Preparation method for phosphatidase D
A phospholipase and barley technology, applied in biochemical equipment and methods, enzymes, hydrolytic enzymes, etc., can solve the problem that barley roots cannot be effectively used
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[0013] The present invention provides a kind of preparation method of phospholipase D, wherein, described preparation method comprises:
[0014] 1) Extracting the barley root with water to obtain the extract M1;
[0015] 2) After centrifuging the extract M1 at least once, take the supernatant M2;
[0016] 3) adding ammonium sulfate solution to the supernatant M2, mixing and salting out, centrifuging, and removing the lower layer to obtain phospholipase D.
[0017] In the present invention, the barley root is leached with water first, and then the leached product is centrifuged at least once to remove the centrifuged supernatant, and then ammonium sulfate solution is added to the supernatant for salting out, so that the barley root is further extracted The phospholipase D is extracted, and then centrifuged again, and the centrifuged precipitate is taken to obtain a product containing phospholipase D.
[0018] The consumption of each material here can be selected according to ...
Embodiment 1
[0027] 1) Extract 10 parts by weight of barley root powder and 100 parts by weight of water at 30°C for 15 hours to obtain extract M1;
[0028] 2) Centrifuge the extract M1 at a rate of 1500r / min for 5min, then centrifuge at a rate of 3000r / min for 3min, and take the supernatant M2;
[0029] 3) Add 50 parts by weight of ammonium sulfate solution (concentration: 30% by weight) to the supernatant M2, mix and salt out for 3 hours, centrifuge at a speed of 3000 r / min for 8 minutes, remove the lower layer of sediment, and prepare an enzyme solution containing phospholipase D A1. (enzyme activity is 97.2U / g)
Embodiment 2
[0031] 1) Extract 10 parts by weight of barley root powder and 300 parts by weight of water at 40°C for 30 hours to obtain extract M1;
[0032] 2) Centrifuge the extract M1 at a rate of 3000r / min for 20min, then centrifuge at a rate of 5000r / min for 10min, and take the supernatant M2;
[0033] 3) Add 200 parts by weight of ammonium sulfate solution (concentration: 30% by weight) to the supernatant M2, mix and salt out for 6 hours, centrifuge at a rate of 6000 r / min for 15 minutes, remove the lower layer of sediment, and prepare an enzyme solution containing phospholipase D A2. (enzyme activity is 93.5U / g)
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