Amino Acids

, Volume 48, Issue 2, pp 357–363

Genetic incorporation of recycled unnatural amino acids

Original Article

Abstract

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.

Keywords

Genetic incorporation Unnatural amino acids Recycle Culture medium 

Supplementary material

726_2015_2087_MOESM1_ESM.docx (222 kb)
Supplementary material 1 (DOCX 222 kb)

Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  1. 1.Department of ChemistrySogang UniversitySeoulRepublic of Korea

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