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Temperature-Sensitive Mutants of Escherichia coli K-12 with Low Activities of the L-Alanine Adding Enzyme and the D-Alanyl-D-Alanine Adding Enzyme¹

^a Laboratory for Microbiology, State University, Catharijnesingel 59, Utrecht, The Netherlands

ABSTRACT

A number of properties of temperature-sensitive mutants in murein synthesis are described. The mutants grow at 30 C but lyse at 42 C. One mutant possesses a temperature-sensitive D-alanyl-D-alanine adding enzyme, has an impaired rate of murein synthesis in vivo at both 30 and 42 C, and contains elevated levels of uridine diphosphate-N-acetyl-muramyl-tripeptide (UDP-MurNAc-L-Ala-D-Glu-m-diaminopimelic acid) at 42 C. The other mutant possesses an L-alanine adding enzyme with a very low in vitro activity at both 30 and 42 C. Its in vivo rate of murein synthesis is almost normal at 30 C but is much less at 42 C. When the murein precursors were isolated after incubation of the cells in the presence of ¹⁴C-L-alanine, they contained only a fraction of the radioactivity that could be obtained from a wild-type strain. A genetic nomenclature for genes concerned with murein synthesis is proposed.

¹ This paper is taken from a thesis to be submitted by E. J. J. Lugtenberg to the Faculty of Science of the State University of Utrecht in partial fulfillment of the requirements for the Ph.D. degree.

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