- Tytuł:
- Properties of recombinant trehalose synthase from Deinococcus radiodurans expressed in Escherichia coli
- Autorzy:
-
Filipkowski, Paweł
Pietrow, Olga
Panek, Anna
Synowiecki, Józef - Powiązania:
- https://bibliotekanauki.pl/articles/1039724.pdf
- Data publikacji:
- 2012
- Wydawca:
- Polskie Towarzystwo Biochemiczne
- Tematy:
-
trehalose
trehalose synthase
Escherichia coli
gene expression
Deinococcus radiodurans - Opis:
- A trehalose synthase gene from Deinococcus radiodurans (DSMZ 20539) containing 1659 bp reading frame encoding 552 amino acids was amplified using PCR. The gene was finally ligated into pET30Ek/LIC vector and expressed after isopropyl β-d-thiogalactopyranoside induction in Escherichia coli (DE3) Rosetta pLysS. The recombinant trehalose synthase (DraTreS) containing a His6-tag at the C-terminus was purified by metal affinity chromatography and characterized. The expressed enzyme is a homodimer with molecular mass of 126.9 kDa and exhibits the highest activity of 11.35 U/mg at pH 7.6 and at 30°C. DraTreS activity was almost unchanged after 2 h preincubation at 45°C and pH 7.6, and retained about 56% of maximal value after 8 h incubation at 50°C. The DraTreS was strongly inhibited by Cu2+, Hg2+, Zn2+, Al3+ and 10 mM Tris. The Km value of maltose conversion was 290.7 mM.
- Źródło:
-
Acta Biochimica Polonica; 2012, 59, 3; 425-431
0001-527X - Pojawia się w:
- Acta Biochimica Polonica
- Dostawca treści:
- Biblioteka Nauki
Artykuł