Earthworm Freeze Dryer

Earthworm Freeze Dryer

Dried earthworms, royal jelly, ginseng, turtles, turtles and other nutritional and health products, using vacuum freeze-drying process, pure and natural.

The method for preparing earthworm freeze-dried powder by vacuum freeze-drying mechanism is characterized in that it comprises the following steps:

Step 1：Preparation of earthworm coarse powder: the live earthworm is starved for one day, the sediment is naturally spitted out, washed, then dried and crushed under the ventilation condition with the temperature between 2035 ℃, and the earthworm coarse powder is made through 100200 mesh sieve;

Step 2: preparation of crude enzyme solution: take earthworm crude powder, homogenize it with 2 times the volume of 0.1mol/l phosphate buffer PBS with pH 7.8, add 0.1mol/lnacl solution, put it in a refrigerator at 4 ℃ overnight, then centrifuge at 10000r / min at 4 ℃ for 15min, discard the precipitation and take the supernatant; The supernatant was kept warm in a water bath at 50 ℃ for 40 min, cooled by running water, then centrifuged at 4 ℃ for 10 min, took the supernatant, added ammonium sulfate to the supernatant to reach 30% saturation, stood for 3 hours, then centrifuged at 4 ℃ for 10 min at 8000 R / min; Collect the supernatant, add ammonium sulfate to reach 70% saturation, stand for 3 hours, 8000r / min, centrifuge at 4 ℃ for 10 min, and collect the precipitation; Dissolve the precipitate in the buffer solution, dialysis the buffer solution with a dialysis bag containing 5kd molecular weight overnight to remove the salt, then centrifuge the dialysate at 8000r / min at 4 ℃ for 10min, and take the supernatant as the crude earthworm plasmin solution;

Step 3： purification of crude enzyme: the earthworm fibrinolytic enzyme solution was passed through Sephadex G75 gel chromatography, pre column with 0.02mol/L and pH as the buffer buffer of 7.8 phosphate. After the column was removed, the above buffer was eluted to no light absorption at 280nm wavelength, and elution liquid appeared 2 protein peaks, the peak of enzyme activity overlapped with first protein peaks, and the overlapping part of the enzyme fluid was collected. Pass through deaecellulose a52 ion exchange column, and balance with PBS of 0.05mol/l and pH 7.8 before passing through the column; Then it was eluted with PBS ion strength linear gradient containing 00.5mol/l NaCl. There were three protein peaks in the eluent, the enzyme activity peak overlapped with the third peak of the protein peak, and the other two protein peaks basically had no enzyme activity. The collected active part was earthworm plasmin pure enzyme;

Step 4： Modification of earthworm plasmin: add earthworm plasmin pure enzyme and activated peg to 15ml, pH 9.2, 0.1mol/l boric acid buffer. The weight ratio of pure enzyme to peg is 1:50. Keep the temperature for 1 hour at 30 ℃, and the reactant dialyzes 0.1mol/l, pH 7.3 phosphate buffer overnight. The dialysate is centrifuged at 10000r / min and 4 ℃ for 10min. Take the supernatant and pass through the column Sephadex G75, Collect the active components, namely earthworm plasmin modifying enzyme;

Step 5: preparation of earthworm fibrinolytic enzyme lyophilized powder: put earthworm fibrinolytic enzyme modified enzyme in the sample tray with a material thickness of no more than 8mm, put it into the drying room of vacuum Earthworm Freeze Dryer for freeze-drying, freeze it at - 35 ℃ for 12 hours, and then raise the temperature to 30 ℃ under a vacuum of 5 Pa and dry it for 12 hours; The freeze-dried powder of earthworm plasmin modified enzyme was obtained by heating to 40 ℃ and vacuum drying for 14 hours. The water content was 7.0%, the protein content was 68%, and the enzyme activity was 1700u / mg.

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Pre-sale:

* As soon as we get your inquiry, we will always reply to your email on time and try our best to recommend you the most suitable freeze dryer. Detailed tech-info will also be provided for you.

* We can do sample test for customers.

After-sale:

* If possible, please do come to our factory and we will teach workers the whole process of freeze drying. The most appropriate time needed for freeze drying will be tested by us.

* We will keep in touch with you, to make sure every parts of the freeze dryer work well. One year warranty, free online service for abroad customers.

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