title: [Enzymatic cyclization of peptides using immobilized sortase A].
abstract: Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
authors.forename: Shu-xiang
authors.initials: SX
authors.lastname: Zhang
authors.forename: Min-zhi
authors.initials: MZ
authors.lastname: Liu
authors.forename: Yan
authors.initials: Y
authors.lastname: Yang
authors.forename: Ke-di
authors.initials: KD
authors.lastname: Cheng
authors.forename: Jian-qiang
authors.initials: JQ
authors.lastname: Kong
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D019881:Aminoacyltransferases / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D003500:Cyclization; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007700:Kinetics; D010455:Peptides / Q000378:metabolism*; D010456:Peptides, Cyclic / Q000096:biosynthesis*; D013211:Staphylococcus aureus / Q000201:enzymology
chemical_list: D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D010455:Peptides; D010456:Peptides, Cyclic; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases