1887

Abstract

strain 121 employs a fructosyltransferase (FTF) to synthesize a fructose polymer [a fructan of the levan type, with (2→6) linkages] from sucrose or raffinose. Purification of this FTF (a levansucrase), and identification of peptide amino acid sequences, allowed isolation of the first levansucrase gene (), encoding a protein (Lev) consisting of 804 amino acids. Lev showed highest similarity with an inulosucrase of 121 [Inu; producing an inulin polymer with (2→1)-linked fructosyl units] and with FTFs from streptococci. Expression of in resulted in an active FTF (LevΔ773His) that produced the same levan polymer [with only 2–3 % (2→1→6) branching points] as 121 cells grown on raffinose. The low degree of branching of the levan is very different from bacterial levans known up to now, such as that of , having up to 30 % branches. Although Lev is unusual in showing a higher hydrolysis than transferase activity, significant amounts of levan polymer are produced both and . Lev is strongly dependent on Ca ions for activity. Unique properties of Lev together with Inu are: (i) the presence of a C-terminal cell-wall-anchoring motif causing similar expression problems in , (ii) a relatively high optimum temperature for activity for FTF enzymes, and (iii) at 50 °C, kinetics that are best described by the Hill equation.

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2004-03-01
2020-08-12
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