Unleashing the potential of noncanonical amino acid biosynthesis to create cells with precision tyrosine sulfation

Yuda Chen, Shikai Jin, Mengxi Zhang, Yu Hu, Kuan-Lin Wu, Anna Chung, Shichao Wang, Zeru Tian, Yixian Wang, Peter G. Wolynes and Han Xiao ()
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Yuda Chen: Rice University
Shikai Jin: Rice University
Mengxi Zhang: Rice University
Yu Hu: Rice University
Kuan-Lin Wu: Rice University
Anna Chung: Rice University
Shichao Wang: Rice University
Zeru Tian: Rice University
Yixian Wang: Rice University
Peter G. Wolynes: Rice University
Han Xiao: Rice University

Nature Communications, 2022, vol. 13, issue 1, 1-11

Abstract: Abstract Despite the great promise of genetic code expansion technology to modulate structures and functions of proteins, external addition of ncAAs is required in most cases and it often limits the utility of genetic code expansion technology, especially to noncanonical amino acids (ncAAs) with poor membrane internalization. Here, we report the creation of autonomous cells, both prokaryotic and eukaryotic, with the ability to biosynthesize and genetically encode sulfotyrosine (sTyr), an important protein post-translational modification with low membrane permeability. These engineered cells can produce site-specifically sulfated proteins at a higher yield than cells fed exogenously with the highest level of sTyr reported in the literature. We use these autonomous cells to prepare highly potent thrombin inhibitors with site-specific sulfation. By enhancing ncAA incorporation efficiency, this added ability of cells to biosynthesize ncAAs and genetically incorporate them into proteins greatly extends the utility of genetic code expansion methods.

Date: 2022
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DOI: 10.1038/s41467-022-33111-4

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