged at 140,000 × g at 4°C. The chromatin was enriched by ultracentrifugation at 15,300 × g for 120 minutes, and the precipitate was resuspended in 5 mL of 10 mM Tris-HCl buffer (pH 8.0). The remaining impurities were removed by centrifugation again at 15,300 × g for 15 minutes at 4°C to obtain a high-purity chromatin membrane suspension with an optical density at 860 nm (OD860) of 50.

[0048] (2) Take 100 mL of anticoagulated whole blood sample containing ethylenediaminetetraacetic acid (EDTA, 150 mg), centrifuge at 3000× g for 20 minutes at 4°C, separate and remove the upper plasma and middle white film layer, collect the lower red blood cell pellet, wash the red blood cells three times with pre-cooled isotonic phosphate buffer (PBS) at pH 7.4 at a volume ratio of 1:3, and centrifuge at 3000× g for 10 minutes after each wash; then slowly add the washed red blood cells to pre-cooled 0.25× hypotonic PBS solution (pH 7.4, pre-equilibrated at 4°C) at a ratio of 1:40 (v / v), incubate at 4°C for 90 minutes to complete hemolysis; the hemolyzed blood is centrifuged at 9000× g for 15 minutes at 4°C, discard the supernatant to obtain the red blood cell membrane pellet, repeat this centrifugation and washing step three times to completely remove residual hemoglobin, and finally obtain white red blood cell membranes;

[0049] (3) Peptide E and peptide K served as membrane fusion functional units, and their sequences were EIAALEKEIAALEKEIAALEKEIAALEK (as shown in SEQ ID NO.1) and KIAALKEKIAALKEKIAALKEKIAALKE (as shown in SEQ ID NO.2), respectively. The peptide chains were covalently linked to the cholesterol anchoring group through a polyethylene glycol segment (EG4) to form lipidated peptide E and peptide K, which were named CPE and CPK, respectively. CPE or CPK complex (10 mM stock solution, solvent was DMSO) was added to the erythrocyte membrane and chromatin membrane suspension (10 mM Tris-HCl buffer, pH 7.4) at a volume ratio of 1% to a final concentration of 100 μM, and incubated in a constant temperature shaker at 30°C (frequency 100 rpm) for 30 minutes. Free CPE was then removed by ultracentrifugation at 140,000 × g for 120 minutes at 4°C. The suspension was centrifuged at 15,300 × g for 120 minutes. g centrifugation for 10 min to remove free CPK, and the washing was repeated twice to finally obtain erythrocyte membrane-K and chromatin-E;

[0050] (4) Based on the quantification of phosphatidylcholine (PC) molar concentration, chromatin-E and erythrocyte membrane-K were mixed in 10 mM phosphate buffer (PBS, pH 7.4, containing 5 mM MgCl2, osmotic pressure 290 mOsm / kg) at a PC molar ratio of 1:1. The total volume of the mixed system was 1 mL, of which the total amount of PC was approximately 2 μmol / mL. The mixed membrane suspension was placed in a constant temperature oscillator (frequency 100 rpm) and incubated at 30°C for 30 minutes to promote membrane fusion and induce the formation of fusion vesicles (chromatin metabolosomes).

[0051] (5) The above-mentioned fusion vesicles are placed under light conditions, so that the ATP synthesized by the chromatin components is utilized by the membrane proteins in the red blood cell membrane components on the fusion vesicle membrane, forming a complete energy cascade system. The energy coupling mechanism of the fusion recombinant membrane system is used to improve the efficiency of the ATP metabolic cycle, which is specifically manifested as: the chromatin membrane (containing light-driven F0F1-ATPase) is connected to the red blood cell membrane (containing Na + / K + ATPase and Ca 2+ ATPase) after the directional fusion of step (4), a transmembrane proton gradient and ion electrochemical potential synergistic transmission system is formed, which uses the proton motive force generated by the chromatin light reaction center to drive F0F1-ATPase to synthesize ATP, and at the same time, Na + / K + ATPase or Ca 2+ ATPase converts the energy released by ATP hydrolysis into transmembrane ion transport work, realizing the energy transfer pathway of ATP generation-consumption closed loop.

[0052] Comparative Example 1: Refer to step (4) of Example 1, except that unmodified original red blood cell membranes and chromatin membranes are mixed in the same PC ratio and processed simultaneously.

[0053] The morphology and particle size of the chromatin membrane and erythrocyte membrane of Example 1 were characterized, and TEM images of the chromatin membrane and erythrocyte membrane were obtained as shown in FIG. Figure 1 As shown in A, the vesicle structure of the chromatin membrane and the red blood cell membrane can be seen; the DLS data results are as follows Figure 1 As shown in B, the particle size distribution and uniformity are characterized; in summary, the successful extraction of chromatin membrane and erythrocyte membrane and the formation of vesicle structure are confirmed.

