Purification and characterization of 4-N-trimethylamino-1-butanol dehydrogenase of Pseudomonas sp. 13CM

Maizom Hassan, Sachiko Morimoto, Hiroyuki Murakami, Tsuyoshi Ichiyanagi, Nobuhiro Mori

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A new enzyme, NAD+-dependent 4-N-trimethylamino-1-butanol dehydrogenase from Pseudomonas sp. 13CM, was purified 526-fold to apparent homogeneity in 5 chromatographic steps. The enzyme had a molecular mass of 45kDa and appeared to be a monomer enzyme. The isoeletric point was found to be 4.8. The optimum temperature was 50°C, and the optimum pHs for the oxidation and reduction reactions were 9.5 and 6.0 respectively. The purified enzyme was further characterized with respect to substrate specificity, kinetic parameters, and amino acid terminal sequence. The Km values for trimethylamino-1-butanol and NAD+ were 0.54 mM and 0.22 mM respectively. In the reduction reaction, the apparent Km values for trimethylaminobutylaldehyde and NADH were 0.67 mM and 0.04 mM, respectively. The enzyme was inhibited by SH reagents, chelating reagents, and heavy metal ions. The N-terminal 12 amino acid residues were sequenced.

Original languageEnglish
Pages (from-to)1439-1446
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume71
Issue number6
DOIs
Publication statusPublished - 2007
Externally publishedYes

Fingerprint

butanol
Butenes
Pseudomonas
Purification
Enzymes
NAD
enzymes
Amino acids
Heavy Ions
Amino Acids
Sulfhydryl Reagents
1-Butanol
amino acids
Molecular mass
substrate specificity
Substrate Specificity
Heavy Metals
Chelation
Heavy ions
metal ions

Keywords

  • Alcohol dehydrogenase
  • Choline
  • Pseudomonas sp.
  • Quaternary ammonium compound

ASJC Scopus subject areas

  • Bioengineering
  • Biotechnology
  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry (miscellaneous)
  • Applied Microbiology and Biotechnology
  • Food Science

Cite this

Purification and characterization of 4-N-trimethylamino-1-butanol dehydrogenase of Pseudomonas sp. 13CM. / Hassan, Maizom; Morimoto, Sachiko; Murakami, Hiroyuki; Ichiyanagi, Tsuyoshi; Mori, Nobuhiro.

In: Bioscience, Biotechnology and Biochemistry, Vol. 71, No. 6, 2007, p. 1439-1446.

Research output: Contribution to journalArticle

Hassan, Maizom ; Morimoto, Sachiko ; Murakami, Hiroyuki ; Ichiyanagi, Tsuyoshi ; Mori, Nobuhiro. / Purification and characterization of 4-N-trimethylamino-1-butanol dehydrogenase of Pseudomonas sp. 13CM. In: Bioscience, Biotechnology and Biochemistry. 2007 ; Vol. 71, No. 6. pp. 1439-1446.
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AU - Mori, Nobuhiro

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