title: Genetic incorporation of recycled unnatural amino acids.
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.
authors.affiliation: Department of Chemistry, Sogang University, Seoul, 121-742, Republic of Korea.
authors.forename: Wooseok
authors.initials: W
authors.lastname: Ko
authors.affiliation: Department of Chemistry, Sogang University, Seoul, 121-742, Republic of Korea.
authors.forename: Sanggil
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Sogang University, Seoul, 121-742, Republic of Korea. Jojokyubong@sogang.ac.kr.
authors.forename: Kyubong
authors.initials: K
authors.lastname: Jo
authors.affiliation: Department of Chemistry, Sogang University, Seoul, 121-742, Republic of Korea. hslee76@sogang.ac.kr.
authors.forename: Hyun Soo
authors.initials: HS
authors.lastname: Lee
mesh_terms: D000596:Amino Acids / Q000378:metabolism*; D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000502:physiology*; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D003470:Culture Media; D011506:Proteins
keywords: Culture medium; Genetic incorporation; Recycle; Unnatural amino acids