Preparation method of thylakoid-containing artificial cells and photosynthesis simulating method

A thylakoid, artificial technology, applied in the fields of biochemistry, biotechnology, and synthetic biology, which can solve the problem of not being able to double-simulate

Inactive Publication Date: 2018-03-30
HARBIN INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the problem in the prior art that the photosynthetic system of the bionic nature cannot be double-simulated from both

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  • Preparation method of thylakoid-containing artificial cells and photosynthesis simulating method

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specific Embodiment approach 1

[0017] Specific implementation mode 1: This implementation mode records a method for preparing artificial cells containing thylakoids, and the steps of the method are as follows:

[0018] Step 1: extract thylakoid particles: extract thylakoid particles from fresh spinach leaves, and store them in a buffer solution for later use;

[0019] Step 2: Preparation of GUV solution: apply the phospholipid solution on the ITO glass electrode, form a layer of phospholipid film after the solvent evaporates, fix the PTFE frame with vacuum grease, add liquid phase inside the PTFE frame, and prepare There is an ITO glass electrode with a layer of phospholipid film to add alternating current to prepare GUV solution;

[0020] Step 3: Preparation of thylakoid-containing artificial cells: mix the GUV solution prepared in step 2 with the solution containing thylakoid particles in step 1, and under the action of osmotic pressure, prepare thylakoid-containing artificial cells with complete structur...

specific Embodiment approach 2

[0021] Specific embodiment two: the preparation method of a thylakoid-containing artificial cell described in specific embodiment one, the specific steps of step one are: clean the spinach leaves, control the water; mix the spinach leaves with B1 buffer solution and Grind the spinach in a blender, wherein the mass volume ratio of the spinach leaf to the B1 buffer solution is 1:4g / mL, and the crushing time is 20min; then filter the mixture with 6-8 layers of gauze, and the filtrate is 6000rpm at 4°C Centrifuge 15min in centrifuge to collect complete and broken chloroplast; Add B2 buffer solution to the sediment and process 20min in the cell disruptor to break the chloroplast membrane, wherein the mass volume ratio of the sediment to B2 buffer solution is 1: 50g / mL; subsequently, centrifuge in a centrifuge at 10000rpm at 4°C for 15min, and add B3 buffer solution to the precipitate, wherein the mass volume ratio of the precipitate to B3 buffer solution is 1:50g / mL, and the finally...

specific Embodiment approach 7

[0026] Embodiment 7: A method for simulating photosynthesis using artificial cells containing thylakoids prepared in Embodiment 1. The specific steps of the method are: draw 14 mL of artificial cell solution containing thylakoids, and the concentration of chlorophyll therein It was 2.6mg / mL, irradiated for 2 h at 20°C and 250W xenon lamp to carry out photosynthesis, after photosynthesis, add 0.7mL of 2wt.% Triton X-100 (polyethylene glycol octylphenyl ether) to the mixed solution ) at 25°C for 10 min to break the phospholipid membrane and release the liquid in the vesicles, then add 10 mL of 10% trichloroacetic acid solution to terminate the redox reaction; then centrifuge at 3000 rpm for 10 min, take 1.5 mL of the supernatant and add 1.5 mL of distilled water and 0.1 mL of ferric chloride solution with a mass concentration of 0.1% were kept in a water bath at 25 °C for 10 min; then, the absorption value at 680 nm was measured with an ultraviolet absorption spectrophotometer.

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Abstract

The invention relates to a preparation method of thylakoid-containing artificial cells and a photosynthesis simulating method, and belongs to the fields of biotechnology, biochemistry and synthetic biology. The method comprises the following steps of extracting thylakoid particles from fresh spinach leaves, and storing the thylakoid particles in a buffer solution for use; smearing a phospholipid solution onto an ITO glass electrode; after the solvent volatilization, forming a layer of phospholipid membrane; fixing a polytetrafluoroethylene framework by vacuum grease; adding liquid phases intothe polytetrafluoroethylene framework; applying alternating current onto the ITO glass electrode for preparing a GUV solution; mixing the GUV solution and a thylakoid solution; preparing the thylakoid-containing artificial cells. The thylakoid-containing artificial cells are prepared through the enlightenment of the natural plant cell structure and function; the model can realize the optical driveelectron transfer process under the artificial sunlight irradiation. Through the model building, the attributes of chloroplasts in nature are doubly simulated from the structure and the function. Thecomplicated cells realize simplified research, so that the development and the progress of the biomimetic chemistry are promoted.

Description

technical field [0001] The invention belongs to the technical fields of biotechnology, biochemistry and synthetic biology, and in particular relates to a preparation method of thylakoid-containing artificial cells and a simulation method of photosynthesis. Background technique [0002] Plant cells in nature contain many compartments, among which chloroplast is the key organelle for photosynthesis in plant cells. There are a large number of thylakoids inside the chloroplast, and the thylakoid membrane contains photosynthetic pigments and electron transport chain components. The conversion to active chemical energy (photoreaction)" is carried out on the thylakoid membrane, so the thylakoid membrane is also called the photosynthetic membrane. At present, there have been some reports on the use of phospholipid components, silica components, etc. to encapsulate thylakoid particles extracted from plants, but they are all in single-chamber spherical shells, which cannot meet the si...

Claims

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Application Information

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IPC IPC(8): C12N5/04
CPCC12N5/04
Inventor 韩晓军宗薇
Owner HARBIN INST OF TECH
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