Bacteria were grown aerobically on a rotary shaker
in 250-ml Erlenmeyer flasks containing a volume of
20 ml of culture medium. Anaerobic growth was in
jars (Torsion Balance Co., Clifton, N.J.) under a mixture
of 90% nitrogen and 10% carbon dioxide gas at
a pressure of 3 psi. Bacteria were always adapted to
experimental conditions of growth for at least five
generations. Cells were grown at 37 C, since growth at
either higher or lower temperatures yielded lower
amounts of asparaginase II.