Solid-Phase Enzymatic Peptide Synthesis to Produce an Antioxidant Dipeptide  

作　　者：Yuyao Shan Mengfan Wang Wei Qi Rongxin Su Zhimin He 

机构地区：[1]State Key Laboratory of Chemical Engineering, School of Chemical Engineering and Technology , Tianjin University [2]Tianjin Key Laboratory of Membrane Science and Desalination Technology [3]Collaborative Innovation Center of Chemical Science and Engineering

出　　处：《Transactions of Tianjin University》2019年第3期276-282,共7页天津大学学报（英文版）

基　　金：supported by National Key Scientific Instrument and Equipment Development Projects of China (No. 2012YQ090194);National High Technology Research and Development Program of China ("863" Program, No. 2013AA102204);National Natural Science Foundation of China (Nos. 21676191, 21476165, 21621004)

摘　　要：Peptide bond synthesis is favorable to the production of bioactive small peptides. However, the abuse of toxic reagents remains an issue for chemical synthesis method, whereas the low product yield and purity limit the widespread use of enzymatic method. In this study, a new solid-phase enzymatic peptide synthesis(SPEPS) strategy was developed to produce an antioxidant tyrosine-alanine dipeptide(Tyr-Ala) by using recombinant carboxypeptidase Y(CPY) as the catalyst. The general SPEPS procedure involves three steps. First, the N-protected acyl donor was covalently attached to solid resin. Second,the peptide bond was condensed between the acyl donor and the nucleophile under the catalysis of CPY. Finally, one-step cleavage was performed to remove the protecting group and cleave the peptides from solid resin. Upon the optimization of reaction conditions, 77.92%(±2.723%) yield of Tyr-Ala with high product purity of 90.971%(±2.695%) was obtained.In addition, the antioxidant activity of Tyr-Ala was determined by ABTS method, indicating that the synthesized Tyr-Ala obtained by SPEPS showed a superior antioxidant capability compared with commercial glutathione.Peptide bond synthesis is favorable to the production of bioactive small peptides. However, the abuse of toxic reagents remains an issue for chemical synthesis method, whereas the low product yield and purity limit the widespread use of enzymatic method. In this study, a new solid-phase enzymatic peptide synthesis(SPEPS) strategy was developed to produce an antioxidant tyrosine-alanine dipeptide(Tyr-Ala) by using recombinant carboxypeptidase Y(CPY) as the catalyst. The general SPEPS procedure involves three steps. First, the N-protected acyl donor was covalently attached to solid resin. Second,the peptide bond was condensed between the acyl donor and the nucleophile under the catalysis of CPY. Finally, one-step cleavage was performed to remove the protecting group and cleave the peptides from solid resin. Upon the optimization of reaction conditions, 77.92%(±2.723%) yield of Tyr-Ala with high product purity of 90.971%(±2.695%) was obtained.In addition, the antioxidant activity of Tyr-Ala was determined by ABTS method, indicating that the synthesized Tyr-Ala obtained by SPEPS showed a superior antioxidant capability compared with commercial glutathione.

关 键 词：SOLID-PHASE ENZYMATIC PEPTIDE synthesis CARBOXYPEPTIDASE Y Tyrosine-alanine Antioxidant activity 

分 类 号：T[一般工业技术]