Biochemical and genetic characterization of osmoregulatory trehalose synthesis in Escherichia coli.
It has been shown previously that Escherichia coli accumulates endogenously synthesized trehalose under osmotic stress. We report here that E. coli contained an osmotically regulated trehalose-phosphate synthase which utilized UDP-glucose and glucose 6-phosphate as substrates. In the wild type, the synthase was induced by growth in glucose-mineral medium of elevated osmotic strength and the synthase itself was strongly stimulated by K+ and other monovalent cations. A laboratory strain which expressed the synthase at a high constitutive level was found. GalU mutants, defective in synthesis of UDP-glucose, did not accumulate trehalose. Two genes governing the synthase were identified and named otsA and otsB (osmoregulatory trehalose synthesis). They mapped near 42 min in the flbB-uvrC region. Mutants with an otsA-lacZ or otsB-lacZ operon fusion displayed osmotically inducible beta-galactosidase activity; i.e., the activity was increased fivefold by growth in medium of elevated osmotic strength. Mutants unable to synthesize trehalose (galU, otsA, and otsB) were osmotically sensitive in glucose-mineral medium. But an osmotically tolerant phenotype was restored in the presence of glycine betaine, which also partially repressed the synthesis of synthase in the wild type and of beta-galactosidase in ots-lacZ fusion mutants.
D003001:Cloning, Molecular; D004187:Disaccharides / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D019298:Glucose-6-Phosphate; D005958:Glucosephosphates / Q000378:metabolism; D009876:Operon; D011188:Potassium / Q000494:pharmacology; D012486:Salmonella typhimurium / Q000378:metabolism; D014198:Trehalase / Q000378:metabolism; D014199:Trehalose / Q000096:biosynthesis* / Q000235:genetics; D014532:Uridine Diphosphate Glucose / Q000378:metabolism; D014882:Water-Electrolyte Balance; D001616:beta-Galactosidase / Q000378:metabolism
D004187:Disaccharides; D005958:Glucosephosphates; D019298:Glucose-6-Phosphate; D014199:Trehalose; D001616:beta-Galactosidase; D014198:Trehalase; D011188:Potassium; D014532:Uridine Diphosphate Glucose