Open AccessCrystallization and preliminary X‐ray diffraction studies of D‐ threo ‐3‐hydroxyaspartate dehydratase isolated from Delftia sp. HT23
Author(s) -
Matsumoto Yu,
Yasutake Yoshiaki,
Takeda Yuki,
Tamura Tomohiro,
Yokota Atsushi,
Wada Masaru
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113023956
Subject(s) - dehydratase , stereochemistry , racemization , orthorhombic crystal system , chemistry , lyase , crystallography , enzyme , crystal structure , biochemistry
D‐ threo ‐3‐Hydroxyaspartate dehydratase (D‐THA DH) isolated from the soil bacterium Delftia sp. HT23 is a novel enzyme consisting of 380 amino‐acid residues which catalyzes the conversion of D‐ threo ‐3‐hydroxyaspartate to oxaloacetate and ammonia. D‐THA DH also catalyzes the dehydration of L‐ threo ‐3‐hydroxyaspartate, L‐ erythro ‐3‐hydroxyaspartate and D‐serine. The amino‐acid sequence of D‐THA DH shows significant similarity to that of two eukaryotic D‐serine dehydratases derived from Saccharomyces cerevisiae and chicken kidney. D‐THA DH is classified into the fold‐type III group of pyridoxal enzymes and is the first example of a fold‐type III dehydratase derived from a prokaryote. Overexpression of recombinant D‐THA DH was carried out using a Rhodococcus erythropolis expression system and the obtained protein was subsequently purified and crystallized. The crystals of D‐THA DH belonged to space group I 4 1 22, with unit‐cell parameters a = b = 157.3, c = 157.9 Å. Single‐wavelength anomalous diffraction data were collected to a resolution of 2.0 Å using synchrotron radiation at the wavelength of the Br K absorption edge.