Production and purification of acetylcholinesterase inhibitor from Pseudomonas sp. 960903

摘要

To screen agent for the treatment of Alzheimer's Disease several strains of bacteria producing acetylcholinesterase inhibitor were isolated from soil. Strain 960903 showed strong acetylcholinesterase inhibitory activity and low butyrylcholinesterase inhibitory activity. The strain 960903 was identified as Pseudomonas sp. Acetylcholincsterase inhibitor was highly achieved in fermentation medium containing soluble starch 3.0 percent, glycerol 1.0 percent, pharmamedia 0.5 percent, KCl 0.3 percent, CaCO_3 0.2 percent, MgSO_4 centre dot 7H_2O 0.05 percent, KH_2PO_4 0.05 percent (pH 6.5) at 30 deg C for 4 days. Acetylcholinesterase inhibitor was purified by Diaion WA-30 (OH~-) column chromatography and cellulose column chromatography. Acetylcholinesterase inhibitor showed the maximum wavelength at 205 nm and was soluble in water, acetic acid, ethanol, methanol and dimethyl sulfoxide. The concentration of 50 percent inhibition (IC_(50)) of inhibitor against acetylcholinesterase was 25 #mu#g/ml. The inhibitor was inactivated on heating at 100 deg C for 15 min and more stable in acidic region than alkaline region.