|
type |
Journal Article |
authors |
Nagano, H.; Shibano, K.; Matsumoto, Y.; Yokota, A.; Wada, M.
|
title |
Isolation and amino acid sequence of a dehydratase acting on d-erythro-3-hydroxyaspartate from Pseudomonas sp. N99, and its application in the production of optically active 3-hydroxyaspartate |
journal |
Biosci Biotechnol Biochem |
Activity |
4.3.1.16 |
Family |
4.3.1.16 |
sel |
selected |
ui |
28290777 |
year |
(2017) |
volume |
81 |
number |
6 |
pages |
1156-1164 |
| |
keywords |
3-hydroxyaspartate; Pseudomonas sp. N99; ammonia-lyase; enzymatic resolution; pyridoxal 5′-phosphate |
abstract |
An enzyme catalyzing the ammonia-lyase reaction for the conversion of d-erythro-3-hydroxyaspartate to oxaloacetate was purified from the cell-free extract of a soil-isolated bacterium Pseudomonas sp. N99. The enzyme exhibited ammonia-lyase activity toward l-threo-3-hydroxyaspartate and d-erythro-3-hydroxyaspartate, but not toward other 3-hydroxyaspartate isomers. The deduced amino acid sequence of the enzyme, which belongs to the serine/threonine dehydratase family, shows similarity to the sequence of l-threo-3-hydroxyaspartate ammonia-lyase (EC 4.3.1.16) from Pseudomonas sp. T62 (74%) and Saccharomyces cerevisiae (64%) and serine racemase from Schizosaccharomyces pombe (65%). These results suggest that the enzyme is similar to l-threo-3-hydroxyaspartate ammonia-lyase from Pseudomonas sp. T62, which does not act on d-erythro-3-hydroxyaspartate. We also then used the recombinant enzyme expressed in Escherichia coli to produce optically pure l-erythro-3-hydroxyaspartate and d-threo-3-hydroxyaspartate from the corresponding dl-racemic mixtures. The enzymatic resolution reported here is one of the simplest and the first enzymatic method that can be used for obtaining optically pure l-erythro-3-hydroxyaspartate. |
last changed |
2017/09/06 10:59 |
|