Unlock instant, AI-driven research and patent intelligence for your innovation.

A kind of enzyme-phospholipid microcapsule preparation method beneficial to membrane enzyme mosaic and its activity research

A technology of phospholipid microcapsules and activity, which is applied in the field of scientific research and experimental research on membrane enzyme activity, can solve the problems of small phospholipid microcapsules, unfavorable enzyme protein embedding, technical complexity, etc., and achieve the effects of improved fluidity, good stability, and large volume

Active Publication Date: 2018-10-12
LINYI UNIVERSITY
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem solved by the present invention is: the previous method for forming artificial phospholipid microcapsules is often technically complex, and the formed phospholipid microcapsules are small, which is not conducive to embedding enzyme protein on the microcapsule membrane

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A method for preparing enzyme-phospholipid microcapsules that is beneficial to the research on the activity of bovine heart mitochondrial inner membrane complex III mainly includes the following steps:

[0035] S1. Preparation of phospholipid microparticles:

[0036] (1) Suspend 40 mg of soybean lecithin washed with acetone in 1 ml of buffer, and add sodium cholate to a final concentration of 2%, and control the temperature at 27°C during the process; the buffer has a pH of 7.0 and a concentration of 0.02 mol / L disodium hydrogen phosphate-sodium dihydrogen phosphate buffer;

[0037] (2) Ultrasonic treatment on ice for 5 minutes: put the solution into a container, and use a 20khz probe-type pulsed ultrasonic wave for treatment; the treatment process is as follows: the probe is inserted 1.3cm below the liquid, the liquid level is kept at 6cm, and the pulse time is 5s, accounting for The air ratio is 40%, the temperature is 8°C, and the sound intensity is 150.22W / cm; duri...

Embodiment 2

[0049] A method for preparing enzyme-phospholipid microcapsules, which is beneficial to the research on the activity of bovine heart mitochondrial inner membrane complex IV, mainly includes the following steps:

[0050] S1. Preparation of phospholipid microparticles:

[0051] (1) Suspend 40 mg of soybean lecithin washed with acetone in 1 ml of buffer, and add sodium cholate to a final concentration of 2%, and control the temperature at 27.5°C during the process; the buffer in which has a pH of 7.4 and a concentration of 50 mM Phosphate buffer;

[0052](2) Ultrasonic treatment on ice for 5 minutes: put the solution into a container, and use 20khz probe-type pulsed ultrasonic treatment; the treatment process is as follows: the probe is inserted 1.3cm below the liquid, the liquid level is kept at 8cm, and the pulse time is 6s, accounting for The air ratio is 55%, the temperature is 17°C, and the sound intensity is 411.28W / cm; during ultrasonic treatment, argon gas must be contin...

Embodiment 3

[0064] A method for preparing enzyme-phospholipid microcapsules, which is beneficial to the research on the activity of bovine heart mitochondrial inner membrane complex IV, mainly includes the following steps:

[0065] S1. Preparation of phospholipid microparticles:

[0066] (1) Suspend 40 mg of soybean lecithin washed with acetone in 1 ml of buffer, and add sodium cholate to a final concentration of 2%, and control the temperature at 28°C during the process; the buffer in which has a pH of 7.3 and a concentration of 100 mM Hepes-KOH buffer;

[0067] (2) Ultrasonic treatment on ice for 5 minutes: put the solution into a container, and use a 20khz probe-type pulsed ultrasonic wave for treatment; the treatment process is as follows: the probe is inserted 1.3cm below the liquid, the liquid level is maintained at 10cm, and the pulse time is 8s, accounting for The air ratio is 70%, the temperature is 26°C, and the sound intensity is 672.34W / cm; during ultrasonic treatment, argon ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
thicknessaaaaaaaaaa
Login to View More

Abstract

The invention discloses a method for preparing enzyme-phospholipid micro-capsules beneficial to research of membrane enzyme inlaying and activity thereof. The method includes the steps of S1), preparation of phospholipid particles, namely 1), suspending soybean phospholipid washed by 40mg of acetone in a 1m1 buffer solution and adding sodium cholate to be 2% in concentration; 2), performing on-ice ultrasound treatment; S2), inlaying of enzyme protein in the phospholipid micro-capsules, namely 1), preparing an enzyme solution; 2), adding a sodium cholate solution to the enzyme solution to be 0.5% in concentration; 3), adding the phospholipid particles obtained in the S1 to enable the mass ratio of the enzyme protein to phospholipid to be 1:40; 4), performing on-ice incubation for 30 minutes; 5), performing on-ice overnight dialysis. The large single-layer micro-capsules formed in the method are large in size, good in stability, good in membrane mobility, favorable for inlaying of the membrane enzyme protein and suitable for research on activity and function of various membrane enzymes.

Description

technical field [0001] The invention relates to the technical field of membrane enzyme activity scientific research experiment research, in particular to an enzyme-phospholipid microcapsule preparation method that is beneficial to membrane enzyme mosaic and activity research. Background technique [0002] Biological cells, especially eukaryotic cells, have a well-developed biomembrane system. Biomembranes are phospholipid bilayers embedded with proteins and carbohydrates. In addition to the plasma membrane (also known as the cell membrane), eukaryotic cells also have membrane systems that separate various organelles, including nuclear membranes, mitochondrial membranes, endoplasmic reticulum membranes, lysosome membranes, Golgi apparatus membranes, chloroplast membranes, vacuolar membranes, Peroxisome membranes, etc. Biofilms are bimolecular-layer lamellar structures with a thickness of about 5-10 nanometers. The biomembrane plays a role in dividing and separating cells a...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68C12Q1/25
CPCC12Q1/25G01N33/68
Inventor 曲元刚
Owner LINYI UNIVERSITY