FIG. 1.

Loss of the PCE dehalogenase produced by S. multivorans subcultured in medium containing pyruvate (40 mM)/fumarate (40 mM) in the absence of PCE. Each 50-ml subculture was inoculated with 5 ml of a grown culture. (a) Immunological analysis of PceA in crude extracts (1 μg protein per lane) after separation by PAGE under denaturing conditions and subsequent blotting to a PVDF membrane; (b) corresponding reductive PCE dehalogenase activity of PceA. For experimental detail, see Materials and Methods. pre-PceA, 55.9-kDa unprocessed form bearing the N-terminal Tat signal peptide; PceA, 52.1-kDa, processed, exported form without the signal peptide.