Gram-scale synthesis of recombinant chitooligosaccharides in Escherichia coli.
Cultivation of Escherichia coli harbouring heterologous genes of oligosaccharide synthesis is presented as a new method for preparing large quantities of high-value oligosaccharides. To test the feasibility of this method, we successfully produced in high yield (up to 2.5 g/L) penta-N-acetyl-chitopentaose (1) and its deacetylated derivative tetra-N-acetyl-chitopentaose (2) by cultivating at high density cells of E. coli expressing nodC or nodBC genes (nodC and nodB encode for chitooligosaccharide synthase and chitooligosaccharide N-deacetylase, respectively). These two products were easily purified by charcoal adsorption and ion-exchange chromatography. One important application of compound 2 could be its utilisation as a precursor for the preparation of synthetic nodulation factors by chemical acylation.
Samain
E
E
Centre de Recherches sur les Macromolécules Végétales (CERMAV-CNRS), Grenoble, France. samain@cermav.grenet.fr
Drouillard
S
S
Heyraud
A
A
Driguez
H
H
Geremia
R A
RA
D000107:Acetylation; D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping
D001426:Bacterial Proteins; D009844:Oligosaccharides; D011994:Recombinant Proteins; C073606:nodB protein, Bacteria; D017351:N-Acetylglucosaminyltransferases; C060771:NodC protein, Rhizobiales; D000581:Amidohydrolases