Genetic incorporation of recycled unnatural amino acids

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• 作者：Wooseok Ko ; Sanggil Kim ; Kyubong Jo ; Hyun Soo Lee
• 关键词：Genetic incorporation ; Unnatural amino acids ; Recycle ; Culture medium
• 刊名：Amino Acids
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：48
• 期：2
• 页码：357-363
• 全文大小：843 KB
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• 作者单位：Wooseok Ko (1)
  Sanggil Kim (1)
  Kyubong Jo (1)
  Hyun Soo Lee (1)
  
  1. Department of Chemistry, Sogang University, Seoul, 121-742, Republic of Korea
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Biochemistry
  Analytical Chemistry
  Biochemical Engineering
  Life Sciences
  Proteomics
  Neurobiology
  
• 出版者：Springer Wien
• ISSN：1438-2199

文摘

The genetic incorporation of unnatural amino acids (UAAs) into proteins has been a useful tool for protein engineering. However, most UAAs are expensive, and the method requires a high concentration of UAAs, which has been a drawback of the technology, especially for large-scale applications. To address this problem, a method to recycle cultured UAAs was developed. The method is based on recycling a culture medium containing the UAA, in which some of essential nutrients were resupplemented after each culture cycle, and induction of protein expression was controlled with glucose. Under optimal conditions, five UAAs were recycled for up to seven rounds of expression without a decrease in expression level, cell density, or incorporation fidelity. This method can generally be applied to other UAAs; therefore, it is useful for reducing the cost of UAAs for genetic incorporation and helpful for expanding the use of the technology to industrial applications.