
Acrasin

Each species of slime mold has its own specific chemical messenger, which are collectively referred to as acrasins. These chemicals signal that many individual cells aggregate to form a single large cell or plasmodium. One of the earliest acrasins to be identified was cAMP, found in the species "Dictyostelium discoideum" by Brian Shaffer, which exhibits a complex swirling-pulsating spiral pattern when forming a pseudoplasmodium.
The term acrasin was descriptively named after Acrasia from Edmund Spenser's Faerie Queene, who seduced men against their will and then transformed them into beasts. Acrasia is itself a play on the Greek akrasia that describes loss of free will.
Extraction.
Methodology for the successful Chemical extraction of Acrasin was first described by Brian Shaffer in 1954 during his tenure at Princeton University in 1954. This involves the pouring of cold methanol over culture plates in which the aggregation of amoebas is general. This was dried in a vacuum at- 10 °C, and the residue was extracted with a small volume of methanol, which was then filtered and once again dried. The resultant product contains high acrasin activity. Schaffer confirmed the stability of the product by exposure to boiling temperature, as well as strong acids and bases.
Purification of acrasin.
Acrasin is a general term for a chemical substance that causes cells to aggregate, or gather. This experiment is to collect and purify a sample of Glorin, which is the acrasin of the P. violaceum species.
GLORIN COLLECTION: P. violaceum were grown and washed in a 1% salt solution. They were then diluted in a 50% salt solution, and 15 mL samples of this solution were placed on petri dishes to allow the P. violaceum to grow and aggregate. After fully aggregated, 40% ethanol was poured over each Petri dish to inhibit acrasinase, and the ethanol was collected and concentrated. (Acrasinase would kill the glorin, so ethanol was needed to inhibit it).
PURIFICATION OF GLORIN: Extract was filtered with water to remove acidic impurities. Whatever was filtered out was mixed with ethanol, and then centrifuged and dried to yield the product. The product was then dissolved in 50% ethanol to prepare for column chromatography.