Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors

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...

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Autores principales: Liu, Li, Wang, Bohao, Li, Sheng, Xu, Fengyuan, He, Qi, Pan, Chun, Gao, Xiangdong, Yao, Wenbing, Song, Xiaoda
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8470325/
https://www.ncbi.nlm.nih.gov/pubmed/34572570
http://dx.doi.org/10.3390/biom11091358
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author Liu, Li
Wang, Bohao
Li, Sheng
Xu, Fengyuan
He, Qi
Pan, Chun
Gao, Xiangdong
Yao, Wenbing
Song, Xiaoda
author_facet Liu, Li
Wang, Bohao
Li, Sheng
Xu, Fengyuan
He, Qi
Pan, Chun
Gao, Xiangdong
Yao, Wenbing
Song, Xiaoda
author_sort Liu, Li
collection PubMed
description 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/(pyl)tRNA(CUA) 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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spelling pubmed-84703252021-09-27 Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors Liu, Li Wang, Bohao Li, Sheng Xu, Fengyuan He, Qi Pan, Chun Gao, Xiangdong Yao, Wenbing Song, Xiaoda Biomolecules Article 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/(pyl)tRNA(CUA) 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. MDPI 2021-09-14 /pmc/articles/PMC8470325/ /pubmed/34572570 http://dx.doi.org/10.3390/biom11091358 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Liu, Li
Wang, Bohao
Li, Sheng
Xu, Fengyuan
He, Qi
Pan, Chun
Gao, Xiangdong
Yao, Wenbing
Song, Xiaoda
Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors
title Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors
title_full Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors
title_fullStr Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors
title_full_unstemmed Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors
title_short Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors
title_sort convenient genetic encoding of phenylalanine derivatives through their α-keto acid precursors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8470325/
https://www.ncbi.nlm.nih.gov/pubmed/34572570
http://dx.doi.org/10.3390/biom11091358
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