Method for improving expression activity of aquaporin in vesicle, and reverse osmosis membrane

A technology of aquaporin and channel protein, which is applied to a method of improving the expression activity of aquaporin in vesicles and the field of reverse osmosis membranes, which can solve the problems of reducing the activity of aquaporin, decreased activity, and insufficient binding force, etc. Achieve the effect of improving water flux and desalination rate, and improving expression activity

Inactive Publication Date: 2017-07-07
宁波日新恒力科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] (1) The number of aquaporins that can be embedded in a vesicle is limited;
[0004] (2) Loss of aquaporin due to incomplete loading of vesicles on aquaporin or insufficient binding force;
[0005] (3) The activity of aquaporin is reduced or even lost in contact with the basement membrane;
[0006] (4) During the process of vesicle binding to the surface of the basement membrane, the difference in the surface morphology of the basement membrane reduces the activity of aquaporins
[0010] However, when aquaporin is directly embedded into the vesicle, the amount of aquaporin itself embedded is small, and the loading of vesicle on aquaporin is not complete or the binding force is not strong enough or the aquaporin is in contact with the basement membrane or the basement membrane The effect of surface morphology on vesicles, etc. In the end, there are very few aquaporins that really play an active role, and the performance of aquaporin reverse osmosis membranes is far from reaching the level where all aquaporins in the membrane can exert their activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Pipette 10mL of 1mg / mL aquaporin solution into a 100mL quantitative flask, add 10ml of glycerol, 1mmol of protein kinase A and 0.5mmol of cyclic AMP kinase, 10mmol of ATP, and calibrate with phosphate buffer with a pH value of 7.5 To the mark, prepare 0.1mg / mL aquaporin stock solution.

[0041] Extract 10ml of the above-mentioned aquaporin stock solution, add 0.2ml of 0.01mol / L calcium chloride solution dropwise, and add phosphoric acid dropwise to adjust the pH value of the solution to 6.5, and add 0.1% polyethylene glycol stearyl ether at a volume concentration of 0.1 ml, centrifuge, remove the supernatant, leave the protein body, and repeat this 4 times.

[0042] Pipette DOPC, add phosphate buffer solution, and oscillate ultrasonically for 30 minutes to prepare a phospholipid vesicle system solution with a concentration of 0.1 mg / mL.

[0043] Mix the aquaporin body with the prepared phospholipid vesicle solution in a ratio of 1:100, prepare phosphorylated aquaporin ...

Embodiment 2

[0045]Pipette 10mL of 1mg / mL aquaporin solution into a 100mL quantitative flask, add 10mL of glycerol, 1mmol of protein kinase A and 0.5mmol of cyclic AMP kinase, 10mmol of ATP, and calibrate with phosphate buffer with a pH value of 7.5 To the mark, prepare 0.1mg / mL aquaporin stock solution. Extract 10ml of the above-mentioned aquaporin stock solution, add 0.2mL of 0.01mol / L calcium chloride solution dropwise, add phosphoric acid dropwise to adjust the pH value of the solution to 6.5, add 0.1% polyethylene glycol stearyl ether with a volume concentration of 0.1mL , centrifuge, remove the supernatant, leave the protein bodies, and repeat this 4 times. Pipette an appropriate amount of DOPC, add phosphate buffer solution, and oscillate ultrasonically for 30 minutes to prepare a phospholipid vesicle system solution with a concentration of 0.1 mg / mL. Mix the aquaporin body with the prepared phospholipid vesicle solution in a ratio of 1:20, prepare phosphorylated aquaporin vesicles...

Embodiment 3

[0047] Pipette 10mL of 1mg / mL aquaporin solution into a 100mL quantitative flask, add 10mL of glycerol, 1mmol of protein kinase C and 0.5mmol of cyclic AMP kinase, 10mmol of ATP, and calibrate with phosphate buffer with a pH value of 7.5 To the mark, prepare 0.1mg / mL aquaporin stock solution. Extract 10ml of the above-mentioned aquaporin stock solution, add 0.2mL of 0.01mol / L calcium chloride solution dropwise, add phosphoric acid dropwise to adjust the pH value of the solution to 6.5, add 0.1% polyethylene glycol stearyl ether with a volume concentration of 0.1mL , centrifuge, remove the supernatant, leave the protein bodies, and repeat this 4 times. Pipette an appropriate amount of DOPC, add phosphate buffer solution, and oscillate ultrasonically for 30 minutes to prepare a phospholipid vesicle system solution with a concentration of 0.1 mg / mL. Mix the aquaporin body with the prepared phospholipid vesicle solution in a ratio of 1:100, prepare phosphorylated aquaporin vesicl...

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PUM

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Abstract

The invention discloses a method for improving the expression activity of aquaporin in a vesicle. The method comprises the following steps: 1, performing phosphorylation on the aquaporin and preparing a phospholipid vesicle system solution; and 2, introducing the phosphorylated aquaporin. The invention also provides an aquaporin reverse osmosis membrane. The aquaporin reverse osmosis membrane is prepared by using the phosphorylated aquaporin vesicle prepared by the above method. The expression activity of the aquaporin is improved through the phosphorylation of the aquaporin; and the phosphorylation modified aquaporin is embedded into the vesicle and applied to the reverse osmosis membrane, so under the condition of achieving water flux and desalinization rate with the same level, the ratio of the used aquaporin to the vesicle is greatly reduced.

Description

technical field [0001] The invention relates to the technical field of water treatment, in particular to a method for increasing the expression activity of aquaporin in vesicles and a reverse osmosis membrane. Background technique [0002] At present, the ultimate goal of the research on aquaporin reverse osmosis membrane is to improve the water flux and desalination rate of the membrane, which is closely related to aquaporin. The loading and activity of aquaporin directly affect the performance of the membrane, often The greater the loading and the higher the activity of aquaporin, the better the performance of the membrane. However, during the actual preparation of aquaporin reverse osmosis membranes, the loading and activity of aquaporins are far below the theoretical level. The reasons are usually as follows: [0003] (1) The number of aquaporins that can be embedded in a vesicle is limited; [0004] (2) Loss of aquaporin due to incomplete loading of vesicles on aquap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00C07K1/107B01D61/02
CPCC07K14/00B01D61/025C07K1/107
Inventor 不公告发明人
Owner 宁波日新恒力科技有限公司
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