Open AccessThermostable NADP + -Dependent Medium-Chain Alcohol Dehydrogenase from Acinetobacter sp. Strain M-1: Purification and Characterization and Gene Expression in Escherichia coli
Author(s) -
Akio Tani,
Yasuyoshi Sakai,
Takeru Ishige,
Nobuo Kato
Publication year - 2000
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.66.12.5231-5235.2000
Subject(s) - escherichia coli , biochemistry , gel electrophoresis , alcohol dehydrogenase , enzyme , biology , nucleic acid sequence , alcohol oxidoreductase , peptide sequence , chemistry , microbiology and biotechnology , gene , nad+ kinase
NADPH-dependent alkylaldehyde reducing enzyme, which was greatly induced byn -hexadecane, fromAcinetobacter sp. strain M-1 was purified and characterized. The purified enzyme had molecular masses of 40 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 160 kDa as determined by gel filtration chromatography. The enzyme, which was shown to be highly thermostable, was most active towardn -heptanal and could usen -alkylaldehydes ranging from C2 to C14 and several substituted benzaldehydes, including the industrially important compounds cinnamyl aldehyde and anisaldehyde, as substrates. ThealrA gene coding for this enzyme was cloned, and its nucleotide sequence was determined. The deduced amino acid sequence encoded by thealrA gene exhibited homology to the amino acid sequences of zinc-containing alcohol dehydrogenases from various sources. The gene could be highly expressed inEscherichia coli , and the product was purified to homogeneity by simpler procedures from the recombinant than from the original host. Our results show that this enzyme can be used for industrial bioconversion of useful alcohols and aldehydes.