3-Hydroxyisobutyrate Dehydrogenase fromPseudomonas putidaE23: Purification and Characterization
- 1 January 1996
- journal article
- Published by Taylor & Francis in Bioscience, Biotechnology, and Biochemistry
- Vol. 60 (12) , 2043-2047
- https://doi.org/10.1271/bbb.60.2043
Abstract
The NAD+-dependent 3-hydroxyisobutyrate dehydrogenase [EC 1.1.1.31] was purified to homogeneity from Pseudomonas putida E23. The enzyme was a tetramer (molecular mass, 120kDa) consisted of identical subunits (molecular mass, 30 kDa). The enzyme was specific for NAD+ (Km, 0.44 mm). The maximal activity was obtained at about pH 10. The enzyme was specific for the l-isomer of 3-hydroxyisobutyrate. In addition to l-3-hydroxyisobutyrate, l-serine, 2-methyl-dl-serine, and 3-hydroxypropionate were substrates. The Km for l-3-hydroxyisobutyrate, l-serine, 2-methyl-dl-serine, and 3-hydroxypropionate were 0.12, 18, 44, and 83 mm, respectively. The enzyme was inhibited by p-chloromercuribenzoate, HgCl2, and AgNO3, but not by EDTA, α,α′-dipyridyl, and o-phenanthroline. The N-terminal 26 amino acid sequence was compared with the sequences deduced from the enzyme genes of rat liver and Pseudomonas aeruginosa.Keywords
This publication has 1 reference indexed in Scilit:
- 2 Evolutionary and Structural Relationships among DehydrogenasesPublished by Elsevier ,1975