| US 7,521,212 B1 | ||
| Method for producing oligopolysaccharides | ||
| Eric Samain, Gieres (France); and Bernard Priem, Grenoble (France) | ||
| Assigned to Centre National de la Recherche Scientifique (CNRS), Paris (France) | ||
| Appl. No. 10/19,954 PCT Filed Jul. 07, 2000, PCT No. PCT/FR00/01972 § 371(c)(1), (2), (4) Date May 24, 2002, PCT Pub. No. WO01/04341, PCT Pub. Date Jan. 18, 2001. |
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| Claims priority of application No. 99 08772 (FR), filed on Jul. 07, 1999. | ||
| Int. Cl. C12P 19/18 (2006.01); C12P 19/26 (2006.01) | ||
| U.S. Cl. 435—84 [435/97; 435/193; 536/55.2; 536/55.3; 536/123; 536/123.1; 536/124; 536/126] | 14 Claims |
| 1. A method for producing an oligosaccharide comprising lactose using a genetically modified cell starting with at least one
internalized exogenous precursor consisting of lactose said method comprising:
(i) obtaining a Lac Z−Y+ E. coli cell that
comprises at least one recombinant gene encoding an enzyme capable of modifying said exogenous precursor or one of the intermediates
in the biosynthetic pathway of said oligosaccharide from said exogenous precursor necessary for the synthesis of said oligosaccharide
from said exogenous precursor, and also the components for expressing said gene in said cell; and
(ii) culturing said cell on a carbon-based substrate in the presence of at least one said exogenous precursor, under conditions
inducing the internalization according to a mechanism of active transport of said exogenous precursor by said cell and the
production of said oligosaccharide by said cell,
wherein said culturing comprises:
(a) a first phase of exponential cell growth ensured by said carbon-based substrate,
(b) a second phase of cell growth limited by said carbon-based substrate which is added continuously, wherein said precursor is added during the second phase.
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