ABSTRACT
Two-dimensional gel electrophoretic analysis of the membrane fraction of
Acetobacter aceti
revealed the presence of several proteins that were produced in response to acetic acid. A 60-kDa protein, named AatA, which was mostly induced by acetic acid, was prepared;
aatA
was cloned on the basis of its NH
2
-terminal amino acid sequence. AatA, consisting of 591 amino acids and containing ATP-binding cassette (ABC) sequences and ABC signature sequences, belonged to the ABC transporter superfamily. The
aatA
mutation with an insertion of the neomycin resistance gene within the
aatA
coding region showed reduced resistance to acetic acid, formic acid, propionic acid, and lactic acid, whereas the
aatA
mutation exerted no effects on resistance to various drugs, growth at low pH (adjusted with HCl), assimilation of acetic acid, or resistance to citric acid. Introduction of plasmid pABC101 containing
aatA
under the control of the
Escherichia coli lac
promoter into the
aatA
mutant restored the defect in acetic acid resistance. In addition, pABC101 conferred acetic acid resistance on
E. coli
. These findings showed that AatA was a putative ABC transporter conferring acetic acid resistance on the host cell. Southern blot analysis and subsequent nucleotide sequencing predicted the presence of
aatA
orthologues in a variety of acetic acid bacteria belonging to the genera
Acetobacter
and
Gluconacetobacter
. The fermentation with
A. aceti
containing
aatA
on a multicopy plasmid resulted in an increase in the final yield of acetic acid.