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1. Diaminopimelate decarboxylase from a soluble extract of Escherichia coli A.T.C.C. 9637 was purified 200-fold by precipitation of nucleic acids, fractionation with acetone and then with ammonium sulphate, adsorption on calcium phosphate gel and chromatography on DEAE-cellulose or DEAE-Sephadex. 2. The purified enzyme showed only one component in the ultracentrifuge, with a sedimentation coefficient of 5.4s. One major peak and three much smaller peaks were observed on electrophoresis of the enzyme at pH8.9. 3. The mol.wt. of the enzyme was approx. 200000. The catalytic constant was 2000mol. of meso-diaminopimelic acid decomposed/min./mol. of enzyme, at 37 degrees . The relative rates of decarboxylation at 25 degrees , 37 degrees and 45 degrees were 0.17:1.0:1.6. At 37 degrees the Michaelis constant was 1.7mm and the optimum pH was 6.7-6.8. 4. There was an excess of acidic amino acids over basic amino acids in the enzyme, which was bound only on basic cellulose derivatives at pH6.8. 5. The enzyme had an absolute requirement for pyridoxal phosphate as a cofactor; no other derivative of pyridoxine had activity. A thiol compound (of which 2,3-dimercaptopropan-1-ol was the most effective) was also needed as an activator. 6. In the presence of 2,3-dimercaptopropan-1-ol (1mm), heavy-metal ions (Cu(2+), Hg(2+)) did not inhibit the enzyme, but there was inhibition by several amino acids with analogous structures to diaminopimelate, generally at high concentrations relative to the substrate. Penicillamine was inhibitory at relatively low concentrations; its action was prevented by pyridoxal phosphate.

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Aims:  To investigate the effect of extrinsic control parameters for ozone inactivation of E. coli in a bubble column. Methods and Results:  Ozone inactivation of Escherichia coli ATCC 25922 in Tryptic Soya Broth was examined. The parameters studied included temperature (ambient, 20, 25 and 30°C), exposure time (up to 30 min), gas flow rate (0·03, 0·06, 0·12, 0·25, 0·5 and 0·75 l min−1) and concentration level (five different levels). The efficacy of ozone treatment was a function of the parameters investigated and optimum control parameters of flow rate (0·12 l min−1), temperature (ambient) and ozone concentration (75 μg ml−1) resulted in a td5 (time required to achieve 5 log reduction) of 20 min. Conclusions:  Optimum control parameters of gas flow rate, ozone concentration and temperature are reported for E. coli inactivation within a bubble column. Significance and Impact of the Study:  In 2001, the FDA approved use of ozone as a direct additive to food and in 2004, issued guidelines for the use of ozone in liquid systems. However, these guidelines highlighted gaps in the literature for ozonation of liquid foods. This study provides useful information regarding optimum extrinsic control parameters for E. coli inactivation in liquid media using a bubble column to ensure microbiological safety.