title: Involvement of surface polysaccharides in the organic acid resistance of Shiga Toxin-producing Escherichia coli O157:H7.
abstract: In general, wild Escherichia coli strains can grow effectively under moderately acidic organic acid-rich conditions. We found that the Shiga Toxin-producing E. coli (STEC) O157:H7 NGY9 grows more quickly than a K-12 strain in Luria-Bertani (LB)-2-morpholinoethanesulphonic acid (MES) broth supplemented with acetic acid (pH 5.4). Hypothesizing that the resistance of STEC O157:H7 to acetic acid is as a result of a mechanism(s) other than those known, we screened for STEC mutants sensitive to acetic acid. NGY9 was subjected to mini-Tn5 mutagenesis and, from 50,000 colonies, five mutants that showed a clear acetic acid-sensitive phenotype were isolated. The insertion of mini-Tn5 in three mutants occurred at the fcl, wecA (rfe) and wecB (rffE) genes and caused loss of surface O-polysaccharide, loss of both O-polysaccharide and enterobacterial common antigen (ECA) and loss of ECA respectively. The other two mutants showed inactivation of the waaG (rfaG) gene but at different positions that caused a deep rough mutant with loss of the outer core oligosaccharide of lipopolysaccharide (LPS) as well as phenotypic loss of O-polysaccharide and ECA. With the introduction of plasmids carrying the fcl, wecA, wecB and waaG genes, respectively, all mutants were complemented in their production of O-polysaccharide and ECA, and normal growth was restored in organic acid-rich culture conditions. We also found that the growth of Salmonella LPS mutants Ra, Rb1, Rc, Rd1, Rd2 and Re was suppressed in the presence of acetic acid compared with that of the parents. These results suggest that the full expression of LPS (including O-polysaccharide) and ECA is indispensable to the resistance against acetic acid and other short chain fatty acids in STEC O157:H7 and Salmonella. To the best of our knowledge, this is a newly identified physiological role for O-polysaccharide and ECA as well as an acid resistance mechanism.
authors.affiliation: Department of Molecular Bacteriology, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Soumitra
authors.initials: S
authors.lastname: Barua
authors.forename: Takafumi
authors.initials: T
authors.lastname: Yamashino
authors.forename: Tadao
authors.initials: T
authors.lastname: Hasegawa
authors.forename: Keiko
authors.initials: K
authors.lastname: Yokoyama
authors.forename: Keizo
authors.initials: K
authors.lastname: Torii
authors.forename: Michio
authors.initials: M
authors.lastname: Ohta
mesh_terms: D019342:Acetic Acid / Q000494:pharmacology; D000818:Animals; D002238:Carbohydrate Epimerases / Q000235:genetics; D002264:Carboxylic Acids / Q000494:pharmacology*; D004251:DNA Transposable Elements; D024901:Drug Resistance, Multiple, Bacterial; D004926:Escherichia coli / Q000187:drug effects* / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics; D020868:Gene Silencing; D005816:Genetic Complementation Test; D005964:Glucosyltransferases / Q000235:genetics; D007658:Ketone Oxidoreductases / Q000235:genetics; D009097:Multienzyme Complexes / Q000235:genetics; D009154:Mutation; D019081:O Antigens / Q000235:genetics / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D012475:Salmonella / Q000187:drug effects / Q000235:genetics; D022621:Shiga Toxin / Q000235:genetics / Q000378:metabolism*; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics
chemical_list: D002264:Carboxylic Acids; D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D019081:O Antigens; D011135:Polysaccharides, Bacterial; C110682:wcaG protein, E coli; D022621:Shiga Toxin; D007658:Ketone Oxidoreductases; D005964:Glucosyltransferases; C098465:WaaG protein, E coli; D017855:Transferases (Other Substituted Phosphate Groups); C083617:wecA protein, E coli; D002238:Carbohydrate Epimerases; C100033:UDP acetylglucosamine-2-epimerase; C083982:wecB protein, E coli; D019342:Acetic Acid