Mimicking the Escherichia coli cytoplasmic environment activates long-lived and efficient cell-free protein synthesis
Mimicking the Escherichia coli cytoplasmic environment activates long-lived and efficient cell-free protein synthesis
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DOI：
10.1002/bit.20026
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发表时间：
2004-04-05
影响因子：
3.8
通讯作者：
Swartz, JR
中科院分区：
文献类型：
--
作者：
Jewett, MC;Swartz, JR
无细胞翻译系统通常利用高能磷酸化合物来再生驱动蛋白质合成所必需的三磷酸腺苷（ATP）。由于昂贵的试剂成本、抑制性副产物（如磷酸盐）的积累和pH变化，这阻碍了该技术以批量形式的广泛使用和实际实施。为了解决这些问题，无细胞蛋白质合成系统已被工程化，其能够使用丙酮酸作为能量来源以产生高产率的蛋白质。“Cytomim”系统在分批反应中合成氯霉素乙酰转移酶（CAT）长达6小时，以产生700 μ g/mL的蛋白质。通过更紧密地复制大肠杆菌细胞质的生理条件，Cytomim系统为蛋白质表达提供了稳定的能量供应，而无需磷酸盐积累、pH变化、外源酶添加或需要昂贵的高能磷酸盐化合物。(C)2004 Wiley Periodicals，Inc.
Cell-free translation systems generally utilize high-energy phosphate compounds to regenerate the adenosine triphosphate (ATP) necessary to drive protein synthesis. This hampers the widespread use and practical implementation of this technology in a batch format due to expensive reagent costs; the accumulation of inhibitory byproducts, such as phosphate; and pH change. To address these problems, a cell-free protein synthesis system has been engineered that is capable of using pyruvate as an energy source to produce high yields of protein. The "Cytomim" system, synthesizes chloramphenicol acetyltransferase (CAT) for up to 6 h in a batch reaction to yield 700 mug/mL of protein. By more closely replicating the physiological conditions of the cytoplasm of Escherichia coli, the Cytomim system provides a stable energy supply for protein expression without phosphate accumulation, pH change, exogenous enzyme addition, or the need for expensive high-energy phosphate compounds. (C) 2004 Wiley Periodicals, Inc.