A WbpL mutant of Pseudomonas putida DOT-T1E strain, which lacks the O-antigenic side chain of lipopolysaccharides, is tolerant to organic solvent shocks
F. Junker et al., A WbpL mutant of Pseudomonas putida DOT-T1E strain, which lacks the O-antigenic side chain of lipopolysaccharides, is tolerant to organic solvent shocks, EXTREMOPHIL, 5(2), 2001, pp. 93-99
Lipopolysaccharides (LPS) are major components of the outer membrane of gra
m-negative bacteria and are considered a defense barrier. To determine if L
PS play a role in resistance to solvents in the solvent-tolerant Pseudomona
s putida DOT-T1E strain, we have generated mutants unable to synthesize the
O-antigen side chain of LPS. The wbpL gene, encoding the enzyme that begin
s the synthesis of the O-antigen side chain of LPS of the solvent-tolerant
strain, was cloned, sequenced, and knocked out in vitro with a cassette enc
oding kanamycin resistance, and a mutant called WbpL0 of the DOT-T1E strain
was generated in vivo by site-directed mutagenesis. The WbpL mutant was co
mpared with the wild-type strain with regard to tolerance to a number of to
xic compounds, including chelating agents, organic acids, detergents, and a
romatic hydrocarbons. It was found that the mutant was as tolerant as the w
ild-type strain to organic acids and aromatic hydrocarbons and more sensiti
ve to ethylenediaminetetraacetic acid and deoxycholate.