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Extracellular Production and Characterization of Streptomyces X-prolyl Dipeptidyl Aminopeptidase

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Abstract

X-prolyl dipeptidyl aminopeptidases (X-PDAPs) are useful in various food industries. In this study, we performed sequence-based screening to obtain a stable X-PDAP enzyme from thermophilic Streptomyces strains. We found three genes that encoded X-PDAP from Streptomyces thermoluteus subsp. fuscus NBRC 14270 (14270 X-PDAP), Streptomyces thermocyaneoviolaceus NBRC 14271 (14271 X-PDAP), and Streptomyces thermocoerulescens NBRC 14273, which were subsequently cloned and sequenced. The deduced amino acid sequences of these genes showed high similarity, with ~80% identity with each other. The isolated X-PDAPs and an X-PDAP from Streptomyces coelicolor were expressed in Streptomyces lividans using a hyperexpression vector: pTONA5a. Among these genes, only 14270 and 14271 X-PDAPs caused overexpression and extracellular production without artificial signal peptides. We also characterized the biochemical properties of purified 14271 X-PDAP. In addition, we found that, in peptide synthesis via an aminolysis reaction, this enzyme recognized d-amino acid derivatives as acyl acceptors, similar to l-amino acid derivatives.

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Acknowledgment

This work was supported by JSPS KAKENHI No. 20550158.

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Authors and Affiliations

  1. Okayama Prefectural Technology Center for Agriculture, Forestry, and Fisheries, Research Institute for Biological Sciences (RIBS), Okayama, 7549-1 Kibichuo-cho, Kaga-gun, Okayama, 716-1241, Japan

    Tadashi Hatanaka, Akihiro Yamasato, Hirokazu Usuki, Yukihiro Yamamoto & Yuya Kumagai

  2. Japan Society for the Promotion of Science (JSPS), Tokyo, Japan

    Hirokazu Usuki

  3. Department of Agricultural, Biological, and Environmental Sciences, Faculty of Agriculture, Tottori University, 4-101 Koyama-minami, Tottori, 680-8553, Japan

    Jiro Arima

Authors
  1. Tadashi Hatanaka
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  2. Akihiro Yamasato
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  3. Jiro Arima
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  4. Hirokazu Usuki
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  5. Yukihiro Yamamoto
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  6. Yuya Kumagai
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Correspondence to Tadashi Hatanaka.

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Hatanaka, T., Yamasato, A., Arima, J. et al. Extracellular Production and Characterization of Streptomyces X-prolyl Dipeptidyl Aminopeptidase. Appl Biochem Biotechnol 164, 475–486 (2011). https://doi.org/10.1007/s12010-010-9149-z

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  • DOI: https://doi.org/10.1007/s12010-010-9149-z

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