The present study aims to detection optimal conditions of production of amylase enzyme from isolate of B. subtillis A4. Nine carbonic sources were represented by starch, maltose, fructose, sucrose, glucose, arabinose, xylose, sorbitol and mannitol) at concentration of 1% for each source. It was found that the best was represented by starch carbonic, which showed higher activity and qualitative activity of 7.647 Unit/ ml and 461.56 Unit/ mg. Ten nitrogen sources were selected, including yeast extract, peptone, trypton, gelatin, urea and meat extract as organic sources Ammonium sulphate, Sodium nitrate, Potassium nitrate and Ammonium chloride as inorganic sources. These sources were added at aconcentration of 0.5% to the production medium. The Peptone source was the best of used nitrogenic sources at concentration of 1%, which showed high activity and qualitative activity at 9.651 Unit/ ml and 506.29 Unit/ mg. The best hydrogen number was 7 there inculcation which showed higher activity and qualitative activity of 14.3 Unit/ ml and 569.38 Unit/ mg. The size of inoculums of 2 × 108 cell was the best which showed high activity and qualitative activity of 16.213 Unit/ ml and 592.9 Unit/ mg.