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    • 22. 发明授权
    • In vitro synthesis of polypeptides by optimizing amino acid metabolism
    • 通过优化氨基酸代谢体外合成多肽
    • US06994986B2
    • 2006-02-07
    • US09948815
    • 2001-09-07
    • James Robert SwartzDong-Myung Kim
    • James Robert SwartzDong-Myung Kim
    • C12P21/06
    • C12N9/1033C07K14/5255C12P21/00
    • Compositions and methods are provided for the enhanced in vitro synthesis of polypeptides. In order to improve the performance of in vitro protein synthesis reactions, metabolic inhibitors, or manipulation of a source organism, is used to diminish or avoid the action of enzymes responsible for undesirable amino acids production or depletion. A homeostatic system may be used for production of ATP, where the required high energy phosphate bonds are generated in situ, e.g. through coupling with an oxidation reaction. The homeostatic energy source will typically lack high energy phosphate bonds itself, and will therefore utilize free phosphate in the reaction mix during generation of ATP. The homeostatic energy source is provided in combination with an enzyme that catalyzes the creation of high energy phosphate bonds and with an enzyme that can use that high energy phosphate bond to regenerate ATP.
    • 提供组合物和方法用于增强多肽的体外合成。 为了改善体外蛋白质合成反应的性能,使用代谢抑制剂或来源生物体的操作来减少或避免负责不良氨基酸产生或消耗的酶的作用。 可以使用稳态系统来生产ATP,其中所需的高能磷酸键是原位生成的。 通过与氧化反应的偶联。 稳态能源本身通常缺乏高能量磷酸键,因此在产生ATP期间将在反应混合物中利用游离磷酸盐。 稳态能量源与催化高能磷酸键的酶的结合,以及能够使用高能磷酸键重新生成ATP的酶组合提供。