We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Purification and in vitro refolding of maize chloroplast transglutaminase over-expressed in Escherichia coli.
- Authors
Carvajal-Vallejos, Patricia K.; Campos, Alexandre; Fuentes-Prior, Pablo; Villalobos, Enrique; Almeida, André M.; Barberà, Eduard; Torné, José María; Santos, Mireya
- Abstract
In contrast to mammalian transglutaminases (TGs), plant members of the superfamily are poorly characterized. In order to produce pure and active TG for its functional and structural studies, variants of maize chloroplast transglutaminase (TGZ, Patent WWO03102128) were sub-cloned into a pET28 vector and overexpressed in Escherichia coli BL21 (DE3) cells. The recombinant proteins were present mainly as insoluble inclusion bodies. The TGZ4p variant with four B-type repeats ( M r∼55 kDa), was affinity purified from urea-solubilized inclusion bodies. TGZ4p was refolded by rapid dilution in a Ca2+- and guanidine-containing buffer. Active TGZ4p shows the general catalytic characteristics described for other TGs.
- Subjects
ESCHERICHIA coli; CHLOROPLASTS; TRANSGLUTAMINASES; PROTEINS; UREA
- Publication
Biotechnology Letters, 2007, Vol 29, Issue 8, p1255
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-007-9377-7