Cloning and characterization of the gene for amylosucrase from Neisseria polysaccharea: production of a linear alpha-1,4-glucan.
The gene for the amylosucrase from Neisseria polysaccharea (ATCC 43768) was cloned by use of a functional expression system in Escherichia coli XL1-Blue. The deduced amino acid sequence of the protein has homology to the sequences of the alpha-amylase class of enzymes, with the highest similarities being found to the sequences of the trehalose synthase from Pimelobacter sp. strain R48 (17) and amylomaltase from Thermotoga maritima (11). However, the regions of highest homology within the alpha-amylase class of enzymes, which are essential for the catalytic activity, are only scarcely found in the sequence of amylosucrase. By using the enzyme isolated from culture supernatants of transformed E. coli cells, it is possible to synthesize linear alpha-1,4-glucans from sucrose, indicating that the enzyme is not capable of producing alpha-1,6-glycosidic linkages on its own.
Büttcher
V
V
Institut für Genbiologische Forschung GmbH Berlin, Germany.
Welsh
T
T
Willmitzer
L
L
Kossmann
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005110:Extracellular Space / Q000201:enzymology / Q000382:microbiology; D005798:Genes, Bacterial; D005936:Glucans / Q000096:biosynthesis* / Q000737:chemistry; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009343:Neisseria / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D021382:Protein Sorting Signals / Q000737:chemistry; D017386:Sequence Homology, Amino Acid
D005936:Glucans; D021382:Protein Sorting Signals; C040088:1,4-glucan; D005964:Glucosyltransferases; C019700:amylosucrase