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(TAC). In a high osmotic medium involving NaCl, however, AZCand DHP showed strong growth inhibition, and these inhibitions were completely reversed by proline. By isolating analog-resistant mutants under this high osmotic condition, we were able to construct prolinehyperproducing strains. Wesupposed that the increase in analog sensitivity was due to the increase in analog content of the cells. This paper deals with changes in the intracellular concentration of proline analog during high osmotic stress. The strain used in this study was S. marcescens strain SP103,2) a proline oxidase-deficient mutant of wild strain 8000.4) The minimal medium was modified Davis-Mingioli medium as described previously.5) Growth experiments were performed with a Hitachi automated recording incubator system as reported previously.5) The intracellular analog or proline was extracted with hot water as described previously.3) In a high osmotic medium, NaCl was added during exponential growth, and the cells were cultured further for 1 hr. Cells were washed with the same osmotic stressed medium. Proline was determined by a microbioassay using Leuconosioc mesenteroides P-60.

Increase in the intracellular concentration of a proline analog during high osmotic stress in Serratia marcescens.