[0054] The chromatin membrane sample of Example 1 was supplemented with 0.2 mM ultrapure ADP and 5 mM Pi, and then illuminated for 3 minutes (860 nm, 30 mW). ATP synthesis was then measured using a luciferin-luciferase assay (FLAA, Sigma) (ultrapure ADP was purchased from Immunochemistry Technologies (306-ADP100-2); free Pi was provided by 1× phosphate buffer (10 mMTris-HCl, 5 mM NaH2PO4, 2.5 mM citric acid, 5 mM MgCl2)). The functional characterization results of ATP synthesis by ATP synthase in the chromatin membrane under conditions of illumination and substrate presence (ADP and Pi) were obtained as shown below. Figure 2 As shown in A; 500 μM ATP was added to the erythrocyte membrane suspension of Example 1, when the ATP-dependent proteins on the erythrocyte membrane (such as Na + / K + ATPase and Ca 2 + When ATPase hydrolyzes ATP, inorganic phosphate (Pi) is released, and inorganic phosphate reacts with malachite green reagent (MAK113, Sigma) to form a stable dark green colorimetric product. The signal intensity is proportional to the ATPase activity. The absorbance of the colorimetric product at 620 nm is measured to obtain the ATP-dependent proteins of the red blood cell membrane (such as Na + / K + ATPase and Ca 2+ ATPase) consumes ATP Figure 2 As shown in B.

[0055] The molecular structures of the lipidated E peptide (CPE) and K peptide (CPK) used in the membrane fusion process in Example 1, including the cholesterol anchor end, the EG4 segment and the E / K peptide motif (EIAALEKEIAALEKEIAALEKEIAALEK and KIAALKEKIAALKEKIAALKEKIAALKE) are as follows: Figure 3 shown.

[0056] E peptide or K peptide is linked to FITC fluorophore through 6-aminohexanoic acid (ACP), named FITC-E and FITC-K respectively. CPE or CPK can be combined with the corresponding fluorescent peptide FITC-K or FITC-E. By observing the changes in the fluorescence signal on the membrane surface, it is proved that CPE and CPK can be effectively anchored to the membrane surface. Figure 4 These are the molecular structural formulas of the two fluorescent peptides used in Example 1 to verify whether CPE and CPK are inserted into the phospholipid membrane.

[0057] Fluorescent peptides were used to verify that the lipopeptide CPE or CPK was integrated into the chromatin or erythrocyte membrane. Figure 5 As shown, specifically, the cholesterol membrane anchors of CPE and CPK spontaneously embedded into phospholipid membranes through hydrophobic interactions, and were named chromatin-E and erythrocyte membrane-K, respectively. Subsequently, complementary fluorescent peptides (FITC-E and FITC-K) were added to induce the formation of E / K coiled-coil structures. Chromatin-E and erythrocyte membrane-K produced membrane-associated fluorescence, confirming that a fusion peptide recognition pair had been successfully constructed on the surfaces of the two membranes. Figure 5 The functionalization effect of pre-modified membranes (chromosome-E and erythrocyte membrane-K) before fusion was verified.

[0058] Fluorescence images characterizing the formation of chromatin-E and erythrocyte membrane-K Figure 6 As shown, chromatin-E and erythrocyte membrane-K were mixed and incubated with fluorescent peptides FITC-K and FITC-E, respectively, and the unbound free peptides were removed. Figure 6 The generation of fluorescent signals on the media indicates that the lipidated peptides CPE and CPK were successfully integrated into the chromatin membrane and erythrocyte membrane (right). In contrast, no binding of the complementary fluorescent peptides was observed in the control group with unmodified lipopeptides (right).

[0059] The schematic diagram of the fusion process and the formation of the energy cascade in step (5) of Example 1 is as follows: Figure 7 As shown, the chromatin-E and erythrocyte membrane-K recognize and fuse through the E / K peptide to form a closed vesicle (chromatin metabosome), in which the chromatin provides light-driven proton gradient to drive ATP synthase to synthesize ATP, and the erythrocyte membrane uses ATP to drive ion pumps to achieve transmembrane transport.

[0060] The morphology and particle size of the pigment metabolite formed by fusion in Example 1 were characterized, and the TEM image was obtained as follows: Figure 8 As shown in A, the vesicle structure of the fusion vesicle is shown; the DLS results are shown in Figure 8 As shown in B, its particle size distribution and uniformity are characterized; in summary, the formation and structural stability of fusion vesicles are verified.

[0061] The structured illumination microscopy (SIM) image of the pigment metabolite of Example 1 is as follows: Figure 9As shown, the red signal is the autofluorescence of the chromatin (left), and the green signal is the DiO probe that labels the red blood cell membrane (middle). The two co-localize in the fusion structure (right), indicating that the two membrane components have been effectively integrated, verifying the coexistence of dual sources of fusion vesicle components.

