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Solid-Phase Enzymatic Peptide Synthesis to Produce an Antioxidant Dipeptide

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Abstract

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.

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Acknowledgements

This study was 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) and National Natural Science Foundation of China (Nos. 21676191, 21476165, 21621004).

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Authors and Affiliations

  1. State Key Laboratory of Chemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, China

    Yuyao Shan, Mengfan Wang, Wei Qi, Rongxin Su & Zhimin He

  2. Tianjin Key Laboratory of Membrane Science and Desalination Technology, Tianjin, 300350, China

    Mengfan Wang, Wei Qi & Rongxin Su

  3. Collaborative Innovation Center of Chemical Science and Engineering, Tianjin, 300350, China

    Wei Qi & Rongxin Su

Authors
  1. Yuyao Shan
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  2. Mengfan Wang
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  3. Wei Qi
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  4. Rongxin Su
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  5. Zhimin He
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Correspondence to Mengfan Wang.

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Shan, Y., Wang, M., Qi, W. et al. Solid-Phase Enzymatic Peptide Synthesis to Produce an Antioxidant Dipeptide. Trans. Tianjin Univ. 25, 276–282 (2019). https://doi.org/10.1007/s12209-018-0174-2

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  • DOI: https://doi.org/10.1007/s12209-018-0174-2

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