Synchronized Chemoenzymatic Synthesis of Monodisperse Hyaluronan Polymers

摘要: The length of the hyaluronan (HA) polysaccharide chain dictates its biological effects in many cellular and tissue systems. Long short HA polymers often appear to have antagonistic or inverse effects. However, no source very defined, uniform with sizes greater than 10 kDa is currently available. We present a method produce synthetic narrow size distributions range ∼16 ∼2 MDa. Pasteurella synthase enzyme, pmHAS, catalyzes synthesis polymer utilizing monosaccharides from UDP-sugar precursors. Recombinant pmHAS will also elongate exogenously supplied oligosaccharide acceptors vitro nonprocessive fashion. As result bypassing slow initiation step vitro, elongation process synchronized presence acceptor; thus all products are similar length. In contrast, without use an acceptor, final difficult predict more polydisperse. desired constructed by controlling reaction stoichiometry (i.e. molar ratio precursors acceptor molecules). modified allows containing tags (e.g. fluorescent, radioactive). this scheme, each molecule has single foreign moiety at reducing terminus. Alternatively, radioactive uniformly labeled native polymers. Overall, reagents monodisperse defined structures should assist elucidation numerous roles health disease.