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Immobilization on magnetic nanoparticles of therecombinant trehalose synthase from Deinococcusgeothermalis
Abstrakt
tIn our study the gene encoding trehalose synthase from Deinococcus geothermalis was cloned and overexpressed inEscherichia coli Rosetta (DE3)pLysS. Wild-type trehalose synthase has been purified from host protein after cell dis-ruption and precipitation at 20% ammonium sulphate saturation. Recombinant trehalose synthase was immobilizedonto glutaraldehyde activated silanized magnetic ferrous-ferric oxide by using covalent binding method. The mor-phology and surface of the obtained particles were characterized using SEM. These images show that all sampleshave a particle size below 30 nm. The obtained immobilized preparation has specific activity of 0.134 U/g supportwhen measured at 40◦C using maltose as substrate. Immobilization process was almost fully completed after 30 minof the reaction at 30◦C. The highest immobilization yield of the enzyme was achieved at glutaraldehyde concentra-tion of 10 mM. No significant differences in optimal pH and temperature were observed upon immobilization. Theimmobilized trehalose synthase exhibited mass transfer limitation, which is reflected by higher KMand activationenergy values. In addition, immobilized trehalose synthase was easily separated from the reaction medium by anexternal magnetic field and retained 82% of its initial activity after successive twelve repeated batches reaction.
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Informacje szczegółowe
- Kategoria:
- Publikacja w czasopiśmie
- Typ:
- artykuł w czasopiśmie wyróżnionym w JCR
- Opublikowano w:
-
FOOD AND BIOPRODUCTS PROCESSING
nr 91,
wydanie 4,
strony 632 - 637,
ISSN: 0960-3085 - Język:
- angielski
- Rok wydania:
- 2013
- Opis bibliograficzny:
- Panek A., Pietrow-Tobiszewska O., Synowiecki J., Filipkowski P.: Immobilization on magnetic nanoparticles of therecombinant trehalose synthase from Deinococcusgeothermalis// FOOD AND BIOPRODUCTS PROCESSING. -Vol. 91, iss. 4 (2013), s.632-637
- Weryfikacja:
- Politechnika Gdańska
wyświetlono 128 razy
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Expression of Deinococcus geothermalis Trehalose Synthase Gene in Escherichia coli and Its Enzymatic Properties
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- + 2 autorów