Properties of recombinant trehalose synthase from Deinococcus radiodurans expressed in Escherichia coli - Publikacja - MOST Wiedzy

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Properties of recombinant trehalose synthase from Deinococcus radiodurans expressed in Escherichia coli

Abstrakt

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 recombinanttrehalose 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 30oC.DraTreS activity was almost unchanged after 2 h preincubation at 45oC and pH 7.6, and retained about 56% of maximal value after 8 h incubation at 50oC. The DraTreS was strongly inhibited by Cu2+, Hg2+, Zn2+, Al3+ and 10mM Tris. The Km value of maltose conversion was 290.7 mM.

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Informacje szczegółowe

Kategoria:
Publikacja w czasopiśmie
Typ:
artykuł w czasopiśmie wyróżnionym w JCR
Język:
angielski
Rok wydania:
2012
Opis bibliograficzny:
Filipkowski P., Pietrow-Tobiszewska O., Panek A., Synowiecki J.: Properties of recombinant trehalose synthase from Deinococcus radiodurans expressed in Escherichia coli// Acta Biochimica Polonica.. -Vol. 59, nr. No.3 (2012), s.425-431
Weryfikacja:
Politechnika Gdańska

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