3 Biotech

, 8:173 | Cite as

Purification and characterization of Stenotrophomonas maltophilia-derived l-amino acid ester hydrolase for synthesizing dipeptide, isoleucyl-tryptophan

  • Md Saddam Hossain
  • Takahiro Tanaka
  • Kazuyoshi Takagi
  • Junji Hayashi
  • Mamoru Wakayama
Original Article


In the present study, we purified α-amino acid ester hydrolase (AEH) from cell-free extracts of the Stenotrophomonas maltophilia strain HS1. The approximately 70-kDa AEH from S. maltophilia HS1 (SmAEH) was homogeneous in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analyses, and was present as a tetramer in gel-filtration experiments. The activity of the SmAEH enzyme was then determined by monitoring the synthesis of the antihypertensive agent dipeptide isoleucyl-tryptophan (Ile-Trp) from isoleucyl methyl ester (Ile-OMe) and tryptophan (Trp). In these experiments, SmAEH had wide substrate specificity for acyl donors, such as Gly-OMe, β-Ala-OMe, Pro-OMe and Trp-OMe and Ile-OMe, and maximal activity were observed under conditions of pH 9.0 and 30 °C. SmAEH also showed the greatest stability at pH 9.0, whereas its activity was reduced by 40% after 10-min incubation at approximately 50 °C. In subsequent activity assays in the presence of various metal ions, Ag+ strongly inhibited enzyme activity. Finally, SmAEH activity was completely inhibited by phenylmethanesulfonyl fluoride (PMSF), suggesting that the protein is a serine protease.


α-Amino acid ester hydrolase Stenotrophomonas maltophilia Antihypertensive dipeptide Ile-Trp 


Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interests.

Supplementary material

13205_2018_1195_MOESM1_ESM.docx (233 kb)
Supplementary material 1 (DOCX 232 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biotechnology, College of Life SciencesRitsumeikan UniversityKusatsuJapan
  2. 2.Department of Applied Chemistry, College of Life SciencesRitsumeikan UniversityKusatsuJapan

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