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Identification of an L-alanine Export System in Escherichia Coli and Isolation and Characterization of Export-Deficient Mutants

Identification of an L-alanine Export System in Escherichia Coli and Isolation and Characterization of Export-Deficient Mutants

Hatsuhiro Hori, Tasuke Ando, Emiko Isogai, Hiroshi Yoneyama, Ryoichi Katsumata

FEMS Microbiol Lett. 2011 Mar;316(2):83-9.

PMID: 21208269

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP1948318	L-Alanyl-L-alanine		1948-31-8	Price

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