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    • 5. 发明申请
    • BIOSYNTHESIS OF POLYKETIDE SYNTHASE SUBSTRATES
    • 聚合酶基因的生物合成
    • WO2002068613A1
    • 2002-09-06
    • PCT/US2002/006399
    • 2002-02-28
    • THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITYKHOSLA, ChaitanPFEIFER, Blaine
    • KHOSLA, ChaitanPFEIFER, Blaine
    • C12N1/21
    • C12N9/1288C12N9/16C12N15/52C12P1/00C12P17/189
    • 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.
    • 描述了催化生产起始物的酶的使用和聚酮化合物的增量剂单元。 此外,描述了可以接受多种起始单元如取代的苯甲酸酯并且可用于产生天然产物的取代衍生物的改性负载模块。 这些酶可用于增强天然产生的聚酮化合物或合理设计的聚酮化合物的产率。 通过使用这些技术,已经在大肠杆菌中完成了完整的聚酮化合物的合成。 通过将生产延迟到指数期后并通过维持培养基中相对恒定的营养水平,可以增强聚酮化合物和其他次级代谢产物的微生物生物。 此外,通过提供硫酯酶II的表达系统,可以增强聚酮化合物的产生。 因此,通过改变宿主生物体并改变培养条件,可以增强次级代谢物的合成。 本发明还导致具有所需特性的宿主生物体用于生产这种聚酮化合物并评估基因改组的结果。