[0062] Schematic diagram of using FRET method to characterize membrane fusion Figure 10 As shown in A, to monitor lipid mixing during fusion, the erythrocyte membrane is simultaneously labeled with a FRET pair (donor DiO and acceptor DiI) to form erythrocyte membrane-K-DiO-DiI. Subsequent fusion with chromatin-E increases the distance between the donor and acceptor, resulting in a decrease in FRET efficiency. The curve of the change in donor fluorescence intensity over time is shown in Figure 10 B shows the kinetic differences between the fusion group and the physical mixed control group (chromatin + erythrocyte membrane).

[0063] Fluorescence co-localization analysis was performed on the pigment metabolite of Example 1, and confocal microscope images were obtained as shown in FIG. Figure 11 As shown in A, the two membrane-derived fluorescent signals co-localized in the fusion group (pigment metabolosome), while they separated in the physical mixing group; the results of flow cytometry quantitative analysis are shown in Figure 11 As shown in B, the proportion of double-positive events in the confirmed fusion group was approximately 80%.

[0064] Figure 12 A is the functional verification of the photogenerated ATP in the fusion system. Figure 12 As shown in A, it is confirmed that the pigment metabolite can effectively respond to light and synthesize ATP in the presence of substrates; the glucose internalization experiment mediated glucose into the vesicles through the red blood cell membrane GLUT1 and participated in the cascade reaction, indicating that the membrane protein function is retained. The results are shown in Figure 12 As shown in B. Figure 13 Shown are the results of ion pump activity characterization.

[0065] Adding ouabain and lanthanum nitrate to inhibit Na + / K + ATPase and Ca 2+ ATPase, measuring the release of free phosphate in the system, thereby verifying the functional activity and contribution of the ion pump in the pigment metabolosome membrane, and obtaining the results such as Figure 13 shown.

[0066] The luciferin-luciferase system was used to monitor the change in ATP concentration over time in the pigment metabolite prepared in Example 1 and the physical mixed control group (pigment body + red blood cell membrane) of Comparative Example 1. The results were as follows: Figure 14 As shown, the pigment metabolosome was observed to have a faster ATP decay rate, indicating that its ATP transfer and consumption efficiency was higher.

[0067] Sustainability verification of pigment metabolomic system Figure 15 As shown, after light-induced ATP production in the pigment metabolosome, re-illumination after a consumption period can still restore some ATP production and consumption dynamics, indicating that the system has the ability of dynamic energy regulation and continuous operation.

[0068] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A fusion membrane system for constructing artificial organelles, characterized in that: include: The first membrane component: the chromatin membrane, which contains photosystem I and FOF1-ATP synthase for light-driven ATP synthesis; Second membrane component: erythrocyte membrane, which contains Na + / K + ATPase, Ca 2+ ATPase and GLUT1 membrane protein, which consume ATP to drive ion transport; Fusion-promoting components: including cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4; The first membrane component and the second membrane component form a fusion vesicle structure under the action of the fusion promoting component.

2. A method for constructing a fusion membrane system for constructing an artificial organelle according to claim 1, characterized in that: The following steps are involved: The plastid membranes were isolated from the photosynthetic bacterium Rhodobacter sphaeroides, washed, resuspended in Tris-HCl buffer, and purified to obtain purified plastid membranes retaining photosystem I and F0F1-ATP synthase. Separate red blood cell membranes from anticoagulated whole blood and process them to obtain purified red blood cell membranes, retaining their Na + / K + ATPase, Ca 2+ ATPase and GLUT1 membrane protein; Cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4 were added to the chromatin membrane and erythrocyte membrane suspensions, respectively, to form two pre-fusion membranes, chromatin-E and erythrocyte membrane-K; The chromatin-E and erythrocyte membrane-K are mixed, incubated, and induced to form fusion vesicles; The fusion vesicles were placed under light conditions to construct an artificial organelle - the pigment metabolosome.

3. The method for constructing a fusion membrane system for constructing an artificial organelle according to claim 2, characterized in that: In the step of purifying to obtain purified chromatin membranes, the purification operations are specifically high-pressure crushing and ultracentrifugation.

4. The method for constructing a fusion membrane system for constructing an artificial organelle according to claim 2, wherein: In the step of obtaining purified red blood cell membranes through treatment, the treatment operations specifically include hypotonic treatment and washing centrifugation.

5. The method for constructing a fusion membrane system for constructing an artificial organelle according to claim 2, wherein: In the step of adding cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4 to the chromatin membrane and erythrocyte membrane suspension respectively, the concentrations of cholesterol-PEG-(EIAALEK)4 and cholesterol-PEG-(KIAALKE)4 are 100 μM respectively.

6. The method for constructing a fusion membrane system for constructing an artificial organelle according to claim 2, characterized in that: In the step of mixing the chromatin-E and the erythrocyte membrane-K, the chromatin-E and the erythrocyte membrane-K are mixed at a phospholipid molar ratio of 1:

1.

7. The method for constructing a fusion membrane system for constructing an artificial organelle according to claim 2, characterized in that: The incubation temperature is 30° C. and the incubation time is 30 min.

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