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
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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 ...
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