PMID- 19193709
OWN - NLM
STAT- MEDLINE
DCOM- 20090625
LR  - 20131121
IS  - 1756-2651 (Electronic)
IS  - 0021-924X (Linking)
VI  - 145
IP  - 5
DP  - 2009 May
TI  - Gene cloning and expression of pyridoxal 5'-phosphate-dependent 
      L-threo-3-hydroxyaspartate dehydratase from Pseudomonas sp. T62, and 
      characterization of the recombinant enzyme.
PG  - 661-8
LID - 10.1093/jb/mvp023 [doi]
AB  - L-threo-3-Hydroxyaspartate dehydratase (L-THA DH, EC 4.3.1.16), which catalyses 
      the cleavage of L-threo-3-hydroxyaspartate (L-THA) to oxalacetate and ammonia, 
      has been purified from the soil bacterium Pseudomonas sp. T62. In this report, 
      the gene encoding L-THA DH was cloned and expressed in Escherichia coli, and the 
      gene product was purified and characterized in detail. A 957-bp nucleotide 
      fragment was confirmed to be the gene encoding L-THA DH, based on the agreement 
      of internal amino acid sequences. The deduced amino acid sequence, which belongs 
      to the serine/threonine dehydratase family, shows similarity to YKL218c from 
      Saccharomyces cerevisiae (64%), serine racemase from Schizosaccharomyces pombe 
      (64%) and Mus musculus (36%), and biodegradative threonine dehydratase from E. 
      coli (38%). Site-directed mutagenesis experiments revealed that lysine at 
      position 53 is an important residue for enzymatic activity. This enzyme exhibited 
      dehydratase activity specific only to L-THA [K(m) = 0.54 mM, V(max) = 39.0 
      micromol min(-1) (mg protein)(-1)], but not to other 3-hydroxyaspartate isomers, 
      and exhibited no detectable serine/aspartate racemase activity. This is the first 
      report of an amino acid sequence of the bacterial enzyme that acts on L-THA.
FAU - Murakami, Tomoko
AU  - Murakami T
AD  - Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido 
      University, Sapporo, Japan.
FAU - Maeda, Takayuki
AU  - Maeda T
FAU - Yokota, Atsushi
AU  - Yokota A
FAU - Wada, Masaru
AU  - Wada M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090204
PL  - England
TA  - J Biochem
JT  - Journal of biochemistry
JID - 0376600
RN  - 0 (Amino Acids)
RN  - 0 (Metals)
RN  - 0 (Nucleotides)
RN  - 0 (Recombinant Proteins)
RN  - 5V5IOJ8338 (Pyridoxal Phosphate)
RN  - EC 4.2.1.- (Hydro-Lyases)
RN  - EC 4.2.1.- (L-threo-3-hydroxyaspartate dehydratase)
RN  - K3Z4F929H6 (Lysine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acids/metabolism
MH  - Base Sequence
MH  - Catalytic Domain
MH  - Cloning, Molecular
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Genes, Bacterial
MH  - Hydro-Lyases/chemistry/*genetics/isolation & purification/metabolism
MH  - Hydrogen-Ion Concentration
MH  - Lysine/metabolism
MH  - Metals/metabolism
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Nucleotides/metabolism
MH  - Phylogeny
MH  - Pseudomonas/*enzymology/genetics
MH  - Pyridoxal Phosphate/*metabolism
MH  - Recombinant Proteins/*metabolism
MH  - Sequence Alignment
MH  - Spectrum Analysis
MH  - Substrate Specificity
MH  - Temperature
EDAT- 2009/02/06 09:00
MHDA- 2009/06/26 09:00
CRDT- 2009/02/06 09:00
PHST- 2009/02/06 09:00 [entrez]
PHST- 2009/02/06 09:00 [pubmed]
PHST- 2009/06/26 09:00 [medline]
AID - mvp023 [pii]
AID - 10.1093/jb/mvp023 [doi]
PST - ppublish
SO  - J Biochem. 2009 May;145(5):661-8. doi: 10.1093/jb/mvp023. Epub 2009 Feb 4.