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Gram-scale synthesis of recombinant chitooligosaccharides in Escherichia coli

1997; Elsevier BV; Volume: 302; Issue: 1-2 Linguagem: Inglês

10.1016/s0008-6215(97)00107-9

1873-426X

Eric Samain, Sophie Drouillard, A. Heyraud, Hugues Driguez, R. Geremia,

Studies on Chitinases and Chitosanases

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.