Biotechnology Letters

, Volume 34, Issue 6, pp 1093–1099 | Cite as

A mutant leucine aminopeptidase from Streptomyces cinnamoneus with enhanced l-aspartyl l-amino acid methyl ester synthetic activity

  • Jiro Arima
  • Mirai Kono
  • Manami Kita
  • Nobuhiro Mori
Original Research Paper
First Online:
Received:
Accepted:

Abstract

l-Aspartyl l-amino acid methyl ester was synthesized using a mutant of a thermostable leucine aminopeptidase from Streptomyces cinnamoneus, D198 K SSAP, obtained in previously. A peptide of high-intensity sweetener, l-aspartyl-l-phenylalanine methyl ester, was selected as a model for demonstrating the synthesis of l-aspartyl l-amino acid methyl ester. The hydrolytic activities of D198 K SSAP toward l-aspartyl-l-phenylalanine and its methyl ester were, respectively, 74-fold and fourfold higher than those of wild type. Similarly, the initial rate of the enzyme for l-aspartyl-l-phenylalanine methyl ester synthesis was over fivefold higher than that of wild-type SSAP in 90% methanol (v/v) in a one-pot reaction. Furthermore, other l-aspartyl l-amino acid methyl esters were synthesized efficiently using D198 K SSAP. Results show that the substitution of Asp198 of SSAP with Lys is effective for synthesizing l-aspartyl l-amino acid methyl ester.

Keywords

Aminopeptidase l-aspartyl-l-phenylalanine methyl ester Peptide bond formation Reverse reaction 
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Notes

Acknowledgment

This research was supported by a grant for young researchers from Nikki-Saneyoshi.

Supplementary material

10529_2012_877_MOESM1_ESM.docx (410 kb)
Supplementary material 1 (DOCX 409 kb)

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Jiro Arima
    • 1
  • Mirai Kono
    • 1
  • Manami Kita
    • 1
  • Nobuhiro Mori
    • 1
  1. 1.Department of Agricultural, Biological, and Environmental Sciences, Faculty of AgricultureTottori UniversityTottoriJapan

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