PREPARATION OF SPHEROPLASTS OF AEROBACTER CLOACAE

Abstract

A method for the preparation of spheroplasts from Aerobacter cloacae has been devised. Washed cells are exposed to the combined action of lysozyme and 1.0 M tris buffer at pH 9.0 in 20% sucrose. Such treatment weakens the cell wall structure but most cells retain their rod shape. Subsequent dilution with three volumes of distilled water converts the rods instantaneously into spheres. The time of exposure to lysozyme required for satisfactory production of spheroplasts can be greatly reduced if the cells are preincubated with the tris buffer.