Biosynthesis of polyketide synthase substrates

Details Biosynthesis of polyketide synthase substrates Biosynthesis of polyketide synthase substrates

C - Chemistry, Metallurgy

12

N

C12N 1/21 (2006.01) C12N 9/00 (2006.01) C12N 9/12 (2006.01) C12N 9/16 (2006.01) C12N 15/52 (2006.01) C12N 15/54 (2006.01) C12N 15/63 (2006.01) C12N 15/81 (2006.01) C12P 1/00 (2006.01) C12P 17/18 (2006.01) C12P 19/62 (2006.01)

Patent

CA 2438798

The use of enzymes which catalyze the production of starter and extender units for polyketides is described. In addition, modified loading modules are described, which can accept a variety of starting units such as substituted benzoates, and which can be used to generate substituted derivatives of natural products. These enzymes may be used to enhance the yield of polyketides that are natively produced or polyketides that are rationally designed. By using these techniques, the synthesis of a complete polyketide has been achieved in E. coli. Production can be enhanced in microbial organisms of polyketides and other secondary metabolites by delaying production until after exponential phase and by maintaining relatively constant nutrient levels in the medium. In addition, polyketide production can be enhanced by providing an expression system for a thioesterase II. Thus, by modifying the host organism and changing the culture conditions, synthesis of a secondary metabolite can be enhanced. The present invention also results in a host organism with desirable characteristics to be used in the production of such polyketides and to assess the results of gene shuffling.

L'invention concerne l'utilisation d'enzymes qui catalysent la production d'unités d'amorçage et d'extension de polycétides. Elle concerne de plus des modules de chargement modifiés, pouvant recevoir diverses unités d'amorçage telles que des benzoates substitués, et qui peuvent servir à produire des dérivés substitués de produits naturels. Ces enzymes peuvent servir à améliorer le rendement de polycétides produits de façon native ou de polycétides mis au point rationnellement. A l'aide de ces techniques, la synthèse d'un polycétide complet a pu être mise en oeuvre chez <i>E. coli</i>. On peut améliorer la production de polycétides et d'autres métabolites secondaires chez des organismes microbiens en reportant celle-ci après une phase exponentielle et en maintenant des taux de nutriments relativement constants dans le milieu. De plus, on améliore la production de polycétides en prévoyant un système d'expression pour une thioestérase II. Une modification de l'organisme hôte et des conditions de culture permet ainsi d'améliorer la synthèse d'un métabolite secondaire. L'invention permet aussi d'obtenir un organisme hôte qui possède des caractéristiques souhaitables en vue de la production de polycétides et de l'évaluation des résultats de réarrangement génique.

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