Evaluation and biosynthetic incorporation of chlorotyrosine into recombinant proteins

• Published in: Biotechnology and bioprocess engineering 2012, 17(4), , pp.679-686
• Main Authors: Ayyadurai, Niraikulam, King Saud University, Riyadh, Saudi Arabia, Deepankumar, Kanagavel, Yeungnam University, Gyeongsan, Republic of Korea, Prabhu, Nadarajan Saravanan, Yeungnam University, Gyeongsan, Republic of Korea, Budisa, Nediljko, Biocatalysis Group, Institute of Chemistry, Berlin Institute of Technology/TU Berlin, Berlin, Germany, Yun, H.D., Yeungnam University, Gyeongsan, Republic of Korea
• Format: Article
• Language: English
• Subjects: Amino acids; Analysis; Binding sites; Bioinformatics; Biotechnology; Chemical engineering; Chemistry; Chemistry and Materials Science; Chlorine; chloro-tyrosine; E coli; Efficiency; Genetic engineering; GFP; homology modeling; Industrial and Production Engineering; noncanonical amino acids; Polymers; protein engineering; Protein folding; Protein synthesis; PROTEINAS; PROTEINE; PROTEINS; Research Paper; Studies; Transfer RNA; Translation; Tyrosine; 생물공학
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-012-0066-6

Recently, non-canonical amino acids (NCAA) incorporation was developed to enhance the functional properties of proteins. Incorporation of NCAA containing chlorine atom is conceptually an attractive approach to prepare pharmacologically active substances, which is a difficult task since chlorine is bulky atom. In this study, we evaluated the efficiency and extent of in vivo incorporation of tyrosine analogue 3-chlorotyrosine [(3-Cl)Tyr] into the recombinant proteins GFP and GFPHS (highly stable GFP). The incorporation of (3-Cl)Tyr into GFP leads to dramatic reduction in the expression level of protein. On the other hand, the incorporation of (3-Cl)Tyr into GFPHS was expressed well as a soluble form. In addition we used bioinformatics tools for the analysis to explore the possible constraints in micro-environment of each natural amino acid residue to be replaced with chlorine atom accommodation into GFPHS. In conclusion, our approaches are reliable and straightforward way to enhance the translation of chlorinated amino acids into proteins.