Structure of the enzymatically synthesized fructan inulin.
Construction, purification and characterization of a fusion protein of maltose-binding protein of Escherichia coli and the fructosyltransferase of Streptococcus mutans is described. With the purified protein, in vitro synthesis of inulin was performed. The obtained polysaccharide was characterized by high-performance size-exclusion chromatography (HPSEC) and static light scattering (SLS) in dilute aqueous and dimethyl sulfoxide solution. For all samples very high molecular weights between 60 x 10(6) and 90 x 10(6) g/mol and a remarkable small polydispersity index of 1.1 have been determined. Small root-mean-square radii of gyration point to a compact conformation in dilute solution. No difference between native and enzymatically synthesized inulin was observed by X-ray powder diffraction and thermoanalysis of solid samples.
Heyer
A G
AG
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Golm, Germany. heyer@mpimp-golm.mpg.de
Schroeer
B
B
Radosta
S
S
Wolff
D
D
Czapla
S
S
Springer
J
J
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002236:Carbohydrate Conformation; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins; D005630:Fructans / Q000737:chemistry; D006602:Hexosyltransferases / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D007444:Inulin / Q000096:biosynthesis / Q000737:chemistry*; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D009391:Nephelometry and Turbidimetry; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013295:Streptococcus mutans / Q000201:enzymology / Q000235:genetics; D014961:X-Ray Diffraction
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D005630:Fructans; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; C461599:maltose transport system, E coli; D007444:Inulin; D006602:Hexosyltransferases; C021451:inulosucrase