A convenient method for genetic incorporation of multiple noncanonical amino acids into one protein in Escherichia coli

2010 ◽  
Vol 6 (4) ◽  
pp. 683 ◽  
Author(s):  
Ying Huang ◽  
William K. Russell ◽  
Wei Wan ◽  
Pei-Jing Pai ◽  
David H. Russell ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Convenient Method ◽  
Noncanonical Amino Acids ◽  
Genetic Incorporation

scholarly journals Genetic Incorporation of Noncanonical Amino Acids Using Two Mutually Orthogonal Quadruplet Codons

2019 ◽  
Vol 8 (5) ◽  
pp. 1168-1174 ◽  
Author(s):  
Erome Daniel Hankore ◽  
Linyi Zhang ◽  
Yan Chen ◽  
Kun Liu ◽  
Wei Niu ◽  
...  
Keyword(s):  
Amino Acids ◽  
Noncanonical Amino Acids ◽  
Genetic Incorporation

Overproduction of noncanonical amino acids by Escherichia coli cells

Microbiology ◽  
2007 ◽  
Vol 76 (6) ◽  
pp. 712-718 ◽  
Author(s):  
E. V. Sycheva ◽  
T. A. Yampol’skaya ◽  
E. S. Preobrajenskaya ◽  
A. E. Novikova ◽  
N. G. Matrosov ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Noncanonical Amino Acids ◽  
Escherichia Coli Cells

scholarly journals Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System

10.3791/54273 ◽  
2016 ◽  
Author(s):  
Emanuel G. Worst ◽  
Matthias P. Exner ◽  
Alessandro De Simone ◽  
Marc Schenkelberger ◽  
Vincent Noireaux ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Translation System ◽  
Noncanonical Amino Acids

A Facile System for Genetic Incorporation of Two Different Noncanonical Amino Acids into One Protein inEscherichia coli

2010 ◽  
Vol 49 (18) ◽  
pp. 3211-3214 ◽  
Author(s):  
Wei Wan ◽  
Ying Huang ◽  
Zhiyong Wang ◽  
William K. Russell ◽  
Pei-Jing Pai ◽  
...  
Keyword(s):  
Amino Acids ◽  
Inescherichia Coli ◽  
Noncanonical Amino Acids ◽  
Genetic Incorporation

Global substitution of hemeproteins with noncanonical amino acids in Escherichia coli with intact cofactor maturation machinery

2017 ◽  
Vol 106 ◽  
pp. 55-59
Author(s):  
Jan-Stefan Völler ◽  
Tuyet Mai Thi To ◽  
Hernan Biava ◽  
Beate Koksch ◽  
Nediljko Budisa
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Noncanonical Amino Acids

scholarly journals Directed evolution of rRNA improves translation kinetics and recombinant protein yield

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Fan Liu ◽  
Siniša Bratulić ◽  
Alan Costello ◽  
Teemu P. Miettinen ◽  
Ahmed H. Badran
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Protein Synthesis ◽  
Bacterial Population ◽  
Protein Yield ◽  
Rrna Sequence ◽  
Evolutionary Trajectory ◽  
Noncanonical Amino Acids ◽  
Rate Limiting

AbstractIn bacteria, ribosome kinetics are considered rate-limiting for protein synthesis and cell growth. Enhanced ribosome kinetics may augment bacterial growth and biomanufacturing through improvements to overall protein yield, but whether this can be achieved by ribosome-specific modifications remains unknown. Here, we evolve 16S ribosomal RNAs (rRNAs) from Escherichia coli, Pseudomonas aeruginosa, and Vibrio cholerae towards enhanced protein synthesis rates. We find that rRNA sequence origin significantly impacted evolutionary trajectory and generated rRNA mutants with augmented protein synthesis rates in both natural and engineered contexts, including the incorporation of noncanonical amino acids. Moreover, discovered consensus mutations can be ported onto phylogenetically divergent rRNAs, imparting improved translational activities. Finally, we show that increased translation rates in vivo coincide with only moderately reduced translational fidelity, but do not enhance bacterial population growth. Together, these findings provide a versatile platform for development of unnatural ribosomal functions in vivo.

scholarly journals Comparative Analyses of the Transcriptome and Proteome of Escherichia coli C321.△A and Further Improving Its Noncanonical Amino Acids Containing Protein Expression Ability by Integration of T7 RNA Polymerase

2021 ◽  
Vol 12 ◽  
Author(s):  
Huawei Yi ◽  
Jing Zhang ◽  
Famin Ke ◽  
Xiurong Guo ◽  
Jian Yang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Protein Expression ◽  
Rna Polymerase ◽  
T7 Rna Polymerase ◽  
Noncanonical Amino Acids ◽  
E Coli ◽  
Growth Defects ◽  
Genetic Code Expansion ◽  
And Function

Incorporation of noncanonical amino acids (ncAAs) into proteins has been proven to be a powerful tool to manipulate protein structure and function, and to investigate many biological processes. Improving the yields of ncAA-containing proteins is of great significance in industrial-scale applications. Escherichia coli C321.ΔA was generated by the replacement of all known amber codons and the deletion of RF1 in the genome and has been proven to be an ideal host for ncAA-containing protein expression using genetic code expansion. In this study, we investigated the transcriptome and proteome profiles of this first codon reassignment strain and found that some functions and metabolic pathways were differentially expressed when compared with those of its parent strain. Genes involved in carbohydrate and energy metabolism were remarkably downregulated. Our results may provide important clues about the growth defects in E. coli C321.ΔA. Furthermore, we improved the yields of ncAA-containing proteins in E. coli C321.ΔA by integrating the T7 RNA polymerase system.

scholarly journals Plasticity and Constraints of tRNA Aminoacylation Define Directed Evolution of Aminoacyl-tRNA Synthetases

2019 ◽  
Vol 20 (9) ◽  
pp. 2294 ◽  
Author(s):  
Ana Crnković ◽  
Oscar Vargas-Rodriguez ◽  
Dieter Söll
Keyword(s):  
Amino Acids ◽  
Directed Evolution ◽  
Active Site ◽  
Nonsense Suppression ◽  
Natural Amino Acid ◽  
Noncanonical Amino Acids ◽  
Trna Synthetases ◽  
Aminoacyl Trna Synthetases ◽  
Genetic Incorporation

Genetic incorporation of noncanonical amino acids (ncAAs) has become a powerful tool to enhance existing functions or introduce new ones into proteins through expanded chemistry. This technology relies on the process of nonsense suppression, which is made possible by directing aminoacyl-tRNA synthetases (aaRSs) to attach an ncAA onto a cognate suppressor tRNA. However, different mechanisms govern aaRS specificity toward its natural amino acid (AA) substrate and hinder the engineering of aaRSs for applications beyond the incorporation of a single l-α-AA. Directed evolution of aaRSs therefore faces two interlinked challenges: the removal of the affinity for cognate AA and improvement of ncAA acylation. Here we review aspects of AA recognition that directly influence the feasibility and success of aaRS engineering toward d- and β-AAs incorporation into proteins in vivo. Emerging directed evolution methods are described and evaluated on the basis of aaRS active site plasticity and its inherent constraints.

Engineering Heterodimeric Kinesins through Genetic Incorporation of Noncanonical Amino Acids

2018 ◽  
Vol 13 (8) ◽  
pp. 2229-2236
Author(s):  
Andrew R. Popchock ◽  
Subhashis Jana ◽  
Ryan A. Mehl ◽  
Weihong Qiu
Keyword(s):  
Amino Acids ◽  
Noncanonical Amino Acids ◽  
Genetic Incorporation
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