title: Method for Enzyme Design with Genetically Encoded Unnatural Amino Acids.
abstract: We describe the methodologies for the design of artificial enzymes with genetically encoded unnatural amino acids. Genetically encoded unnatural amino acids offer great promise for constructing artificial enzymes with novel activities. In our studies, the designs of artificial enzyme were divided into two steps. First, we considered the unnatural amino acids and the protein scaffold separately. The scaffold is designed by traditional protein design methods. The unnatural amino acids are inspired by natural structure and organic chemistry methods, and synthesized by either organic chemistry methods or enzymatic conversion. With the increasing number of published unnatural amino acids with various functions, we described an unnatural amino acids toolkit containing metal chelators, redox mediators, and click chemistry reagents. These efforts enable a researcher to search the toolkit for appropriate unnatural amino acids for the study, rather than design and synthesize the unnatural amino acids from the beginning. After the first step, the model enzyme was optimized by computational methods and directed evolution. Lastly, we describe a general method for evolving aminoacyl-tRNA synthetase and expressing unnatural amino acids incorporated into a protein.
authors: Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
authors: C
authors: C
authors: Hu
authors: Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China. Electronic address: jwang@ibp.ac.cn.
authors: J
authors: J
authors: Wang
mesh_terms: D000596:Amino Acids / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry* / Q000235:genetics; D002625:Chemistry, Organic / Q000379:methods; D057930:Click Chemistry / Q000379:methods; D019295:Computational Biology; D019020:Directed Molecular Evolution; D004798:Enzymes / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*
chemical_list: D000596:Amino Acids; D004798:Enzymes; D000604:Amino Acyl-tRNA Synthetases
keywords: Enzyme design; Genetic code expansion; Nature inspired; Synthesize; Unnatural toolkit