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Biosynthesis, Characterization, and Bioactivity of L-Alanyl-L-tyrosine in Promoting Melanin Synthesis

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Abstract

L-Alanyl-L-tyrosine (L-Ala-Tyr) is a dipeptide formed by the condensation of L-alanine methyl ester and L-tyrosine. After entering the body, it can be rapidly broken down to release tyrosine. In this study, L-Ala-Tyr was successfully prepared by using α-ester acyltransferase as biocatalyst and alanine methyl ester (L-Ala-OMe) and tyrosine (L-Tyr) as acyl donor and nucleophile, respectively. The dipeptide yield was increased from 15 to 50% by optimizing the conditions: boric acid-borax (0.2 mol/L), 30°C, pH 9.5, 2:1 acyl donor to nucleophile ratio, DES (ChCl/urea), and 15%(v/v) water content. The catalytic product is then isolated and purified. The structure of the product was identified by high-performance liquid chromatography, mass spectrometry, proton nuclear magnetic resonance, and carbon spectroscopy. Its biological activity was preliminarily determined by the B16-F10 mouse melanoma cell model. The results showed that the purity of L-Ala-Tyr prepared by the separation and purification method of this study was 96.8%, and the mass spectrometry and nuclear magnetic resonance spectroscopy showed that the structure of the peptide was consistent with the expected structure. In addition, the preliminary physiological activity identification results show that L-Ala-Tyr has no toxic effect on cells in the concentration range of 100–800 μmol·L−1, and at the optimal concentration, compared with the positive control 8-methoxypsoralen, it can promote the production of melanin.

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Data Availability

All further relevant data generated or analyzed during this study are included in this published article and its supplementary files.

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Funding

This study was supported by the Anhui Provincial Natural Science Foundation (grant no. 2108085MC120) and Anhui Province Key Research and Development Plan Project 2023s07020013.

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Author notes

  1. Yuna Fan and Jinao Wei  contributed equally to this work.

Authors and Affiliations

  1. School of Food and Biological Engineering, Hefei University of Technology, Tunxi Road No. 193, Hefei, 230009, Anhui, China

    Yuna Fan, Jinao Wei, Zhiwei Li, Jingwen Yang, Xueqin Hu & Hongbin Zhang

Authors
  1. Yuna Fan
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  2. Jinao Wei
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  3. Zhiwei Li
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  4. Jingwen Yang
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  5. Xueqin Hu
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Contributions

YN Fan: software, validation, formal analysis, investigation, writing (original draft), and visualization. JA Wei: software and formal analysis. ZW Li: software. JW Yang: methodology. HB Zhang: conceptualization and supervision. XQ Hu: conceptualization, formal analysis, resources, data curation, writing (review and editing), visualization, and funding acquisition.

Corresponding authors

Correspondence to Xueqin Hu or Hongbin Zhang.

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This study does not contain any studies with human participants or animals performed.

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Key points

• This study provides a method to synthesize dipeptide by using α-ester acyltransferase in DES.

• Through process optimization, the conversion rate of the substrate was improved.

• The physiological activity of Ala-Tyr was proved by the melanocyte experiment.

Supplementary Information

ESM 1

(DOCX 183 kb)

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Fan, Y., Wei, J., Li, Z. et al. Biosynthesis, Characterization, and Bioactivity of L-Alanyl-L-tyrosine in Promoting Melanin Synthesis. Appl Biochem Biotechnol (2023). https://doi.org/10.1007/s12010-023-04713-5

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