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Updated: Oct 18, 2025

Genetic Incorporation of Biosynthesized L-dihydroxyphenylalanine DOPA and Its Application to Protein Conjugation
Published on: August 24, 2018
Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors
Li Liu1, Bohao Wang1, Sheng Li1
1Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China.
Abstract:
The activity and function of proteins can be improved by incorporation of non-canonical amino acids (ncAAs). To avoid the tedious synthesis of a large number of chiral phenylalanine derivatives, we synthesized the corresponding phenylpyruvic acid precursors. Escherichia coli strain DH10B and strain C321.ΔA.expΔPBAD were selected as hosts for phenylpyruvic acid bioconversion and genetic code expansion using the MmPylRS/pyltRNACUA system. The concentrations of keto acids, PLP and amino donors were optimized in the process. Eight keto acids that can be biotransformed and their coupled genetic code expansions were identified. Finally, the genetic encoded ncAAs were tested for incorporation into fluorescent proteins with keto acids.
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