《Fully productive cell-free genetic code expansion by structure-based engineering of Methanomethylophilus alvus pyrrolysyl-tRNA synthetase》 was written by Seki, Eiko; Yanagisawa, Tatsuo; Kuratani, Mitsuo; Sakamoto, Kensaku; Yokoyama, Shigeyuki. Computed Properties of C11H22N2O4 And the article was included in ACS Synthetic Biology on April 17 ,2020. The article conveys some information:
Pyrrolysyl-tRNA synthetase (PylRS)/tRNAPyl pairs from Methanosarcina mazei and Methanosarcina barkeri are widely used for site-specific incorporations of non-canonical amino acids into proteins (genetic code expansion). In this study, we achieved the full productivity of cell-free protein synthesis for difficult, bulky non-canonical amino acids, such as Nε-((((E)-cyclooct-2-en-1-yl)oxy)carbonyl)-L-lysine (TCO*Lys), by using Methanomethylophilus alvus PylRS. First, based on the crystal structure of M. alvus PylRS, the productivities for various non-canonical amino acids were greatly increased by rational engineering of the amino acid-binding pocket. The productivities were further enhanced by using a much higher concentration of PylRS over that of M. mazei PylRS, or by mutating the outer layer of the amino acid-binding pocket. Thus, we achieved full productivity even for TCO*Lys. The quantity and quality of the cell-free-produced antibody fragment containing TCO*Lys were drastically improved. These results demonstrate the importance of full productivity for the expanded genetic code. The experimental part of the paper was very detailed, including the reaction process of H-Lys(Boc)-OH(cas: 2418-95-3Computed Properties of C11H22N2O4)
H-Lys(Boc)-OH(cas: 2418-95-3) belongs to amino acids. Amino acids are not generally considered to be electrochemically active because products of the oxidation accumulate on the electrode surface and prevent it from participating in any further electrochemical processes.Computed Properties of C11H22N2O4
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