Separation of Amylase with Protein Gel (SDS-Gel)

Application of different protein samples after 80% ammonium-sulfate precipitation and dialysis. S (Standard): Fermentas Prestrained protein marker (#Sm0671), amount applicated: 5µl;
LB: full medium LB;
M9: minimal medium M9;
M: Methionine; E:Ethionine;
N: Norleucine.

20 µl of each substance were applicated on the Gel (+ 5µl 5 x SDS buffer). Amylase bands are marked with a red dashed box.

In addition to the catalase, the Bacillus subtilis also segregates amylase E into the culture medium.  It has a mass of approx. 70kDa and is noted with a red box. Because of the low levels in respect to the over-all protein level of the individual probes, a comparison of the different amounts of amylase is hardly possible. This could be made possible with specific ethanol glycogen precipitation and is currently being attempted at the Max-Planck-Institute. The Gel does show however, that a synthetic amylase with Ethionine (or Norleucine) is present in the probes.