title: Identification of an L-alanine export system in Escherichia coli and isolation and characterization of export-deficient mutants.
abstract: An Escherichia coli strain that exhibits a double auxotrophy for L-alanine and D-alanine was constructed. During growth in the presence of the dipeptide L-alanyl-L-alanine (Ala-Ala), this was fully consumed with concomitant extracellular accumulation of l-alanine in a twofold molar concentration compared with the dipeptide. This finding indicates that the strain not only can hardly degrade L-alanine but has an export system(s) for L-alanine. To obtain access to the system, we chemically mutagenized the L-alanine-nonmetabolizing strain and isolated mutants with increased Ala-Ala sensitivity. Two such mutants accumulated L-alanine up to 150-190 mM in the cytoplasm with a reduced rate of L-alanine export relative to the parent strain in the presence of Ala-Ala. Furthermore, when chloramphenicol was added together with Ala-Ala, the parent strain accumulated L-alanine in the cytoplasm to a level similar to that observed in the mutants in the absence of chloramphenicol. In contrast, the intracellular l-alanine level in the mutants did not change irrespective of chloramphenicol treatment. From these results, we conclude that E. coli has an inducible l-alanine export carrier, together with a second, as yet unidentified, mechanism of alanine export.
authors.affiliation: Laboratory of Animal Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Hatsuhiro
authors.initials: H
authors.lastname: Hori
authors.forename: Tasuke
authors.initials: T
authors.lastname: Ando
authors.forename: Emiko
authors.initials: E
authors.lastname: Isogai
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yoneyama
authors.forename: Ryoichi
authors.initials: R
authors.lastname: Katsumata
mesh_terms: D000409:Alanine / Q000378:metabolism*; D001692:Biological Transport; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation
chemical_list: D029968:Escherichia coli Proteins; D000409:Alanine