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    IN VITRO MACROMOLECULE BIOSYNTHESIS METHODS USING EXOGENOUS AMINO ACIDS AND A NOVEL ATP REGENERATION SYSTEM
    1.
    发明公开
    IN VITRO MACROMOLECULE BIOSYNTHESIS METHODS USING EXOGENOUS AMINO ACIDS AND A NOVEL ATP REGENERATION SYSTEM 有权
    体外MAKROMOLEKüLBIOSYNTHESEVERFAHREN使用外源性氨基酸和ATP再生系统

    公开(公告)号：EP1177311A1

    公开(公告)日：2002-02-06

    申请号：EP00923078.0

    申请日：2000-03-15

    申请人： THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY

    发明人： SWARTZ, James, R. ,  KIM, Dong-Myung

    IPC分类号： C12P21/06

    CPC分类号： C12N9/1033 ,  C07K14/5255 ,  C12P21/00

    摘要： 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.

    CONTROL OF METABOLIC FLUX IN CELL-FREE BIOSYNTHETIC SYSTEMS
    5.
    发明公开
    CONTROL OF METABOLIC FLUX IN CELL-FREE BIOSYNTHETIC SYSTEMS 审中-公开
    代谢流动的在无细胞系统合成控制

    公开(公告)号：EP3004335A1

    公开(公告)日：2016-04-13

    申请号：EP14807322.4

    申请日：2014-06-05

    申请人： Greenlight Biosciences Inc. ,  The Board of Trustees of The Leland Stanford Junior University

    发明人： SWARTZ, James, R.

    IPC分类号： C12N9/00

    CPC分类号： C12Q3/00 ,  C12N9/00

    摘要： Methods are provided for controlling metabolic flux rate in a cell-free system comprising a complex set of enzymes, to produce a desired product of a pathway of interest. In the methods of the invention, measurements of metabolic performance parameters are taken by continuous monitoring or intermittent monitoring. Based on the metabolic performance parameters, the system is modified by one or more steps comprising: (i) altering enzyme levels in the cell-free system; (ii) altering feed rate of a substrate that controls redox flux or carbon flux to the cell-free system; (iii) altering 02 addition to the cell-free system; (iv) controlling efficiency of electron transport system by altering leakage across a membrane; wherein enzymes present in the pathway of interest catalyze production of a desired product.