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    • 75. 发明公开
    • BIOSYNTHESIS OF POLYKETIDE SYNTHASE SUBSTRATES
    • 多肽合成酶底物的生物合成
    • EP1381672A1
    • 2004-01-21
    • EP02725059.6
    • 2002-02-28
    • THE BOARD OF TRUSTEES OFTHE LELAND STANFORD JUNIOR UNIVERSITY
    • KHOSLA, ChaitanPFEIFER, Blaine
    • C12N1/21C12N9/00C12N15/63C12N15/54C12N15/81
    • 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.
    • 描述了催化生产聚酮化合物的起始剂和增量剂单元的酶的使用。 另外,描述了修改的装载模块,其可以接受多种起始单元,例如取代的苯甲酸酯,并且可以用于生成天然产物的取代衍生物。 这些酶可用于提高天然产生的聚酮化合物或合理设计的聚酮化合物的产量。 通过使用这些技术,已经在大肠杆菌中完成了合成完整的聚酮化合物。 在聚酮化合物和其他次级代谢产物的微生物中通过延迟生产直到指数期后和通过保持培养基中相对恒定的营养水平可以提高产量。 另外,通过提供硫酯酶II的表达系统可以增强聚酮化合物的生产。 因此,通过改变宿主生物体并改变培养条件,可以增强次级代谢产物的合成。 本发明还导致具有所需特征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。