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A genomic clone bank ofAcetobacter polyoxogenes NBI1028 constructed inEscherichia coli by use of the expression vector pUC18 was screened with antibody raised against membrane-bound aldehyde dehydrogenase (ALDH; 75 kilodaltons [kDa]) fromA. polyoxogenes NBI1028. A clone that synthesized a 41-kDa protein cross-reactive with anti-ALDH antibody was isolated. For cloning of the full-length ALDH structural gene, a cosmid gene bank was screened by Southern blot hybridization with the cloned DNA as a probe, and subcloning from the positive cosmid clone was performed with shuttle vector pMV24. Plasmid pAL25, containing the full-length ALDH structural gene, was isolated and expressed in bothE. coli andAcetobacter aceti to produce a fused protein (78 kDa) with a short NH2 -terminal β-galactosidase peptide. pAL25 conferred ALDH production on a mutant ofA. aceti lacking the enzyme activity. Transformation ofA. aceti subsp.xylinum NBI2099 with pAL25 caused 2- and 1.4-fold increases in the production rate and in the maximum concentration of acetic acid in submerged fermentation, respectively.

applied and environmental microbiology

Cloning of the Membrane-Bound Aldehyde Dehydrogenase Gene of <i>Acetobacter polyoxogenes</i> and Improvement of Acetic Acid Production by Use of the Cloned Gene

Cloning of the Membrane-Bound Aldehyde Dehydrogenase Gene of Acetobacter polyoxogenes and Improvement of Acetic Acid Production by Use of the Cloned Gene