Reference : Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysacchar...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/11889
Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysaccharea amylosucrase
English
Albenne, C. [> > > >]
Skov, L. K. [> > > >]
Mirza, O. [> > > >]
Gajhede, M. [> > > >]
Feller, Georges mailto [Université de Liège - ULg > Département des sciences de la vie > Labo de biochimie >]
D'Amico, Salvino [Université de Liège - ULg > > GIGA-Research >]
Andre, G. [> > > >]
Potocki-Veronese, G. [> > > >]
van der Veen, B. A. [> > > >]
Monsan, P. [> > > >]
Remaud-Simeon, M. [> > > >]
2-Jan-2004
Journal of Biological Chemistry
Amer Soc Biochemistry Molecular Biology Inc
279
1
726-734
Yes (verified by ORBi)
International
0021-9258
Bethesda
[en] Amylosucrase from Neisseria polysaccharea is a remarkable transglucosidase from family 13 of the glycosidehydrolases that synthesizes an insoluble amylose-like polymer from sucrose in the absence of any primer. Amylosucrase shares strong structural similarities with alpha-amylases. Exactly how this enzyme catalyzes the formation of alpha-1,4-glucan and which structural features are involved in this unique functionality existing in family 13 are important questions still not fully answered. Here, we provide evidence that amylosucrase initializes polymer formation by releasing, through sucrose hydrolysis, a glucose molecule that is subsequently used as the first acceptor molecule. Maltooligosaccharides of increasing size were produced and successively elongated at their nonreducing ends until they reached a critical size and concentration, causing precipitation. The ability of amylosucrase to bind and to elongate maltooligosaccharides is notably due to the presence of key residues at the OB1 acceptor binding site that contribute strongly to the guidance ( Arg(415), subsite +4) and the correct positioning (Asp(394) and Arg(446), subsite +1) of acceptor molecules. On the other hand, Arg(226) (subsites +2/+3) limits the binding of maltooligosaccharides, resulting in the accumulation of small products (G to G3) in the medium. A remarkable mutant (R226A), activated by the products it forms, was generated. It yields twice as much insoluble glucan as the wild-type enzyme and leads to the production of lower quantities of by-products.
http://hdl.handle.net/2268/11889

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