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Chemical protein synthesis enabled engineering of saccharide oxidative cleavage activity in artificial metalloenzymes

Int J Biol Macromol. 2023-11; 
Jindi Luo, Chunmao He
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Proteins, Expression, Isolation and Analysis … The gene encoding the fusion protein was synthesized and codon-optimized for E. coli expression (Genscript Inc., Nanjing, China). The synthetic gene was cloned into the pTXB1 … Get A Quote

摘要

Chemical protein (semi-)synthesis is a powerful technique allowing the incorporation of unnatural functionalities at any desired protein site. Herein we describe a facile one-pot semi-synthetic strategy for the construction of a type 2 copper center in the active site of azurin, which is achieved by substitution of Met121 with unnatural amino acid residues bearing a strong ligand N,N-bis(pyridylmethyl)amine (DPA) to mimic the function of typical histidine brace-bearing copper monooxygenases, such as lytic polysaccharide monooxygenases (LPMOs) involved in polysaccharide breakdown. The semi-synthetic proteins were routinely obtained in over 10-mg scales to allow for spectroscopic measurements (UV-Vis, CD, and EPR... More

關(guān)鍵詞

Artificial metalloenzyme, Chemical protein synthesis, Saccharide oxidative cleavage
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