RRC ID 48708
著者 Hirato Y, Tokuhisa M, Tanigawa M, Ashida H, Tanaka H, Nishimura K.
タイトル Cloning and characterization of d-threonine aldolase from the green alga Chlamydomonas reinhardtii.
ジャーナル Phytochemistry
Abstract d-Threonine aldolase (DTA) catalyzes the pyridoxal 5'-phosphate (PLP)-dependent interconversion of d-threonine and glycine plus acetaldehyde. The enzyme is a powerful tool for the stereospecific synthesis of various β-hydroxy amino acids in synthetic organic chemistry. In this study, DTA from the green alga Chlamydomonas reinhardtii was discovered and characterized, representing the first report to describe the existence of eukaryotic DTA. DTA was overexpressed in recombinant Escherichia coli BL21 (DE3) cells; the specific activity of the enzyme in the cell-free extract was 0.8 U/mg. The recombinant enzyme was purified to homogeneity by ammonium sulfate fractionation, DEAE-Sepharose, and Mono Q column chromatographies (purified enzyme 7.0 U/mg). For the cleavage reaction, the optimal temperature and pH were 70 °C and pH 8.4, respectively. The enzyme demonstrated 90% of residual activity at 50 °C for 1 h. The enzyme catalyzed the synthesis of d- and d-allo threonine from a mixture of glycine and acetaldehyde (the diastereomer excess of d-threonine was 18%). DTA was activated by several divalent metal ions, including manganese, and was inhibited by PLP enzyme inhibitors and metalloenzyme inhibitors.
巻・号 135
ページ 18-23
公開日 2017-3-1
DOI 10.1016/j.phytochem.2016.12.012
PII S0031-9422(16)30298-9
PMID 28038776
MeSH Aldehyde-Lyases / metabolism Amino Acid Sequence Chlamydomonas reinhardtii / enzymology Chlamydomonas reinhardtii / metabolism* Cloning, Molecular Escherichia coli / genetics Glycine / metabolism Glycine Hydroxymethyltransferase / metabolism* Pyridoxal Phosphate / metabolism Stereoisomerism Substrate Specificity Threonine
IF 3.044
引用数 3
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY PLANT SCIENCES
リソース情報
藻類 NIES-2237