Unlike bacterial and mammalian cells, carrot cells are able to tolerate N-phosphonoacetyl-L-aspartate (PALA), a potential inhibitor of pyrimidine biosynthesis, by detoxifying the compound. Anion-exchange chromatography showed that detoxified PALA was less negatively charged than PALA, and allowed detoxified PALA to be isolated. Incubation of detoxified PALA with a low-specificity carboxylic-ester hydrolase fully restored the ability to inhibit aspartate transcarbamoylase, the target enzyme, indicating that the detoxification involves the formation of carboxylic ester. G.1.c. analysis of the alcohol products of enzymic hydrolysis, and of their ratio to PALA, showed that the detoxification produced a mixture of mono- and di-carboxylic esters and of methyl and ethyl esters. The detoxification mechanism showed considerable specificity towards PALA, since the analogous carboxy groups of succinate were not modified in the same way. Succinate was depleted much more slowly, no succinate esters could be detected, and the presence of a 10-fold excess of succinate did not inhibit the esterification rate of PALA. The possible significance of these results is discussed.
