Applied Biochemistry and Biotechnology

, Volume 189, Issue 1, pp 305–317 | Cite as

Amino Acids as Additives against Amorphous Aggregation: In Vitro and In Silico Study on Human Lysozyme

  • Naghmeh Saadati-Eskandari
  • Latifeh NavidpourEmail author
  • Parichehreh Yaghmaei
  • Azadeh Ebrahim-HabibiEmail author


The effect of 16 amino acids (AA) with various physicochemical properties was investigated on human lysozyme (HL) heat-induced amorphous aggregation. UV-Visible spectrophotometry was used to monitor the kinetics of aggregation in the absence and presence of AA, and transmission electron microscopy (TEM) images were taken from the aggregates. To conduct in silico experiments, Autodock vina was used for docking of AA into protein (via YASARA interface), and FTmap information was checked for an insight onto putative binding sites. Prediction of aggregation-prone regions of lysozyme was made by AGGRESCAN and Tango. Among all tested AA, phenylalanine had the best anti-aggregation effect, followed by lysine. In addition, based on in silico tests, Trp 109 and Val 110 of lysozyme are suggested to be of importance in the aggregation process of the enzyme. In conclusion, phenylalanine, arginine, and lysine were found to affect the nucleation phase of lysozyme aggregation and could be considered as suitable stabilizing structures for this enzyme.


Aggregation Lysozyme Amino acids Phenylalanine, transmission Electron microscopy 


Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest. Experiments were done in vitro with purchased recombinant enzyme.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

12010_2019_3010_MOESM1_ESM.docx (654 kb)
ESM 1 (DOCX 653 kb)


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biology, Faculty of Basic Sciences, Science and Research BranchIslamic Azad UniversityTehranIran
  2. 2.Department of Medicinal Chemistry, Faculty of PharmacyTehran University of Medical SciencesTehranIran
  3. 3.Biosensor Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences InstituteTehran University of Medical SciencesTehranIran
  4. 4.Endocrinology and Metabolism Research Center, Endocrinology and Metabolism Clinical Sciences InstituteTehran University of Medical SciencesTehranIran

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