Production of O-acetylated and sulfated chitooligosaccharides by recombinant Escherichia coli strains harboring different combinations of nod genes.
High cell density cultivation of recombinant Escherichia coli strains harboring the nodBC genes (encoding chitooligosaccharide synthase and chitooligosaccharide N-deacetylase, respectively) from Azorhizobium caulinodans has been previously described as a practical method for the preparation of gram-scale quantities of penta-N-acetyl-chitopentaose and tetra-N-acetylchitopentaose (Samain, E., Drouillard, S., Heyraud, A., Driguez, H., Geremia, R.A., 1997. Carbohydr. Res. 30, 235-242). We have now extended this method to the production of sulfated and O-acetylated derivatives of these two compounds by coexpressing nodC or nodBC with nodH and/or nodL that encode chitooligosaccharide sulfotransferase and chitooligosaccharide O-acetyltransferase, respectively. In addition, these substituted chitooligosaccharides were also obtained as tetramers by using nodC from Rhizobium meliloti instead of nodC from A. caulinodans. These compounds should be useful precursors for the preparation of Nod factor analogues by chemical modification.
Centre de Recherche sur les Macromolécules Végétales (CERMAV-CNRS), Grenoble, France. samain@cermav.cnrs.fr
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Samain
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Chazalet
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Geremia
D000107:Acetylation; D000581:Amidohydrolases / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D004926:Escherichia coli / Q000235:genetics*; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D009844:Oligosaccharides / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011995:Recombination, Genetic; D013464:Sulfuric Acids / Q000737:chemistry
D001426:Bacterial Proteins; D009844:Oligosaccharides; D013464:Sulfuric Acids; C073606:nodB protein, Bacteria; D017351:N-Acetylglucosaminyltransferases; C060771:NodC protein, Rhizobiales; D000581:Amidohydrolases