Gene cloning and characterization of aldehyde dehydrogenase from a petroleum-degrading bacterium, strain HD-1

Naoko Okibe, Kei Amada, Shin Ichi Hirano, Mitsuru Haruki, Tadayuki Imanaka, Masaaki Morikawa, Shigenori Kanaya

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The hd-ald gene encoding aldehyde dehydrogenase (hd-ALDH) from an mixotrophic petroleum-degrading bacterium, strain HD-1 was cloned and sequenced, hd-ALDH (506 amino acids) is a member of the NAD+-dependent aldehyde dehydrogenase group. The hd-ald gene was expressed in Escherichia coli, and the recombinant enzyme was purified and characterized biochemically and enzymatically. The molecular weight of the enzyme was estimated to be 55,000 by SDS-PAGE, and 224,000 by gel filtration chromatography, suggesting that it acts as a tetramer. The CD spectrum suggests that the helical content of the enzyme is 10%. hd-ALDH was active on various aliphatic aldehyde substrates. The K(m) values of the enzyme were 6.4 μM for acetaldehyde, 4.2 μM for hexanal, 2.8 μM for octanal, and 0.84 μM for decanal, whereas the k(cat) values for these substrates were nearly equal (51-64 min-1). These results indicate that hd-ALDH acts preferentially on long-chain aliphatic aldehydes.

Original languageEnglish
Pages (from-to)7-11
Number of pages5
JournalJournal of Bioscience and Bioengineering
Volume88
Issue number1
DOIs
Publication statusPublished - Jan 1 1999
Externally publishedYes

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Aldehyde Dehydrogenase
Cloning
Petroleum
Aldehydes
Organism Cloning
Bacteria
Enzymes
Crude oil
Genes
Gene encoding
Acetaldehyde
Substrates
Chromatography
NAD
Escherichia coli
Gel Chromatography
Amino acids
Polyacrylamide Gel Electrophoresis
Gels
Molecular Weight

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Gene cloning and characterization of aldehyde dehydrogenase from a petroleum-degrading bacterium, strain HD-1. / Okibe, Naoko; Amada, Kei; Hirano, Shin Ichi; Haruki, Mitsuru; Imanaka, Tadayuki; Morikawa, Masaaki; Kanaya, Shigenori.

In: Journal of Bioscience and Bioengineering, Vol. 88, No. 1, 01.01.1999, p. 7-11.

Research output: Contribution to journalArticle

Okibe, Naoko ; Amada, Kei ; Hirano, Shin Ichi ; Haruki, Mitsuru ; Imanaka, Tadayuki ; Morikawa, Masaaki ; Kanaya, Shigenori. / Gene cloning and characterization of aldehyde dehydrogenase from a petroleum-degrading bacterium, strain HD-1. In: Journal of Bioscience and Bioengineering. 1999 ; Vol. 88, No. 1. pp. 7-11.
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